Biological Control Of Diseases Research Articles

The Influence of Temperature on the Growth, Sporulation, Colonization, and Survival of Trichoderma spp. in Grapevine Pruning Wounds

Trichoderma is a genus of fungi used for the biological control of plant diseases and a large number of its bio-formulates are available in the market. However, its efficacy under field conditions remains unclear, especially for the protection of grapevine plants against Grapevine Trunk Diseases (GTDs). These diseases are caused by a complex of fungal pathogens whose main point of entrance into the affected plants is through pruning wounds. In this research, different Trichoderma native strains have been evaluated according to their ability to grow at different temperatures and their capacity to colonize pruning wounds in adverse climatic conditions. Strains from section Trichoderma have adapted to cooler conditions. On the other hand, strains from clade Harzianum/Virens grow at higher temperatures. However, differences can also be found between strains inside the same clade/section. Native strains were able to colonize more than 70% of vine pruning wounds in winter conditions. The Trichoderma strain T154 showed a significantly higher re-isolation degree from vine plants and its concentration was optimized for spraying onto vine plants. In conclusion, Trichoderma native strains are better adapted to survive in a changing environment, and they could give better protection to grapevine plants in co-evolution with each specific vineyard.

Biological control of damping-off and root rot disease caused by Rhizoctonia solani on cucumber plants

Biological control of damping-off and root rot disease caused by Rhizoctonia solani on cucumber plants

Differences of bacterial and fungal communities in the tree and rhizosphere of the healthy and twig blight-diseased bayberry

Twig blight is a serious disease of bayberry, which limits bayberry production. In order to prevent and manage the disease, we used high-throughput sequencing technology to analyze bacterial and fungal community richness and diversity in various organs of the tree, bulk and rhizosphere soil of healthy and diseased bayberry. The results showed significant differences in richness and diversity of bacteria and fungi in bulk soil, rhizosphere soil, roots, trunks, barks, and leaves between twig blight diseased trees and healthy trees. In bulk soil, the richness and diversity of bacteria significantly decreased, while that of fungi significantly increased. In barks of diseased trees, the richness and diversity of bacteria significantly increased, and those of fungi significantly decreased. The relative abundance of dominant bacteria and fungi in various organs, bulk soil, and root surface soil changed significantly at phylum, class, and genus levels in the diseased trees. The relative abundance of Pseudomonas sp. on the trunks, roots, and root surface soils of diseased trees significantly decreased, and Fusarium sp. of the diseased root surface and bulk soils also significantly decreased, while the relative abundance of Penicillium sp. on the diseased root surface and bulk soils significantly increased. Pestalotiopsis sp., from the same genus as the twig blight pathogen, was less abundant in the roots but more abundant in the leaves, trunks, barks as well as root surface soils and bulk soils of the diseased bayberry trees than those of the health trees. The relative abundance of Pestalotiopsis sp. was positively correlated with those of most of the fungi. Our results might provide useful theoretical basis for the development of ecological improvement and healthy-tree cultivation technology, and biological control of bayberry twig blight disease.

​Promoting Rhizosphere Microorganisms and Biopesticides: A Review

Microbes are associated inside and outside tissue parts of producers like plants, lichen, algae and phototrophic bacteria. Biofertilizer bacteria, fungi, algae and protozoa provide the suitable environment condition, nutrition constituents and inhibit harmful microorganism from rhizosphere zone. Biofertilizer and biocontrol microbes play key roles for improved soil texture, secreting, hormones, antibiotics and various stress signal compounds, enhancement plant parts elongation and prevent pathogen microbial population. Plant growth promotion rhizosphere microorganisms can improve crop tolerance for the non- living stresses such as drought, heat and salinity and living stress such as soil borne pathogens, over load of the microbial population liable to turn out to be more incessant as various atmosphere design keep on creating. Plant disease major issue for loss of productivity and unsafe for living organism due to chemical pesticides, insecticides, fungicides, bactericides and chemical fertilizer. Due to its high potential as an alternative/complement to these pesticides, biological disease control is now generally recognized and constitutes a low cost-efficient eco-friendly biofertilizer, biocontrol, biopesticide and bioinsecticide.

