Bacillus spp. as Bio-factories for Antifungal Secondary Metabolites: Innovation Beyond Whole Organism Formulations.

Several fungi act as parasites for crops causing huge annual crop losses at both pre- and post-harvest stages. For years, chemical fungicides were the solution; however, their wide use has caused environmental contamination and human health problems. For this reason, the use of biofungicides has been in practice as a green solution against fungal phytopathogens. In the context of a more sustainable agriculture, microbial biofungicides have the largest share among the commercial biocontrol products that are available in the market. Precisely, the genus Bacillus has been largely studied for the management of plant pathogenic fungi because they offer a chemically diverse arsenal of antifungal secondary metabolites, which have spawned a heightened industrial engrossment of it as a biopesticide. In this sense, it is indispensable to know the wide arsenal that Bacillus genus has to apply these products for sustainable agriculture. Having this idea in our minds, in this review, secondary metabolites from Bacillus having antifungal activity are chemically and structurally described giving details of their action against several phytopathogens. Knowing the current status of Bacillus secreted antifungals is the base for the goal to apply these in agriculture and it is addressed in depth in the second part of this review.

Biological Control with Trichogramma in China: History, Present Status, and Perspectives.

Trichogramma species make up one of the most commonly used groups of natural enemies for biological control programs worldwide. Given the major successes in using Trichogramma to control economically important lepidopterous pests on agricultural crops in China, the biology and ecology of these wasps have been intensively studied to identify traits that contribute to successful biological control. Since the 1960s, improved mass production of Trichogramma and better augmentative release methods to suppress agricultural pests have been achieved. We review the history of research and development; current knowledge on biodiversity and bio-ecology of the species used; and achievements in mass-rearing methods, release strategies, and current large-scale applications in China. In addition, we discuss potential issues and challenges for Trichogramma research and applications in the future.

Success of microbial genes based transgenic crops: Bt and beyond Bt.

Transgenic technology could hold the key to help farmers to fulfill the ever increasing fast-paced global demand for food. Microbes have always wondered us by their potentials and thriving abilities in the extreme conditions. The use of microorganisms as a gene source in transgenic development is a promising option for crop improvement. The aforesaid approach has already for improving the characteristics of food, industrial, horticulture, and floriculture crops. Many transgenic crops containing microbial genes have been accepted by the farmers and consumers worldwide over the last few decades. The acceptance has brought remarkable changes in the status of society by providing food safety, economic, and health benefits. Among transgenic plants harboring microbial genes, Bacillus thuringiensis (Bt) based transgenic were more focused and documented owing to its significant performance in controlling insects. However, other microbial gene-based transgenic plants have also reserved their places in the farmer's field globally. Therefore, in this review, we have thrown some light on successful transgenic plants harboring microbial genes other than Bt, having application in agriculture. Also, we presented the role of microbial genetic element and product thereof in the inception of biotechnology and discussed the potential of microbial genes in crop improvement.

Essential Oil-Based Bioherbicides: Human Health Risks Analysis.

In recent years, the development of new bio-based products for biocontrol has been gaining importance as it contributes to reducing the use of synthetic herbicides in agriculture. Conventional herbicides (i.e., the ones with synthetic molecules) can lead to adverse effects such as human diseases (cancers, neurodegenerative diseases, reproductive perturbations, etc.) but also to disturbing the environment because of their drift in the air, transport throughout aquatic systems and persistence across different environments. The use of natural molecules seems to be a very good alternative for maintaining productive agriculture but without the negative side effects of synthetic herbicides. In this context, essential oils and their components are increasingly studied in order to produce several categories of biopesticides thanks to their well-known biocidal activities. However, these molecules can also be potentially hazardous to humans and the environment. This article reviews the state of the literature and regulations with regard to the potential risks related to the use of essential oils as bioherbicides in agricultural and horticultural applications.

Mycoviruses in Biological Control: From Basic Research to Field Implementation.

