Bacteria For Biological Control Research Articles

Do biological control agents adapt to local pest genotypes? A multiyear test across geographic scales.

Parasite local adaptation has been a major focus of (co)evolutionary research on host-parasite interactions. Studies of wild host-parasite systems frequently find that parasites paired with local, sympatric host genotypes perform better than parasites paired with allopatric host genotypes. In contrast, there are few such tests in biological control systems to establish whether biological control parasites commonly perform better on sympatric pest genotypes. This knowledge gap prevents the optimal design of biological control programs: strong local adaptation could argue for the use of sympatric parasites to achieve consistent pest control. To address this gap, we tested for local adaptation of the biological control bacterium Pasteuria penetrans to the root-knot nematode Meloidogyne arenaria, a global threat to a wide range of crops. We measured the probability and intensity of P. penetrans infection on sympatric and allopatric M. arenaria over the course of 4 years. Our design accounted for variation in adaptation across scales by conducting tests within and across fields, and we isolated the signature of parasite adaptation by comparing parasites collected over the course of the growing season. Our results are largely inconsistent with local adaptation of P. penetrans to M. arenaria: in 3 of 4 years, parasites performed similarly well in sympatric and allopatric combinations. In 1 year, however, infection probability was 28% higher for parasites paired with hosts from their sympatric plot, relative to parasites paired with hosts from other plots within the same field. These mixed results argue for population genetic data to characterize the scale of gene flow and genetic divergence in this system. Overall, our findings do not provide strong support for using P. penetrans from local fields to enhance biological control of Meloidogyne.

Biological control potential of worrisome wheat blast disease by the seed endophytic bacilli.

Crop production often faces challenges from plant diseases, and biological control emerges as an effective, environmentally friendly, cost-effective, and sustainable alternative to chemical control. Wheat blast disease caused by fungal pathogen Magnaporthe oryzae Triticum (MoT), is a potential catastrophic threat to global food security. This study aimed to identify potential bacterial isolates from rice and wheat seeds with inhibitory effects against MoT. In dual culture and seedling assays, three bacterial isolates (BTS-3, BTS-4, and BTLK6A) demonstrated effective suppression of MoT growth and reduced wheat blast severity when artificially inoculated at the seedling stage. Genome phylogeny identified these isolates as Bacillus subtilis (BTS-3) and B. velezensis (BTS-4 and BTLK6A). Whole-genome analysis revealed the presence of genes responsible for controlling MoT through antimicrobial defense, antioxidant defense, cell wall degradation, and induced systemic resistance (ISR). Taken together, our results suggest that the suppression of wheat blast disease by seed endophytic B. subtilis (BTS-3) and B. velezensis (BTS-4 and BTLK6A) is liked with antibiosis and induced systemic resistance to wheat plants. A further field validation is needed before recommending these endophytic bacteria for biological control of wheat blast.

Contribution of Native Plasmids of Pantoea vagans C9-1 to Epiphytic Fitness and Fire Blight Management on Apple and Pear Flowers and Fruits.

Pantoea vagans C9-1 (C9-1) is a biological control bacterium that is applied to apple and pear trees during bloom for suppression of fire blight, caused by Erwinia amylovora. Strain C9-1 has three megaplasmids: pPag1, pPag2, and pPag3. Prior bioinformatic studies predicted these megaplasmids have a role in environmental fitness and/or biocontrol efficacy. Plasmid pPag3 is part of the large Pantoea plasmid (LPP-1) group that is present in all Pantoea spp. and has been hypothesized to contribute to environmental colonization and persistence, while pPag2 is less common. We assessed fitness of C9-1 derivatives cured of pPag2 and/or pPag3 on pear and apple flowers and fruit in experimental orchards. We also assessed the ability of a C9-1 derivative lacking pPag3 to reduce populations of E. amylovora on flowers and disease incidence. Previously, we determined that tolerance to stresses imposed in vitro was compromised in derivatives of C9-1 lacking pPag2 and/or pPag3; however, in this study, the loss of pPag2 and/or pPag3 did not consistently reduce the fitness of C9-1 on flowers in orchards. Over the summer, pPag3 contributed to survival of C9-1 on developing apple and pear fruit in two of five trials, whereas loss of pPag2 did not significantly affect survival of C9-1. We also found that loss of pPag3 did not affect C9-1's ability to reduce E. amylovora populations or fire blight incidence on apple flowers. Our findings partially support prior hypotheses that LPP-1 in Pantoea species contributes to persistence on plant surfaces but questions whether LPP-1 facilitates host colonization.

