Biological control of plant diseases – What has been achieved and what is the direction?

The global sustainability agenda is increasing the demand for reduction in inputs into agricultural production while maintaining profitable yield of quality products. Plant diseases are a major constraint for both yield and product quality, but often tools for their control are ineffective or lacking. Biological control using antagonistic microorganisms has long been a subject of research resulting in a wide range of products that are now available and marketed in specific territories around the world. These preparations are often niche products with narrow uses. The research effort is intense both to develop new biological control agents (BCAs) and to obtain knowledge of the mechanisms underlying biological disease control. The prospects for biological control are promising. As a minimum, BCAs supplement other sustainable disease management practices such as disease resistance, and present opportunities for controlling diseases for which other approaches are ineffective or unavailable. We can realistically expect increasing use of BCAs to control crop diseases in ways that will benefit the environment. This review paper arose from a webinar held by the British Society for Plant Pathology as part of the International Year of Plant Heath (IYPH2020), at which many of the 300 participants posed or discussed interesting questions. This review is based on that input and the panel members at the webinar are all included as co-authors in this review.     

Combined use of biocontrol agents to manage plant diseases in theory and practice

Effective use of biological control agents (BCAs) is a potentially important component of sustainable agriculture. Recently, there has been an increasing interest among researchers in using combinations of BCAs to exploit potential synergistic effects among them. The methodology for investigating such synergistic effects was reviewed first and published results were then assessed for available evidence for synergy. Correct formulation of hypotheses based on the theoretical definition of independence (Bliss independence or Loewe additivity) and the subsequent and statistical testing for the independence-synergistic-antagonistic interactions have rarely been carried out thus far in studies on biocontrol of plant diseases. Thus, caution must be taken when interpreting reported "synergistic" effects without assessing the original publications. Recent theoretical modeling work suggested that disease suppression from combined use of two BCAs was, in general, very similar to that achieved by the more efficacious one, indicating no synergistic but more likely antagonistic interactions. Only in 2% of the total 465 published treatments was there evidence for synergistic effects among BCAs. In the majority of the cases, antagonistic interactions among BCAs were indicated. Thus, both theoretical and experimental studies suggest that, in combined use of BCAs, antagonistic interactions among BCAs are more likely to occur than synergistic interactions. Several research strategies, including formulation of synergy hypotheses in relation to biocontrol mechanisms, are outlined to exploit microbial mixtures for uses in biocontrol of plant diseases.     

The role of bacillus-based biological control agents in integrated pest management systems: plant diseases

ABSTRACT Bacillus-based biological control agents (BCAs) have great potential in integrated pest management (IPM) systems; however, relatively little work has been published on integration with other IPM management tools. Unfortunately, most research has focused on BCAs as alternatives to synthetic chemical fungicides or bactericides and not as part of an integrated management system. IPM has had many definitions and this review will use the national coalition for IPM definition: "A sustainable approach to managing pests by combining biological, cultural, physical and chemical tools in a way that minimizes economic, health and environmental risks." This review will examine the integrated use of Bacillus-based BCAs with disease management tools, including resistant cultivars, fungicides or bactericides, or other BCAs. This integration is important because the consistency and degree of disease control by Bacillus-based BCAs is rarely equal to the control afforded by the best fungicides or bactericides. In theory, integration of several tools brings stability to disease management programs. Integration of BCAs with other disease management tools often provides broader crop adaptation and both more efficacious and consistent levels of disease control. This review will also discuss the use of Bacillus-based BCAs in fungicide resistance management. Work with Bacillus thuringiensis and insect pest management is the exception to the relative paucity of reports but will not be the focus of this review.     

Challenges facing arthropod biological control: identifying traits for genetic improvement of predators in protected crops

Biological control is an efficient pest control method but there are still limitations that are hindering its wider adoption. Genetic improvement of biological control agents (BCAs) can help to overcome these constraints, but the choice of key attributes for better performance that need to be selected is still an open question. Several characteristics have been suggested but the harsh reality is that selective breeding of BCAs has received a lot of attention but resulted in very little progress. Identifying the appropriate traits to be prioritized may be the first step to reverse this situation. In our opinion, the best way is to look at the factors limiting the performance of key BCAs, especially generalist predators (pesticide compatibility, prey‐density dependence, non‐suitable crops, and extreme environmental conditions), and according to these challenges, to choose the attributes that would allow BCAs to overcome those limitations. The benefits of selection for higher resistance to toxins, whether artificially applied (pesticides) or plant produced (plant defenses); increased fitness when feeding on non‐prey food (supplemented or plant‐derived); and better adaptation to extreme temperature and humidity are discussed. In conclusion, genetic improvement of BCAs can bring about new opportunities to biocontrol industry and users to enhance biocontrol resilience. © 2020 Society of Chemical Industry     