The effect of different plant activators and biological preparete on the biological control of bacterial speck disease in tomato.

Amaç: Pseudomonas syringae pv. tomato (Okabe) Young, Dye and Wilkie, domates bitkisinde bakteriyel benek olarak bilinen hastalığa neden olur. Hastalığın mücadelesinde bakırlı preparatlar kullanılmakla birlikte, son yıllarda kullanılan bakırlı preparatlara karşı hastalık etmeninin dayanıklılık geliştirdiği bilinmektedir. Bu bağlamda sürdürülebilir tarım uygulamaları kapsamında kimyasallara alternatif çevre dostu olan mücadele yöntemlerinin kullanımı önem arz etmektedir. Bu çalışmada, domates bakteriyel benek hastalığının mücadelesinde dayanıklılığı uyaran bitki aktivatörleri ile biyolojik preparatın kullanım olanaklarının araştırılması amaçlanmıştır.Yöntem ve Bulgular: Organik ve geleneksel üretimlerde kullanılabilen beş bitki aktivatörü (Crop-Set, ISR-2000, KingBo, Sergomil L60, Turf-Set) ve bir biyolojik preparat olan Serenade® SC (%1.34 Bacillus subtilis QST 713)’nin bakteriyel benek hastalığına karşı etkisi plastik sera koşullarında araştırılmıştır. Elde edilen verilere göre uygulamaların yapraklardaki hastalık belirtilerini %16-50, gövdedeki belirtileri %25-50 oranında azalttığı saptanmıştır. Yapılan değerlendirmeler sonucunda, yapraklardaki benek ve gövdedeki kara leke belirtilerini baskılamada tüm uygulamalar başarılı bulunmuş olmakla birlikte, uygulamalar arasında Turf-Set, ISR-2000 ve Sergomil en başarılı uygulamalar olarak belirlenmiştir. Söz konusu uygulamalar bitki kuru ağırlığında %4-56 oranında artış sağlamıştır.Genel Yorum: Yapraktaki ve gövdedeki hastalığı baskılama ile bitki büyümesine olan etki değerlendirildiğinde Turf-Set ve ISR-2000 uygulamaları en etkili uygulamalar olarak belirlenmiştir.Çalışmanın Önemi ve Etkisi: Çalışmadan elde edilen sonuçlar, bitki aktivatörlerinin özellikle organik tarımda kimyasallara alternatif mücadele yöntemi olarak kullanılma potansiyeline sahip olduğunu göstermektedir.

Molecular identification and biological control of Ralstonia solanacearum from wilt of papaya by natural compounds and Bacillus subtilis: An integrated experimental and computational study

Ralstonia solanacearum is a harmful pathogen that causes severe wilt disease in several vegetables. In the present study, we identified R. solanacearum from wilt of papaya by 16S rRNA PCR amplification. Virulence ability of R. solanacearum was determined by amplification of approximately 1500 bp clear band of hrpB gene. Further, in-vitro seed germination assay showed that R. solanacearum reduced the germination rate up to 26.21%, 34% and 33.63% of cucumber, bottle guard and pumpkin seeds, respectively whereas shoot and root growth were also significantly decreased. Moreover, growth inhibition of R. solanacearum was recorded using antibacterial compound from medicinal plant and antagonistic B. subtilis. Petroleum ether root extract of Rauvolfia serpentina showed highest 22 ± 0.04 mm diameter of zone of inhibition where methanolic extract of Cymbopogon citratus and ethanolic extract of Lantana camara exhibited 20 ± 0.06 mm and 20 ± 0.01 mm zone of inhibition against R. solanacearum, respectively. In addition, bioactive compounds of B. subtilis inhibited R. solanacearum growth by generating 17 ± 0.09 mm zone of inhibition. To unveil the inhibition mechanism, we adopted chemical-protein interaction network and molecular docking approaches where we found that, rutin from C. citratus interacts with citrate (Si)-synthase and dihydrolipoyl dehydrogenase of R. solanacearum with binding affinity of −9.7 kcal/mol and −9.5 kcal/mol while quercetin from B. subtillis interacts with the essential protein F0F1 ATP synthase subunit alpha of the R. solancearum with binding affinity of −6.9 kcal/mol and inhibit the growth of R. solanacearum. Our study will give shed light on the development of eco-friendly biological control of wilt disease of papaya.

