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Pros and cons of using green biotechnology to solve food insecurity and achieve sustainable development goals

• Review Article
• Published: 07 February 2021
• Volume 6 , article number  29 , ( 2021 )

Cite this article

• Hebatallah Ahmed Nasser   ORCID: orcid.org/0000-0002-2801-4506 1 ,
• Mohamed Mahmoud 2 ,
• Mahmoud M. Tolba 3 ,
• Rasha Ali Radwan 4 ,
• Nayra Mahmoud Gabr 5 ,
• Aliaa Aly ElShamy 1 ,
• Mohamed Salah Yehya 6 ,
• Axel Ziemke 7 &
• Mohamed Yousri Hashem 8  

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The overall aim of sustainable development is to facilitate economic growth strategies that do not cause long-term environmental damage. To achieve this, a number of goals must be realized, including food security to prevent hunger. Globally, food security is threatened by climate change, for example global warming. Recently, green biotechnology has attracted considerable interest, as it offers techniques that could ensure food security despite the devastating consequences of climate change, such as droughts and floods. However, applying green biotechnology to agriculture and food production also poses some potential health risks and environmental hazards. This review examines the potential of green biotechnology as a possible solution to food insecurity resulting from climate change. It summarizes the pros and cons of green biotechnology with respect to achieving the United Nations Sustainable Development Goals. The review concludes that despite the economic advantages of green biotechnology, especially its capacity to enhance crop productivity and resistance to pests, some aspects of green biotechnology do not permit sustainable agricultural and medical development due to their risks to life on land or to human health and wellbeing.

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Abbreviations

Bacillus thuringiensis

Carbon dioxide

Deoxyribonucleic acid

Deeper Rooting 1

Environmental Protection Agency

Food and Drug Administration

Green fluorescent protein

Genetically improved

Genetically modified

Galanthus nivalis agglutinin

Human immunodeficiency virus type 1

Insulin-like growth factor 1

Intellectual property rights

Nonsteroidal anti-inflammatory drugs

Potential of hydrogen

Parts per million

Ribonucleic acid

United Nations Sustainable Development Goals

United States Department of Agriculture

World Health Organization

World Trade Organization

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Nasser, H.A., Mahmoud, M., Tolba, M.M. et al. Pros and cons of using green biotechnology to solve food insecurity and achieve sustainable development goals. Euro-Mediterr J Environ Integr 6 , 29 (2021). https://doi.org/10.1007/s41207-020-00240-5

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Biotechnology has emerged as a full fledged multifaceted discipline in advanced research and development. Green Biotechnology being arguably the most intensely debated and because it is the most visible of all the fields of Biotechnology. It entails the deliberate manipulation of plants, animals or microbes to produce effective and efficient environmentally sustainable novel and desirable products.

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Growing a circular economy with fungal biotechnology: a white paper

• Vera Meyer   ORCID: orcid.org/0000-0002-2298-2258 1 ,
• Evelina Y. Basenko   ORCID: orcid.org/0000-0001-8611-5447 2 ,
• J. Philipp Benz   ORCID: orcid.org/0000-0001-5361-4514 3 ,
• Gerhard H. Braus   ORCID: orcid.org/0000-0002-3117-5626 4 ,
• Mark X. Caddick   ORCID: orcid.org/0000-0002-5489-6557 2 ,
• Michael Csukai   ORCID: orcid.org/0000-0003-0615-7802 5 ,
• Ronald P. de Vries   ORCID: orcid.org/0000-0002-4363-1123 6 ,
• Drew Endy   ORCID: orcid.org/0000-0001-6952-8098 7 ,
• Jens C. Frisvad   ORCID: orcid.org/0000-0002-0573-4340 8 ,
• Nina Gunde-Cimerman   ORCID: orcid.org/0000-0002-9464-3263 9 ,
• Thomas Haarmann   ORCID: orcid.org/0000-0001-6033-8140 10 ,
• Yitzhak Hadar   ORCID: orcid.org/0000-0001-7510-7847 11 ,
• Kim Hansen   ORCID: orcid.org/0000-0002-2670-4677 12 ,
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• Nancy P. Keller   ORCID: orcid.org/0000-0002-4386-9473 14 ,
• Nada Kraševec   ORCID: orcid.org/0000-0002-0925-4272 15 ,
• Uffe H. Mortensen   ORCID: orcid.org/0000-0002-7794-7273 8 ,
• Rolando Perez 7 ,
• Arthur F. J. Ram   ORCID: orcid.org/0000-0002-2487-8016 16 ,
• Eric Record   ORCID: orcid.org/0000-0002-7545-9997 17 ,
• Phil Ross 18 ,
• Volha Shapaval   ORCID: orcid.org/0000-0003-2674-1328 19 ,
• Charlotte Steiniger   ORCID: orcid.org/0000-0001-5329-2391 1 ,
• Hans van den Brink 20 ,
• Jolanda van Munster   ORCID: orcid.org/0000-0003-0412-7001 21 ,
• Oded Yarden   ORCID: orcid.org/0000-0002-4722-535X 11 &
• Han A. B. Wösten   ORCID: orcid.org/0000-0002-0399-3648 22  

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Fungi have the ability to transform organic materials into a rich and diverse set of useful products and provide distinct opportunities for tackling the urgent challenges before all humans. Fungal biotechnology can advance the transition from our petroleum-based economy into a bio-based circular economy and has the ability to sustainably produce resilient sources of food, feed, chemicals, fuels, textiles, and materials for construction, automotive and transportation industries, for furniture and beyond. Fungal biotechnology offers solutions for securing, stabilizing and enhancing the food supply for a growing human population, while simultaneously lowering greenhouse gas emissions. Fungal biotechnology has, thus, the potential to make a significant contribution to climate change mitigation and meeting the United Nation’s sustainable development goals through the rational improvement of new and established fungal cell factories. The White Paper presented here is the result of the 2nd Think Tank meeting held by the EUROFUNG consortium in Berlin in October 2019. This paper highlights discussions on current opportunities and research challenges in fungal biotechnology and aims to inform scientists, educators, the general public, industrial stakeholders and policymakers about the current fungal biotech revolution.

Introduction

The term ‘biotechnology’ was coined by the Hungarian Karl Ereky [ 1 ] in 1919, the same year that Pfizer became the first company to commercialise a product manufactured by the controlled fermentation of a mould. The product Pfizer made was citric acid for commercial use as a flavouring agent, acidifier and chelating agent in food, beverages, and the pharma and chemical industries. It was produced biotechnologically with the mould Aspergillus niger [ 2 ], and 100 years later, citric acid is still produced with this filamentous fungus and has formed a fast growing multibillion Euro market for convenience foods and beverages [ 2 , 3 ]. Many organic acids, enzymes, life-saving antibiotics and drugs are produced by filamentous fungi, and a lot of our foods and beverages would not exist at all without their fermentative capacities [ 4 ]. It is undisputed that filamentous fungal-based biotechnology is of pivotal importance to our daily lives. Many companies around the globe are leveraging the power of filamentous fungi, with major players in Europe that include: AB Enzymes, BASF, Bayer, Chr. Hansen, Dyadic International, DSM, DuPont, Kerry Group, Marlow Foods, Novozymes, Puratos, Syngenta and Roal Oy.

In a more recent endeavour, biologists, chemists, bioinformaticians, bioengineers, process engineers and material scientists have collaborated to turn by-products and waste from agriculture and forestry into composite materials with the help of mushroom-forming fungi. The vision offered by this group is disruptive: soon, we will sit on mushroom-created furniture, we will live in houses made from fungal hyphae, plastics and insulation materials in cars, trains, planes and spacecrafts will be composed of fungal composites and many of our textiles will be derived from fungi. In the not so distant future, new fungal-based products will be introduced into the market that offer similar or superior characteristics compared to classic petroleum-derived products, with a reduced or even negative carbon footprint and full biodegradability. Another vision is not so far-fetched at all: products made with animal leather will be superseded in quality and price by those made with pure fungal mycelium. Such replacements will have substantial implications for the livestock, textile and fashion industries.

Fungi growing as a yeast morphology have impacted society for millennia and have been instrumental for the production of bread, beer and wine. However, fungal species capable of the filamentous growth form, which are the focus of this White Paper, have additional beneficial properties such as the productions of a diverse array of metabolites, enzymes and materials. Indeed, Scientific American stated in 2019 that: “The mycelium revolution is upon us” [ 5 ]. A 100 years after the birth of fungal biotechnology, this platform technology is undergoing a renaissance by providing sustainable solutions for diverse industries and markets [ 6 ]. In the following, we will summarize current and anticipated products made from moulds and mushrooms and related new avenues of research. We will also highlight recommendations discussed during the EUROFUNG 2nd Think Tank consortium for ways to drive innovation in this renaissance through strategic and infrastructural measures for basic and applied science on filamentous fungi.

The lifestyle of filamentous fungi—moulds and mushrooms

The life of a filamentous fungus usually starts with a spore, which has a diameter of only a few microns (µm) (Fig.  1 ). The spore starts to swell in a humid and nutrient-rich environment and germinates. A germ tube is formed that elongates to eventually form a thread-like, filamentous cell, called a hypha. After the hypha grows and elongates for some time, it forms a network of interconnected hyphal threads called a mycelium. When nutrients become limited in the substrate within which the mycelium lives, the mycelium starts to explore the air and space in order to form reproductive structures. Ascomycetes (moulds) can form conidiophores that produce asexual spores at their ends and/or fruiting bodies that produce sexual spores inside them. Basidiomycetes are known for their ability to form fruiting bodies to generate sexual spores. Some of these fruiting bodies are colloquially called mushrooms. These fruiting bodies consist of mycelia which are more densely packed and different in their composition compared to the rather loose substrate mycelia, which forms three-dimensional net-like structures resembling the global system of interconnected computer networks.

The fungal life cycle

Filamentous fungi are both invisible and visible. The diameter of fungal hyphae range from 2 to 10 µm, and a fungal mycelium consists of a network of mm- to cm-long hypha. The mycelia of mushroom-forming fungi can colonise large surface areas, as illustrated by an individual of the honey mushroom Armillaria bulbosa (common name honey mushroom in English and Hallimasch in German) which has colonised > 1000 hectares of forest soil, making it the largest and oldest organism on Earth [ 7 ]. Mycelia of mushroom-forming fungi can also be grown on various by-products and waste streams from forestry and agriculture. The efficiency of colonisation and biomass formation is determined by the composition and physical properties of the substrate, the environmental growth conditions (temperature, humidity and pH) and the genetic make-up of the fungus. A strain of Schizophyllum commune (common name split gill), for instance, in which two regulatory genes for mushroom formation are inactivated, produces threefold more biomass of fungal tissue when compared to the parental strain [ 8 ].

Filamentous fungi have evolved to become superbly efficient decomposers and have developed the ability to feed on and break down organic matter and polymeric substances [ 9 ]. Polysaccharides from plant biomass make up most of the biomass on Earth (450 out of a total of 550 gigatons of carbon; [ 10 ]) and represent the major carbon sources that drives fungal growth. In order to digest, humans must first ingest. Filamentous fungi first digest and then ingest. The fungal cells that infiltrate a substrate secrete enzymes, such as cellulases, amylases, pectinases, inulases, proteases and lipases, into the surrounding medium and hydrolyse (break down) plant polysaccharides (e.g. cellulose, starch, pectin, inulin), proteins and lipids. The degradation products of polysaccharides are monosaccharides, such as glucose, and oligosaccharides, which are subsequently taken up into the hyphae with the help of specific sugar transporters. This hydrolytic capacity for plant biomass, coupled with an extraordinary high secretion capacity for enzymes (up to 100 g/L are reported; [ 11 ]), forms the basis for the success of high-performance enzyme-producing cell factories, such as A. niger, A. oryzae, Trichoderma reesei and Thermothelomyces thermophilus (all being ascomycetes). Their enzyme products are exploited by a diverse array of major industries, including food and feed, detergent, pulp and paper, fuel, pharmaceutical and chemical (Fig.  2 ; [ 4 ]).

Industries profiting from the metabolic capacities of filamentous fungi

Notably, the predicted gene set for plant polysaccharide-degrading enzymes in these fungi is between 100 and 250, whereas only 30 can be found in the baker’s yeast Saccharomyces cerevisiae . The success of S. cerevisiae, which cannot grow on plant polymeric substances [ 12 ], or the bacterium Escherichia coli as platform organisms to produce biofuel (ethanol), jet fuel (terpenoids), fine and commodity chemicals is only guaranteed by the filamentous fungal cell factories mentioned above that provide S. cerevisiae and E. coli with simple carbon sources during the fermentation process. Hence, filamentous fungal cell factories play a central role in the sustainable production of biofuels and chemicals.

The product portfolio of filamentous fungi—an overview

The product range of filamentous fungi is not limited to citric acid and enzymes (Table  1 ). In fact, the natural metabolic capacities of filamentous fungi are extraordinarily diverse and unmatched in nature. Organic acids, chemicals, antibiotics and other drugs, proteins and enzymes, meat alternatives, vitamins, polyunsaturated fatty acids and even composite materials and vegan leathers are existing fungal products. Table  2 highlights some currently traded pharmaceuticals derived from filamentous fungi.

Note that T. thermophilus was formerly named Myceliophthora thermophila and that the P. chrysogenum strain used for penicillin production was recently reidentified as P. rubens [ 14 , 15 ]. While this review focuses on the Dikarya lineage, that is the ascomycetes and basidiomycetes, five species in this list are found in another lineage, the Mucoromycota.

Some examples of current developments with fungal cell factories that will help to consolidate bio-economies and human health are summarized as follows.

Aspergillus niger is currently being explored by academia as a potential producer of other organic acids beyond citric acid, such as itaconate and galactarate. Itaconate could replace petroleum-based polyacrylic acid, which is a precursor for the polymer industry (absorbent polymers, polyester resins, synthetic latex) and galactarate could replace the current petroleum-based polyethylene terephthalate (PET) used for plastic production [ 17 , 18 ].

A. oryzae , used to produce Asian food and beverages for over a 1000 years, has recently gained interest as a producer of malate, which has multiple applications in the food (acidulant, flavour enhancer), chemical (polyester resins) and pharmaceutical (acidulant) industries [ 19 ]. While several microbial cell factories from bacterial, yeast or filamentous fungal origin have been genetically engineered during the last few years to produce malate, A. oryzae is among the strains with the greatest yields [ 20 ].

A. terreus is exploited as a cell factory to produce two molecules: the organic acid itaconate, which is of interest for the polymer industry [ 21 ], and lovastatin, which is a cholesterol-lowering drug. Lovastatin has been marketed under the tradename Mevacor since the late 1980 s [ 22 ]. It, furthermore, serves as a starter molecule for manufacturing semisynthetic statins, such as simvastatin (trade name Zocor), which is the second leading statin on the market [ 23 ]. Current research is focused on re-routing lovastatin biosynthesis to one of its biosynthetic intermediates, monacolin J, which is the preferred precursor for Zocor synthesis [ 24 ]. Notably, a genetically engineered A. oryzae strain can reach monacolin J concentrations of about 5.5 g/L, which is considerably higher than has been achieved in heterologous hosts, such as S. cerevisiae (75 mg/L) or Pichia pastoris (renamed Komagataella phaffii ; 600 mg/L) [ 25 ].

P. chrysogenum ( P. rubens ) is an antibiotics producer and the main cell factory used to produce penicillin and semisynthetic derivatives, with production levels up to 55 g/L [ 26 ]. Notably, the biosynthetic route for penicillin has recently been genetically reprogrammed towards pravastatin, which constitutes an interesting alternative to Zocor for the pharma industry [ 27 ].