Work of the EPPO/IOBC Panel on biological control agents

EPPO started work on biological control agents (BCAs) in 1996, and the joint EPPO/IOBC Panel was established in 1997. The history of the Panel is provided and EPPO Standards developed by the Panel are described. These Standards are: PM 6/1 ‘First import of exotic biological control agents for research under contained conditions’, PM 6/2 ‘Import and release of non‐indigenous biological control agents’ and PM 6/3 ‘List of biological control agents widely used in the EPPO region’. The last of these has been annually updated as a ‘Positive List’ of BCAs for which EPPO recommends its member countries to use a simplified procedure for import and releases. EPPO activities in biological control have been focused on the safety aspects of the introduction of invertebrate BCAs. However, the scope of EPPO's work in this area is now under review, and a number of issues are being considered by the Working Party on Phytosanitary Regulations. These include the remit of the Panel and Standards in relation to micro‐organisms, the evaluation of potential environmental benefits as well as potential risks from releases and the potential for use of BCAs against regulated pests and those recommended for regulation.     

Biosuppression of Botrytis cinerea in grapes

There is increasing interest in the use of biological control agents (BCAs) and plant resistance stimulants to suppress botrytis bunch rot in grapes, caused by Botrytis cinerea . Numerous different filamentous fungi, bacteria and yeasts have been selected as potential BCAs for control of grey mould based upon demonstrated antagonism towards B. cinerea. Biological suppression of the pathogen arises via competition for nutrients and space, the production of inhibitory metabolites and/or parasitism. Preformed and inducible grapevine defence mechanisms also contribute to disease suppression by preventing or delaying pathogenic infection. Furthermore, various biotic and abiotic agents can stimulate grapevine defence mechanisms and so elevate resistance to B. cinerea infection. Biosuppression of B. cinerea in vineyards, using BCAs and resistance stimulants, has been inconsistent when compared with that observed in controlled glasshouse or laboratory conditions. This may be attributable, in part, to the innate variability of the field environment. Research to improve field efficacy has focused on formulation improvement, the use of BCA mixtures and combinational approaches involving BCAs and plant resistance stimulants with complementary modes of action.     

Regulation and use of biological control agents in Hungary

A wide range of biological control agents (BCAs) have been authorized and used in Hungary for the control of pests (including pathogens). BCAs are key elements of both integrated pest management and organic farming for different crops. Authorization of BCAs depends on the type of ‘active substance’. Micro‐organisms can be authorized under the EU Regulation (EC) No. 1107/2009 as plant protection products (PPP), while macro‐organisms are not within its scope. Plant protection tools (traps, mating disruption) are registered at a national level in Hungary, although mass traps and mating disruption pheromones are considered to be PPP. Plant extracts can be authorized as PPP, but some of them can be approved as a ‘basic substance’ and in this case they do not need further authorization. In Hungary, the vast majority of macro‐organisms used as BCAs have been mainly used in greenhouses and polytunnels for the control of thrips, whiteflies, aphids, leaf mining flies, phytophagous mites and lepidopteran pests in vegetable and ornamental crops. Recently a non‐indigenous chalcid wasp, Torymus sinensis, has been successfully introduced for the control of the oriental chestnut gall wasp, Dryocosmus kuriphilus, on chestnut (Castanea sativa) on different sites.     