Efficacy of variable timing of Pythium oligandrum applications on red clover grown under field conditions

Biological control of root diseases of red clover (Trifolium pratense L.) could be effective but optimization of its application is needed to balance treatment efficacy with costs. Our objectives were to evaluate the effects of different Pythium oligandrum application treatments on root morphological development, plant root disease score and forage yield in a three-year field experiment. Red clover cv. Start was managed under a three-cut management in two harvest years with four treatments: (i) untreated control, (ii) multiple applications of P. oligandrum (intensive treatment), (iii) a single spring application, and (iv) a single autumn application. Plants from each plot were evaluated for root morphology traits and scored for root diseases in each autumn. Forage yield, compressed stand height and stem density were evaluated in each cut. All P. oligandrum treatments increased root dry matter and root potential index, but only the intensive treatment significantly improved forage yield (by 31%) in comparison with the control. This effect could be largely associated with plant growth stimulation resulting in increased stand height before cutting. Among single application treatments, autumn timing significantly reduced plant root disease score and showed higher stand height than spring application. One application later in the season seems likely to be effective for forage growth stimulation, but probably at least two applications are needed to produce maximum effect. Despite some drought stress during the first two years, this field study confirmed an effect of P. oligandrum on improved root development and a positive yield response, thus showing a high potential of this biological control treatment under field conditions.

Biological Control of Fungal Diseases by Trichoderma aggressivum f. europaeum and Its Compatibility with Fungicides.

Our purpose was to evaluate the ability of Trichoderma aggressivum f. europaeum as a biological control agent against diseases from fungal phytopathogens. Twelve isolates of T. aggressivum f. europaeum were obtained from several substrates used for Agaricus bisporus cultivation from farms in Castilla-La Mancha (Spain). Growth rates of the 12 isolates were determined, and their antagonistic activity was analysed in vitro against Botrytis cinerea, Sclerotinia sclerotiorum, Fusarium solani f. cucurbitae, Pythium aphanidermatum, Rhizoctonia solani, and Mycosphaerella melonis, and all isolates had high growth rates. T. aggressivum f. europaeum showed high antagonistic activity for different phytopathogens, greater than 80%, except for P. aphanidermatum at approximately 65%. The most effective isolate, T. aggressivum f. europaeum TAET1, inhibited B. cinerea, S. sclerotiorum, and M. melonis growth by 100% in detached leaves assay and inhibited germination of S. sclerotiorum sclerotia. Disease incidence and severity in plant assays for pathosystems ranged from 22% for F. solani to 80% for M. melonis. This isolate reduced the incidence of Podosphaera xanthii in zucchini leaves by 66.78%. The high compatibility by this isolate with fungicides could allow its use in combination with different pest management strategies. Based on the results, T. aggressivum f. europaeum TAET1 should be considered for studies in commercial greenhouses as a biological control agent.

