Biological Control Of Diseases Research Articles

Microbial community and chemical composition of cigar tobacco (Nicotiana tabacum L.) leaves altered by tobacco wildfire disease

AbstractTobacco wildfire disease caused by Pseudomonas syringae pv. tabaci is one of the most destructive foliar bacterial diseases occurring worldwide. However, the effect of wildfire disease on cigar tobacco leaves has not been clarified in detail. In this study, the differences in microbiota and chemical factors between wildfire disease‐infected leaves and healthy leaves were characterized using high‐throughput Illumina sequencing and a continuous‐flow analytical system, respectively. The results demonstrated significant alterations in the structure of the phyllosphere microbial community in response to wildfire disease, and the infection of P. syringae pv. tabaci led to a decrease in bacterial richness and diversity. Furthermore, the content of nicotine, protein, total nitrogen, and Cl− in diseased leaves significantly increased by 47.86%, 17.46%, 20.08%, and 72.77% in comparison to healthy leaves, while the levels of total sugar and reducing sugar decreased by 59.59% and 70.0%, respectively. Notably, the wildfire disease had little effect on the content of starch and K+. Redundancy analysis revealed that Pseudomonas, Staphylococcus, Cladosporium, and Wallemia displayed positive correlations with nicotine, protein, total nitrogen, Cl− and K+ contents, while Pantoea, Erwinia, Sphingomonas, Terrisporobacter, Aspergillus, Alternaria, Sampaiozyma, and Didymella displayed positive correlations with total sugar and reducing sugar contents. Brevibacterium, Brachybacterium, and Janibacter were found to be enriched in diseased leaves, suggesting their potential role in disease suppression. Co‐occurrence network analysis indicated that positive correlations were prevalent in microbial networks, and the bacterial network of healthy tobacco leaves exhibited greater complexity compared to diseased tobacco leaves. This study revealed the impact of wildfire disease on the microbial community and chemical compositions of tobacco leaves and provides new insights for the biological control of tobacco wildfire disease.

The Prospect of Hydrolytic Enzymes from Bacillus Species in the Biological Control of Pests and Diseases in Forest and Fruit Tree Production.

Plant diseases and insect pest damage cause tremendous losses in forestry and fruit tree production. Even though chemical pesticides have been effective in the control of plant diseases and insect pests for several decades, they are increasingly becoming undesirable due to their toxic residues that affect human life, animals, and the environment, as well as the growing challenge of pesticide resistance. In this study, we review the potential of hydrolytic enzymes from Bacillus species such as chitinases, β-1,3-glucanases, proteases, lipases, amylases, and cellulases in the biological control of phytopathogens and insect pests, which could be a more sustainable alternative to chemical pesticides. This study highlights the application potential of the hydrolytic enzymes from different Bacillus sp. as effective biocontrol alternatives against phytopathogens/insect pests through the degradation of cell wall/insect cuticles, which are mainly composed of structural polysaccharides like chitins, β-glucans, glycoproteins, and lipids. This study demonstrates the prospects for applying hydrolytic enzymes from Bacillus sp. as effective biopesticides in forest and fruit tree production, their mode of biocidal activity and dual antimicrobial/insecticidal potential, which indicates a great prospect for the simultaneous biocontrol of pests/diseases. Further research should focus on optimizing the production of hydrolytic enzymes, and the antimicrobial/insecticidal synergism of different Bacillus sp. which could facilitate the simultaneous biocontrol of pests and diseases in forest and fruit tree production.

Whole genome sequencing provides evidence for Bacillus velezensis SH-1471 as a beneficial rhizosphere bacterium in plants