Mycoviruses from plant pathogens can induce hypovirulence (reduced virulence) in their host fungi and have gained considerable attention as potential biocontrol tools. An increasing number of mycoviruses that induce fungal hypovirulence, from a wide variety of taxonomic groups, are currently being reported. Successful application of these viruses in disease management is greatly dependent on their ability to spread in the natural populations of the pathogen. Mycoviruses generally lack extracellular routes of transmission. Hyphal anastomosis is the main route of horizontal mycovirus transmission to other isolates, and conidia of vertical transmission to the progeny. Transmission efficiencies are influenced by both the fungal host and the infecting virus. Interestingly, artificial transfection methods have shown that potential biocontrol mycoviruses often have the ability to infect a variety of fungi. This expands their possible use to the control of pathogens others than those where they were identified. Mycovirus research is also focused on gaining insights into their complex molecular biology and the molecular bases of fungus-virus interactions. This knowledge could be exploited to manipulate the mycovirus and/or the host and generate combinations with enhanced properties in biological control. Finally, when exploring the use of mycoviruses in field conditions, the pathogen life style and the characteristics of the disease and crops affected will deeply impact the specific challenges to overcome, and the development of biocontrol formulations and delivery methods.

Microbial biopesticides for invertebrate pests and their markets in the United States.

Microbial pesticides based on bacteria, fungi and viruses or their bioactive compounds have long been developed as alternatives for synthetic pesticides to control invertebrate pests. However, concern for environmental and human health from excessive reliance on chemical pesticides, changes in residue standards, and increased demand for organically grown produce has contributed to a considerable growth in their use in recent years. There are currently 356 registered biopesticide active ingredients in the U.S., including 57 species and/or strains of microbes or their derivatives, labelled for use against pestiferous insects, mites and nematodes. Strains of Bacillus thuringiensis for Lepidoptera remain the most popular products, but newer bacterial strains and their metabolites have been developed against a wider range of arthropods for use on fruit, vegetable and ornamental crops. Currently, ten fungal species/strains are registered against thrips, whiteflies, aphids, or other sucking pests and plant parasitic nematodes in greenhouse, nursery and field crops, while five nucleopolyhedroviruses and three granuloviruses are registered for Lepidoptera in field and greenhouse grown vegetables and ornamentals, tree fruit and nuts, forestry, and stored products. Many of these products are organic listed and most have 4 h or less reentry and no pre-harvest restrictions. Investment by multinational companies, advances in screening, industrial fermentation and storage of new microorganisms, are increasing the market share for microbials. Here, we summarize the market for microbial-based pesticides labelled for invertebrates in the U.S. We cover current uses and recent advances that further advance their use in additional markets in the coming decades.

Microbial control of phytophagous invertebrate pests in South Africa: Current status and future prospects.

Invertebrate pests pose a significant threat to food security on the African continent. In response, South Africa has become one of the largest importers of chemical pesticides in sub-Saharan Africa, with several hundred active ingredients registered. To address the over-reliance on such chemicals, the South African Department of Agriculture, Forestry and Fisheries (DAFF) has eliminated or restricted several pesticides since the late 1970s. The recent launch of the South African National Bio-Economy Strategy and establishment of the South African Bioproducts Organisation (SABO), together with new guidelines for registration of biopesticides in 2015, also support this endeavour. Concurrently, entomopathogen-related research and bioproduct development has increased over the past decade. Currently, 31 products (seven manufactured locally) are registered under the Fertilizers, Farm Feeds, Agricultural Remedies and Stock Remedies Act 36 of 1947. Commercially important microbes include Beauveria bassiana (Cordycipitaceae), Metarhizium anisopliae (Clavicipitaceae), Cydia pomonella granulovirus, Cryptophlebia leucotreta granulovirus, Helicoverpa armigera nucleopolyhedrovirus (Baculoviridae) and Bacillus thuringiensis subsp. kurstaki and B. thuringiensis subsp. aizawai (Bacillaceae). Both parasitic and entomopathogenic nematodes (EPNs) show potential for development as bioinsecticides with one commercial EPN product, based on Heterorhabditis bacteriophora (Heterorhabditidae), registered under the Act. Rapid scientific progression, supported by a favourable legislative environment, should facilitate further advances in microbial control of phytophagous invertebrate pests in South Africa.