Use of efficient microorganisms in agriculture

The interest in the use of microorganisms in agricultural practices has increased significantly in recent years, both in the promotion of plant growth and in the biological control of plant pests and diseases. This literature review work aimed to address information on the use of isolation, multiplication, use and storage methodologies for efficient microorganisms (Effective Microorganisms, EM) in agriculture. These microorganisms have important functions for their hosts, as they have symbiotic interactions with them, and are capable of protecting plants from attack by insects, diseases and herbivorous mammals through the production of toxins. The use of EM in agriculture aims to accelerate the natural composition of organic matter and promote the balance of microbial flora contributing to plant development. EMs are potential substitutes for chemical products, and can thus favor the preservation of the environment. They are collected from fertile forest soils through simple and inexpensive methodologies, consisting of a tool with potential to be used both by family farmers and on a small and large scale. They can be used in different ways, the main ones being in soil, plant, water and animals. The use of EM is an accessible and low-cost technique, in addition to being easy to prepare within the property itself, contributing to the sustainability of agricultural systems.

Mycoparasitic nature of Egyptian Trichoderma isolates and their impact on suppression Fusarium wilt of tomato

BackgroundPlant disease administration is difficult due to the soil-borne nature of the phytopathogens. Biological control of plant disease is a safe mode to avoid the problems related to fungal diseases that affect crops productivity.ResultsTwenty-three Trichoderma isolates were isolated from soil, surrounding healthy tomato roots from different regions in the Egyptian Governorate of Menoufia. Using a dual culture method to test the efficiency of Trichoderma isolates, the most effective isolate identified as Trichoderma atrovirde with percentage inhibition against Fusarium oxysporum f. sp. lycopersici (92.11%) and scanning electron microscope examination documented the mycoparasitic nature of T. atrovirde to F. oxysporum. Treatments with 10% filtrate T. atrovirde improved the growth aspects of tomato plants than the control plants or infected only, as well as the increase in phenol content (15.09 ug. g-1dry weight) and decreased disease incidence percentage (8%) than the plants infected only (60%).ConclusionsThis study clearly demonstrated that T. atrovirde had a significant inhibition against F. oxysporum. Greenhouse assays displayed the protective role of T. atrovirde inoculation directly against pathogen or indirectly related to the defense mechanism in the plant. So, this study recommends using T. atrovirde for biological control of wilt disease in tomato plants.

Reaping the Benefits of Microorganisms in Cropping Systems: Is the Regulatory Policy Adequate?

Within food plant cropping systems, microorganisms provide vital functions and ecosystem services, such as biological pest and disease control, promotion of plant growth and crop quality, and biodegradation of organic matter and pollutants. The beneficial effects of microorganisms can be achieved and/or enhanced by agricultural management measures that target the resident microbial biodiversity or by augmentation with domesticated and propagated microbial strains. This study presents a critical review of the current legislation and regulatory policies pertaining to the utilization of plant-beneficial microorganisms in the European Union (EU). For augmentative approaches, the nature of the intended effect and the product claim determine how a microbiological product is categorized and regulated, and pre-market authorization may be mandatory. Typically, microbial products have been incorporated into frameworks that were designed for evaluating non-living substances, and are therefore not well suited to the specific properties of live microorganisms. We suggest that regulatory harmonization across the sector could stimulate technical development and facilitate implementation of crop management methods employing microorganisms. Possible scenarios for regulatory reform in the longer term are discussed, but more investigation into their feasibility is needed. The findings of this study should serve as a catalyst for more efficient future use of plant-beneficial microorganisms, to the benefit of agriculture as well as the environment.

In vitro efficacy of biologically active compounds derived from Navicula arenaria against soil borne phytopathogenic Macrophomina phaseolina and Fusarium oxysporum