Bacillus is widely used in agriculture due to its diverse biological activities. We isolated a Bacillus velezensis SH-1471 from the rhizosphere soil of healthy tobacco, which has broad-spectrum antagonistic activity against a variety of plant pathogenic fungi such as Fusarium oxysporum, and can be colonized in the rhizosphere of a variety of plants. This study will further explore its mechanism by combining biological and molecular biology methods. SH-1471 contains a ring chromosome of 4,181,346 bp with a mean G + C content of 46.18%. We identified 14 homologous genes related to biosynthesis of resistant secondary metabolite, and three clusters encoded potential new antibacterial substances. It also contains a large number of genes from colonizing bacteria and genes related to plant bacterial interactions. It also contains genes related to environmental stress, as well as genes related to drug resistance. We also found that there are many metabolites in the strain that can inhibit the growth of pathogens. In addition, our indoor pot test found that SH-1471 has a good control effect on tomato wilt, and could significantly improve plant height, stem circumference, root length, root weight, and fresh weight and dry weight of the aboveground part of tomato seedlings. Therefore, SH-1471 is a potential biological control strain with important application value. The results of this study will help to further study the mechanism of SH-1471 in biological control of plant diseases and promote its application.

Biological control of anthracnose fruit rot disease (Colletotrichum spp.) and fruit yield improvement of jujube (Zizyphus jujuba Miller var. inermis Rehder) using Bacillus velezensis CE 100

Colletotrichum spp. are notorious phytopathogens that cause anthracnose rot disease in jujube fruits, have a complex lifestyle (with necrotrophic, hemibiotrophic, quiescent, and endophytic phases), they spread rapidly mainly through the production of spores and are increasingly becoming resistant to various fungicides which makes them difficult to control. This study investigated the inhibition of spore germination and mycelial growth of Colletotrichum spp. to suppress anthracnose rot disease in jujube fruits through the activity of cell wall degrading enzymes, and improvement of jujube fruit characteristics and yield via the production of indole-3-acetic acid (IAA) and gibberellin acid (GA3) by Bacillus velezensis CE 100. The bacterial culture broth exhibited a strong chitinase, β-1,3-glucanase and protease and the crude fraction suppressed the germination of spores by more than 95.0 % at a concentration of 100 µL/mL, and substantially reduced mycelial growth of Colletotrichum spp. The microscopic analysis revealed that the treatments degraded the cell walls and caused severe structural distortion of the spores and hyphae of Colletotrichum spp. Field application of B. velezensis CE 100 on the jujube tree crown and in the rhizosphere lowered the rate of anthracnose disease incidence by 2.8-fold and disease severity index (DSI) by 4.4-fold, compared to the control. In addition, B. velezensis CE 100 produced IAA (up to 15.6 mg/mL) and GA3 (up to 1.3 mg/mL), and the field application of the bacterial culture on the jujube tree crown and in the rhizosphere consequently improved the average fruit characteristics (fruit length, fruit diameter and fresh weight by 1.2-, 1.4- and 1.5-fold), chlorophyll index (photosynthetic rate by 1.1-fold), and fruit yield (by 2.1-fold) of jujube, compared to the control group. Thus, the inoculation of B. velezensis CE 100 has the potential effectiveness as a bio-fungicide against anthracnose rot disease in jujube fruits, and as a bio-fertilizer to improve fruit characteristics and yield.

Enhancing biological control of apple rot: Unveiling the antifungal potential and mechanism of Bacillus amyloliquefaciens HZ-12′s lipopeptide

Rot caused by fungal pathogen Aspergillus niger is an important postharvest disease of apples. To control the apple rot resulting from A. niger, five Bacillus strains with antagonism against A. niger were screened by Oxford cup diffusion method. Among them, Bacillus amyloliquefaciens HZ-12 had the most prominent antifungal activity, and its control effect reached 89.0 % on apple rot caused by A. niger. Moreover, the antifungal substances from B. amyloliquefaciens HZ-12 were analyzed, and the lipopeptide was suggested to be the key substance due to strong antifungal stability and surface activity. The lipopeptide extract was confirmed to damage the cell wall and membrane of A. niger, leading to leakage of intracellular nucleic acids and cell death. Further genetic engineering by insertion of a strong promoter significantly improved the iturin A yield by 4.2-fold, and corresponding antifungal effect of B. amyloliquefaciens was obviously increased. This study provided a new strain for biological control of apple rot disease, and also indicated the lipopeptide as antifungal and its corresponding antifungal mechanism.