Bacillus mexicanus sp. nov., a biological control bacterium isolated from the common bean (Phaseolus vulgaris L.) crop in Sinaloa, Mexico.

Strain FSQ1T was isolated from the rhizosphere of the common bean (Phaseolus vulgaris L.) crop sampled in a commercial field located in the Gabriel Leyva Solano community, which belongs to the Guasave municipality (state of Sinaloa, Mexico). Based on its full-length 16S rRNA gene sequence, strain FSQ1T was assigned to the genus Bacillus (100 % similarity). This taxonomic affiliation was supported by its morphological and metabolic traits. Strain FSQ1T was a Gram-stain-positive bacterium with the following characteristics: rod-shaped cells, strictly aerobic, spore forming, catalase positive, reduced nitrate to nitrite, hydrolysed starch and casein, grew in the presence of lysozyme and 2 % NaCl, utilized citrate, grew at pH 6.0-8.0, produced acid from glucose, was unable to produce indoles from tryptophan, and presented biological control against Sclerotinia sclerotiorum. The whole-genome phylogenetic results showed that strain FSQ1T formed an individual clade in comparison with highly related Bacillus species. In addition, the maximum values for average nucleotide identity and from Genome-to-Genome Distance Calculator analysis were 91.57 and 44.20 %, respectively, with Bacillus spizizenii TU-B-10T. Analysis of its fatty acid content showed the ability of strain FSQ1T to produce fatty acids that are not present in closely related Bacillus species, such as C18 : 0 and C20 : 0. Thus, these results provide strong evidence that strain FSQ1T represents a novel species of the genus Bacillus, for which the name Bacillus mexicanus sp. nov. is proposed. The type strain is FSQ1T (CM-CNRG TB51T=LBPCV FSQ1T).

Postharvest preservation effect of composite biocontrol agent on tomatoes

Biological control has been highly sought after as an effective means of controlling postharvest diseases in tomatoes. In order to explore effective biocontrol bacteria for postharvest biological control, this experiment isolated and screened 3 bacterial strains with potential for biocontrol. These 3 strains were Bacillus velezebsis (WZ-37), Bacillus subtilis subsp.subtilis (WXCDD105), and Bacillus amyloliquefaciens (SS). The enhancer for different biocontrol bacteria were also screened, and the effectiveness of composite biocontrol agents was evaluated. The results showed that these strains had a good inhibitory effect on the occurrence of tomato postharvest gray mold disease, significantly reducing the disease incidence rate while enhancing fruit quality and preservation. Among them, WZ-37 showed the most outstanding biocontrol effect, while the composite biocontrol effect of SS+WZ-37 was the best. Each strain has its corresponding optimal enhancer, and the addition of enhancer can increase the concentration of biocontrol bacteria and improve the biocontrol effect. In this study, three synergists, 0.01% B1, 0.01% Thr, and 0.01% Met, were screened to have the best effect on our strains. After spraying the biocontrol agents, the water loss rate of the postharvest fruits was significantly decreased, and a biofilm formed on the surface of tomatoes effectively reduced the occurrence of diseases. Therefore, these strains have high applied value and promotional prospects, and can be used for postharvest disease biological control of tomato.

Screening for Novel Beneficial Environmental Bacteria for an Antagonism-Based Erwinia amylovora Biological Control.

Erwinia amylovora, the bacterial species responsible for fire blight, causes major economic losses in pome fruit crops worldwide. Chemical control is not always effective and poses a serious threat to the environment and human health. Social demands for eco-sustainable and safe control methods make it necessary to search for new biocontrol strategies such as those based on antagonists. A bacterial collection from different fire blight-free Mediterranean environments was tested for antagonistic activity against Spanish strains of E. amylovora. Antagonistic assays were carried out in vitro in culture medium and ex vivo in immature loquat and pear fruits. Results revealed that 12% of the 82 bacterial isolates tested were able to inhibit the growth of several strains of the pathogen. Some of the isolates also maintained their antagonistic activity even after chloroform inactivation. Selected isolates were further tested ex vivo, with several of them being able to delay and/or reduce fire blight symptom severity in both loquats and pears and having activity against some E. amylovora strains. The isolates showing the best antagonism also produced different hydrolases linked to biocontrol (protease, lipase, amylase, and/or DNAse) and were able to fix molecular nitrogen. Based on this additional characterization, four biocontrol strain candidates were further selected and identified using MALDI-TOF MS. Three of them were Gram-positive bacteria belonging to Bacillus and Paenarthrobacter genera, and the fourth was a Pseudomonas strain. Results provide promising prospects for an improvement in the biological control strategies against fire blight disease.