Effect of reduced risk pesticides on greenhouse vegetable arthropod biological control agents

BACKGROUND: Arthropod biological control agents (BCAs) are commonly released for greenhouse vegetable insect pest management. Nevertheless, chemicals remain a necessary control tactic for certain insect pests and diseases and they can have negative impacts on BCAs. The compatibility of some formulated reduced risk insecticides (abamectin, metaflumizone and chlorantraniliprole) and fungicides (myclobutanil, potassium bicarbonate and cyprodinil + fludioxonil) used, or with promise for use, in Canadian greenhouses with Orius insidiosus (Say), Amblyseius swirskii (Athias‐Henriot) and Eretmocerus eremicus (Rose & Zolnerovich) was determined through laboratory and greenhouse bioassays. RESULTS: Overall, the insecticides and fungicides were harmless as residues to adult BCAs. However, abamectin was slightly to moderately harmful to O. insidiosus and A. swirskii in laboratory bioassays, whereas metaflumizone was slightly harmful to E. eremicus. CONCLUSIONS: In general, these products appear safe to use prior to establishment/release of these adult BCAs. Copyright © 2010 Society of Chemical Industry     

木霉几丁质酶及其对植物病原真菌的拮抗作用

 木霉(Trichoderma spp.)作为重要的植病生防因子(biocontrol agents)一直受到普遍关注。木霉菌株产生的包括几丁质酶在内的细胞壁降解酶,在木霉重寄生中起重要作用。本文论述木霉几丁质酶的种类、诱导产生、理化特性及其对植物病原真菌的拮抗作用,并对木霉几丁质酶及其基因的生防应用进行讨论。     

Selection of biological control agents for controlling soil and seed-borne diseases in the field

Different screening methods for selection of biological control agents (BCAs), for controlling soil and seed-borne diseases, are discussed. The shortcomings of laboratory methods focused on mechanism of action are discussed and we conclude that these methods should be used with caution if candidates with multifactorial or plant mediated mechanisms of control are to be obtained. In vitro screens may be useful for specific groups of microorganisms, thus, screens for antibiotics may be relevant for Streptomyces spp., and promising results have been obtained using soil plating or precolonized agar methods to screen for mycoparasitism and competitive saprophytic ability. Experience with screening in the Nordic programme Biological control of seed borne diseases in cereals is summarized. Research in the four participating countries – Finland, Sweden, Norway and Denmark – followed the same paradigm: that of obtaining antagonists, well adapted to different Nordic environments, and developing them as effective BCAs. Potential antagonists were isolated from different sources and in planta screening methods were developed in order to optimize selection of antagonists effective against a range of seed borne pathogens. Screens in the laboratory or greenhouse were followed by screening in the field. The different screening procedures are compared and evaluated.     

Biological Control of Plant Diseases: The European Situation

The most common approach to biological control consists of selecting antagonistic microorganisms, studying their modes of action and developing a biological control product. Despite progress made in the knowledge of the modes of action of these biological control agents (BCAs), practical application often fails to control disease in the fields. One of the reasons explaining this failure is that the bio-control product is used the same way as a chemical product. Being biological these products have to be applied in accordance with their ecological requirements. Another approach consists of induction of plant defence reactions. This can be done by application of natural substances produced by or extracted from microorganisms, plants, or algae. Since they do not aim at killing the pathogens, these methods of disease control are totally different from chemical control. Although promising, these methods have not been sufficiently implemented under field conditions. A third approach consists of choosing cultural practices that might decrease the incidence or severity of diseases. These methods include the choice of an appropriate crop rotation with management of the crop residues, application of organic amendments and the use of new technology such as the biological disinfestation of soils. Biological control practices need an integrative approach, and more knowledge than chemical control.     

The regulatory landscape for biological control agents

A biological control agent (BCA) has been defined as ‘an organism [used] to reduce the population density of another organism’. However, an organism which is a BCA may, in addition, fall within the regulatory definition of an invasive alien species, a plant pest, a quarantine plant pest, a harmful organism, a plant protection product, or any combination of these. Therefore the regulatory landscape which has to be navigated by applicants wishing to release BCAs is complex. Examples are given of this complexity, drawing on relevant international standards and European Union legislation. A clearer understanding of the regulatory landscape across the EPPO region is necessary for agreeing routes through it to facilitate rapid adoption of biological control options, where these are available, and to ensure that potential risks and benefits are assessed in a coherent and consistent way.     