Bacterial and Fungal Endophytes of Grapevine Cultivars Growing in Primorsky Krai of Russia

In this study, the biodiversity of endophytic bacteria of cultivated grape varieties from the vineyards of Primorsky Krai, Russia, was analyzed for the first time. Far Eastern grape varieties with a high level of stress resistance are a unique object of research as they are cultivated in cold and humid climates with a short summer season. Grapevine endophytic microorganisms are known as promising agents for the biological control of grapevine diseases and agricultural pests. Using genomic approaches, we analyzed the biodiversity of the endophytic bacteria and fungi in the most common grape varieties of Primorsky Krai, Russia: Vitis vinifera × Vitis amurensis cv. Adele (hybrid No. 82-41 F3), Vitis riparia × V. vinifera cv. Mukuzani (pedigree unknown), two cultivars Vitis labrusca × V. riparia cv. Alfa, and Vitis Elmer Swenson 2-7-13 cv. Prairie Star for the first time. The main representatives of the endophytic microorganisms included 16 classes of bacteria and 21 classes of fungi. The endophytic bacterial community was dominated by Gammaproteobacteria (31–59%), followed by Alphaproteobacteria (9–34%) and, to a lesser extent, by the classes Bacteroidia (9–22%) and Actinobacteria (6–19%). The dominant fungal class was Dothideomycetes (43–77%) in all samples analyzed, with the exception of the grapevine cv. Mukuzani from Makarevich, where Malasseziomycetes was the dominant fungal class. In the samples cv. Alfa and cv. Praire Star, the dominant classes were Tremellomycetes and Microbotriomycetes. A comparative analysis of the endophytic communities of the cultivated grape varieties and the wild grape V. amurensis Rupr. was also carried out. We found that 18–20% of the variance between the endophytic communities accounted for the differences between the cultivated and wild grapevines, while the factors “plant location” and “individual plants” accounted for 50–56% and 3–10% of the variance, respectively. The results of this study can be used to develop new means of biocontrol in vineyards to protect plants from abiotic stresses and pathogens.

Screening, Identification, and Growth Promotion of Antagonistic Endophytes Associated with Chenopodium quinoa Against Quinoa Pathogens.

Fungal disease is one of the important reasons for crop yield reduction. Isolation of important endophytes with biocontrol and growth-promoting effects is of great significance for the exploitation of beneficial microbial resources and the biological control of crop fungal diseases. In this study, endophytes from roots, stems, and leaves of quinoa at different growth and development stages were isolated and purified; then the antagonistic activity and growth-promoting characteristics of antagonistic endophytes were determined. Finally, the antagonistic endophytes were identified by morphological characteristics and ITS/16S rRNA sequence analysis. Our results showed that 122 endophytic fungi and 371 endophytic bacteria were isolated from quinoa, of which three endophytic fungi and seven endophytic bacteria were screened that had inhibitory activity against quinoa pathogenic fungi. Most of the antagonistic strains could produce indole-3 acetic acid and had the ability to dissolve organic phosphorus. In addition, the bacterial suspension of endophytic bacteria had the ability to promote the seed germination and plant growth of quinoa. The endophytic fungi with antagonistic activity were identified as Penicillium raperi and P. pulvillorum; the endophytic bacteria were identified as Bacillus paralicheniformis, B. tequilensis, and B. velezensis, respectively. The strains of quinoa endophytes in this study can provide rich microbial resources and a theoretical basis for biological control of plant fungal diseases and agricultural production.

The Characterization of the Inhibitory Substances Produced by Bacillus pumilus LYMC-3 and the Optimization of Fermentation Conditions