Navicula arenaria Donkin 1861 is a marine microalga belongs to class of bacillariophycea. Recently, microalgae and their products are used as biological control of diseases caused by phytopathogenic fungi. This is considered environmentally ecofriendly method to overcome the plant damage caused by soil borne pathogenic fungi and thereby economic loss. Therefore, this study aimed to investigate in vitro the antifungal activity of N. arenaria isolate PS 31 extracellular and intracellular metabolites against two taxa of soil borne phytopathogenic fungi; Macrophomina phaseolina and Fusarium oxysporum. N. arenaria PS 31 hexane extract was the most effective extract on growth inhibition of both phytopathogenic fungi. There is no significant difference between miconazole and N. arenaria hexane extract of 5.6 mg/ml on growth inhibition of F. oxysporum. The inhibitory effect of hexane extract at 5.6, 4.2 and 2.8 mg/ml and ethyl acetate extract at 35% (v/v) was higher than controls. Ethyl acetate extract was effective on growth inhibition of M. phaseolina (29.67%). GC-MS analysis of N. arenaria hexane fraction revealed the presence of potent antifungal compounds such as Phenol, 2,2'-methylenebis[6-(1,1-dimethylethyl)-4-methyl-, di-n-octyl phthalate, cholestane-3,5-diol, 5-acetate,(3.beta.,5.alpha.), Cholestan-3-ol,(3.beta.,5.beta.)- and beta.-Sitosterol. These results suggest that N. arenaria hexane extract can be used in biological control of plant diseases caused by M. phaseolina and F. oxysporum.

Assessing Aerated Vermicompost Tea Combined with Microbial Biological Control Agents for Suppression of Fusarium and Rhizoctonia.

Biological control of plant diseases is important in organic greenhouse vegetable production, where fungicide use is limited. Organic producers use microbially diverse substrates, including composts, as media for plant growth. Previous research into the impact of vermicompost on the efficacy of applied biocontrol agents is limited. An invitro assay was developed to test the efficacy of two biological control agents in a competitive microbial background. Suppression of the pathogen Fusarium oxysporum f. sp. radicis-cucumerinum by Clonostachys rosea f. catenulata (Gliocladium catenulatum strain J1446 [Prestop]) and Bacillus subtilis strain QST 713 (Rhapsody), was assessed on agar media amended with aerated vermicompost tea (ACT). Pathogen growth was reduced more by C. rosea than ACT alone, and C. rosea was equally effective when combined with ACT. In contrast, B. subtilis reduced pathogen growth less than ACT and, when combined, reduced pathogen growth no more than ACT alone. Both biocontrol agents were similarly tested with ACT against F. oxysporum f. sp. radicis-cucumerinum and Rhizoctonia solani on cucumber and radish. Additive, neutral, and antagonistic responses, depending on host, pathogen, and biocontrol agent, were observed. ACT alone provided more consistent disease suppression on cucumber compared with B. subtilis or C. rosea. In combination, disease suppression was most often better than each biocontrol alone but not better than ACT alone. ACT had antagonistic or additive interactions with C. rosea in the radish/R. solani pathosystem, depending on the experiment. The specific and general suppression of plant diseases by biological control agents in microbially rich environments is variable and warrants further study.

Prospects of Biological Control of Plant Diseases in Ghana: A Mini-Review

Prospects of Biological Control of Plant Diseases in Ghana: A Mini-Review

Biological control of important fungal diseases of potato and raspberry by two Bacillus velezensis strains.

Stem canker and black scurf caused by Rhizoctonia solani are the important diseases in potato, while spur blight caused by Didymella applanata is a major disease in red raspberry. In Western Siberia, both crops are grown predominantly in small-scale farming that requires maximal usage of biological products for plant protection instead of chemicals. We evaluated two promising Bacillus velezensis strains BZR 336 g and BZR 517 isolated in the south of Russia (45°1′N, 38°59′E) for their biological control potentials against the potato and red raspberry diseases under the more severe weather conditions of Western Siberia (55°1′N, 82°55′ E). We tested two techniques to apply biocontrol agents: (1) coating the seeds (potato tubers) and (2) spraying over the plants (raspberry canes). In each case, we estimated B. velezensis strains on two plant cultivars differed by the disease resistance. The degree of B. velezensis influence on disease incidence and severity depended on the bacterial strain, the protected plant, and its cultivar. We also demonstrated that two B. velezensis strains significantly stimulated plant growth of potato, which contributed to the plant productivity on both cultivars. The BZR 336 g strain affected the potato productivity more than the BZR 517 strain. Under the influence of both bacterial strains, raspberry yield was significantly higher compared to the control on the susceptible cultivar. These findings indicated that two southern B. velezensis strains had proved their efficacy as biological control agents in the control of the serious fungal infection of potato and raspberry plants under the more severe ecological conditions of Western Siberia. For the first time, we demonstrated B. velezensis strains potential for use as biological control agents against R. solani on potato, and against D. applanata on red raspberry.