T. reesei and T. thermophilus (formally Myceliophthora thermophila ) are of commercial interest because of their ability to secrete cellulases and hemicellulases. These enzymes are key to converting lignocellulosic biomass into biofuel. Lignocellulose is composed of cellulose, hemicellulose and lignin and is a by-product of agriculture (e.g. straw, bagasse, corn stover) and forestry (e.g. sawdust). The thermostability of (hemi)cellulases from T. thermophilus are of special interest for current lignocellulose degradation processes, as high process temperatures are preferred in biorefineries to reduce the viscosity and increase the solubility of lignocellulosic biomass [ 28 ]. Genetically optimised hypersecreting strains have been established during the last two decades which achieve cellulase titres of 100 g/L [ 11 ]. These are the highest titres ever reported for protein secretion and exceed by 10–10,000-fold what can be achieved nowadays with bacterial, yeast or mammalian cell factories. Protein secretion titres in these cell factories are usually in the order of mg/L to only a few g/L [ 29 , 30 , 31 ]. Notably, T. thermophilus has been genetically reprogrammed to generate platform chemicals, such as fumarate, directly from renewable feedstocks to replace petroleum-based processes, which is of interest for the manufacturing of synthetic resins, biodegradable polymers (up to 17 g/L) and malate (up to 200 g/L) [ 32 , 33 ]. The fungus has also been optimised by Dyadic International as a producer for biologics, including vaccines, therapeutic enzymes, proteins and biosimilars [ 34 ].

Filamentous fungi as meat replacements

The use of fungi as a source of food predates recorded history [ 35 ]. These have been predominantly the mushrooms from supermarket shelves and foraging expeditions. The fruiting bodies of these fungi have a wide variety of tastes and textures and some of these are considered very meat-like, for example “Chicken of the Woods” ( Laetiporus sulphurous ) and “Beefsteak fungus” ( Fistulina hepatica ). There has recently been a move to create meat-like products from fungal mycelium grown in fermenters, rather than the solid fruiting bodies. This has allowed the introduction of ascomycetes, traditionally used as flavour modifiers in such foods as blue cheese, to enter the food chain as convincing meat substitutes. The longest established of these companies is Marlow Foods, using Fusarium venenatum under the trade name Quorn™, but several other companies have recently shown an interest in this area, including Mycorena ( A. oryzae ; [ 36 ]), Sustainable Bioproducts ( F. oxysporum ; [ 37 ]) and MycoTechnology, using the basidiomycete Lentinula edodes [ 38 ]. The hyphae of filamentous fungi, when aligned and organised, provide a structure that looks like and has the mouth feel of meat, particularly chicken, due to the similarity of fibre size (Fig.  3 ). The high amino acid and fibre content, and low saturated fat, combined with the high digestibility of fungal protein (see Additional file 1 : Table S1), make this an exceptionally healthy food [ 39 ].

Unique textural attributes of Quorn™. Electron microscopic images of protein fibres from spun soya and chicken and hyphal filaments of F. venenatum . Bar, 100 µm

The fungi for food replacements are grown in fermenters on simple salts and glucose as the carbon source. Quorn™ uses air lift pressure cycle fermenters (Fig.  4 ). These are 30 m tall and have an operating volume of approximately 150 m 3 , which offer several operational and control benefits over conventional fermenters, including low shear stress allowing longer (and, therefore, higher quality) hyphae.

The Quorn™ fermentation process. A continuous supply of medium is fed into the fermenter and the broth is harvested simultaneously. The harvested broth is heated to a temperature that destroys proteases but leaves RNAses active, allowing the RNA content of the mycelium to be reduced to less than 2%, which is a regulatory requirement. Once the broth has been heat-treated, the mycelium is spun down to form a paste, which is mixed with binders and flavouring agents before being shaped, cooked and frozen. The supernatant from the paste is currently sent for treatment as wastewater, but active research at Marlow Foods is looking into how the 1.5% solids in the waste can be recovered as a food grade co-product

Glucose can be provided from several sources, currently by enzymatic digestion of starch from wheat or maize using the enzyme glucoamylase from A. niger , but lignocellulose is being considered for the future. As the protein from the original grain is retained for other uses and not fed to F. venenatum , the metric of the amount of protein per hectare, used to compare many animal proteins, is misleading when applied to fungi. The use of protein in animal feeds, whether animals or insects, acts to concentrate (with losses) what is present in the original, whereas fungi add protein to a protein-free feedstock. Furthermore, production of fungal protein is highly water-efficient in comparison to animal protein, requiring about one-tenth that of beef and half that of chicken. Quorn Foods was the first global meat alternative brand to achieve third party certification of its carbon footprint figures [ 40 ]. However, there is considerable potential to reduce the ecological impact of fermented F. venenatum further, and, as such, Marlow Foods are pursuing a research programme involving multiple external partners looking at water reduction, carbohydrate choice, fermenter optimisation, co-product valorisation and more.

Filamentous fungi as biomaterials

Fungi thrive by decomposing and consuming dead plants by breaking down the plant’s cellulose, lignin and other sugars, then rearranging these molecules into their own biomass to grow. Their cell wall is secured by chains of chitin and glucans, which, like cellulose and keratin, are naturally forming polymers found in the toughest of organic tissues. Chitin is the same ingredient that creates the durable and flexible exoskeletons of insects and shellfish [ 41 ]. During the colonisation of substrates, hyphae bind the organic particles together, while degrading them simultaneously. A composite material is obtained, consisting of a bulk of organic substrate bound together by the hyphal network, by inactivating the fungus before the substrate is degraded (e.g. by drying or by heat inactivation). Pure fungal materials are obtained by complete degradation of the substrate or removing the fungal skin from the substrate. Both pure and composite mycelium can be used for different applications [ 42 ].

Stopping fungal growth during colonisation of the substrate results in materials with properties similar to that of expanded polystyrene or other foams (Fig.  5 ; [ 43 , 44 ]). Such materials can be used as packaging material or for heat or acoustic insulation. The properties of both composite and pure mycelium materials are dependent on the substrate, the type of fungus, the growth conditions and post-processing. Heat pressing, for instance, improves the homogeneity, strength and stiffness of the mycelium composite material, shifting its performance from foam-like to cork- and wood-like (Fig.  5 ; [ 45 ]). A range of mycelium composite materials with different properties can be produced [ 46 ]. Future studies should focus on the coating of mycelium materials to reduce water uptake and volatile organic compound escape. The genetic engineering of strains, however, may make coating superfluous.

Material properties that can be achieved with fungal mycelia. a Mycelium composites. b Mycelium textiles. The pictures depicted are reproduced from [ 45 ], which has been published under a Creative Commons Attribution licence (CC BY, http://creativecommons.org/licenses/by/4.0/ )

Mushroom-based materials have the potential for uses in place of leather, textiles and some plastics. The strength of pure mycelium of a wild-type S. commune strain is similar to that of natural materials, such as wood and leather [ 47 ]. By contrast, a material can be obtained by deleting a gene that encodes a protein called hydrophobin, that pearls off water droplets from fruiting bodies, with a strength similar to that of thermoplastics. The difference in the material properties between these strains is explained by a better packing of the hyphae in the deletion strain, resulting in an increased density of the mycelium. In addition, the absence of the hydrophobin in this strain results in a mycelium that is no longer water-repellent, consequently, it absorbs more water and dries more slowly than the wild-type [ 47 ]. Thus, mycelium-based materials can provide desirable mechanical properties which can also be bio-performative, for example, by buffering water availability. Future studies, therefore, should focus on not only the mechanical but also bioactive properties, making mycelium a potential source of new innovative materials with advanced properties. Future studies should also reveal whether mycelia can be utilised to develop materials with properties similar to the material families of the elastomers, composites, metals and ceramics (Fig.  5 ).

The flexibility and aesthetics of mushroom materials were first appreciated by artists and designers who used them to grow living art works [ 48 , 49 ]. Similar to cement and plaster, mycelia will bind, harden and set into a variety of solidified configurations. These designers and artists soon discovered that materials grown from this dense living matrix could be used to make advanced composites, foams and performance plastics, and the commercial potential soon followed. Companies that are designing and engineering mycelium materials include MycoWorks [ 50 ], Ecovative Design [ 51 ], NEFFA [ 52 ] and MOGU [ 53 ], to name but a few. Research programmes at MycoWorks proved that fine mycelium materials can be grown into the texture of sheet urethanes, animal skins and foams, with surfaces that are velvety and fluffy, leathery and rubbery, or beetle-shell brittle and shiny. After a mycelium-based object has been grown, it can be cut, processed and machined like many other materials. Demand for these materials has been driven by concerns for stabilizing the price of supply chains into the future. One of the first markets for which fungal mycelium can provide a solution is the fashion industry, where there is both a consumer and market demand for alternatives to the animal skins and plastics used to make apparel. A sensual and strong material for luxury fashion has been developed by moulding the body of the mycelium into a sheet and processing it in many of the same ways as animal leather (Fig.  6 ).

MycoWorks’ fungal analogues for composites and leather. a Analogues for synthetic wood composites and expanded polystyrene foams. Mushrooms are very sensitive to their surroundings, and it is possible by altering subtle factors to make their tissue express a range of variably determined physical characteristics. While these materials can be grown into building components for construction and interior architectures, they can also be grown with delicately tuneable qualities. The strength, durability and biodegradable nature of mushroom-based materials suggest many ways in which fungi may be used. When the material is processed with traditional industrial wrapping and laminating equipment, it is possible to create functional materials. b Analogues for animal leather. The MycoWorks technology is able to tune fine mycelium leather to have material advantages similar to animal skin, becoming supple, elastic and strong, with excellent return, drape, compression and insulation. This mycelium leather, launched in early 2020 as Reishi™, has been designed as a drop-in material for existing leather processing machine tools, where it can be cured, finished and manufactured using well-honed industrial techniques and formulas

Notably, mushroom-based materials can absorb and dissipate a variety of energetic forces, ranging from sounds to seismic waves to ballistics [ 54 ]. They are naturally flame retardant, good thermal insulators and can be grown as flexible or rigid as one desires [ 50 , 55 ]. While incredibly strong and durable, these materials can readily be broken down into constituent minerals and dispersed easily back into the world. Materials grown through a process of fermentation and decomposition require far less energy, water and other resources than conventional manufacturing.

Filamentous fungi and a wood-based bio-economy

In order to shift away from a fossil-based economy and mitigate climate change triggered by the continuous industrial overproduction of CO 2 , many countries around the world, including those of the European Union, support the transformation to a renewable, bio-based and resource-efficient economy. Wood is considered to play a key role in the efforts towards this goal, due to the large potential of forests and wood products to sequester carbon [ 56 , 57 ]. The term “sustainable” originates from forestry, initially coined by a German mining administrator, Hans Carl von Carlowitz, more than 300 years ago, who oversaw the timber supply of silver mines and realised that regrowth did not match the consumption rates [ 58 ]. Although the concept of “sustainability” has substantially broadened over time, it became a guiding concept in forestry, and recent surveys are demonstrating that European forests are still showing positive net annual increments [ 59 ] and are, therefore, actively storing carbon—a prerequisite for any economy that strives to become net carbon-neutral.

Current wood utilisation includes three major streams: energy, wood products, and pulp and paper, with about 60% going towards materials and 40% into energy in Europe. In this way, about 20% of the total carbon in use is annually sequestered into long-life materials [ 60 ]. However, in order to keep carbon stored within materials for as long as possible, a circular economy will be beneficial, including both recycling and reuse of wood products in a cascading fashion [ 61 ] before a final utilisation for energy services. Since an increased demand for and scarcity of wood can be expected with the transition to a bio-based economy, such resource efficiency will become even more important in the future. While high-quality wood would, therefore, be optimally used as timber for solid products initially, later utilisation steps in a virtual cascade would focus on the inherent fibres, polymers and monomeric wood constituents, such as sugars and lignans. This is where filamentous fungi have their highest potential to assist due to their natural ability of selectively and efficiently degrading and/or modifying all of these major wood constituents.

Due to the natural ability of fungi as heterotrophic organisms to efficiently degrade lignocellulosic plant biomass and convert the constituent sugars into energy-rich molecules, filamentous fungi or fungal enzyme cocktails have a high potential to be employed in the upgrading of wood in biorefinery applications towards second-generation biofuels or chemicals [ 62 ]. Up to now, however, mostly straws are used as substrate for these processes (e.g. the Sunliquid ® process by Clariant), with only a few examples of wood conversion (e.g. bioethanol from Borregaard); wood being much more recalcitrant, due to generally higher lignin contents among other things [ 63 ]. Considering all of the above, an integration of biotechnological processes using fungal platforms into pulp and paper mills which are already performing (chemical-based) wood separation, such as Kraft pulping, can be envisioned as having substantial technological and economic advantages [ 64 ].

Such “wood-based biorefineries” already exist—mostly in Scandinavia and eastern Europe [ 65 ]. Nevertheless, most of these are far from being true multi-product factories. Optimally, biorefineries would integrate biomass conversion processes and equipment to produce bioenergy and a range of products, including pulp, bio-based chemicals (e.g. for food and pharma), biofuels, electricity and heat. Regarding efficiency, the ability to use multiple feedstocks, such as those originating from pulping processes (pulpwood, extracts from effluents, fractions of pulping liquors), by-products from sawmills (e.g. wood shavings), or recovered materials (e.g. recycled paper and wood/fibre waste), would be advantageous. Many of these substrates are unsuitable for chemical upgrading due to their low purities and would have to undergo costly cleaning steps. Filamentous fungi, on the other hand, have the potential to circumvent these problems, since many are surprisingly robust against inhibitors and can selectively transform desired target compounds out of complex mixtures, including lignin-derived molecules [ 66 ].

In summary, wood-based biorefineries can play a central role in the transition towards sustainable economies, with the ability to generate commercial resources with a smaller carbon footprint of resources and emissions than traditionally produced materials (such as concrete). However, which product systems are the most promising ones regarding sustainability and efficiency must be assessed. Filamentous fungi can be of great value in these efforts and help to move more wood from energy into material use and, thus, to higher value creations. Nevertheless, substantial challenges and knowledge gaps can be identified that need to be addressed to be successful in the mid- to long-term:

Research in wood decomposition is lagging behind that for more readily degradable biomass, such as agricultural residues and straws.

Considering the benefits of using recovered (pre-used) wood products, pulp mill side streams and forestry/wood industry residues, the research and adaptation of fungal cell factories should include these to a much greater extent than done today.

Climate change is slowly altering the tree composition, and it can be expected that hardwoods, such as beech and oak, will replace the more drought- and pathogen-sensitive softwoods, such as the currently predominating spruce, in the future. These types of wood differ in their composition, with clear implications for fungal degradation [ 67 , 68 ]. Directed or engineered fungal adaptation and the optimisation of fungal enzyme cocktails will be needed to address these differences in order to optimise utilisation.

Lignin is the most underutilised of all wood polymers due to its complex structure and difficulties in processing. It forms an amorphous network of phenyl propane units coupled to hemicellulose via ester linkages. It is the second most abundant natural material on Earth after cellulose. The worldwide production of lignin is approximately 100 million tons annually [ 69 ]. One of the main challenges of lignin valorisation lies in its diverse structure and poor solubility. Most of the lignin obtained (Kraft lignin/black liquor) is incinerated directly in the pulping plants to recover energy. Only about 2% of the technical lignin globally available (~ 5 million tons) is currently utilised. Basidiomycetes are unique in their vast abilities to modify and/or mineralise lignin, and these organisms should play a central role in developing the utility of this highly promising polyphenol. The potential of lignin as a material reservoir will be achieved through much more rigorous characterisation of the structure/function predictability achieved through these fungal transformations.