Sulfur-oxidizing bacteria (SOB) and sulfate-reducing bacteria (SRB) in oil reservoir and biological control of SRB: a review.

Sulfur-oxidizing bacteria (SOB) and sulfate-reducing bacteria (SRB) inhabit oilfield production systems. Sulfur oxidation driven by SOB and dissimilatory sulfate reduction driven by SRB play important roles in sulfur cycle of oil reservoirs. More importantly, hydrogen sulfide produced by SRB is an acidic, flammable, and smelly toxic gas associated with reservoir souring, corrosion of oil-production facilities, and personnel safety. Effective control of SRB is urgently needed for the oil industry. This depends on an in-depth understanding of the microbial species that drive sulfur cycle and other related microorganisms in oil reservoir environments. Here, we identified SOB and SRB in produced brines of Qizhong block (Xinjiang Oilfield, China) from metagenome sequencing data based on reported SOB and SRB, reviewed metabolic pathways of sulfur oxidation and dissimilatory sulfate reduction, and ways for SRB control. The existing issues and future research of microbial sulfur cycle and SRB control are also discussed. Knowledge of the distribution of the microbial populations, their metabolic characteristics and interactions can help to develop an effective process to harness these microorganisms for oilfield production.

Screening of Bacillus subtilis HAAS01 and Its Biocontrol Effect on Fusarium wilt in Sweet Potato

Fusarium wilt, a disease caused by Fusarium oxysporum f.sp batatas (Fob) is an important disease in sweet potato production. Using endophytic bacteria for biological control of sweet potato diseases is one of the important ways. A Bacillus subtilis with antagonistic effect on Fusarium wilt of sweet potato was isolated from soil by confrontation culture. According to the biological characteristics, 16S rDNA sequence analysis, and physiological and biochemical analysis, the Bacillus subtilis HAAS01 was named. A pot experiment was conducted for the biological control experiment of strain HAAS01, and the endogenous hormone content, antioxidant enzyme activity, soluble protein content, and related gene expressions of sweet potato plants were detected. The results showed that the HAAS01 strain could promote the production of endogenous hormones and resist the infection of plant diseases together with defensive enzymes and upregulation of related gene expressions. In summary, Bacillus subtilis HAAS01 was effective in controlling Fusarium wilt of sweet potato and has potential for application and development.

Diversity of bacterial strains in biochar-enhanced Amazon soil and their potential for growth promotion and biological disease control in tomato

ABSTRACT The use of bacteria in growth promotion and biological control of plant diseases can minimize environmental contamination caused by the indiscriminate use of pesticides and chemical fertilizers. We aimed to evaluate growth promotion and biological control of Corynespora cassiicola in tomato seedlings mediated by beneficial bacteria isolated from a non-rhizospheric Amazon soil containing different amounts of biochar, and to identify to which groups of bacteria the strains belong. We obtained 200 strains of bacteria from experimental plots containing biochar doses of 0, 40, 80 and 120 t ha-1. Of these, 53 strains were selected by root colonization tests. Based on growth promotion parameters, 25 strains were screened, identified by molecular characterization and evaluated for indoleacetic acid (IAA) production, phosphate solubilization and biological control. The best dose of biochar for colony formation was 40 t ha-1, and a regression model indicated 34 t ha-1 as the optimal dose. The production of IAA was observed in 18 (75%) strains, and two (8%) strains were able to solubilize phosphate. The efficiency in root growth promotion was up to 125%, and the percentage of plant protection ranged from 50 to 59%. Molecular characterization showed that the bacteria used in this study belong to the genera Bacillus and Lysinibacillus.