Preparation of Nanomaterial Wettable Powder Formulations of Antagonistic Bacteria from Phellodendron chinense and the Biological Control of Brown Leaf Spot Disease.

Brown leaf spot disease caused by Nigrospora guilinensis on Phellodendron chinense occurs in a large area in Dayi County, Chengdu City, Sichuan Province, China each year. This outbreak has severely reduced the production of Chinese medicinal plants P. chinense and caused substantial economic losses. The bacterial isolate JKB05 was isolated from the healthy leaves of P. chinense, exhibited antagonistic effects against N. guilinensis and was identified as Bacillus megaterium. The following fermentation medium and conditions improved the inhibitory effect of B. megaterium JKB05 on N. guilinensis: 2% glucose, 0.1% soybean powder, 0.1% KCl, and 0.05% MgSO4; initial concentration 6 × 106 cfu/ml, and a 42-h optimal fermentation time. A composite of 0.1% nano-SiO2 JKB05 improved the thermal stability, acid-base stability and ultraviolet resistance by 16%, 12%, and 38.9%, respectively, and nano-SiO2 was added to the fermentation process. The best formula for the wettable powder was 35% kaolin, 4% polyethylene glycol, 8% Tween, and 2% humic acid. The following quality test results for the wettable powder were obtained: wetting time 87.0 s, suspension rate 80.33%, frequency of microbial contamination 0.08%, pH 7.2, fineness 95.8%, drying loss 1.47%, and storage stability ≥83.5%. A pot experiment revealed that the ability of JKB05 to prevent fungal infections on P. chinense increased considerably and achieved levels of control as high as 94%. The use of nanomaterials significantly improved the ability of biocontrol bacteria to control this disease.

Biological prevention and control of pitaya fruit canker disease using endophytic fungi isolated from papaya.

Pitaya fruit canker is an important disease in pitaya production. Facilitating resistance through the application of biological control principles is a promising alternative to traditional control strategies. This study evaluated the induced resistance of Penicillium rolfsii, numbered Y17 isolated from papaya leaves in pitaya fruit, and evaluated the activity of the defense enzymes, total antioxidant capacity (T-AOC), and malondialdehyde (MDA) content of the treated fruit. The results demonstrate that treatment with Y17 effectively induced resistance of pitaya fruit to canker disease caused by Neoscytalidium dimidiatum, with an inhibition rate of 70.87%. In addition, Y17 notably improved the activities of peroxidase, catalase, and polyphenol oxidase as well as the T-AOC of the treated samples. Y17 treatment reduced the MDA content in these fruits. Taken together, our results suggest that Y17 treatment could trigger pitaya fruit defense responses and effectively induce resistance to fruit canker disease.

Streptomyces consortia-mediated plant defense against Fusarium wilt and plant growth-promotion in chickpea