Filamentous fungi to mitigate plastic pollution

Plastics are widely used in the global economy because they are inexpensive, versatile, lightweight and durable. Millions of tons of plastics accumulate in the environment annually due to their stability, poor recycling and low circular use, and they represent an ecological threat to nature and human health. At least 350–400 million tons of plastics are being produced annually worldwide [ 70 ], while its production keeps increasing by an average of 7% per annum. In 2014, for example, 311 million tons were produced, among these, 59 million tons in Europe. About 64% of the total European plastic demand is concentrated in five countries: Germany, Italy, France, the United Kingdom and Spain. The main types of plastics are polyethylene, polypropylene, polyvinyl chloride and polystyrene [ 71 ]. In 2014, the plastic waste in Europe amounted to 25.8 million tons that were recycled (30%), subjected to incineration for energy recovery (40%) or sent to landfill (30%). Efficient recycling and incineration are only practiced in a few countries, while in most countries, more than 50% of plastic waste is landfilled [ 71 ]. The latter undergo photo oxidation, degradation and mechanical disruption that often give rise to small fragments, i.e. microplastic particles [ 72 ]. Microplastics, plasticizers and plastic additives as well as added monomers and oligomers can enter surface and/or ground water and accumulate in the environment exerting toxic effects [ 73 ]. Approximately 5 to 13 million tons of plastic waste enters the Earth’s oceans every year, as the intentional or unintentional final destination [ 74 ]. When this waste is ingested by marine animals, it accumulates in the food chain [ 72 ]. The most abundant type of primary microplastics in water environments are fibres [ 75 ] that originate from washing synthetic textiles. Replacing synthetic textiles with mycelium-based textiles can be a means to greatly reduce plastic pollution. In addition, fungal materials could replace plastics in a variety of other applications (see above).

Current plasticizers are used, among other things, to improve the workability of concrete, the construction material most used in the world. Plasticizers enable the use of less water by keeping the same viscosity of the concrete, thereby producing a highly workable material that hardens into a strong final product. Plasticizers are typically not covalently bound to the polymers, which, over time, may lead to environmental contamination via leaching [ 73 ]. A bio-based alternative to the current fossil-based concrete plasticisers would be lignin functionalized to a higher solubility by the action of fungal or bacterial laccases or other oxidative enzymes. The very hydrophobic lignin is insoluble in water and alcohol but soluble in alkaline solutions. Fungal laccases are prevalent in asco- and basidiomycetes and are typically most active at low pH values of 3–5.5, which is a drawback for their use in functionalizing lignin. Accordingly, laccases have to be identified or engineered that have a shifted optimum pH towards an alkaline pH. One recent successful approach at AB Enzymes has been the engineering of a laccase from the ascomycete Melanocarpus   albomyces using a directed evolution approach, which has led to threefold improved kinetics at pH 9.8 [ 76 ]. Notably, the principle applicability of laccase-functionalized lignin has been shown, but immediate commercial implementation is hindered by the general availability and inexpensive cost of fossil-based plasticizers.

To date, very little is known about the biodegradability of petroleum-derived polymers and plastics with fungi and only a few reports are available in the literature. Many basidiomycetes are generally known for their ability to degrade polycyclic aromatic hydrocarbons and are applied in bioremediation processes of contaminated soils and liquids [ 77 , 78 , 79 , 80 ]. So far, plastic-degrading fungi have been successfully isolated from weathered plastic waste obtained from marine habitats and terrestrial waste treatment facilities. The main fungal genera that were isolated and used for lab-scale plastic degradation, primarily of PET and polyurethane so far are: Acremonium , Alternaria , Aspergillus , Aureobasidium , Cladosporium , Debaryomyces , Emericella , Fusarium , Gliocladium , Mucor , Nectria , Neonectria , Penicillium , Phoma, Plectosphaerella , Rhizopus and Trichoderma [ 81 , 82 , 83 , 84 ]. In addition, enzymes produced by basidiomycetes ( Agrocybe aegerita, Auricularia auricular - judae, Bjerkandera adusta, Nematoloma frowardii, Pycnoporus cinnabarinus, Stropharia rugosoannulata ) have been identified and used for plastic degradation. Among the most promising enzymes used for PET and polyurethane treatment have been esterases, cutinases, lipases and lignin-modifying and unspecific macromolecule-depolymerising and hydrolytic enzymes, such as different peroxidases, laccases, glucose oxidases and cytochromes P450. However, no distinct degradation effects for virgin and pre-treated plastics have been achieved solely by the use of pure enzymes [ 70 ]. Hence, currently ongoing research is focused on mycelial-based degradation (whole cell degradation) of pure or mixed cultures in combination with purified enzyme cocktails.

The Janus face of filamentous fungi

The fungal kingdom is huge: six million species are estimated to exist on Earth [ 85 ], of which only a small proportion is known. Most are saprophytes and feed on dead plants and animals, and only a few dozen are exploited in biotechnology as cell factories. Leveraging their metabolic capacities and flexibilities will be key to achieving the circular economy outlined above (Fig.  2 ).

However, many filamentous fungi pose a threat to plant, animal and human life. As comprehensively covered in the first white paper of the EUROFUNG consortium published in 2016 [ 4 ], a multitude of plant-pathogenic fungi destroy staple crops annually, which would be enough to feed 600 million people [ 86 ]. Human-pathogenic fungi compromise the life of 1.2 billion people worldwide and kill about 1.5–2 million annually, exceeding the fatalities caused by malaria or tuberculosis [ 87 ]. Regarding more information on the huge impact of fungal pathogens on human health and food security and the related challenges, the interested reader is referred to the EUROFUNGs white paper from 2016 [ 4 ], a review covering the top 10 most feared fungi published in 2018 [ 88 ] and to the One Health report published by the American Academy of Microbiology in 2019 [ 89 ].

Meeting the agricultural production required to feed the increasing world population [ 90 ] is likely to be made more difficult by climate change. This problem is exacerbated by a wide range of plant diseases caused by the diverse group of fungal and newly emerging pathogens. A list of the ten most important fungal pathogens of plants published in 2012 [ 91 ] is outdated. If this list had been collated more recently it would have undoubtedly also included the basidiomycete Phakopsora pachyrhizi , which causes Asian soybean rust and threatens the sustainability of the millions of hectares of soybean grown in Brazil each year, despite only being detected in Brazil for the first time in 2001 [ 92 ]. Soybean production is now only viable in Brazil because of the availability of effective fungal control agents. However, even with the solutions available from the agricultural industry, crop losses remain high in soybean and a wide range of other crops [ 93 ]. In many cases, the losses seen are due to fungi that have developed resistance to existing control strategies [ 94 , 95 , 96 ]. The speed with which many species of fungi evolve and become resistant to existing control strategies means that there is a constant need to identify agents with novel modes of action. However, the development of novel fungicides by companies active in this field, such as Bayer Crop Science and Syngenta, requires a huge effort and major investment (Fig.  7 ), taking more than 10 years from the identification of a new screening ‘hit’ to market [ 97 ].

From science to market—the fungicide discovery and approval pipeline. Historically, the biological activity profile (fungicidal potency and spectrum) was used to filter hit compounds to lead molecules. Nowadays, regulatory requirements regarding human and ecotoxicological safety mean that proxy assays for these regulatory requirements are now utilised as early in the discovery pipeline as possible as part of the selection criteria for compound progression. It will typically take 10–12 years for a new fungicide to pass through the various stages of research (lead generation, early and late lead evaluation, optimisation and candidate confirmation) before promotion into evaluation and, finally, product development. After reaching the market, a significant amount of investment is still required for product lifecycle management (e.g. product monitoring feeds into improvements in formulation)

The screening of hundreds of thousands of random chemical inputs has historically produced a very low yield of hits, therefore, hypothesis-driven input selection methods are increasingly being utilised. However, the current target assessment and prioritization approaches are still heavily reliant on information from ‘model’ fungal systems, such as S. cerevisiae and Neurospora crassa , rather than on good quality data produced from fungal pathogens that result in the highest crop losses. It is remarkable that almost all plant pathogens diverged from the fungal ‘model’ systems millions of years ago and have very diverse modes of pathogenicity [ 98 , 99 , 100 ]. Therefore, the reliance on these classic ‘model’ systems is deeply unsatisfactory and a significantly better understanding of the genetics and metabolism of fungal plant pathogens is likely to help deliver novel fungal control strategies. In this respect, it is worth mentioning that the EUROFUNGs white paper from 2016 stressed that the term ‘model’ is misleading and advocated a shift in focus towards ‘reference’ strains [ 4 ].

Some ubiquitous filamentous fungi which are not pathogenic and usually thrive on dead plant material are still dangerous for human health when consuming crop-based products. Filamentous fungi produce a plethora of so-called secondary metabolites (SMs), of which a subset, the mycotoxins, are harmful. These can be produced by fungi growing either on the crops, some without disturbing plant growth, or post-harvest. The SMs are synthesised by accessory biosynthetic pathways thought to be important for a range of processes, including spore development, spore survival and fungal communication. Recent advances promote the view that fungal mycotoxins are fitness factors that help to establish a niche for protection from competing microbes [ 101 , 102 ].

Mycotoxins are toxic, carcinogenic and/or mutagenic fungal SMs and can be present in high concentrations in food and feed products. Mycotoxins have significant economic effects on a wide number of crops, most commonly but not exclusively contaminating grain crops (maize, wheat, barley), peanuts and tree nuts, beets, grapes and coffee. The Food and Agriculture Organization has estimated that 25% of the world’s crops are contaminated with unacceptable levels of mycotoxins. Economic losses are not easily quantified, but based on the reports that exist, losses are staggering. Aflatoxin, for example, is the most problematic mycotoxin, with estimated annual losses due to contamination ranging from $52 million to $1.68 billion regarding the US corn crop alone [ 103 ]. A relatively small number of ascomycete genera— Aspergillus, Penicillium, Fusarium, Claviceps and Alternaria —produce most of the common mycotoxins. Major mycotoxins include aflatoxins, citrinin, cyclopiazonic acid, ergot alkaloids, fumonisins, ochratoxin, patulin, trichothecenes and zearalenone [ 104 ]. All these metabolites have adverse effects on vertebrate cells depending on the amount ingested and the age and sex of the consumer. Aflatoxin B 1 is the mycotoxin most studied and the mechanism by which it causes liver cancer is well understood [ 105 ]. Trichothecenes, which interfere with protein synthesis in animals and humans, are common contaminates of wheat and a huge problem across all wheat-growing regions in the world. Patulin is of especial concern as it is frequently found in apples and products derived from apples that are popular children’s foods, such as apple sauce and juice [ 104 ]. It has also been proposed that SMs cause adverse human health effects after exposure by inhalation (e.g. exposure to moulds indoors; [ 106 ]).

Although the molecular genetics of many mycotoxins is well understood [ 104 ], efforts to control the contamination of food and feed with these small molecules is still to be realised. Aside from the challenges of controlling mycotoxin contamination using traditional efforts, such as breeding plants for resistance and/or detoxifying contaminated products, several emerging issues have complicated the ability to tackle the mycotoxin problem. Four significant issues to be considered in relation to mycotoxin amelioration are: (i) masked mycotoxins, (ii) climate change, (iii) emerging mycotoxins and (iv) co-occurrence of mycotoxins. Masked mycotoxins refer to plant derivatives of fungal mycotoxins. There is concern and data to support that these hybrid molecules can contribute to toxicity either directly or indirectly through the release of the parent mycotoxin in animal digestive tracks [ 107 ]. Research groups worldwide are now examining how climate change might impact mycotoxin synthesis for diverse fungi and crops. Several studies suggest increases in mycotoxin contamination although predictive modelling approaches need to be refined [ 108 , 109 , 110 ]. Emerging mycotoxins refers to toxic molecules that occur frequently in products but are not consistently monitored (e.g. eniatins, alternariol, emodin and sterigmatocystin; [ 111 ]). The frequency and occurrence of these less common mycotoxins should ideally be subject to risk assessment. Finally, many mycotoxins co-occur in the same crop, yet, worldwide regulations only consider the toxicology of a single mycotoxin [ 112 ]. Little is known whether combinations of mycotoxins affect the consumer in an antagonistic, additive or synergistic manner. Despite the wealth of published works on the topic of mycotoxins, substantial gaps remain in the knowledge of how to reduce contamination, what quantities are important in health, whether co-occurrence is a topic to consider and how climate change will impact future risks of these fungal metabolites.

Measures to advance science on filamentous fungi

Exploiting and fighting fungi for a sustainable, resource-saving bio-economy that operates in a circular manner demands considerable improvements in research and development on scientific and community levels:

Science: (i) improved understanding of fungal biology in a diverse set of species and strains, including lab (reference) strains, industrially exploited strains and ecologically relevant species; (ii) improved high-throughput technologies and tools to study the dynamics of fungal growth, product formation and pathogenicity and their adaptation to changing environmental conditions; (iii) investment in the standardisation of measures, methods and automation tools; and (iv) investment in open access infrastructures for the storage, analysis and reuse of systems biology data, including biofoundries.

Community: (i) overcoming fragmentation of the research community studying filamentous fungi; (ii) increasing the number of scientists devoting their lifetime to the study of fungi; (iii) the improved intersectoral and interdisciplinary education and training at undergraduate and graduate levels; and (iv) the engagement of scientists and educators in public outreach and the dissemination and engagement of the general public in fungal science.

The science on filamentous fungi has always been lagging behind that on many other microorganisms, such as yeast and bacteria. Four main reasons for this delay can be specified: firstly, genetic transformation of filamentous fungi is less efficient and more time-consuming compared to unicellular bacteria and yeast [ 113 ]. Secondly, genome sequences of filamentous fungi only became available a decade after the genomes of the ‘model’ unicellular fungus S. cerevisiae (1996; [ 114 ]) and the ‘model’ bacterium E. coli (1997; [ 115 ]) were released to the public. Thirdly, the genomes of filamentous fungi contain far more genes. A filamentous fungal genome usually carries between 9000 and 14,000 genes, whereas E. coli has about 4000 and S. cerevisiae about 6000 genes. Finally, there are a few million fungal species, consequently, the research communities studying a particular filamentous fungus are considerably small and fragmented. Each filamentous fungus is an individual of its own, different from all the other fungi, although its outlook is covered by the universal filamentous form. One of the largest of these scientific communities, studying A. niger , consists of only about 30 research labs worldwide [ 2 ], whereas more than 1800 research labs studying S. cerevisiae are registered at the Saccharomyces Genome Database [ 116 ].

Some substantial measures for basic and applied science on filamentous fungi are highlighted in the following. These need to be addressed in the short- to mid-term in order to fully leverage the power of these microorganisms for a circular economy.

Community engagement and sustainable infrastructures for high-quality fungal omics data

Biology has entered the digital age in many ways, most obviously with the rapid growth of large datasets, including genomes, transcriptomes, proteomes, phenomes and metabolomes. The exploitation of these data underpins the future of biology but requires effective infrastructure, supported and facilitated by easily accessible online technologies delivering high-value analyses. However, this revolution presents major technological challenges and resource implications. This is particularly stark for the fungal research community, due to the diversity of both the organisms and fields of research. On top of this, a vast amount of biological information and data is not in a searchable, digital form but is buried throughout our rich and varied literature.