Microbial insecticides in Iran: History, current status, challenges and perspective.

Biological control programs with arthropods have been in practice in Iran since the 1930s. However, development and registration of microbial biopesticides is much more recent. Currently, about 15 biopesticide products are registered or pending registration for commercial use on various crops. Products based on Bacillus thuringiensis subsp. kurstaki are most widely sold for lepidopteran pest control, followed by B. thuringiensis subsp. isralensis against dipteran pests in vector control programs. Additionally, mycoinsecticides based on Lecanicillium lecanii and Beauveria bassiana, against various arthropod pests, and a mycofungicide based on Trichoderma harzianum for controlling soilborne diseases are also registered. In Iran, the national Plant Protection Organization (PPO) manages regulation of microbial pesticides and the Pesticide Supervision Board within the PPO oversees registration of all pesticides. Currently, two Iranian companies produce microbial pesticides and two more companies are approved to start production in the near future. We review the history of microbial control of arthropod pests in Iran with examples of sustainable agricultural practices, the current status of the market and registration procedures for microbial pesticides, along with the challenges and opportunities for the advancement of microbial control in Iran.

Microbial biopesticides for insect pest management in India: Current status and future prospects.

The biopesticide industry in India is undergoing rapid change, reflecting increased global trade in agricultural commodities, a changing regulatory environment and evolving consumer preferences. Currently biopesticides comprise ≈ 5% of the Indian pesticide market, with at least 15 microbial species and 970 microbial formulations registered through the Central Insecticides Board and Registration Committee (CIBRC). As of 2017, over 200 products based on entomopathogenic fungi (Beauveria bassiana, B. brongniartii, Metarhizium anisopliae s.l., Lecanicillium lecanii and Hirsutella thompsonii) and nematicidal fungi (Purpureocillium lilacinum and Pochonia chlamydosporia) are registered for use against various arthropods and plant parasitic nematodes. Regarding bacteria, over 30 products based on Bacillus thuringiensis (Bt) subsp. kurstaki are registered against bollworms, loopers and other lepidopterans, while 12 based on Bt subsp. israelensis and three with Bt subsp. sphaericus have been used against mosquitoes. Two viruses are registered, namely Helicoverpa armigera nucleopolyhedrovirus (22 products) and Spodoptera litura nucleopolyhedrovirus (5 products) for use against bollworms and armyworms. Four entomopathogenic nematode species are sold in Indian market. These include long-lasting wettable powder formulations of Heterorhabditis indica developed by the ICAR-National Bureau of Agricultural Insect Resources, Bengaluru which have been distributed on a large scale to control white grubs and other sugarcane pests. Biopesticide research on the subcontinent is at a relatively early stage, but evolving rapidly, and focusing on indigenous entomopathogens. Despite onerous regulation, quality-control issues and limited large-scale production facilities, investment in domestic fermentation technologies, improved delivery systems, and promotion of biological control through private and public initiative will increase the share of microbial biopesticides in the country.

Microbial biopesticides for control of invertebrates: Progress from New Zealand.

Biopesticides are needed for control of endemic and invasive pests impacting New Zealand's primary sectors including pests that are emerging as a result of climate change and farming intensification. Products developed in New Zealand are usually based on endemic strains of microorganisms, including new species/strains with novel modes of action. For example, Invade and BioShield were developed using endemic strains of the bacterium Serratia entomophila, for use in New Zealand only. To date, most of these home-grown products have either struggled for market share or have remained in small niche markets. However, the number of products registered for use has been steadily increasing in response to consumer demand. Factors limiting past use of biopesticides in New Zealand include market size, registration costs and limited efficacy over a range of climatic zones. Many promising new agents are currently under development as biopesticides with international applications and the launch of several new start-up companies suggests a brighter future for biopesticide use in New Zealand.