Bacillus pumilus plays a very important role in the biological control of plant diseases. In this study, we conducted indoor biocontrol experiments using the LYMC-3 strain of B. pumilus to assess its efficacy against poplar canker and tea gray blight. Simultaneously, we explored the optimal fermentation conditions for the production of inhibitory substances against Phomopsis macrospora and Pestalotiopsis vesicolor. Subsequently, the inhibitory substances were extracted crudely and subjected to a preliminary investigation of their properties. The results of the indoor biocontrol experiments showed that the LYMC-3 strain exhibited disease prevention rates of 44.12% and 48.59% against poplar canker and tea gray blight, respectively. The fermentation optimization experiment showed that a culture medium containing 10 g/L of glucose, 15 g/L of tryptone, and 5 g/L of sodium chloride favored the production of inhibitory substances via the LYMC-3 strain. Among the crude extracts obtained, the non-protein extracts demonstrated the most obvious inhibitory effects. Furthermore, the inhibitory substances exhibited high polarity, excellent thermal stability, and high sensitivity to strong acids and bases. This study establishes a theoretical foundation for the purification and identification of inhibitory substances using the LYMC-3 strain. Additionally, it contributes to elucidating biological control mechanisms and facilitates the application of the LYMC-3 strain in production.

Establishment of Biocontrol Agents and Their Impact on Rhizosphere Microbiome and Induced Grapevine Defenses Are Highly Soil-Dependent

With a reduction in available chemical treatments, there is an increased interest in biological control of grapevine trunk diseases. Few studies have investigated the impact of introducing beneficial microorganisms in the rhizosphere on the existing indigenous soil microbiome. In this study, we explored the effect of two biocontrol agents (BCAs), Trichoderma atroviride SC1 (Ta SC1) (Vintec; Certis Belchim) and Bacillus subtilis PTA-271 (Bs PTA-271), on the grapevine rhizosphere bacterial and fungal microbiome as well as plant defense expression using high-throughput amplicon sequencing and quantitative real-time polymerase chain reaction (PCR), respectively. Additionally, we quantified both Ta SC1 and Bs PTA-271 in the rhizosphere over time using droplet digital PCR. The fungal microbiome was more affected by factors such as soil type, BCA treatment, and sampling time compared with the bacterial microbiome. Specifically, Ta SC1 application produced negative impacts on fungal diversity, whereas application of BCAs did not affect bacterial diversity. Interestingly, the survival and establishment of both BCAs showed opposite trends depending on the soil type, indicating that the physicochemical properties of soils have a role in BCA establishment. Fungal co-occurrence networks were less complex than bacterial networks but highly impacted by Ta SC1 application. Soils treated with Ta SC1 presented more complex and stable co-occurrence networks, with a higher number of positive correlations. Induced grapevine defenses also differed according to the soil, being more affected by BCA inoculation on sandy soil. The findings of this research emphasize the complex relationships among microorganisms in the rhizosphere and highlight the significance of taking into account various factors, such as soil type, sampling time, and BCA treatment, and their influence on the structure and dynamics of microbial communities.

Postharvest biocontrol of green mold (Penicillium digitatum) in citrus by Bacillus velezensis strain S161 and its mode of action

Green mold caused by Penicillium digitatum is a major postharvest disease of citrus that can result in significant losses during storage. Synthetic chemical fungicides typically used to control this disease have raised potential concerns regarding environmental pollution, human health, and the development of fungicide resistance. Biological control of postharvest diseases has been demonstrated to be an effective alternative to chemical control. In the present study, we investigated the postharvest biocontrol efficacy of Bacillus velezensis strain S161 against P. digitatum and its potential mechanisms of action. Results indicated that a cell-free supernatant (CFS) obtained from liquid culture of S161 strongly inhibited the mycelial growth, spore germination, and germ tube elongation of P. digitatum in vitro. The CFS effectively protected citrus fruit against P. digitatum, providing 95.6 % control. Further investigations revealed that the CFS degraded and ruptured hyphal membranes and spores and induced the excessive accumulation of reactive oxygen species (ROS) in hyphal cells resulting in oxidative damage. CFS exposure also resulted in the downregulation of genes associated with spore germination, growth, reproduction, and virulence. antiSMASH analysis of biosynthetic gene clusters (BGCs) in B. velezensis indicated that B. velezensis possesses several core clusters, encoding the synthesis of surfactin, bacillomycin D, fengycin, bacilysin, bacillibactin, macrolactin, difficidin, bacillaene, and amylocycicin. Further analysis using Liquid Chromatography Tandem Mass Spectrometry (LC-MS) revealed the presence of four of these metabolites (surfactin, iturin, fengycin, and bacillibactin) in the CFS of B. velezensis. Additional bioassays conducted in sealed plates revealed that volatile organic compounds (VOCs) produced by S161 also inhibited the growth of P. digitatum. A total of 24 VOCs were found to be produced by S161 using headspace solid-phase microextraction/gas chromatography-mass spectrometry (HS-SPME/GC-MS). The identified VOCs were mainly ketones and alcohols. Collectively, our results indicate that B. velezensis strain S161 is an effective biocontrol agent of P. digitatum on citrus and inhibits pathogen germination and growth through a range of mechanisms, including the downregulation of genes associated with spore germination and growth, induction of oxidative stress, and the antimicrobial activity of VOCs.