Omics data is very cost-effective, but its utility is largely dependent on making the data accessible and usable for the wider research community. Every effort should be made to develop the future of genomics-driven fungal research in order that everyone can have full access to reliable genomic information. Consequently, accessing and representing diverse aspects of biology, ranging from phenotypes and subcellular localisation to protein interactions or the geographical location of isolates, in a searchable format is not easily achieved or readily automated. Therefore, a lot of the work required is time-consuming and requires dedicated personnel with biological expertise and information technology skills to apply appropriate classifications and ontologies. New solutions are needed for a large amount of multispecies omics data comparison. Addressing these issues requires co-ordination between researchers, publishers and databases, so that, at the time of publication, key information is deposited in a logical way within these resources. This will require the community to act proactively to facilitate the integration of data, consistency in the use of terminology and gene names, and journals to consider these issues when defining their requirements for data deposition and developing practices to facilitate text mining [ 117 ].

Fungal genome sequences are hosted at several fungal databases (e.g. FungiDB [ 118 ], JGI MycoCosm [ 119 ], Ensembl Fungi [ 120 ]), many more genomes are deposited in the NCBI, ENA or DDBJ, and some are hosted by private resources. Free, open access to data is generally accepted as a key priority, which, consequently, requires significant long-term funding for well maintained and resourced, actively developing databases to serve their user communities. Many funding organisations have contributed to setting up and maintaining bioinformatic resources, but the required long-term commitment is difficult to obtain, and with an inevitable cliff edge each time renewal of funding is sought, the reality is that key resources have been lost and will be lost in the future. A consequence of the funding issues is a general drive to centralise, with a number of organism-specific resources having been closed down or frozen (e.g. AspGD), due to a loss of funding. How to address this global issue is not easy and needs both engagement and commitment from funding agencies and/or governments, alongside concerted efforts from the community to shape policy. Alternative funding models must also be considered. However, most proposals potentially endanger the ethos of full open access.

Early genome sequences were limited in quality due mainly to the sequencing technologies used; more recent higher quality and better assembled genome sequences suffer from poor annotation. In the early days, consortia were formed to improve upon the automated gene calling and annotation by manual curation and verification of genes. Nowadays, very few genomes make it past the initial draft annotation [ 4 ]. Consequently, errors are likely to occur, and these will be perpetuated and propagated in the annotation of other genomes, influencing downstream experimental analysis. Additionally, comparison of outputs from different bioinformatic pipelines using the same datasets will generally not give exactly the same interpretation—the outputs represent probabilities and models are, thus, not absolute. The resulting differences inevitably complicate the comparison of genomes and other types of data. Lack of awareness or technical understanding can also lead to the misinterpretation and misinformation of any programme or bioinformatic output. This underlines the need for improved training and development, not only for dedicated bioinformaticians but even more for biologists in general. Currently, there is a serious lack of qualified personnel with adequate formal training. Consequently, as a minimum, there is a need for appropriate training regarding accepted rules for omics data handling, enabling fungal biologists to efficiently contribute their domain-specific expertise using ontologies, terminologies and platforms that minimise the need for professional curation and maximise value to the broader user community.

Databases, such as FungiDB and MycoCosm, have a challenge in choosing the organisms and datasets they should host and on where the particular reference genomes, their limited annotation and curation efforts should focus. Consequently, the active engagement with and involvement of researchers is essential in making these decisions. Most researchers inevitably regard their own work as a priority and there is a danger that those with the ‘sharpest elbows’ will prevail. Therefore, it is important that communities ensure they are well represented and informed of decisions, proposed priorities and opportunities.

Opportunities and challenges in getting value from fungal omics data

The integration of databases provides opportunities for shared resources and infrastructures which have inevitable cost-savings. ClinEpiDB [ 121 , 122 ], for example, accommodates a diverse range of large-scale epidemiological datasets for enteric, respiratory and parasitic pathogens but no fungal data. The common infrastructure used by ClinEpiDB and FungiDB could potentially facilitate the integration of epidemiological datasets for fungal pathogens and, as such, allow room for innovation. These resources, allowing the mapping of disease and genomic data, could be potentially developed more broadly for fungal researchers interested in plant disease, ecology and the migration of key pathogens or drug resistance. Additionally, this may facilitate new developments to include the characterisation of transcriptional changes associated with host pathogen interactions [ 123 ] or between microbiomes [ 124 ].

A major challenge for both the database providers and their users is that data is not static nor is the interpretation of data generally limited to a single definitive result. Genomes, for example, may be re-sequenced using new technologies and additional datasets, such as proteomics or RNA-Seq, are constantly being added. These changes and additions, alongside modifications to bioinformatic pipelines, will facilitate improved gene calling and annotation. Consequently, gene models may be removed or split, misannotated open reading frames revised, differential splicing revealed and untranslated regions, defining the transcription start sites and mature 3′ ends, clarified. These changes can result in difficulties. It is challenging, for example, to map all N. crassa knockout mutants to the current gene models hosted by different databases. Researchers, therefore, need to be aware that genomes are not set in stone but evolve with data and technical advances and the databases must make these changes clear and traceable. Updating repository records is paramount to ensure that all databases have access to the most recent information and, as such, provide the best possible service to their users.

Another confounding factor when harnessing genome data of filamentous fungi is the fact that between 40 and 50% of the genes in a filamentous fungal genome are ‘hypothetical’ and lack functional predictions [ 4 ]. In addition, only 2–10% of the genes with predicted functions have been experimentally studied in filamentous fungi and, thus, have a verified function [ 125 , 126 ]. This renders in silico reconstructed genome-wide metabolic models considerably incomplete, i.e. with gaps, dead-end reactions and dead-end metabolites. Even for the ‘model’ yeast S. cerevisiae , 21% of its predicted genes (i.e. about 1400 genes) still have dubious functional predictions in 2019, which is 22 years after the release of its genome sequence and despite a research community with more 1800 labs worldwide [ 127 ]. One powerful approach to overcome this limitation is the interrogation of gene expression networks based on hundreds of transcriptomic datasets available for filamentous fungi. This wealth of data has recently been harnessed to improve gene annotations and assign gene function predictions to 65% of genes predicted in the A. niger genome [ 128 ], which now can be accessed at FungiDB.

Notably, ecologically important pathogens and endophytic fungi have traditionally received very little support due to the absence of dedicated funding. Those managing the bioinformatics resource need to work with the researchers to define the types of data and criteria that apply when selecting datasets for inclusion so that the process is rigorous, not biased or arbitrary, and that appropriate computational workflows are developed in a timely way. This will also help the researchers to consider relevant criteria when designing their experiments and specific initiatives may be developed to address particular problems. A key issue in confirming gene models, for example, is identification of the 5’ and 3’ ends of transcripts. Most RNA-Seq approaches are poor at defining transcript ends, but specific approaches can be applied that address this directly. In this way, the structural annotation of key genomes could be significantly improved for the wider community. Awareness of such issues may help the research community optimise the utility of their data.

Other on-line resources provide bioinformatic pipelines to assist and minimise the expertise required. One example is Companion [ 129 ], which provides a genome production pipeline. The most recent version has proved effective in the production of usable fungal genomes based on the de novo analysis of 25 previously characterised reference genomes. The future development and continued support of this and other such initiatives should help the lab-based researchers complete a good first draft of a genome annotation efficiently without major resource implications. Furthermore, recent developments in the Apollo pipeline [ 130 ], integrated within databases such as VEuPathDB [ 131 ], allows researchers to inspect and modify genome features efficiently and can also be utilised in private workspaces. Data can remain private to a specific work group and be transferred to public databases when appropriate. Consequently, the research community can fully engage to develop shared resources. As these contributions can be attributed to their author, this can be acknowledged by the community and potentially lead to micropublications, incentivising participation and rewarding those who contribute.

Last but not least, the rapid evolution of sequencing and other high-throughput technologies requires active discussions to improve policies and infrastructure for data sharing, integration and analysis in order to resolve the bottlenecks and challenges faced. The changes will not happen overnight, but effective communication should involve all parties concerned, including the on-line database providers, researchers, funding agencies and policymakers. In order for these amazing resources to be used to their full potential for high impact research, these different parties must actively strive to ensure we have effective data sharing policies that will ultimately result in long-term sustainability, accessibility and utility.

Improved tools and technologies to study fungal biology

The molecular and analytical toolkit available for filamentous fungi has improved substantially during the 4 years since the first white paper was published in 2016 [ 4 ], demonstrating the high innovation potential of the filamentous fungal research community. Genome editing via CRISPR has become routine and is nowadays run in a multiplex manner for many reference strains [ 132 , 133 ]. Fourier-transformed infrared spectroscopy has become a next-generation phenotyping technology to identify fungal strains and their metabolic products [ 134 ], while the power of surface analysis tools, such as mass spectrometry-based imaging techniques, has been exploited to identify changes that are brought about by filamentous fungi and their enzymes when they attack and modify insoluble lignocellulose materials [ 135 ]. “Ready-to-use-microfluidics” have been implemented to study the dynamics of fungal growth and fungal interaction with soil bacteria [ 136 ], X-ray microcomputed tomography has been successfully applied to study spatial distribution of hyphae within mycelia and diffuse mass transport therein [ 137 , 138 ], and gene and protein networks have been developed to assist our understanding of filamentous fungal biology holistically [ 128 , 139 ].

Hence, the technological foundation is sound and powerful and no longer represents a critical hurdle for gaining knowledge on fungal biology. As of now, high-throughput methods for automated cloning, cultivation, mining and screening of fungal strains are, unfortunately, not yet state-of-the-art.

Standardisation as a driver for engineering fungal biology

Standardisation is a key for success. Standard reference methods, measures, substrates and materials ensure reliable fabrication and reproducible production characteristic of advanced global industries, such as the semiconductor and forest products industries [ 140 , 141 , 142 ]. Engineering biology, broadly called synthetic biology, is similarly benefitting from standardisation [ 143 , 144 ]. The mycelium materials research community and industry are already benefitting from ASTM and ISO standards developed for products in the forest, textile and acoustic absorbance industries. Accelerating the development and adoption of standard reference materials, measures and methods across the mycelial materials community and beyond could enable advancements in basic science on filamentous fungi and engineering of fungal-based bioproduction analogous to the advancements in the semiconductor industry. New options for sharing mycelium materials that reduce transaction costs, allow for redistribution and enable commercial production could provide the incentive for the broad adoption of standard references, measures and methods that support reliable fabrication and reproducible science [ 145 , 146 ].

Reliable, standardised measurements are critical to reproducible science [ 147 ]. Metrology formalizes reliable measurements, and advances in metrology for biological systems are greatly benefitting basic science and the engineering of biology [ 148 , 149 ]. Characterising organisms, substrates and production environments in real-time across orders of magnitude in the scale of space and time could accelerate the development of advanced fungal products while, simultaneously, benefitting basic science and the engineering of biological systems. As an example, standard metrological materials, such as well-characterised 1 cm 3 units of mycelium material, and metrological methods could help to co-ordinate practitioners across locations and resource settings, reduce manufacturing costs, reduce time to market and disseminate best practices [ 150 ].

The confluence of standard approaches and automation tools characteristic of modern semiconductor foundries have cultivated the semiconductor industry into one of the most significant in human history. Recent advancements in the automated construction of synthetic organisms are making biofoundries economically viable and they are increasingly being recognized by the life sciences research community as an essential infrastructure for basic and applied research [ 151 , 152 ]. Companies such as Gingko Bioworks and Ecovative Design have invested substantial capital in biofoundries for constructing organisms and there is an emerging ecosystem of “cloud laboratories” that aim to provide practitioners access to automated labs without the need for large capital investments [ 153 , 154 , 155 , 156 ]. Biofoundries with capacities for constructing filamentous fungal organisms could build on recent work to synthesize the genomes of the bacteria E. coli, Caulobacter crescentus and the yeast S. cerevisiae , with the ultimate goal of enabling advancements in synthetic filamentous fungal organism construction analogous to the construction of JCVI-sc3.0 and Sc2.0 [ 157 , 158 , 159 , 160 ]. Work to insulate biological systems from evolutionary drift can enable the deployment of engineered biology for applications outside the research laboratory or commercial bioreactors, while mitigating biosafety and biosecurity risks [ 161 ]. Such is the promise of distributed fungal-based bioproduction that biofoundries, for example, for mycelium materials in support of basic science, engineering and production, strategically placed across the planet, could ramify into impacts surpassing that of the semiconductor industry.

Concluding remarks

We would not be able to live the life we are living without the help of moulds and mushrooms from nature. Fungi are our present and they will shape our future. They are champions in recycling and material transformation; their biosynthetic capacities are unmatched in the microbial world. We should do our best to harness their abilities! Fundamental and applied science on fungi offers solutions for the shift from our current petroleum-based economy into a bio-based circular economy, opens new avenues for food security as demand increases from a growing human population, and provides us with new concepts on how to ensure human, animal and plant health in the future. Science on fungi discussed in this white paper already contributes to 10 out of 17 UN development goals (Fig.  8 ) and their role will become even more important for the future of mankind.

Fungal biotechnology has the potential to make a significant contribution meeting 10 out of 17 United Nation’s sustainable development goals through the rational improvement of filamentous fungal cell factories

The economist Peter Drucker (1909–2005) once said: “The best way to predict your future is to create it.” Stronger mutual collaborations between scientists, engineers, artists, designers and industrial stakeholders, and vivid communication with the general public and policymakers will ensure that the inter- and transdisciplinary science on fungi will create a path towards innovative breakthroughs. As Peter Drucker suggests, this will create a sustainable economic future based on fungal cell factories for years to come.

Availability of data and materials

Not applicable.

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Protein digestibility corrected amino acid (PDCAA) scores.

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Meyer, V., Basenko, E.Y., Benz, J.P. et al. Growing a circular economy with fungal biotechnology: a white paper. Fungal Biol Biotechnol 7 , 5 (2020). https://doi.org/10.1186/s40694-020-00095-z

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Green Biotechnology: Ensuring the sustainability of the science

September 13, 2021

Liam Hopkins

When you look back at the history of scientific progress, you will see that it has always been driven by the need to solve problems.

Despite all the progress that has been made in the past century, we are still facing the same problems. So, what can scientists do to ensure the sustainability of their research?

The answer lies in the field of green biotechnology.

By focusing on the environmental impact of biotechnology, researchers are developing new ways to make biotechnology more sustainable. We will look at how green biotechnology promotes the sustainability of our food, agriculture, and healthcare systems.

What is Biotechnology

Biotechnology is a branch of applied science that focuses on exploiting biomolecular, genetic, and cellular systems and using living organisms to manipulate or develop products and technologies beneficial to human life. Some scientifically developed products using biotechnology include genetically altered food, hormones, healthcare-focused products, etc. 

Though biotechnology is flourishing in the healthcare and pharmaceutical sectors, organic food industries, genomic, and biofuel sectors are also adopting this thriving scientific technology for better production. 

Understanding biotechnology better:

• This specific discipline focuses on analysing the evaluation process and characteristics of living organisms at the molecular level. It involves chemistry, physics, biology, statistics, and different technologies to evaluate microorganism life-cycle and functionalities. 
• This flourishing technology strives to develop medicines and therapeutics to increase the human lifespan. 
• Though a promising sector to improve our quality of life, investing in biotechnology is prone to risks. It may need a lot of money to initiate and complete research on a specific product, say medicine, but the success rate may be low, or you may never get expected results. 