Comparisons of single versus multiple bacterial species on biological control of anthurium blight

ABSTRACT Effects of single versus multiple biological control agents (BCAs) on suppression of bacterial blight of anthurium were studied using a bioluminescent strain (V108LRUH1) of Xanthomonas campestris pv. dieffenbachiae. When five BCAs (GUT3, GUT4, GUT5, GUT6, and GUT9) were coinoculated in various combinations with V108LRUH1 into filter-sterilized guttation fluids of anthurium plants, a mixture of all five strains or four strains without GUT9 was most inhibitory to V108LRUH1. None of the individual BCAs inhibited V108LRUH1 in the guttation fluid. When BCAs were sprayed at congruent with10(8) CFU/ml on the foliage of a susceptible cultivar 1 day prior to inoculation with V108LRUH1, GUT6 alone and any mixtures containing GUT6 were highly effective in suppressing wound invasion and subsequent leaf infection by V108LRUH1. When tested on several cultivars that differed in susceptibility to the disease, the mixture of five strains or four strains without GUT9 consistently suppressed leaf infection regardless of the cultivars. In some cultivars, BCAs completely suppressed both wound and hydathode invasion by V108LRUH1, resulting in no infection in many leaves. These results indicate that application of bacterial mixtures provides anthurium cultivars with bacterial communities suppressive to X. campestris pv. dieffenbachiae. The results also suggest that selecting an effective mixture of BCAs first and then removing ineffective strains may be a better general approach to finding the most effective BCAs than finding individual strains and combining them.     

我国植物病害生物防治的现状及发展策略

植物病害生物防治是利用有益微生物和微生物代谢产物对农作物病害进行有效防治的技术与方法,本文介绍了我国在生物农药资源筛选评价体系、基因工程、发酵代谢工程等方面取得的可喜进展和研制出的多种高效植物病害生物防治药物;提出未来5～10年,植物病害生物防治学科领域主要工作是集成现代科学技术,进一步明确不同生防因子控制植物病害的分子机制,加强国际合作与交流,建立有关领域共享技术的合作研究平台,提高植物病害生物防治的基础与应用基础研究水平;指出植物病害的生物防治是保障食品和环境安全的重要措施,对国家可持续农业的健康发展具有重大的战略意义。     

Interactions of air temperature,relative humidity and biological control agents on grey mold of bean

The interactions ofBotrytis cinerea and seven biological control agents (BCAs) were examined in controlled environments to determine the influence of selected relative humidities (RH) (90,95, and 100%) and air temperatures (20,24 and 28 C) on grey mold of bean. All main effects and interactions were significant (P0.05) among the 72 treatments. In the control, lesions of grey mold developed under all environmental conditions but were largest at 24 C×95 and 100% RH, and 28 C×95% RH. Interactions of environment, BCAs and grey mold were complex.Alternaria alternata, Drechslera sp.,Myrothecium verrucaria, Trichoderma viride, Gliocladium roseum and an unidentified pink yeast were all highly dependent on environment for biological control efficacy, and changes of 4 C or 5% RH were associated with variability in disease suppression that ranged from 15 to 100%. Efficacy ofEpicoccum purpurascens appeared independent of environment and this BCA suppressed disease by 100% in all of the environmental treatments. Suppression of grey mold by many of the BCAs was most effective under environmental conditions least conducive to disease. Therefore, evaluations of potential BCAs in environmental conditions that are marginal for disease can overestimate their efficacy in field environments. Assessments of biological control efficacy in various environments can be used to more accurately assess the potential of BCAs.     

Selection of bacterial antagonists for the biological control of Rhizoctonia solani in oilseed rape (Brassica napus)

Spore-forming bacteria isolated from oilseed rape (OSR) roots were tested for biological control activity against Rhizoctonia solani damping-off of OSR, using a simple slurry coat formulation. From a total of 239 strains which were tested, Bacillus subtilis 205 was selected as the most promising biocontrol agent (BCA). A known isolate of B. subtilis (NCIMB 12376), which had previously shown activity against R. solani in cotton, was used as a standard comparison throughout. In an in vivo screening, pre-emergence damping-off disease control by B. subtilis 205 was excellent, with plant stands being almost as high as non-diseased controls. However, post-emergence survival was less good, although the surviving plants exhibited significantly reduced hypocotyl rot. Growth-room trials which assessed the effect of ambient temperature on the biological control efficacy of B. subtilis strains 12376 and 205 showed that isolate 205 was more effective at lower temperatures. Disease control by isolate 205 at 15°C/10°C (day/night) suggests that this bacterium could operate under autumn sowing field conditions. Further investigations also indicated that the addition of iron (III) to the BCA formulation may enhance disease control by both bacilli. The potential of bacterial BCAs for the control of seedling diseases is discussed.     