Entomopathogenic nematology in Latin America: A brief history, current research and future prospects.

Since the 1980s, research into entomopathogenic nematodes (EPNs) in Latin America has produced many remarkable discoveries. In fact, 16 out of the 117 recognized species of EPNs have been recovered and described in the subcontinent, with many more endemic species and/or strains remaining to be discovered and identified. In addition, from an applied perspective, numerous technological innovations have been accomplished in relation to their implementation in biocontrol. EPNs have been evaluated against over 170 species of agricultural and urban insects, mites, and plant-parasitic nematodes under laboratory and field conditions. While much success has been recorded, many accomplishments remain obscure, due to their publication in non-English journals, thesis dissertations, conference proceedings, and other non-readily available sources. The present review provides a brief history of EPNs in Latin America, including current findings and future perspectives.

Current status and perspectives of fungal entomopathogens used for microbial control of arthropod pests in Brazil.

Entomopathogenic fungi play a central role in Brazil's biopesticide market. Approximately 50% of registered microbial biopesticides comprise mycoinsecticides and/or mycoacaricides consisting of hypocrealean fungi, with most based on Metarhizium anisopliae sensu stricto (s. str.) and Beauveria bassiana s. str. These fungi are mainly used to control spittlebugs in sugarcane fields and whiteflies in row crops, respectively, with annual applications surpassing three million hectares. Research also emphasizes the potential of fungal entomopathogens to manage arthropod vectors of human diseases. Most registered fungal formulations comprise wettable powders or technical (non-formulated) products, with relatively few new developments in formulation technology. Despite the large area treated with mycoinsecticides (i.e., approx. 2 million ha of sugarcane treated with M. anisopliae and 1.5 million ha of soybean treated with B. bassiana), their market share remains small compared with the chemical insecticide market. Nevertheless, several major agricultural companies are investing in fungus-based products with the aim at achieving more sustainable IPM programs for major pests in both organic and conventional crops. Government and private research groups are pursuing innovative technologies for mass production, formulation, product stability and quality control, which will support cost-effective commercial mycoinsecticides. Here, we summarize the status of mycoinsecticides currently available in Brazil and discuss future prospects.

Widespread adoption of Bt cotton and insecticide decrease promotes biocontrol services.

Over the past 16 years, vast plantings of transgenic crops producing insecticidal proteins from the bacterium Bacillus thuringiensis (Bt) have helped to control several major insect pests and reduce the need for insecticide sprays. Because broad-spectrum insecticides kill arthropod natural enemies that provide biological control of pests, the decrease in use of insecticide sprays associated with Bt crops could enhance biocontrol services. However, this hypothesis has not been tested in terms of long-term landscape-level impacts. On the basis of data from 1990 to 2010 at 36 sites in six provinces of northern China, we show here a marked increase in abundance of three types of generalist arthropod predators (ladybirds, lacewings and spiders) and a decreased abundance of aphid pests associated with widespread adoption of Bt cotton and reduced insecticide sprays in this crop. We also found evidence that the predators might provide additional biocontrol services spilling over from Bt cotton fields onto neighbouring crops (maize, peanut and soybean). Our work extends results from general studies evaluating ecological effects of Bt crops by demonstrating that such crops can promote biocontrol services in agricultural landscapes.

Recombinant entomopathogenic agents: a review of biotechnological approaches to pest insect control.