History of Biotechnology

Since the invention of civilisation, biotechnology has been present. From the Egyptians using yeast for baking bread to the Chinese using fermentation for brewing: all are anyway the utilisation of this technology. Though today’s advanced biotechnology got invented in the late 19th century, classic biotechnology was there even before our knowledge about chromosomes, DNA, and genes. 

Agricultural and food sectors started utilising this emerging technology at the commence of the 20th century. In the mid-century, after the revolution in genetic science research and the invention of the manipulation method of DNA, biotechnological research started thriving. DNA sequencing has become a common practice in manipulating different microorganisms and utilising them and their derivatives in inventing new medicines, crops, and therapeutics. 

Some milestones this flouring technology achieved include:

• In 1919 : The term ‘biotechnology’ was first invented by Karl Ereky , a Hungarian agricultural engineer.
• In 1928: Alexander Fleming , a Scottish scientist, and bacteriologist used biotechnology for developing penicillin , a life-saving antibiotic. 
• In 1943: Oswald Theodore Avery , a Canadian-American medical scientist, invented DNA that encodes all genetic information of organisms for living, development, and reproducibility.
• In 1995: The double helix pattern of DNA was invented by Francis Crick and James Watson .
• In 1969: In Vitro Synthesis of Enzymes was made possible. Again, thanks to biotechnology.
• In 1983: Genetic engineers developed the world’s first genetically engineered crop plant, and it was tobacco .
• In 1997: The engineers were able to present Dolly , a sheep, as the world’s first mammal cloned from mammary gland cells. 
• In 2001: The International Human Genome Sequencing Consortium published the first draft of the human genome sequencing.
• In 2010: The J. Craig Venter Institute created the world’s first artificial/synthetic cell.
• In 2013: The scientists of the USA developed the first bionic eye. 
• In 2020: Biotechnology facilitates the vaccine development process against the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2).

Biotechnology Types

Red biotechnology (bio-pharmaceutical).

This branch of biotechnology concentrates on improving the human lifespan through developing new drugs, regenerative therapies, vaccines, molecular diagnostics techniques, antibiotics, and artificial organs using genetic manipulation. 

It aims to evaluate how cell proteins function and how diseases attack cells when cell proteins cannot function properly. 

White biotechnology

A branch of biotechnology solely concentrates on improving industrial production of bio-products like biofuels, detergents, chemicals, etc., manipulating molds, fungus, bio-catalysts (enzymes), and more through biological processes. 

Blue Biotechnology

It is the technique of utilising molecular biology for exploiting living aquatic organisms and their derivatives to develop essential goods, knowledge, and services. 

Yellow Biotechnology

It is the oldest branch of biotechnology that aims to study insects, their cells, and different microorganisms to produce various ingredients for developing more nutritious foods and use them for industrial purposes. An example of this technology is the research run on developing cooking oil low in saturated fat. 

Green Biotechnology

Green or agricultural biotechnology, also termed agritech, is a thriving field of biotechnology that facilitates the analysis and invention of genetically altered plants, crops, animals, and transgenic plants and improves their production and nutritional quality, evading long-term environmental damage.

It highlights the method of implanting foreign genes into economically valuable plants, exploiting microorganisms to develop genetically modified fertile and resistant seeds, and increasing agricultural production. 

Today, around 13+ million farmers use green biotechnology techniques for fighting off pests and producing plants with high yields throughout the world. 

Green Biotechnology: Benefits, Tools and Applications

Over the last few decades, green biotechnology has brought about radical changes in the agricultural and environmental sectors. It is a scientifically proven method of improving food production and promoting the environment and economic sustainability. 

Let’s have a look at the tools and applications of this flourishing technology. 

Green biotechnology has revolutionised the research on new vaccine development. It is especially beneficial for under-developed and developing countries where mass vaccination can be highly expensive. 

After relentless efforts, genetic engineers could invent genetically modified plants and crops that carry antigenic proteins from pathogens. These antigens get activated and develop immunity against the infectious viruses once ingested. 

An excellent implementation of green biotechnology in developing vaccines is the anti-lymphoma vaccine, a vaccine against cancer. This vaccine is derived from tobacco. In this process, the plant is genetically engineered to carry RNA extracted from malignant B-cells. When injected into the body, this vaccine can develop an immune response against cancer in the patient’s body. 

Bio-fertiliser

Green biotechnology was the cornerstone in developing bio-fertiliser, an essential element to ensure quality and quantity production in the agricultural sector. 

Artificial multiplication of microorganisms such as blue/green algae, fungi, bacteria, etc., is the principle of bio-fertiliser. 

They can boost plant production by improving microbial processes in the soil. When applied, they colonise the rhizosphere of the crops and plants and ensure fast growth. 

Bio-fertilisers are essential for better photophosphorylation, nitrogen fixation, and making more nutrients available in the soil. 

Rhizobium is suitable for leguminous crops, while azotobacter works well in boosting the growth of grains like cotton, maize wheat, potato, mustard, and other vegetables. 

These fertilisers are environment-friendly, improve soil condition, and do not overload the soil ecosystem. 

Plant and Animal Reproduction

Inventing the genetic modification processes for plant and animal reproduction is another cutting-edge success of green biotechnology. 

This branch of biotechnology has facilitated animal and plant reproduction through enhancing their growth, boosting feed and nutrition utilisation, and decreasing waste via more effective resource utilisation. 

The traditional breeding approaches like cross-breeding, wide crosses and embryo rescue, cross-pollination, translocation breeding, etc., require a lot of time. Technologies such as embryo transfer, artificial insemination, etc., are proven to be very successful in manipulating and regulating plant and animal breeding and getting expected results. Thanks to agritech that has driven genetic engineers to invent these advanced genome-based breeding methods. 

These processes can be started at the molecular level and involve removing, overexpressing, or introducing new genes in the hosting plant or animal. 

Green biotech has helped researchers invent these efficient ways of boosting food production, especially in developing and underdeveloped countries, and ensure agricultural sustainability. 

Antibiotics

Green biotechnology also facilitates various effective yet less-risky antibiotic production from plants and microorganisms. They are suitable for both animals and humans. 

These antibiotics are sometimes developed by manipulating fungi or other pathogens to get benefits over bacterial growth. They are orally fed, which means you can add them to an animal’s food directly. Hence, unlike traditional antibiotics, they are cost-effective yet equally efficient. 

As these antibiotics are plant-based, they can be produced in a large amount at a time. Unlike mammalian cell antibiotics, these antibiotics are less risky and easy to purify. But their overdose can lead to the development of antibiotic-resistant bacterial strains. 

Molecular Breeding

Molecular breeding is the latest trend among biotech industries for producing different hybrid plants. It is the process of locating the genome regions linked to desirable phenotypic traits under water stress and then combining genes to invent new varieties and hybrids. Molecular breeding is an indispensable tool of modern green biotech for enhancing the quality of various crop species. 

Based on the genotypic assays, this DNA marker-assisted technique manipulates genes and, by identifying and improving the traits of interest in plants, it boosts commercial plant hybridisation. 

The plants and crops developed in this process are of high quality, resistant to severe climate conditions, and thus ensures agricultural sustainability. 

Genetic engineers and scientists are adopting and utilising green biotechnology for improving human health and increasing their lifespan. 

They are trying to develop genetically manipulated food and crops high in nutritional value to help humans develop immunity and combat different diseases. Success in this attempt is Golden Rice, a beta carotene-rich synthesised rice as an excellent source of vitamin A, a lack of which can lead to irreversible blindness. This fortified rice is developed by cloning three genes: one bacterium gene and two daffodil genes. 

As golden rice contains these bio-synthesized beta-carotene genes, it can successfully fulfill the deficiency of essential nutrients and vitamins, especially in people who eat rice as their staple food. 

On the flip side, their parental strains can only synthesise beta-carotene naturally in the photosynthetic parts. 

Biofuel is another achievement of green biotechnology that works great to reduce the carbon footprint of the earth. Biotechnology is the basis of advanced biofuels that work like gasoline but offer better fuel economy and conversion rates. 

These biofuels are obtained from renewable feedstocks and will meet approximately one-fourth of the world’s demand for transport fuels by 2050. 

These next-gen feedstocks comprise wooden waste and biomass, plant residuals, algae, municipal wastes, etc. 

Pesticide-resistant Crops

Pesticides can pollute water, soil, turf and harm the overall cultivation environment. Besides killing bugs or weeds, these chemically formed products can be lethal to other living organisms like fish, birds, beneficial beetles, and non-target crops. 

The use of pesticides can significantly decrease/kill pollinators like bees that can drastically reduce crop production. Thanks to green biotechnology, which has enabled scientists to invent genetically modified plants and crops that are resistant to pesticides. So the farmers can specifically eliminate harmful pests without causing any adverse effects on other beneficial insects, plants, and the environment. 

The first herbicide-tolerant crop was soybeans. 

Pest-resistant Crops

Genetically modified (GM) pest-resistant crops are specially engineered crops resistant to specific pests. These pests are harmful to optimal plant growth. 

The foreign gene introduced in the host seed to be pest-resistant was first found in a bacteria (Bacillus thuringiensis). Hence it is called a Bt crop. Some of the commercially produced Bt crops are corn, potatoes, and cotton. 

The principle of this technology is to infuse a protein in essential crops that is resistant to harmful insects but harmless for the human body. 

Green technology does not focus on attaining agricultural sustainability and producing more food and crops for combatting nutrition deficiency and starvation only; it also aims to utilise genetic processes like gene removal, transfer, and identification to manipulate floral traits: shape, scent, nectar, colour, etc. 

These technologies focus on developing new varieties and improving flower production using micropropagation, self-/cross-pollination, mutation, tissue culture, etc. 

With the help of agriculture biotechnology, genetic engineers could invent 50 new breeds of ornamental and floral plants by now. 

Micropropagation

It is a biotechnical process that employs cell culture and tissue culture to produce vegetated plants and crops. In this artificial propagation technique, a large number of plants can be propagated using asexual means, and every plant is genetically indistinguishable from the original plant. This process is excellent for multiplying the number of endangered plant species. 

Bioremediation

Over the last few decades, environmental pollution has become an alarming topic. It is the outcome of unplanned industrialisation, unsafe agricultural practices, and more. 

Thanks to green biotechnology that has enabled scientists to invent bioremediation, the process of using living organisms, like bacteria and microbes, to exclude toxins and pollutants from the environment. It is a natural cleaning method that promotes the biodegradation of contaminants into non-toxic/less-toxic elements. 

The aim of using the microorganisms is that they break down or immobilise the chemical-based toxins by extracting energy from them to live. Bioremediation is becoming more popular and adopted worldwide because of its eco-friendly features.

• It is a natural process with no or fewer environmental effects.
• Most applications are in-suite that require no transportation.
• Kills pollutants from groundwater and soil in no time
• It is a cost-effective approach to detoxify, mineralise or degrade contaminants from the environment. 

Biotic Strain Confrontation

Green biotechnology has revolutionised the cultivation process by enabling scientists to manipulate plant genes to withstand severe weather conditions, like drought, salinity, etc. 

Genetic engineers could invent biotic strain confrontation, the process of detecting genes that enables plants to grow and endure highly saline environments. The theory behind this process is the up or down-regulation of records that help researchers alter the genetic traits in plants to make them drought-resistant. 

It is undoubtedly a surpassing invention that can help developing countries grow more crops and food and combat starvation. 

Green Biotechnology: A Step Towards Sustainability

As we have already mentioned, green biotechnology can bring agricultural sustainability. Let’s look at some examples on how this branch helps the world produce more food for the increasing population without/ less impacting the ecosystem. 

• Green technology is the foremost step towards sustainable development. It aims to foster sustainable economic growth by ensuring food security without causing long-term environmental damage. 
• Agri-Tech is becoming more popular in underdeveloped and developing countries for feeding the growing population. It is reported that around 12 million farmers from 23 countries used green technologies for producing canola, corn, cotton, and soybean. Among the participating countries, 11 were developed countries, and 12 were developing ones, where approximately 11 million farmers belong to small farms. It is an excellent step towards the green revolution.
• We know how climate change is affecting the world’s agriculture and leading us to food insecurity. As GMO (Genetically Modified Organisms) crops are resistant to insects and pesticides, farmers worldwide can produce more crops and food, bring economic solvency, and cultivate sustainably. For instance, the use of pesticides in cultivation was reduced by 70 million pounds only in the USA in 2005.
• Producing GM food is inevitable. With the increasing population, the world will face a severe food crisis as the food demand will increase by 70% by 2050. Hence, we must move to biotech food production to meet the increasing food demand. 
• According to USDA , farmers are more interested in green technologies than traditional cultivation processes for producing soybean, corn, and cotton.  
• Green biotechnology aims to produce biopolymers from renewable sources to lower the carbon footprint. Using biofuels is an excellent step to minimise environmental pollution by decreasing the use of fossil fuels. 
• Moreover, the world is facing a severe shortage of non-renewable resources like coal and natural gas. And if we keep on using fossil fuels without finding alternative fuel sources, they will come to an end by 2060. The use of biotechnology and biofuels is the latest trend to check the ever-increasing fuel price worldwide.

Risks in Using Green Biotechnology

Though green biotechnology has tremendously helped us achieve agricultural sustainability to a great extent by increasing food production and decreasing environmental pollution, it has some potential risks. 

• The extensive use and production of GMO species may somehow decrease the biodiversity in plants. Agriculturists suspect the traditional plant species may become rare over time if biotech food dominates. 
• The impact of transgenic plants on ‘non-target species’ is still unknown and under research. Some ecologists say that the microspores from these plants (Bt corn) can harm other beetles like caterpillars, while some are concerned about the insect lifespan that would live on these plants. 
• A potential threat of using green technology lies in infusing antibiotic-resistant genes into plants to examine the desirable attributes. These genes can sometimes develop harmful fungi and bacteria that are resistant to a specific antibiotic. It means the antibiotic-resistant germs do not respond to the antibiotics meant to kill them. Infections caused by these fungi and bacteria are difficult to treat and require long-term medication. 

Should We Embrace Green Biotechnology?

We have already mentioned, green biotechnology is the adoption of scientific approaches and tools to alter genetic traits in microorganisms for producing better products with favourable features in sectors like food, biofuels, vaccines, and more. 

Now, let’s assume that these scientific practices are harmful to the human and environment. 

But how would the world feed the ever-growing population if the food production is not multiplied according to the population increase? Biotech food is the only solution here. 

With the alarming impact of global warming, employing agritech is the call of the time to produce crops and plants that can endure severe climate changes. 

Agriculture is one of the leading causes of today’s abrupt climate change and greenhouse gas emissions. Due to unplanned deforestation, the use of CFC-contained sprays and fertiliser, etc., the world’s temperature might increase by 2-3 degrees in the next 50-years. Unlike traditional cultivation processes, biotech plants contribute positively to lessening CO2 emissions by decreasing dependence on pesticides and fertiliser and improving yield with less arable lands. 

But it is imperative to produce GMO products following the regulations and rules strictly to minimise the associated risks.  

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A Review Green biotechnology - a help to the environment

About Authors: Kiran K.Vaghasiya*, Alpesh J.Shiroya Bhagwan Mahavir College Of  Biotechnology , Surat *[email protected]

Abstract Green biotechnologydeals with the use of environmentally-friendly solutions as an alternative to traditional agriculture, horticulture, and animal breeding processes. An example is the designing of transgenic plants that are modified for improved flavor, for increased resistance to pests and diseases, or for enhanced growth in adverse weather conditions. Genetically enhanced crops are one tool that could contribute to a more harmonious balance between food production and our surrounding environment.  The overall message is that biotech plants can, and already do, contribute positively to reducing CO2 emissions and anticipating the impact of climate change on food scarcity. This will increase as they are more widely adopted. This document aims to provide background information about the role green biotech currently plays, and can play in future, in helping to combat climate change.