Field application of allelopathic bacteria to control invasion of little seed canary grass in wheat.

Development of successful biological weed control can help to resolve various environmental challenges created by the chemical and mechanical weed control. The current study is aimed at investigating the potential of allelopathic bacteria (hereinafter as AB) for biological weed control in wheat rather than the traditional areas of plant allelopathy, phyto-pathology, and insect biocontrol agents. Eleven strains of AB were obtained that were inhibitory to little seed canary grass of which 5 also inhibited wheat in our previous studies. The remaining strains indicated the potential for biological control of this weed in wheat. Five efficient strains were selected for this purpose to conduct pot and field trials. Seeds of little seed canary grass were sown together in potted soils with wheat seeds inoculated with AB strains. A subsequent field trial was conducted at a site selected based on chronic infestations of canary grass. Seeds of the weed and inoculated wheat were co-seeded directly in field soil. For inoculation, these strains were formulated in sterilized peat and applied to seeds. In pot trials, the invasion of little seed canary grass in wheat reduced wheat grain yield up to 59.9%. Four strains suppressed the weed which resulted in recovery wheat grain yield losses from 20.1 to 66.9%. The field trial showed that the invasion of little seed canary grass reduced wheat grain yields up to 53.9%. Four strains suppressed the weed which resulted into recovery of grain wheat grain yield losses 34.3 to 64.3%. These findings were consistent with improvement of other agronomic, physiological, and chemical parameters of the crop where the four strains of AB (L9, T42, 7O0, and O010) were applied.

Field Performance of Allelopathic Bacteria for Biological Weed Control in Wheat: Innovative, Sustainable and Eco-Friendly Approach for Enhanced Crop Production

Application of allelopathic bacteria (AB) for weed suppression may be helpful to solve various environmental challenges posed by conventional weed control techniques. In our earlier studies, around 400 strains of rhizobacteria of five weeds and wheat were isolated, screened for production of phytotoxic substances, and tested for phytotoxic activity on wild oat and little seed canary grass, and possible effects on wheat under laboratory conditions. We obtained 13 strains inhibitory to wild oat (Avena fatua L.) and 11 to little seed canary grass (Phalaris minor Retz.). Five of these (13 and 11) strains also suppressed wheat (Triticum aestivum L.) while others either stimulated or remained ineffective on wheat in separate bioassays. The success of any weed biocontrol technique, however, depends on its response under field conditions. Therefore, the present study was conducted to investigate biological weed control of the five most efficient strains of AB under natural conditions in pot and field trials. Wheat was artificially invaded with wild oat in the pot trial through seeding. Wheat of the field trial was artificially invaded with wild oat and little seed canary through seeding. The selected strains belonged to pseudomonads (Pseudomonas putida, P. fluorescence, P. aeruginosa, and P. alcaligenes) and their inocula were prepared using sterilized peat. The inoculated seeds of wild oat and wheat were sown together in a pot trial. The inoculated seeds of wild oat, little seed canary grass, and wheat were sown together in the field experiment. The field was selected based on chronic infestation of these weeds. However, weed invasion was ensured by adding seeds of weeds (inoculated with the respective strains of AB, according to treatment plan). A severe invasion of wild oat was observed in the pot trial, which reduced the grain yield of infested wheat up to 60.8%. The effectiveness of applied strains controlled 22.0–76.3% loss of grain yield of infested wheat. Weed invasion in the field trial reduced the grain yield of the crop up to 56.3% and effectiveness of the applied strains controlled 29.0–60.7% loss of grain yield of infested wheat. The study of other agronomic, physiological, and chemical parameters of the crop and weeds supported these findings. Harnessing the potential of these strains exhibited in our studies may be helpful to introduce an innovative, sustainable, and eco-friendly weed control technique for production of wheat.

The Effect of Biochars and Endophytic Bacteria on Growth and Root Rot Disease Incidence of Fusarium Infested Narrow-Leafed Lupin (Lupinus angustifolius L.).