Development of Trichoderma Formulation and Application to Control Durian Anthracnose Disease

Anthracnose is a plant fungal disease that damages both quantitative and qualitative crop yields. Biological control of plant disease is an alternative method enabling reduced use of chemicals. The objective of this research was to develop a formulation of Trichoderma for controlling plant anthracnose disease. The efficacy of T. asperellum MSU007 in controlling Colletotrichum sp. by dual culture technique showed that T. asperellum MSU007 was able to inhibit the mycelium growth of Colletotrichum sp. with 50.76 - 74.85 % inhibition. Three formulations of T. asperellum MSU007 were developed. The viability of T. asperellum MSU007 in the formulation after drying at 45 °C showed that the viabilities of formulation 2 and 3 were not significantly different at 5.26×108 and 5.30×108 cfu/g, respectively, while that of formulation 1, the viability of T. asperellum MSU007 was reduced to 3.73×106 cfu/g. After storage at room temperature for 30 days, formulations 2 and 3 did not result in significantly different viability of T. asperellum MSU007 at 4.40×108 and 4.90×108 cfu/g, respectively, and after 90 days of storage at room temperature, it was found that only formulation 3 did not decrease T. asperellum MSU007 viability at 4.33×108 cfu/g. The efficacy of Trichoderma bioproduct in inhibiting anthracnose pathogens on durian leaves was tested. It was found that spraying formulation 3 after inoculation of Colletotrichum sp. resulted in the least anthracnose severity at 15.67 ± 4.04 %, which was significantly different from other treatments including the use of carbendazim. However, it is necessary to further study the efficacy of Trichoderma formulation in the inhibition of durian anthracnose pathogens in field conditions in order to be able to use this bioproduct in future disease control. HIGHLIGHTS This research developed an effective Trichoderma bioproduct to control the durian anthracnose causative agent Colletotrichum spp. which has sufficient potential to be developed and applied in the control of this disease, replacing the use of chemicals in the future. GRAPHICAL ABSTRACT

Herbicides may threaten advances in biological control of diseases and pests.

Advances in agriculture include integrated methods of controlling pests, diseases, and weeds with biocontrollers, which are constantly increasing, along with herbicides. The objective is to present a systematic review of the main reports of herbicide effects on non-target organisms used in applied biological control and those naturally occurring in the ecosystems controlling pests. The categories were divided into predatory and parasitoid arthropods. Three hundred and fifty reports were analyzed, being 58.3% with parasitoids and 41.7% with predators. Lethal or sublethal effects of herbicides on reproduction, predation, genotoxicity, and abundance of biological control organisms have been reported. Two hundred and four reports of the impact of herbicides on parasitoids were analyzed. The largest number of reports was with parasitoids of the genus Trichogramma, with wide use in managing pests of the herbicide-tolerant transgenic plants. Most tests evaluating effects on parasitism, emergence, and mortality of natural enemies subjected to herbicides are with parasitoids of Lepidoptera eggs with a high diversity and use in managing these pests in different crops. Additive and synergistic effects of molecules increase the risks of herbicide mixtures. Herbicide use for weed management must integrate other control methods, as the chemical can impact natural enemies, reducing the biological control of pests.

An assembled bacterial community associated with Artemisia annua L. causes plant protection against a pathogenic fungus.