Reference Id: PHARMATUTOR-ART-1461

INTRODUCTION Biotechnology is any technological application that uses biological systems, living organisms, or derivatives thereof, to make or modify products or processes for specific use. [1 ] Today, the known applications for biotechnology can be seen as a spectrum.

White(Grey)Biotechnology White biotechnology is applied to industrial processes. An important example is bioremediation by microbes where microbes are utilized to clean up toxic or hazardous industrial wastes in the environments, such as PCBs. A second example is the use of microbes to produce products for industrial use, such as the subtilisin enzymes now widely used in laundry detergents.

BlueBiotechnology Blue Biotechnology is aquatic use of biological technology.

RedBiotechnology Red biotechnology refers to medical applications of biotechnology, such as antibiotics and pharmaceuticals that are based on recombinant DNA technology.

Multicolored Biotechnology Biotechnology is often interdisciplinary, and so many applications may be classified in more than one color category. For example, production of biodiesel fuel from agricultural or waste materials could be considered to be both white and green, or white and blue, biotechnology.

GreenBiotechnology Green biotechnology refers to biological techniques to plants with the aim of improving the nutritional quality, quantity and production economics. such as production of disease-resistant or UV-resistant plants, or plants that have superior qualities, by means of genetic modification. Other examples include production of biofuels, such as ethanol or methane, from crops such as corn, or even from marine algae grown at land-based production facilities.

In 1996, the first genetically modified crops were cultivated in the USA. In 2009, 14 million farmers in 25 countries used GM crops, the overwhelming majority of whom (13 million) were small-scale farmers in developing and emerging countries. The annual global acreage has increased to more than 134 million hectares worldwide [2 ] , Green biotechnology pays off economically. This can be seen in the rising number of farmers who opt for GM crops. GM seed tends to be more expensive – but in return, it reduces expenses in other areas, such as the cost of pesticides, machines and labor. But above all: yields generally increase considerably, because plants' own mechanisms protect them from harmful insects and more effective weed management reduces harvest losses which used to be considered inevitable. In 2010, after long political delay, another GM crop was  approved for cultivation   for the first time since 1998: the Amflora potato, with a modified starch composition exclusively processed in the starch industry. [3 ]

Green biotechnology involves the use of environmentally friendly solutions as an alternative to traditional industrial agriculture, horticulture and animal breeding processes.

• use of bacteria to facilitate the growth of plants
• development of pest-resistant grains
• engineering of plants to express pesticides
• use of bacteria to assure better crop yields instead of pesticides and herbicides
• production of superior plants by stimulating the early development of their root systems
• use of plants to remove heavy metals such as lead, nickel, or silver, which can then be extracted ("mined") from the plants
• genetic manipulation to allow plant strains to be frost-resistant
• use of genes from soil bacteria to genetically alter plants to promote tolerance to fungal pathogens
• use of bacteria to get plants to grow faster, resist frost and ripen earlier.

Green revolution Worldwide agricultural productivity has benefited from two green revolutions that have brought crop varieties, allowing higher yields and able to tolerate stress and resist pests and diseases. [4 ]

The first green revolution The first green revolution—from the early 1960s to 1975—introduced new varieties ofwheat, rice, and maize that doubled or tripled yields. The new varieties were highly susceptible to pest infestation and thus required extensive chemical spraying. But they were also responsive to high rates of fertilizer application under irrigation. But they were also responsive to high rates of fertilizer application under irrigation. So, large- and medium-scale farmers in regions with adequate irrigation facilities, easy access to credit, sufficient ability to undertake risks, and good market integration adopted the new varieties [5 ] But these requirements meant that the new technology bypassed most poor African farmers [6 ] [7 ]

The second green revolution The second green revolution—from 1975 to the 1990s—sought to consolidate lessons from the first by developing crops with a wider range of traits desirable for less well endowed areas and smallholder farmers. These traits included tolerance to stress and resistance to pests and diseases.

The third green revolution The third green revolution is the biotechnology or gene revolution. Biotechnology offers possibilities for further amplifying the achievements of the first and second green revolutions.  Four areas in which biotechnology is likely to have significant impact . [8 ]

• Improving genome management (through use of molecular markers for quantitative trait improvement, introgression of new germ plasm into breeding lines, genetic diversity analysis, and parental selection). • Enhancing genetic analysis (through introduction of new genes, directed mutagenesis, optimization of gene expression, and gene discovery). • Quickening the pace of conventional plant research (through new biotechnological Techniques —conventional breeders must rely on phenotypic evaluation, which does not always accurately indicate the information present in a plant’s genome). • Improving agricultural yields.

Green Biotechnology – introduction in three waves [9 ] Green biotechnology covers the whole spectrum from more advantageous and simplified cultivation (input-traits), through improved quality of the plants for animal feed or food purposes (output-traits) down to production and the extraction of new, non-plant contents (molecular pharming).

The goals of breeding genetically modified plants correspond to those of conventional plant breeding: on the one hand quantitative (increase in yield) and qualitative improvements (taste, colour of the blooms, shelf-life, raw materials), and, on the other hand, an improvement in resistance against biotic (fungi, pests, viruses, bacteria, nematode worms) and a-biotic stress factors (cold, heat, wet, drought, salt content). In addition, the plant can also be used as a “bioreactor” to produce enzymes, antibodies, recombinant proteins or pharmaceutical active ingredients (molecular pharming).

1 input traits : The expression “input-traits” refers to characteristics, which lead to an improvement in the properties of a crop from a farming point of view.

Generally, this involves resistance genes, which are introduced into a crop with the use of genetic engineering methods. These resistance genes allow tolerance to herbicides or protect from fungi, pests certain insect, disease and other harmful organisms.

Genetic modification means that resistance against specific harmful products has now been built in to crops such as maize, rape, soya and cotton.

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Herbicide resistance crops : Herbicide resistanceis the inherent ability of a species to survive and reproduce following exposure to a dose of herbicide normally lethal to its wild type. [10 ] maize, oilseed rape and sugar beet – can give the farmer much more flexibility in controlling weeds. [11 ]   Herbicide  resistance is being used worldwide in cotton, potato, maize, soya, tobacco and wheat crops.

The resistance of the crop to herbicides means that prophylactic applications of herbicides, The advantage is that the soil is not bare, and therefore erosion through wind and water can be prevented. Genetically modified, herbicide-resistant plants have so far been bred to withstand the non-selective herbicides which currently dominate the market so that their behaviour in the environment is already well known. With the active ingredient glyphosate has been available on the market for 25 years,whereas glufosinate in the product  has been sold worldwide since 1984. In both products, the active ingredient is absorbed through the green parts of the plant. These non-selective herbicides act by blocking enzymes. However, transgenetic plants, in which additional, extraneous genes have been transplanted, can neutralise the herbicidal effect. In the case of, the active ingredient glufosinate blocks the activity of a plant enzyme, so that toxic ammonia (NH3) accumulates both in the cultivar and the weeds. By contrast, if the cultivar now includes an additional gene (derived from a fungus), For example, tolerance to the herbicide glufosinate  is conferred by the bacterial gene bar , which metabolizes the herbicide into a non-toxic compound the herbicide glufosinate is inactivated against it. [12 ]    As a result, there is no accumulation of ammonia, so that the genetically modified plant can survive. However, the weed is killed off.

Insect resistance crops : Insect resistance is the second most frequently used commercial trait in genetically modified crops after herbicide resistance. To date, the  insect resistant transgenic plants that are commercially available are those expressing genes which code for Bacillus thuringiensis (Bt) that produces a protein toxic to certain insects (of Lepidoptera , Coleoptera and Diptera families).

Bt Endotoxins and their Genes Initially, Bt toxins were classified into 14 distinct groups and 4 classes classification based on their host range. . [13 ]     These are: · CryI (active against Lepidoptera [“Cry” stands for “crystalline” reflecting the crystalline appearance of the d-endotoxin; “Cry” is used to denote the protein whereas “ cry ” denotes the respective gene]), · CryII ( Lepidoptera and Diptera ), · CryIII ( Coleoptera ) and · CryIV ( Diptera . The amount of area being cultivated with these crops is rapidly increasing [14 ]    and other genes coding for new Bt-toxins, lectins, proteinase or α -amylase inhibitors, and other insecticidal products have been successfully engineered in plants [15 ] [16 ]    Some of these plants are being tested at the field scale, such as peas ( Pisum sativum ) expressing the gene coding for common bean α -amylase inhibitors ( α AIs) [17 ]    

In order to make a cultivar resistant to pests, a gene of the soil bacterium Bacillus thuringiensis (Bt) is transplanted. This genetic modification protects the plants from pests  by producing a toxin with the help of the Bt gene, which destroys the pest. As a result, insecticides are not required and loss of yield through pest damage can be prevented. Genetically modified maize, cotton and potato varieties are now being grown with inbuilt Bt genes worldwide. [11 ]

fungas resistance crops : Fungal diseases have been one of the principal causes of crop losses ever since humans started to cultivate plants. [18 ]

Transgenics with antifungal molecules : Antifungal compounds include antifungal proteins fromplants and lower organisms and metabolites like phytoalexins. Under certain conditions, both microorganisms and plantsproduce low mo1 wt, antimicrobial substances. In plants, such compounds known as phytoalexins are often synthesized locally and accumulate after exposure to pathogens and/or stresses. In many cases, a correlation has been found between the concentration of phytoalexins and resistance to specific pathogens. Recently, Háin and coworkers (1990) transferred a gene encoding stilbene synthase into tobacco. [19 ]

ANTIFUNGAL PROTEINS Proteins with the ability to inhibit the growth of fungi in vitro are abundantly present in the plant kingdom. Whether they are involved in the defense against fungal infections in vivo is not known. This idea is supported by examples of transgenic tobacco plants that show enhanced resistance against the fungus Rhizoctonia solani, which is brought about by the constitutive expression of genes encoding proteins shown to have in vitro antifungal activity. [20 ]

Pathogenesis-related proteins : VanLoon and Van Kammen showed that a set of proteins is induced in tobacco plants after tobacco mosaic virus infection10. These proteins were described as pathogenesis-related (PR) proteins. [21 ]

The first report on developing fungus-resistant transgenics came in 1991. Broglie et al . constitutively expressed bean chitinase gene in tobacco and Brassica napus and the plants showed enhanced resistance to Rhizoctonia solani .Chitinases and /3 -1,3-Glucanases are Pathogenesis-related proteins . [22 ]

Plant ribosome-inactivating proteins (RIPs)  : Plant ribosome inactivating proteins (RIPs) have N -glycosidase activity and they remove an adenine residue from 28S rRNA. [23 ]

RIPs do not affect ribosomes of plants in which they are produced and show various degrees of specificity toward ribosomes of other plants. Funga1 ribosomes can be targets of RIPs as well. In in vitro assays a barley RIP has a lower antifungal activity than chitinases or P-1,3-glucanases from barley. However, a strong synergy is observed when barleyRIP is mixed with either of these two hydrolases [24 ] Recently, a barley RIP gene under the control of a wound-inducible promoter was introduced into tobacco. RIprogeny showed an increased resistance to R. solani . [25 ]

Small cystein-rich proteins : In addition to PR proteins, there are other plant proteins which have antifungal activities. A number of small cystein-rich proteins form a separate group of antifungal polypeptides. Some of these are chitin-binding proteins, plant defensins and thionins.

These include (a) recently identified seed proteins from Raphanus sativus [26 ]   Amaranthus caudatus, and Mirabilis jalapa; (b) hevein, a lectin from Urtica dioica; and (c) thionins, which are antimicrobial peptides occumng in seeds and leaves of both mono- and dicotyledonous plants [27 ]

Phytoalexins : Phytoalexins are antimicrobial low molecular weight secondary metabolites produced in plants following pathogen attack and are believed to have a role in plant defense [28 ] Hain and coworkers introduced the gene encoding stilbene synthase from grape vine [29 ]  ( Vitis vinifera ) into tobacco plants.The expression of stilbene synthase (or resveratrol synthase) gene resulted in the production of resveratrol, a stilbene-type phytoalexin. Such transgenics showed enhanced resistance to B . cinerea . Similar transgenic plants were developed in rice, tomato, barley and wheat and were shown to have increased resistance to Magnaporthe grisea , P . infestans and B . cinerea respectively [30 ] [31 ] [32 ]

Virus resistance crops : Viruses cause many diseases in plants and lead to loss of yield. There are no direct ways of fighting virus infections in plants using conventional crop protection.The method generally used is to fight the insects that transmit the viruses(stinging or sucking insects, such as aphids), instead of the viruses themselves by chemical means. In a number of crops, transgenics resistant to an infective virus have been developed by introducing a sequence of the viral genome in the target crop by genetic transformation. Virus-resistant transgenics have been developed in many crops by introducing by some proteins.these are follows

Coat protein : The use of viral CP as a transgene for producing virus resistant plants is one of the most spectacular successes achieved in plant biotechnology. Numerous crops have been transformed to express viral CP and have been reported to show high levels of resistance in comparison to untransformed plants first reported resistance against TMV in transgenic tobacco expressing the TMV CP gene [33 ] . The resistance was manifested as delayed appearance of symptoms as well as a reduced titre of virus in the infected transgenic plants, as compared to the controls. The resistance against TMV using TMV CP in tobacco was also reported to be effective against other tobamoviruses whose CP was closely related to that of TMV but not effective against viruses which were distantly related to TMV [34 ]

Movement protein : Movement proteins (MP) are essential for cell-to-cellmovement of plant viruses. These proteins have been shown to modify the gating function of plasmodesmata, thereby allowing the virus particles or their nucleoprotein derivatives to spread to adjacent cells. This phenomenon was first used to engineer resistance against TMV in tobacco by producing modified MP which are partially active as a transgene. [35 ] . [36 ] . In contrast to the single MP gene in tobamoviruses, viral movement is mediated by a set of three overlapping genes, known as the triple-gene-block (TGB) in potex-, carla- and hordeiviruses [37 ] .

Satellite RNA: Besides using the genomic components of an infectious virus, a strategy exploiting the use of satellite RNA associated with certain viruses received great attention. Some strains of CMV encapsidate satellite RNA (sat RNA) in addition to the tripartite messenger sense, single-stranded RNA genome. transgenic tobacco plants expressing multiple or partial copies of CMV sat-RNA showed attenuated symptoms when challenged with CMV [38 ] .

The virus resistance crops is produced by transplanting a gene that carries the genetic information for the virus’s membrane protein. If the plant itself produces the viral membrane protein, then the virus can no longer get in, and the plant has become resistant to the virus.