The effects of biochar on plant growth vary depending on the applied biochar type, study site environmental conditions, microbial species, and plant–microbial interactions. The objectives of the present study were therefore to assess 1) the response of growth parameters of lupin and root disease incidence to the application of three biochar types in a loamy sandy soil, and 2) the role of endophytic bacteria in biological control of root rot disease incidence in lupin after the amendment of soil with different biochar types. As biochar types we tested (i) hydrochar (HTC) from maize silage, (ii) pyrolysis char from maize (MBC), and (iii) pyrolysis char from wood (WBC) at three different concentrations (1%, 2%, and 3% of char as soil amendments). There were no significant effects in lupin shoot and root growth in soils amended with WBC at any of the concentrations. MBC did not affect plant growth except for root dry weight at 2% MBC. HTC char at 2% concentration, significantly increased the root dry weight of lupin by 54–75%, and shoot dry weight by 21–25%. Lupin plants grown in soil amended with 2% and 3% WBC and MBC chars showed 40–50% and 10–20% disease symptoms, respectively. Plants grown in soil without biochar and with HTC char were healthy, and no disease incidence occurred. Pseudomonas putida L2 and Stenotrophomonas pavanii L8 isolates demonstrated a disease reduction compared to un-inoculated plants under MBC and WBC amended soil that was infested with Fusarium solani.

Forest tree associated bacteria for potential biological control of Fusarium solani and of Fusarium kuroshium, causal agent of Fusarium dieback

Although the use of crop-associated bacteria as biological control agents of fungal diseases has gained increasing interest, the biotechnological potential of forest tree-associated microbes and their natural products has scarcely been investigated. The objective of this study was to identify bacteria or bacterial products with antagonistic activity against Fusarium solani and Fusarium kuroshium, causal agent of Fusarium dieback, by screening the rhizosphere and phyllosphere of three Lauraceae species. From 195 bacterial isolates, we identified 32 isolates that significantly reduced the growth of F. solani in vitro, which mostly belonged to bacterial taxa Bacillus, Pseudomonas and Actinobacteria. The antifungal activity of their volatile organic compounds (VOCs) was also evaluated. Bacterial strain Bacillus sp. CCeRi1-002, recovered from the rhizosphere of Aiouea effusa, showed the highest percentage of direct inhibition (62.5 %) of F. solani and produced diffusible compounds that significantly reduced its mycelial growth. HPLC-MS analyses on this strain allowed to tentatively identify bioactive compounds from three lipopeptide groups (iturin, surfactin and fengycin). Bacillus sp. CCeRi1-002 and another strain identified as Pseudomonas sp. significantly inhibited F. solani mycelial growth through the emission of VOCs. Chemical analysis of their volatile profiles indicated the likely presence of 2-nonanone, 2-undecanone, disulfide dimethyl and 1-butanol 3-methyl-, which had been previously reported with antifungal activity. In antagonism assays against F. kuroshium, Bacillus sp. CCeRi1-002 and its diffusible compounds exhibited significant antifungal activity and induced hyphal deformations. Our findings highlight the importance of considering bacteria associated with forest species and the need to include bacterial products in the search for potential antagonists of Fusarium dieback.

Evaluation of Bacteria from Soil and Rhizosphere as Herbicidal Candidates of Some Broadleaf Weeds

The use of bacteria in biological weed control is an alternative ecofriendly way to tackle weed problems and reduce the risk of herbicide resistance. In this study, we obtained four main active bacterial isolates under the genera Pseudomonas, Bacillus and Xanthomonas from Wadi El Natroun region. All the tested bacterial isolates caused high significant reductionsin seed germination and seedling growth of Convolvulus arvensis and Portulaca oleracea. Bioassaying ethyl acetate crude extracts of these isolates showed that Pseudomonas sp. (isolate 1) was the most active against seedling stage of Portulaca oleracea. The cultural filtrate of the same isolate caused 100% reduction when assayed on seed germination, shoot and root length of Convolvulus arvensis and Portulaca oleracea. At the highest concentration (40mg/ml) of the crude ethyl acetate extract, the reduction percentage in total biomass fresh weights of Portulaca oleracea and Convolvulus arvensis seedlings reached 71.27 and 39.37%, respectively. The EC50 values (concentration that inhibited growth by 50%) were 1.3 and 1.64mg/ml for Portulaca oleracea and Convolvulus arvensis, respectively. Comparative 16S rRNA gene sequencing analysis and biochemical characterization demonstrated that isolate FS15 was member of the genus Pseudomonas and belongs to the Pseudomonas aeruginosa group with Pseudomonas aeruginosa strain NR 113599.1 as the closest relative (99.5% sequence similarity). In conclusion, Pseudomonas aeruginosa has high potential to be developed as natural bacterial herbicide and may be used in broadleaf weed control.