The microorganisms associated with a plant influence its growth and fitness. These microorganisms accumulate on the aerial and root surfaces of plants, as well as within the plants, as endophytes, although how the interaction between microorganisms protects the plant from pathogens is still little understood. In the current study, the impact of assembled the bacterial communities against the pathogenic fungus to promote Artemisia annua L. growths was investigated. We established a model of bacterium-fungus-plant system. Eight bacterial strains and a fungal pathogen Globisporangium ultimum (Glo) were isolated from wild A. annua roots and leaves, respectively. We assembled the six-bacteria community (C6: Rhizobium pusense, Paracoccus sp., Flavobacterium sp., Brevundimonas sp., Stenotrophomonas sp., and Bacillus sp.) with inhibition, and eight-bacteria community (C8) composing of C6 plus another two bacteria (Brevibacillus nitrificans and Cupriavidus sp.) without inhibition against Glo in individually dual culture assays. Inoculation of seedlings with C8 significantly reduced impact of Glo. The growth and disease suppression of A. annua seedlings inoculated with C8 + Glo were significantly better than those of seedlings inoculated with only Glo. C8 had more inhibitory effects on Glo, and also enhanced the contents of four metabolites in seedling roots compared to Glo treatment only. Additionally, the inhibitory effects of root extracts from A. annua seedlings showed that Glo was most sensitive, the degree of eight bacteria sensitivity were various with different concentrations. Our findings suggested that the non-inhibitory bacteria played a vital role in the bacterial community composition and that some bacterial taxa were associated with disease suppression. The construction of a defined assembled bacterial community could be used as a biological fungicide, promoting biological disease control of plants.

BdCV1-Encoded P3 Silencing Suppressor Identification and Its Roles in Botryosphaeria dothidea, Causing Pear Ring Rot Disease.

Pear ring rot disease is an important branch disease, caused by Botryosphaeria dothidea. With the discovery of fungal viruses, the use of their attenuated properties for biological control provides a new strategy for the biological control of fungal disease. RNA silencing is a major antiviral defense mechanism in plants, insects, and fungi. Viruses encode and utilize RNA silencing suppressors to suppress host defenses. Previous studies revealed that Botryosphaeria dothidea chrysovirus 1 (BdCV1) exhibited weak pathogenicity and could activate host gene silencing by infecting B. dothidea. The aim of our study was to investigate whether BdCV1 can encode a silencing suppressor and what effect it has on the host. In this study, the capability of silencing inhibitory activity of four BdCV1-encoded proteins was analyzed, and the P3 protein was identified as a BdCV1 RNA silencing suppressor in the exotic host Nicotiana benthamiana line 16c. In addition, we demonstrated that P3 could inhibit local silencing, block systemic RNA silencing, and induce the necrosis reaction of tobacco leaves. Furthermore, overexpression of P3 could slow down the growth rate and reduce the pathogenicity of B. dothidea, and to some extent affect the expression level of RNA silencing components and virus-derived siRNAs (vsiRNAs). Combined with transcriptomic analysis, P3 had an effect on the gene expression and biological process of B. dothidea. The obtained results provide new theoretical information for further study of interaction between BdCV1 P3 as a potential silencing suppressor and B. dothidea.

Streptomyces silvisoli sp. nov., a polyene producer, and Streptomyces tropicalis sp. nov., two novel actinobacterial species from peat swamp forests in Thailand.