Three strains of Streptomyces griseus (CAI-24, CAI-121 and CAI-127) and one strain each of Streptomyces africanus (KAI-32) and Streptomyces coelicolor (KAI-90) were reported by us as biocontrol agents against Fusarium wilt, caused by Fusarium oxysporum f. sp. ciceri (FOC), and as plant growth-promoters (PGP) in chickpea. In the present study, the combined effect of these Streptomyces strains as a consortium were assessed for their biocontrol potential against Fusarium wilt and PGP in chickpea. Based on their compatibility, biocontrol ability and PGP performance, two consortia were assembled, consortium-1 having all the five strains of Streptomyces sp. and consortium-2 having the two promising strains (CAI-127 and KAI-32). Under greenhouse conditions, consortium-1 and consortium-2 were found to reduce the Fusarium wilt disease incidence by 55% and 74%, while under field conditions, these were by 86% and 96% in year-1 and by 54% and 69% in year-2, respectively, when compared to the positive control (only FOC treated). Shoot samples treated with consortia + FOC contained significantly enhanced antioxidant enzymes such as superoxide dismutase, catalase, ascorbate peroxidase, guaiacol peroxidase, glutathione reductase and phenylalanine ammonia-lyase, when compared to the positive control (only FOC treated) or the negative control samples (neither FOC nor consortia treated). When the consortia were evaluated for their PGP traits under field conditions in two chickpea cultivars, JG11 and ICCV2, and in two consecutive years, nodule number was found to enhance up to 25%, nodule weight up to 49%, leaf area up to 37%, leaf weight up to 43%, root weight up to 23%, shoot weight up to 35%, seed weight up to 30%, seed number up to 29%, total dry matter up to 22% and grain yield up to 22% over the un-inoculated control plants. This study had demonstrated that the selected consortium of Streptomyces spp. has a greater potential for biological control of Fusarium wilt disease and PGP in chickpea.

Comparative Analysis of Mitochondrial Genome Features among Four Clonostachys Species and Insight into Their Systematic Positions in the Order Hypocreales.

The mycoparasite fungi of Clonostachys have contributed to the biological control of plant fungal disease and nematodes. The Clonostachys fungi strains were isolated from Ophiocordyceps highlandensis, Ophiocordyceps nigrolla and soil, which identified as Clonostachys compactiuscula, Clonostachys rogersoniana, Clonostachys solani and Clonostachys sp. To explore the evolutionary relationship between the mentioned species, the mitochondrial genomes of four Clonostachys species were sequenced and assembled. The four mitogenomes consisted of complete circular DNA molecules, with the total sizes ranging from 27,410 bp to 42,075 bp. The GC contents, GC skews and AT skews of the mitogenomes varied considerably. Mitogenomic synteny analysis indicated that these mitogenomes underwent gene rearrangements. Among the 15 protein-coding genes within the mitogenomes, the nad4L gene exhibited the least genetic distance, demonstrating a high degree of conservation. The selection pressure analysis of these 15 PCGs were all below 1, indicating that PCGs were subject to purifying selection. Based on protein-coding gene calculation of the significantly supported topologies, the four Clonostachys species were divided into a group in the phylogenetic tree. The results supplemented the database of mitogenomes in Hypocreales order, which might be a useful research tool to conduct a phylogenetic analysis of Clonostachys. Additionally, the suitable molecular marker was significant to study phylogenetic relationships in the Bionectriaceae family.

Novel and rapid agar plate methods for in vitro assessment of bacterial biocontrol isolates' antagonism against multiple fungal phytopathogens.

Biological control of plant diseases with antagonistic bacteria is a promising alternative to conventional chemical control strategies. In vitro screening for inhibition of mycelial growth of phytopathogenic fungi by bacterial isolates is the first step in selecting putative bacterial biocontrol agents. Dual culture plate assay is the most common method involved in this first-line selection process. However, it needs independent agar plates to test antagonism by a specific bacterial isolate against each of the fungal phytopathogen. Two modified in vitro antagonism tests are proposed here. Antagonistic activity of a putative biocontrol bacterial strain against four different fungal phytopathogens could be assessed in a single agar plate simultaneously. A comparison of the new methods with conventional dual culture plate assay was also done. The proposed methods are easy to perform and results of antagonism are obtained rapidly. Results of fungal inhibition were qualitatively comparable with that generated through dual culture plate assay. Quantity of resources such as agar medium and plates required for the modified antagonistic assays is several folds less than that required for dual culture plate assay.