2 output traits ( Agronomic traits ) : theplant is modified to withstandimproved qualitative contents, or so-called output-traits. These are quality or product features of the plant. They can be improve amino acid or oil composition, increase shelf life, eliminate undesirable antigens or supply additional vitamin and minerals. The goal of quality or output-traits is thus to improve the quality of agricultural products. This is important in so far as these products can be used directly as food or as the starting point for the food and animal feed industry, or serve as industrial raw materials. Output-traits will therefore offer advantages primarily to processing companies and consumers. Specific amino acids, which are used as building blocks in proteins, are essential in food, as they cannot be produced by the body itself and therefore have to be absorbed from food. Maize, for example, as a low content of the amino acids lysine and methionine. Attempts are being made to increase the nutritional value of important basic foods and animal feed such as maize, soya beans and rape, with the help of genetic engineering by increasing the essential amino acid contents. [9 ]

3 molecular farming : Plant molecular farming is the use of genetically modified plants to produce pharmaceutical products or industrial chemicals. [39 ] If plants are used not only for food or animal feed, but also as „bio factories“ for the production of specific active ingredients, such as antibodies, vaccines, recombinant proteins or pharmaceuticals, then the term “molecular farming” is used. [9 ] In principle, genetic engineering can be used to introduce any proteins into plants as required.. A  broad range of plant species used for the PMF including alfa alfa, Arabidopsis,banana carrot ,maize, rice, potato,sugar cane, tomato tobacco,wheat etc.

Somatotropin & Glucocerebrosidase drug obtained from Tobacco, Cholera vaccines from  Potato, Malaria vaccines from Tobacco [39 ]

Contribution of green biotechnology to the environments in helping to combat climate change : Climate change threatens all elements essential for life, such as water, food, health, environment and land. The temperatures could increase by 2 to 3 degrees in the next fifty years, leading to a catastrophic scenario at the end of the century with a 5 to 8 degree increase forecast, if nothing is done. This is a dramatic scenario whose most obvious symptom would be the change in weather conditions; more heat waves, storms and floods caused by melting glaciers (which could affect more than 30% of the world’s agricultural lands) [40 ]

The three major contributions of greenbiotechnologyto the mitigation of the impact of climate change are: A. Greenhouse gas reduction B. Crops adaptation C. Protection and increase yield with less surface [40 ]

Greenhouse Gas Reduction Agriculture is a major source of greenhouse gas emissions [41 ] . Agricultural practices - such as deforestation, cattle feedlots and fertilizer use - currently account for about 25% of greenhouse gas emissions. Agriculture accounts for 14% of CO2 emission. Agriculture is also a major source of methane (CH4) and nitrous oxide (N2O), with latest estimates showing that it accounts for 48% of methane emissions and 52% of N2O emissions

Solutions: 1. Less fuel consumption on farm : GMOs can help decrease the necessity and frequency of spraying. Genetic engineering crops have highly helped farmers to adopt less intensive agricultural methods such as reduced tillage or no tillage. For example, GM Insect Resistant Crops have been developed to resist insects and to use fewer insecticide treatments. This consequently leads to a reduction of fuel used by farmers when they spray pesticides on their fields, which means a saving in carbon dioxide emissions. Barfoot and Brookes [42 ] ,  study  indicates that in terms of greenhouse gases, each litre of tractor diesel consumed contributes an estimated 2.75 kg of CO2 into the atmosphere. According to a more recent report from Barfoot and Brookes [143 ] , The adoption of reduced tillage or no tillage systems in respect of fuel use results in reductions of carbon dioxide emissions of 89.44 kg/ha and 40.43 kg/ha respectively.

Reduced fertilizer use : Nitrous oxide has a global warming potential of 296, meaning it has a global warming potential (GWP) about 300 times greater than carbon dioxide — this means that 1 pound of nitrous oxide is counted as 296 pounds of CO2. In addition, nitrous oxides stay in the atmosphere for more than 100 years. The formation and release of N2O from agricultural fields happens when nitrogen fertilizer applied to crops interacts with common soil bacteria. It is estimated that nitrogen fertilizer accounts for one-third of the GHGs produced by agriculture [44 ] ,  Reduced fertilizer use will mean less nitrogen pollution of ground and surface waters.

The company, Arcadia Biosciences, has developed GM rice and canola that uses Nitrogen  more efficiently, so the plants need less fertilizer. Arcadia's Nitrogen Use Efficiency (NUE) technology produces plants with yields that are equivalent to Conventional varieties but which require significantly less nitrogen fertilizer because they use it more efficiently. [45 ] This technology has the potential to reduce the amount of Nitrogen fertilizer that is lost by farmers every year due to leaching into the air, soil and waterways. In addition to environmental pressures, nitrogen costs can represent a Significant  portion of a farmer's input costs and can significantly impact farmer profitability.

B. Crops adaptation [46 ] Solutions must be developed to adapt crops to new conditions, such as other types of soils or harsher conditions as drought and salinity. Climate change poses a real challenge in terms of available agricultural land and fresh water use. The agricultural sector uses a huge amount of available fresh water - 70% of the water currently consumed by humans is used in agriculture, and this is likely to increase as temperatures rise. Moreover, 24.7 million acres of farmland worldwide are lost each year due to salinity caused by unsustainable irrigation techniques. In a warmer climate, plants will react to stresses, such as drought, by consuming large quantities of energy which is normally used for growth and seed Production.

Green biotechnology offers solutions that could help farmers facing this challenge. This is absolutely essential for farmers in developing countries as well as in some developed countries  where yields would be affected substantially. Genetic modification is already being used to develop crops tolerant to drought conditions. Indeed, a number of crop varieties have been developed which are stress tolerant. Tolerance to abiotic stress like water shortage and salinity is complex but promising results have been obtained in model plants and are being transferred to important food species in field conditions. Maize, wheat and rice would be the first important crops benefiting from these emerging technologies. [47 ] The first biotech maize varieties with drought tolerance are expected to be commercialized by around 2011 and the trait has already been incorporated in several other crops [48 ]

C. Protection and increase yield with less surface It is likely that with temperature rises and desertification, land area available for farming will be reduced. Moreover, populations are rapidly increasing. By 2025, there will be 2.5 billion more people than today. This population boom will result in a 35% increase in demand for food supplies. According to a United Nations report, farmers will need to at least double their production over the next 25 years to feed all these people [49 ] More than two-fifths of the 55% increase in the world's meat consumption between 1997 and 2020 is expected to occur in China, according to the International Food Policy Research Institute [50 ] . The major aim of agricultural biotechnology is to enhance agricultural productivity and maximise the productive capacity of our diminishing resources.

Sustainable agricultural practices One of the early successes of biotechnology has been the ability to insert genes from a naturally occurring soil bacterium, Bacillus thuringiensis ( Bt ), into maize, cotton, and other crops to impart internal protection from insect feeding. For many farmers, Bt crops are proving to be a valuable tool for integrated pest management programs by giving growers new pest control choices. This is the case for fungal diseases which are a real problem for farmers in maize and cereal cultivation. Not only do they cause yield losses,some fungi can also produce toxic substances, the so-called “mycotoxin”. There are over  300 different known toxins, each with specific effects. For example, the level of fumonisin, one of those 300 toxins, is associated with oesophageal cancer and neural tube defects. Bt maize is a powerful tool to reduce the level of fumonisin which could have significant benefits in developing countries, especially where unprocessed maize is a key part of the diet. [51 ] In France,

Orama, a trade association in the agricultural sector, found out that the average gain of Bt maize was about 9.2 quintal per hectare compared to conventional maize by studying

13 plots of Bt maize field in France in 2005. [52 ] .

Increased yield to address tomorrow’s challenges Science and technology must spearhead agricultural production in the next 30 years at a pace faster than the Green Revolution did during the past three decades, Dr. Diouf DrJacques Diouf, Director-General Food and Agriculture Organization of the United Nations(FAO) asserted. Farmers need to produce more food than ever before. Each year, globalpopulation grows by more than 73 million . This is only slightly less than adding a population the size of Germany’s each year. As a result, world population is expected to reach 7 billion by 2013 and 8 billion by 2028. And, as people in developing countries attain higher levels of education and income, the demand for higher-quality food increases. The combined effect of population gains and income gains around the world is projected to increase the demand for food 55 percent by 2030. Biotechnology and advances in breeding are helping agriculture achieve higher yields and meet the needs of an expanding population with limited land and water resources. Production of primary food and feed crops — maize, wheat, rice and oilseeds — has increased by 21 percent [53 ].

The applications of green biotechnology : Green biotechnology which is more commonly known as Plant Biotechnology is a rapidly expanding field within Modern biotechnology. Use of environment friendly and cost effective alternatives to industrial chemicals such as bio fuels, bio fertilizers and bio pesticides are not only resulting in enhanced crop output, improvement in health and safety standards, these new products are also leading to less environment pollution and use of green technology.

The applications of green biotechnology can be found in two distinct areas: Agricultural Biotechnology : refers to the application of biotechnology techniques in crop improvement. Today Agricultural biotechnology encompasses the following main areas of research and application: Plant tissue culture: A technique that allows whole plants to be produced from minute amounts of plant parts like the roots, leaves or stems or even just a single plant cell under laboratory conditions. [54 ].

Plant genetic engineering: The selective, deliberate transfer of beneficial gene(s) from one organism to another to create new improved crops. The American Society of Plant Biologists (ASPB) submits this statement supporting the continued, responsible use of new technologies, such as recombinant DNA technology (hereafter referred to as "genetic engineering” or “GE"), which can add effective tools to those needed to combat hunger and maintain a healthy environment.

The use of GE to modify plants represents a significant advance in plant science, Modified crops resulting from plant biotechnology are also expected to provide major health benefits to people throughout the world. Examples include enhancing the vitamin and mineral content of staple foods  . [55 ] eliminating common food allergens, [56 ] [57 ] developing higher protein quality and quantity in widely consumed crops [58 ] and modifying plants to contain vaccines against many illnesses [59 ]

GE plants are also expected to be useful in nonfood applications, such as phytoremediation [60 ] where plants remove contaminating pollutants from soils and water resources and serve as biofactories to create compounds presently made using nonrenewable resources, e.g., industrial oils and fuels.

There are following method of gene transfer 1. Direct methods -protoplast microinjection -particle bombardment -protoplast polyethyleneglycol (PEG) method -protoplast electroporation

2. Agrobacterium -mediated transformation Agrobacterium tumefaciens Agrobacterium rhizogenes (Hairy Root)

Environmental Biotechnology: Environmental Biotechnology refers to the application of biotechnological processes in the protection and restoration of environmental quality. Currently, environmental biotechnology is being applied in cleaning up pollution, waste water treatment, air purification and waste gases treatment, as it is the case of bioremediation, the use of biological systems to clean up air pollution and contaminated soil or water. Industry is developing more and more processes in this prevention area to reduce environmental impact.

Biofertilizer : To reduce the impact of excess chemical fertilizers in the field of agriculture the biofertilizer is a potential tool, biologically fixed nitrogen is such a source which can supply an adequate amount of Nitrogen to plants and other nutrients to some extent. Many free living and symbiotic bacteria which fix atmospheric Nitrogen were used as biofertiliser material as a substitute for Nitrogen fertilizer. In general two types of biofertiliser are used 1. Bacterial Biofertilizer 2. Algal Biofertilizer

Conclusion : Biotechnology offers rich opportunities to increase agricultural productivity. It accelerates plant and animal breeding efforts. It offers solutions to previously intractable problems. Need to develop appropriate national policies and identify key national priorities for biotechnology, bearing in mind the potential biological risks and the needs of poor people who rely on agriculture for their livelihoods. And the international community needs to loosen the arrangements for access to proprietary technology—enabling developing countries to provide poor farmers with improved seeds while protecting them from inappropriate restrictions on propagating their crops.

Acknowledgment : Dr. R. Krishnamurthy Director of bhagwan mahavir college of M.sc. biotechnology, vesu- surat.

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Green Nanotechnology: Advancement in Phytoformulation Research

1 Pharmaceutical Sciences Laboratory, Faculty of Science and Engineering, Åbo Akademi University, 20520 Turku, Finland; if.oba@lasnabk (K.K.B.); if.oba@kahbarpn (N.P.); if.oba@hnesorej (J.M.R.)

Surya P. Gautam

2 CT Institute of Pharmaceutical Sciences, Jalandhar 144020, India; moc.liamy@matuagayrus

Kuldeep K. Bansal

Neeraj prabhakar, jessica m. rosenholm.

The ultimate goal of any scientific development is to increase well-being and human health. Novel strategies are required for the achievement of safe and effective therapeutic treatments beyond the conventional ones, and society needs new requirements for new technologies, moving towards clean and green technology development. Green nanotechnology is a branch of green technology that utilizes the concepts of green chemistry and green engineering. It reduces the use of energy and fuel by using less material and renewable inputs wherever possible. Green nanotechnology, in phytoformulations, significantly contributes to environmental sustainability through the production of nanomaterials and nanoproducts, without causing harm to human health or the environment. The rationale behind the utilization of plants in nanoparticle formulations is that they are easily available and possess a broad variability of metabolites, such as vitamins, antioxidants, and nucleotides. For instance, gold (Au) nanoparticles have attracted substantial attention for their controllable size, shape, and surface properties. A variety of copper (Cu) and copper oxide (CuO) nanoparticles have also been synthesized from plant extracts. Titanium dioxide and zinc oxide nanoparticles are also important metal oxide nanomaterials that have been synthesized from a number of plant extracts. International and domestic laws, government and private-party programs, regulations and policies are being carefully reviewed and revised to increase their utility and nurture these nanoscale materials for commercialization. Inspiring debates and government initiatives are required to promote the sustainable use of nanoscale products. In this review, we will discuss the potential of the utilization of plant extracts in the advancement of nanotechnology.

1. Introduction

Nanotechnology is cited as a key technology of the 21st century and has generated a great deal of excitement world-wide, but it has been slowed down because of the poor understanding of hazards associated with nanotechnology and fewer policies to manage new risks. Researchers, however, continue to move ahead, engaging themselves to conquer the challenges ranging from managing, producing, funding, regulatory, and technical aspects. Green nanotechnology is a branch of green technology that utilizes the concepts of green chemistry and green engineering, where the word “green” refers to the use of plant products ( Figure 1 ). It reduces the use of energy and fuel by using less material and renewable inputs wherever possible. Furthermore, nanotechnological products, processes, and applications are expected to contribute significantly to environmental and climate protection by saving raw materials, energy, and water, as well as by reducing greenhouse gases and hazardous waste. Increased energy efficiency, reduced waste and greenhouse gas emission, and decreased consumption of non-renewable raw materials are the main advantages of green nanotechnology. Green nanotechnology offers a great opportunity to stop the adverse effects before they occur [ 1 , 2 ].

Branches of green nanotechnology.

Green nanotechnology does not ascend de novo; rather, it forms on the principles of green chemistry and engineering. Apart from such obvious areas as the development of solar cells, biofuels, and fuel cells, green nanotechnology applications might involve the use of nanomaterials in clean production processes that synthesize nanoparticles, using sunlight or by recycling industrial waste products into nanomaterials. There is some “truly” green nanotechnology, i.e., fully growing nanomaterials in plants—however, they will never reach the scale required for the industrial production of nanomaterials. In order to make a conclusive observation, green nanotechnology needs a full process assessment like other industrially manufactured products [ 3 , 4 ].

2. Herbal Approach for Developing Nanoparticles

The activity of herbal medicines depends on the overall function of active components, as all the constituents provide synergistic action and, thus, improve the therapeutic value. Each active constituent is related to each other and they all play significant roles ( Table 1 ). On the other hand, the insoluble character of most of the drugs of herbal origin leads to lower bioavailability and, because of this, systemic clearance is increased and frequent administration or a higher dose is required—all of which renders the drug a low-class drug for therapeutic use.

Herbal drug-loaded nanoparticles.