Two novel actinobacterial strains, designated RB6PN23T and K1PA1T, were isolated from peat swamp soil samples in Thailand and characterized using a polyphasic taxonomic approach. The strains were filamentous Gram-stain-positive bacteria containing ll-diaminopimelic acid in their whole-cell hydrolysates. Phylogenetic analysis of their 16S rRNA gene sequences revealed that strain RB6PN23T was most closely related to Streptomyces rubrisoli (99.1 % sequence similarity) and Streptomyces ferralitis (98.5%), while strain K1PA1T showed 98.8 and 98.7% sequence similarities to Streptomyces coacervatus and Streptomyces griseoruber, respectively. However, the average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH) values were below the species-level thresholds (95-96 % ANI and 70 % dDDH). The genomes of strains RB6PN23T and K1PA1T were estimated to be 7.88 Mbp and 7.39 Mbp in size, respectively, with DNA G+C contents of 70.2 and 73.2 mol%. Moreover, strains RB6PN23T and K1PA1T encode 37 and 24 putative biosynthetic gene clusters, respectively, and in silico analysis revealed that these new species have a high potential to produce unique natural products. Genotypic and phenotypic characteristics confirmed that strains RB6PN23T and K1PA1T represented two novel species in the genus Streptomyces. The names proposed for these strains are Streptomyces silvisoli sp. nov. (type strain RB6PN23T=TBRC 17040T=NBRC 116113T) and Streptomyces tropicalis sp. nov. (type strain K1PA1T=TBRC 17041T=NBRC 116114T). Additionally, a giant linear polyene compound, neotetrafibricin A, exhibiting antifungal activity in strain RB6PN23T, was identified through HPLC and quadrupole time-of-flight MS analysis. The crude extract from the culture broth of strain RB6PN23T exhibited strong antifungal activity against Fusarium verticillioides, Fusarium fujikuroi and Bipolaris zeicola. This finding suggests that strain RB6PN23T could be a promising candidate for biological control of fungal diseases.

Cinnamaldehyde Inhibits Postharvest Gray Mold on Pepper Fruits via Inhibiting Fungal Growth and Triggering Fruit Defense.

Gray mold infected with Botrytis cinerea frequently appears on fruits and vegetables throughout the supply chain after harvest, leading to economic losses. Biological control of postharvest disease with phytochemicals is a promising approach. CA (cinnamaldehyde) is a natural phytochemical with medicinal and antimicrobial activity. This study evaluated the effect of CA in controlling B. cinerea on fresh pepper fruit. CA inhibited B. cinerea growth in vitro significantly in a dose- (0.1-0.8 mM) and time-dependent (6-48 h) manner, with an EC50 (median effective concentration) of 0.5 mM. CA induced the collapse and breakdown of the mycelia. CA induced lipid peroxidation resulting from ROS (reactive oxygen species) accumulation in mycelia, further leading to cell leakage, evidenced by increased conductivity in mycelia. CA induced mycelial glycerol accumulation, resulting in osmotic stress possibly. CA inhibited sporulation and spore germination resulting from ROS accumulation and cell death observed in spores. Spraying CA at 0.5 mM induced a defense response in fresh pepper fruits, such as the accumulation of defense metabolites (flavonoid and total phenols) and an increase in the activity of defense enzymes (PAL, phenylalanine ammonia lyase; PPO, polyphenol oxidase; POD, peroxidase). As CA is a type of environmentally friendly compound, this study provides significant data on the activity of CA in the biocontrol of postharvest gray mold in peppers.

Advances in research and application of Trichoderma for inducing resistance against root rot diseases in root and rhizome of Chinese medicinal materials

Root rot is a microbial disease that is difficult to control and can result in serious losses in the planting of most Chinese medicinal materials. As high as 87.6% of roots or rhizomes of Chinese medicinal materials are susceptible to root rot, which seriously affects the cultivation development of Chinese medicinal materials. Trichoderma fungi, possessing biological control functions, can induce plants to improve their resistance to microbial diseases, promote plant growth, and effectively reduce the losses caused by various microbial diseases on cultivation. At present, Trichoderma is rarely used in the cultivation of Chinese medicinal materials, so it has great application potential for the prevention and control of root rot diseases in farmed Chinese medicinal materials. Based on the above situation, after comparison and discussion, it is believed that compared with chemical control and physical control, biological control of root rot diseases of Chinese medicinal materials is more efficient and meets the development needs of Chinese medicinal materials ecological planting in China. This paper reviewed the progress in the research and application of Trichoderma in the control of root rot diseases in the root and rhizome of farmed Chinese medicinal materials in the past 10 years and found that most of the current research on the biological control of root rot diseases in Chinese medicinal materials was mostly limited to the verification of the inhibitory effect of Trichoderma strains on the growth of the pathogenic microbes. Studies on the induction effect of Trichoderma on Chinese medicinal materials are not in depth. Studies on the responding mechanisms of most Chinese medicinal materials to Trichoderma are highly absent. Moreover, there are few reports on field experiments, which indicates that there is a long way to go before Trichoderma is widely applied in the farming practice of Chinese medicinal materials. To sum up, this paper aimed to link the present and the future and advocated further relevant research and more experiments on the application of Trichoderma in the farming of Chinese medicinal materials.