Although the use of chemical pesticides has decreased in recent years, it is still a common method of pest control. However, chemical use leads to challenging problems. The harm caused by these chemicals and the length of time that they will remain in the environment is of great concern to the future and safety of humans. Therefore, developing new pest control agents that are safer and environmentally compatible, as well as assuring their widespread use is important. Entomopathogenic agents are microorganisms that play an important role in the biological control of pest insects and are eco-friendly alternatives to chemical control. They consist of viruses (non-cellular organisms), bacteria (prokaryotic organisms), fungi and protists (eukaryotic organisms), and nematodes (multicellular organisms). Genetic modification (recombinant technology) provides potential new methods for developing entomopathogens to manage pests. In this review, we focus on the important roles of recombinant entomopathogens in terms of pest insect control, placing them into perspective with other views to discuss, examine and evaluate the use of entomopathogenic agents in biological control.

Insect pathogens as biological control agents: Back to the future.

The development and use of entomopathogens as classical, conservation and augmentative biological control agents have included a number of successes and some setbacks in the past 1years. In this forum paper we present current information on development, use and future directions of insect-specific viruses, bacteria, fungi and nematodes as components of integrated pest management strategies for control of arthropod pests of crops, forests, urban habitats, and insects of medical and veterinary importance. Insect pathogenic viruses are a fruitful source of microbial control agents (MCAs), particularly for the control of lepidopteran pests. Most research is focused on the baculoviruses, important pathogens of some globally important pests for which control has become difficult due to either pesticide resistance or pressure to reduce pesticide residues. Baculoviruses are accepted as safe, readily mass produced, highly pathogenic and easily formulated and applied control agents. New baculovirus products are appearing in many countries and gaining an increased market share. However, the absence of a practical in vitro mass production system, generally higher production costs, limited post application persistence, slow rate of kill and high host specificity currently contribute to restricted use in pest control. Overcoming these limitations are key research areas for which progress could open up use of insect viruses to much larger markets. A small number of entomopathogenic bacteria have been commercially developed for control of insect pests. These include several Bacillus thuringiensis sub-species, Lysinibacillus (Bacillus) sphaericus, Paenibacillus spp. and Serratia entomophila. B. thuringiensis sub-species kurstaki is the most widely used for control of pest insects of crops and forests, and B. thuringiensis sub-species israelensis and L. sphaericus are the primary pathogens used for control of medically important pests including dipteran vectors. These pathogens combine the advantages of chemical pesticides and MCAs: they are fast acting, easy to produce at a relatively low cost, easy to formulate, have a long shelf life and allow delivery using conventional application equipment and systemics (i.e. in transgenic plants). Unlike broad spectrum chemical pesticides, B. thuringiensis toxins are selective and negative environmental impact is very limited. Of the several commercially produced MCAs, B. thuringiensis (Bt) has more than 50% of market share. Extensive research, particularly on the molecular mode of action of Bt toxins, has been conducted over the past two decades. The Bt genes used in insect-resistant transgenic crops belong to the Cry and vegetative insecticidal protein families of toxins. Bt has been highly efficacious in pest management of corn and cotton, drastically reducing the amount of broad spectrum chemical insecticides used while being safe for consumers and non-target organisms. Despite successes, the adoption of Bt crops has not been without controversy. Although there is a lack of scientific evidence regarding their detrimental effects, this controversy has created the widespread perception in some quarters that Bt crops are dangerous for the environment. In addition to discovery of more efficacious isolates and toxins, an increase in the use of Bt products and transgenes will rely on innovations in formulation, better delivery systems and ultimately, wider public acceptance of transgenic plants expressing insect-specific Bt toxins. Fungi are ubiquitous natural entomopathogens that often cause epizootics in host insects and possess many desirable traits that favor their development as MCAs. Presently, commercialized microbial pesticides based on entomopathogenic fungi largely occupy niche markets. A variety of molecular tools and technologies have recently allowed reclassification of numerous species based on phylogeny, as well as matching anamorphs (asexual forms) and teleomorphs (sexual forms) of several entomopathogenic taxa in the Phylum Ascomycota. Although these fungi have been traditionally regarded exclusively as pathogens of arthropods, recent studies have demonstrated that they occupy a great diversity of ecological niches. Entomopathogenic fungi are now known to be plant endophytes, plant disease antagonists, rhizosphere colonizers, and plant growth promoters. These newly understood attributes provide possibilities to use fungi in multiple roles. In addition to arthropod pest control, some fungal species could simultaneously suppress plant pathogens and plant parasitic nematodes as well as promote plant growth. A greater understanding of fungal ecology is needed to define their roles in nature and evaluate their limitations in biological control. More efficient mass production, formulation and delivery systems must be devised to supply an ever increasing market. More testing under field conditions is required to identify effects of biotic and abiotic factors on efficacy and persistence. Lastly, greater attention must be paid to their use within integrated pest management programs; in particular, strategies that incorporate fungi in combination with arthropod predators and parasitoids need to be defined to ensure compatibility and maximize efficacy. Entomopathogenic nematodes (EPNs) in the genera Steinernema and Heterorhabditis are potent MCAs. Substantial progress in research and application of EPNs has been made in the past decade. The number of target pests shown to be susceptible to EPNs has continued to increase. Advancements in this regard primarily have been made in soil habitats where EPNs are shielded from environmental extremes, but progress has also been made in use of nematodes in above-ground habitats owing to the development of improved protective formulations. Progress has also resulted from advancements in nematode production technology using both in vivo and in vitro systems; novel application methods such as distribution of infected host cadavers; and nematode strain improvement via enhancement and stabilization of beneficial traits. Innovative research has also yielded insights into the fundamentals of EPN biology including major advances in genomics, nematode-bacterial symbiont interactions, ecological relationships, and foraging behavior. Additional research is needed to leverage these basic findings toward direct improvements in microbial control.