In phytoformulation research, developing nanotechnology-based dosage forms, e.g., solid lipid nanoparticles (SLNs), polymeric nanoparticles (nanospheres and nanocapsules), proliposomes, liposomes, nanoemulsions, etc., has a great number of advantages for herbal drugs. These include enhancement of solubility and bioavailability, improvement of stability, suppression of toxicity, improvement of pharmacological activity, sustained delivery, improving tissue macrophage circulation, and defense against physical and chemical degradation. Therefore, problems associated with plant medicines can be overcome with nano-sized drug delivery systems (NDDS) of herbal drugs, having a potential future for enhancing their activity. Hence, including nanocarriers as an NDDS in conventional medicine systems would be necessary to combat more chronic diseases like diabetes, cancer, asthma, and others, with the aid of herbal drugs [ 5 , 6 , 7 ].

3. Nanoparticles Synthesized from Plant Extracts

3.1. gold and silver nanoparticles.

Au nanoparticles have gained substantial attention due to their controllable size, shape, and surface properties [ 15 ]. Because of these unique properties, gold nanoparticles have been studied for potential applications in areas such as biosensors, hyperthermia therapy [ 16 ], antibacterial drugs, genetic engineering, and delivery platforms for therapeutics. Environmentally friendly sources of Au nanoparticles are achieved by employing plants, as they are biological factories via green chemistry-based techniques. The study of nanoparticle syntheses also discovered that a variety of shapes, including rod-shaped, irregular, decahedral, icosahedral, and hexagonal, could be produced, depending on the pH of the reaction medium. Furthermore, a leaf extract of eucalyptus macrocarpa could be used to synthesize gold nanoparticles ( Table 2 ). The results from this study show that spherical particles with a size ranging from 20 to 80 nm were obtained as the main product [ 17 ].

A selection of nanoparticles synthesized by various plants.

In addition to synthesizing pure metal nanoparticles by plants in this way, several authors have also reported alloying Au and Ag to investigate the properties of the resulting bimetallic nanoparticles. This bimetallic nanoparticle synthesis comprises a competitive reduction process between two aqueous solutions, each of which contain a different metallic ion precursor that is reacted with a plant extract. In the case of bimetallic nanoparticles, gold has a larger reduction potential than silver, so gold will form first and create the core of the resulting core–shell structure. Subsequently, the reduction of Ag ions, in the same way, results in Ag coalescing on the core to form the shell. There are some plants that have been effectively used to synthesize bimetallic (Au-Ag) nanoparticles, including cashew nut, neem, and West Indies mahogany [ 29 , 30 , 31 ].

3.2. Copper and Copper Oxide Nanoparticles

A variety of copper (Cu) and copper oxide (CuO) nanoparticles have been synthesized from plant extracts. Cu nanoparticles of magnolia leaf extract have been biologically synthesized to develop stable nanoparticles sized 40 to 100 nm. Furthermore, the activity of Cu nanoparticles has shown potential antibacterial activity against cells of Escherichia coli [ 32 ]. Syzygium aromaticum (clove) extracts have been used in the synthesis of Cu nanoparticles with a spherical to granular morphology and a mean particle size of 40 nm [ 33 ]. Cu nanoparticles have been synthesized by using the stem latex of Euphorbia nivulia, that is, common milk hedge. These nanoparticles are stabilized by peptides and terpenoids that are present in latex. Furthermore, these nanoparticles are reported to be toxic to human adenocarcinomic alveolar basal epithelial cells [ 34 ].

3.3. Palladium and Platinium Nanoparticles

Satishkumar et al. [ 35 ] developed palladium nanoparticles from C. zeylanicum (cinnamon) bark extract. During synthesis, the temperature, concentration, and reaction pH of the bark extract changed, but morphology and particle size (15 to 20 nm) were not influenced. Using Annona squamosa (custard apple) peel extract, palladium nanoparticles were also synthesized, ranging in size from 75 to 85 nm [ 36 ]. Camellia sinensis (tea) and Coffea arabica (coffee) extracts have been used to synthesize palladium nanoparticles with sizes ranging from 20 to 60 nm and a cubic crystal symmetry in the center [ 37 ]. Song et al. [ 38 ] reported the first platinum nanoparticles of Diospyros kaki (persimmon) leaf extract, having sizes of 2 to 12 nm. Lately, particle size- and shape-controlled biological synthesis of platinum nanoparticles has also been reported. Plant wood for the nanometer scale has been used for this purpose [ 39 ]. For instance, Coccia et al. [ 40 ] reported isolated lignin from red pine ( Pinus resinosa ) for producing palladium and platinum nanoparticles.

3.4. Titanium Dioxide and Zinc Oxide Nanoparticles

These important metal oxide nanomaterials have been synthesized from a number of plant extracts. For instance, Roopan et al. [ 41 ] established that TiO 2 nanoparticles could be effectively synthesized from Annona squamosa peel; meanwhile, from Nyctanthes arbor-tristis leaf extracts, round particles were found, ranging in size from 100 to 150 nm [ 42 ]. Eclipta prostrata leaf extracts could also produce particles, with a size range of 36 to 68 nm [ 43 , 44 ]. Velayutham et al. synthesized TiO 2 nanoparticles using Catharanthus roseus leaf extract, ranging in size from 25 to 110 nm and irregularly shaped. The suspension of TiO 2 revealed that they were both larvicidal and adulticidal against Bovicola ovis (sheep louse) and Hippobosca maculate (hematophagous fly) [ 45 ]. The antioxidant and antibacterial properties of TiO 2 nanoparticles, synthesized using an extract from Psidium guajava , were evaluated against Pseudomonas aeruginosa, Staphylococcus aureus, Proteus mirabilis, Aeromonas hydrophila , E. coli , and other pathogens. In addition, the antioxidant and antibacterial properties of TiO 2 nanoparticles were evaluated in nanometer scale-up and in bulk [ 46 ].

3.5. Indium Oxide (In 2 O 3 ), Iron Oxide, Lead, and Selenium Nanoparticles

Using a variety of plants, a number of new types of metal oxide and metal nanoparticles were biologically synthesized. Indium oxide nanoparticles were synthesized from aloe vera leaf extract ( aloe barbadensis Miller ). After primary synthesis, for the production of nanoparticles, the precipitates were thermally treated at 400–600 °C. The size of the synthesized spherical nanoparticles ranged from 5 to 50 nm and the size was dependent on the reaction temperature [ 47 ]. In a number of environmental remediation technologies, iron nanoparticles are very important. Thus, a number of studies have focused on green chemistry to synthesize these iron (Fe) nanoparticles. For instance, to synthesize Fe nanoparticles, sorghum bran aqueous extracts have been used. Recently, Pattanayak et al. [ 48 ] synthesized spherical iron nanoparticles that had particle sizes of 100 nm from Azadirachta indica (neem) leaf extract. A while ago, Shah et al. synthesized iron nanoparticles from different plant extracts, such as Cymbopogon citratus (lemon grass tea), Datura innoxia, Tridax procumbens, Calotropis procera, Tinospora cordifolia , and Euphorbia milii . Stem extract was used to synthesize the smallest spherical nanoparticles, from 13 nm, the widest size range (43–342 nm) of nanoparticles synthesized from Cymbopogon citratus leaf extract [ 49 ]. Lead (Pb) and selenium (Se) are two other significant nanoparticles that have been synthesized biologically. Joglekar et al. [ 50 ] were able to synthesize spherical Pb nanoparticles form Jatropha curcas latex, having sizes ranging from 10 to 12.5 nm. Lately, Sasidharan et al. [ 51 ] have been able to synthesize spherical selenium (Se) nanoparticles from citrus reticulata peel extract, with a size of 70 nm.

4. Green Synthesis of Metal Nanoparticles

For a long time, it has been known that plants have the potential for biological reduction of metallic ions and hyper-accumulation [ 52 , 53 ]. Because of such remarkable properties, plants have been considered a more environmentally friendly biological method for synthesis of metallic nanoparticles, and also useful for detoxification applications [ 54 ]. Plant extracts contain various bioactives, such as alkaloids, proteins, phenolic acids, sugars, terpenoids, and polyphenols, which have been found to have an important role in first reducing and then stabilizing the metallic ions, as shown in Figure 2 .

Biological synthesis of nanoparticles by using plant extracts.

The shape and size of nanoparticles mainly depend on the variation in composition and concentration of active biomolecules of different plants, and their interaction with the aqueous metal ions. Especially in chemical and biological synthesis of nanoparticles, the aqueous metal ion precursors from metal salts are reduced, which results in colour change of the reaction mixture and provides a quantitative indication of nanoparticle formation. More importantly, the nanoparticles synthesized from reducing agents may show general toxicity, engendering serious concern for developing environmentally friendly processes. The process of the formation of nanoparticles begins by mixing a metal–salt solution with a sample of plant extract. During the synthesis of nanoparticles, biochemical reduction of the salt solution starts immediately and the change in colour of the reaction mixture indicates the formation of nanoparticles. During synthesis, initially there is an activation period process in which metal ions are converted to zero-valent state from their mono or divalent oxidation states, so that the nucleation of such reduced metal atoms takes place [ 55 ]. Furthermore, the process of nanoparticle synthesis is followed by the integration of smaller neighbouring particles to form larger nanoparticles, which are thermodynamically stable, and, subsequently, the metal ions are reduced biologically. In this way, growth progresses and nanoparticles aggregate to form a variety of shapes such as spheres, cubes, triangles, rods, wires, hexagons, and pentagons. In the final stage of the process, the ability of plant extract to stabilize the nanoparticle finally determines its stable morphology. Significantly, the quality, size, and morphology of the nanoparticles are influenced by properties of the plant extracts; mainly its concentration, reaction time, metal salt concentration, reaction solution pH, and temperature [ 56 , 57 ].

5. Green Nanotechnology: Risk Aspects

The invention of nanotechnology application in the field of green technology has raised concerns regarding the nanomaterial impact on health and safety of workers. This urgently requires scientific, technological, and governmental efforts to manage such kinds of risks for the workforce. This means detecting genuine risks derived from nanomaterial exposure in the workplace, that is, “risk assessment”: To plan the control measures of “risk management” and, finally, to communicate the plan. Overall, all these steps of risk aspects are critical and will be discussed in the following section, which aims to protect the worker from harm and provide all the benefits of green nanotechnology for society ( Figure 3 ).

Risk aspects of green nanotechnology.

6. Risk Assessment

Risk assessment of nanomaterials includes the same processing steps that are used in the risk assessment of other types of materials/chemicals [ 58 ]. These include hazard identification, hazard characterization, dose-response relationships, and assessment of exposure for the different scenarios. Unfortunately, the risk assessment process of nanomaterials still suffers from a deficiency of toxicological data for a variety of nanomaterials. Furthermore, the definition of vital health effects, such as genotoxicity, pulmonary toxicity, or carcinogenicity in conditions of long-term and low-dose exposure, are approaching realistic scenarios that require attention [ 59 ]. For the characterization of occupational nanomaterial risks, the exposure assessment remains a fundamental condition. Efforts should be made to overcome practical barriers that are related to the novelty of green-nanomaterial exposure scenarios, the variations in how to detect and classify nanomaterials, and the queries about the metrics for health-related sampling. That aside, it is also important to carry out biological monitoring studies, which are important to define possible biomarkers of nanomaterials exposure, and effects that are prospectively tested, validated and used in occupational risk evaluation [ 60 ].

7. Risk Management

The aim of risk assessment is to provide computable predictions of risks assisting their evidence-based management [ 61 ]. To effectively manage the potential risks related to green nanotechnology, a plan of risk management including a hierarchy of controls should be emphasized [ 62 ]. The first step of planning is to determine potential exposure to workers, measuring and identifying how this exposure may differ depending on the job task. After identification, potential worker exposure should be managed by using the hierarchy of controls that starts with hazard elimination, adopting a green chemistry by substitution with a non-hazard, and an introduction of engineering controls, including enclosed systems. Administrative control should follow these steps, including training programs by which companies communicate with workers to find out if they have the information to sufficiently understand the routes and nature of potential nanomaterial exposure in the workplace, adequate job procedures, possible risks, preventive and protective measures, and the policies adopted. In this context, it should be important to improve insufficient or inadequate information for workers present on safety data sheets. Risk management, including the use of personal protective equipment (PPE), including respiratory and eye protection, gloves, and lab coats, is the final step for exposure control.

8. Risk Communication

Risk communication is a crucial part of green nanotechnology, relating to the healthy origination and sustainable development of general public transparency. In terms of making available complex technical and health information, risk communication should be made effective, in language that should be accessible and understandable to the general population. Importantly, regulatory scientists, researchers, workforce representatives, industry, and governmental authorities should be engaged in a dialogical progressive communication of the potential green nanotechnology risks, with the objective to form adequate perceptions and attitudes. This is tremendously important to ensure that the spread as well as promotion by mass media involves appropriate information regarding the benefits and challenges of green nanotechnology, protecting public opinion from both unrealistic prospects and excessive consciousness in this regard.

9. Conclusions

As green nanotechnology becomes more commercialized, it will have the potential to become an industry with very strong green credentials. As a general conclusion, it can be said that green nanotechnology involves challenging work in the pharmaceutical industry. Ultimately, however, it improves the quality of life, promotes environmentally friendly commitments, as well as ethical values in the field of nanotechnology.

Acknowledgments

We would like to thank Pharmaceutical Sciences Laboratory, Åbo Akademi University for providing the necessary support.

Author Contributions

Conceptualization, A.V. and S.P.G.; Validation, A.V., and S.P.G.; Formal analysis, A.V., K.K.B. and N.P.; Resources, A.V. and J.M.R.; Writing—original draft preparation, A.V.; Supervision, J.M.R.

Academy of Finland, grant numbers 309374 (J.M.R.) and 309794 (K.K.B., J.M.R.); Sigrid Jusélius Foundation (N.P.).

Conflicts of Interest

The authors declare no conflict of interest.

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ORIGINAL RESEARCH article

Effect of novel polyethylene insert configurations on boneimplant micromotion and contact stresses in total ankle replacement prostheses: a finite element analysis provisionally accepted.

• 1 Zhangjiagang Fifth People's Hospital, China
• 2 Dongying People’s Hospital, China
• 3 Jingxian Hospital, China
• 4 Affiliated Hospital of Zunyi Medical University, China
• 5 First Affiliated Hospital, School of Medicine, Zhejiang University, China

The final, formatted version of the article will be published soon.

Artificial ankle replacement (TAR) has been in existence for decades. TAR has preserved the function of the affected limb for osteoarthritis patients and has become an alternative to ankle arthrodesis. However, TAR has the disadvantage of a high revision rate, which has caused certain obstacles to the promotion of this technology.Considering that the main cause of TAR failure is prosthesis loosening, in order to prolong the TAR survival rate, the reduction of prosthesis micromotion becomes the focus of scholars. In our research this time, we followed the design idea of elastic optimization and developed 3 new inserts. In the new inserts, flexible materials are embedded in ordinary UHMWPE materials, which increases the elasticity of the inserts and can absorb more kinetic energy can reduce the bone-implant interface micromotion and joint surface contact stress, which provides a valuable reference for the development of a new generation of artificial ankle prosthesis. I hope this paper is suitable for Frontiers in Bioengineering and Biotechnology.We deeply appreciate your consideration of our manuscript, and we look forward to receiving comments from the reviewers. If you have any queries, please don't hesitate to contact me at the address below.

Keywords: Total ankle replacement, artificial ankle, insert, elasticity improvement, Finite Element Analysis

Received: 17 Jan 2024; Accepted: 01 Apr 2024.

Copyright: © 2024 Xu, Gong, Hu, Bian, Jin and Li. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY) . The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

* Correspondence: Dr. Zhi Xu, Zhangjiagang Fifth People's Hospital, Zhangjiagang, China

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