Risikofolgeabschätzung für den Einsatz mikrobieller Antagonisten: Gibt es Effekte auf Nichtzielorganismen?

Biological control of phytopathogenic fungi using antagonistic microorganisms is an environmentally friendly alternative in plant protection. However, possible non-target effects of the applied antagonists on ecologically important soil-microbes need to be considered. Therefore, the aim of this study was to analyse the effect of biological control agents (BCAs) on non-target microbes in the field. Whereas the bacterial BCAs Serratia plymuthica HRO-C48 and Streptomyces sp. HRO-71 were applied to control the pathogen Verticillium dahliae on strawberry and potato, the bacterial strains Pseudomonas trivialis 3Re2-7, P. fluorescens L13-6-12, S. plymuthica 3Re4-18 and the fungal antagonists Trichoderma reesei G1/8 and T. viride G3/2 were introduced to control Rhizoctonia solani on lettuce and potato. As the analysed BCAs belong to different microbial groups like grampositive (HRO-71) and gramnegative (HRO-C48, L13-6-12, 3Re2-7, 3Re4-18) bacteria or the ascomycota (G1/8, G3/2) and originated from different micro-habitats like the rhizosphere or the endorhiza, general conclusion could be drawn from our results. After BCA treatment we did not observe any long-term effect on the plant-associated microbes in any tested pathosystem. Therefore, no sustainable risks could be seen for the indigenous micro-organisms. Our new findings may help to improve the development as well as the registration procedures of future microbial plant protection products.     

The <Emphasis Type="Italic">Caenorhabditis elegans</Emphasis> assay: a tool to evaluate the pathogenic potential of bacterial biocontrol agents

Bacterial biocontrol agents (BCAs) open up the possibility of controlling plant pathogens in an environmentally friendly way. Although they are naturally occurring microbes, some of them can cause diseases in humans. For successful registration it is necessary to test potentially adverse effects on the human health of at-risk candidates. Existing pathogenicity assays are cost-intensive, time-consuming and furthermore they are often inappropriate for facultative pathogens. We developed a new, fast and inexpensive bioassay on the basis of the nematode Caenorhabditis elegans, which is a well-accepted model organism to study bacterial pathogenicity. A selection of eight strains from clinical and environmental origin as well as potential and commercial BCAs from the genera Bacillus, Pseudomonas, Serratia and Stenotrophomonas were screened for their potential to kill the nematode in an in vitro agar plate assay. Furthermore, the motility and reproductive behaviour of nematodes exposed to strains were tested in comparison with those fed by the human pathogen Pseudomonas aeruginosa QC14-3-8 (positive control) and the negative control Escherichia coli OP50. Commercial as well as potential biocontrol strains did not display any adverse effects in all tests. In contrast, the C. elegans assay showed slight effects for clinical and environmental Stenotrophomonas strains. Results showed that the nematode C. elegans provides a model system to indicate the pathogenic potential of BCAs in a very early stage of product development.     

Conclusions from the workshop on evaluation and regulation of biological control agents

This paper reports on the Workshop on Evaluation and Regulation of Biological Control Agents (BCAs) held in Budapest on the 23–24 November 2015. Conclusions and recommendations are summarized and explained, and some of the ways in which the recommendations are being taken forward by the joint EPPO IOBC Panel on Biological Control are described. The key conclusions are that properly authorized releases of BCAs are beneficial for crop protection and the environment, with little evidence of adverse effects. Different countries apply different types of regulation in different ways to determine whether such releases can be authorized. A more harmonized approach could increase opportunities for use of biological control and ensure that potential risks are addressed in a more coherent and consistent way.     

天然产物防治果蔬采后病害研究进展

果蔬采后病害的防治正朝着安全、环保的方向发展,各种生物防治方法成为研究的热点,天然产物用于控制果蔬采后病害的研究也受到了越来越多的关注。本文综述了风味化合物、精油、乙酸、茉莉酸类化合物、芥子油甙、蜂胶、植物提取物、壳聚糖、醋酸瓜类萎蔫醇等天然产物在果蔬采后病害防治中的研究、开发与应用,为未来的商业化开发提供新的思路。