Engineering plants for aphid resistance: current status and future perspectives.

KEY MESSAGE: The current status of development of transgenic plants for improved aphid resistance, and the pros and cons of different strategies are reviewed and future perspectives are proposed. Aphids are major agricultural pests that cause significant yield losses of crop plants each year. Excessive dependence on insecticides for aphid control is undesirable because of the development of insecticide resistance, the potential negative effects on non-target organisms and environmental pollution. Transgenic plants engineered for resistance to aphids via a non-toxic mode of action could be an efficient alternative strategy. In this review, the distribution of major aphid species and their damages on crop plants, the so far isolated aphid-resistance genes and their applications in developments of transgenic plants for improved aphid resistance, and the pros and cons of these strategies are reviewed and future perspectives are proposed. Although the transgenic plants developed through expressing aphid-resistant genes, manipulating plant secondary metabolism and plant-mediated RNAi strategy have been demonstrated to confer improved aphid resistance to some degree. So far, no aphid-resistant transgenic crop plants have ever been commercialized. This commentary is intended to be a helpful insight into the generation and future commercialization of aphid-resistant transgenic crops in a global context.

Microbial management of arthropod pests of tea: current state and prospects.

Sustainable tea cultivation will rely increasingly on alternatives to conventional chemical insecticides for pest management that are environment-friendly and reduce the amount of pesticide residues in made tea. Entomopathogens can provide effective control, conserve biodiversity, and serve as alternatives to chemical insecticides under several conditions. Due to their specificity for insects, these pathogens including viruses, bacteria, and fungi are ideal candidates for incorporation in the integrated pest management strategies for tea where their effects on other natural enemies will be minimal. Biological and ecological characteristics of several dominant natural entomopathogenic microorganisms have been well documented throughout the tea-growing countries particularly China, Japan, and India. But research to convert them to microbial insecticide formulations for tea pest control by evolving suitable techniques for production, standardization, formulation, and application has not progressed well except in Japan and China to some extent. Increased use of microbial control will depend on a variety of factors including improvements in the pathogens' virulence, formulation, delivery, etc. and an increased awareness of their attributes by growers and the general public. In this review, we provide an overview of microbial control of the key insect pests of tea and also the scope for future studies for their better utilization.