Unravelling the potential of susceptibility genes in plant disease management: Present status and future prospects

The increasing global population requires an equivalent increase in food production to meet the global food demand. Crop production is challenged by various biotic and abiotic stresses, which decrease crop yield and production. Thus, proper disease management for crops ensures global food security. Various chemical, physical, and biological disease control methods have been devised and used for plant protection. However, due to the low efficiency of these methods, modern research has shifted to genetic engineering approaches. The recent advances in molecular techniques have revealed the molecular mechanisms controlling the plant’s innate immune system and plant-pathogen interactions. Earlier studies revealed that the pathogens utilize the susceptibility (S) genes in hosts for their sustainability and disease development. The resistance achieved by suppressing the S genes expression provides resistance against pathogens. Exploiting S genes for imparting/enhancing disease resistance would offer a more durable and effective alternative to conventional disease control methods. Therefore, the present review highlights the potential of this novel tool for inducing disease resistance in plants.

Inhibitive and prophylactic efficacy of lactic acid bacteria from Apis mellifera (Hymenoptera: Apidae) in combating Paenibacillus infections

Given the increasing interest in alternative methods of prevention and biological control of bacterial diseases in honey bees, the development of new approaches based on probiotic bacteria for honey bee growth, immune function and defense is important. Honey bees have a rich microbiota that varies with seasonal and geographic changes, developmental stages, diet, and social lifestyle. Nevertheless, lactic acid bacteria are an integral part of the honey bee microbiota. In this study, strains of Apilactobacillus, Leuconostoc and Enterococcus isolated from healthy honey bees were identified and their usefulness as an alternative product to control and prevent American and European foulbrood was investigated. The strains were identified by cultural and molecular analysis. The antimicrobial activities against Paenibacillus larvae and Paenibacillus alvei were tested by agar plug and well diffusion method. The inhibitory effect of the obtained 72-h cell-free supernatants on Paenibacillus larvae spore germination was determined by the Bioscreen C method. It turned out that the strains whose prophylactic effect against Paenibacillus larvae spores in larvae was tested were not largely able to prevent infection. Apilactobacillus and Leuconostoc strains did not affect the health of the larvae, and infection was somewhat clinically eliminated in larvae colonized by Apilactobacillus and Enterococcus. The fact that the strains exhibit potent antimicrobial activity and do not reduce larval viability and live weight is a promising finding with regard to their use for treatment, particularly in the early stages of American foulbrood infection.

Isolation, characterization, and evaluation of putative new bacteriophages for controlling bacterial spot on tomato in Brazil.

Bacterial spot is a highly damaging tomato disease caused by members of several species of the genus Xanthomonas. Bacteriophages have been studied for their potential use in the biological control of bacterial diseases. In the current study, bacteriophages were obtained from soil and tomato leaves in commercial fields in Brazil with the aim of obtaining biological control agents against bacterial spot. Phage isolation was carried out by co-cultivation with isolates of Xanthomonas euvesicatoria pv. perforans, which was prevalent in the collection areas. In a host range evaluation, none of the phage isolates was able to induce a lytic cycle in all of the bacterial isolates tested. In in vivo tests, treatment of susceptible bacterial isolates with the corresponding phage prior to application to tomato plants led to a reduction in the severity of the resulting disease. The level of disease control provided by phage application was equal to or greater than that achieved using copper hydroxide. Electron microscopy analysis showed that all of the phages had similar morphology, with head and tail structures similar to those of viruses belonging to the class Caudoviricetes. The presence of short, non-contractile tubular tails strongly suggested that these phages belong to the family Autographiviridae. This was confirmed by phylogenetic analysis, which further revealed that they all belong to the genus Pradovirus. The phages described here are closely related to each other and potentially belong to a new species within the genus. These phages will be evaluated in future studies against other tomato xanthomonad strains to assess their potential as biological control agents.