Growth promotion and mildew suppressive effect of phylloplane bacteria of mulberry (Morus spp.)

Studies were conducted to evaluate growth promotion and mildew ( Phyllactinia corylea ) suppressive effect of phylloplane bacteria of mulberry ( Morus spp.) in vitro and in vivo and the effective bacteria were identified. Among 18 bacteria, 4 isolates showed highly significant (P 70 per cent suppression of conidial germination. The highest reduction was showed by the isolate Pb-5 (13.87%) by suppressing 86.13 per cent conidial germination followed by Pb-4 (17.23%) with 82.17 per cent. Similarly, six isolates significantly increased the seed germination. Significantly high (P 60% reduction of disease severity with the application of B.megaterium (63.42%) and B.cereus (60.73%). The study suggests exploration of either of these bacteria for biological control of mildew in mulberry.

Does soil warming affect the interaction between Pasteuria penetrans and Meloidogyne javanica in tomato plants?

A system to grow tomato plants infected by Meloidogyne javanica under constant temperatures of 18, 21, 24, 27 and 30 °C was developed and used to assess how temperature and the application of the biological control bacterium Pasteuria penetrans affected plant growth, the nematode population and endospore production. Each plant was inoculated with 300 second‐stage juveniles (J2) with four or five spores of P. penetrans attached to their cuticles or with 300 nematodes without P. penetrans. Increasing soil temperature increased tomato growth, the number of endospores per female, and the number of galls of M. javanica at the end of 38 days. Increasing temperatures up to 27 °C also increased the number of egg masses produced by M. javanica. Presence of P. penetrans reduced the numbers of galls and egg masses at all temperatures by up to 52.2% and 61.4% at 27 and 30 °C, respectively. Pasteuria penetrans reduced the M. javanica population even at soil temperatures of 18 and 21 °C. However, temperatures of 27 and 30 °C enhanced nematode control and the production of P. penetrans endospores is faster. The system developed in this work is simple and efficient for growing plants under constant temperatures and can be used for different purposes.

Screening of a biological control bacterium to fight avocado diseases: From agroecosystem to bioreactor

Abstract Avocado diseases remain to be a major constraint for the development of this agribusiness, which is largely led by Latin American countries. In this study, a collection of 667 native avocado bacterial isolates was established and screened for antagonistic activity and bioactive secondary metabolite production against two important avocado pathogens (Phytophthora cinnamomi and Colletotrichum gloeosporioides). The isolate ARP5.1 demonstrated the highest potential for in vitro control of the phytopathogens and was identified as Serratia sp. To enhance bioactive metabolite production by ARP5.1, different combinations of agitation speeds and aeration rates were evaluated on a stirred tank bioreactor. The activity of the crude extracts was tested and cell growth kinetics, oxygen and glucose consumption, and production of secondary metabolites were determined at the fermentation conditions of higher inhibitory activity. Results evidenced oxygen as critical factor for the biosynthesis of metabolites of interest. In addition, shear stress and glucose were limiting factors in metabolite production. Prodigiosin, serratamolide and haterumalide NC were identified in the extracts by Liquid Chromatography coupled to High-Resolution Mass Spectrometry. However, the presence of haterumalide NC might explain the high activity of the crude extract against P. cinnamomi. We concluded that this bacterium might be a promising candidate for the control of avocado pathogens in field and a good source of bioactive metabolites.

Impact of Chemical and Biological Fungicides Applied to Grapevine on Grape Biofilm, Must, and Wine Microbial Diversity.

This study was aimed to measure the impact of the application of a bio-fungicide against Botrytis cinerea on the microbiota involved in the alcoholic fermentation (AF) of Tempranillo Rioja wines. For this purpose, a bio-fungicide composed of the biological control bacterium Bacillus subtilis QST713 was applied to the vineyard. The microbial diversity was analyzed from grape biofilm to wine. Impact on microbial diversity was measured employing indexes assessed with the software PAST 3.10 P.D. Results were compared to non-treated samples and to samples treated with a chemical fungicide mainly composed by fenhexamid. Overall, the impact of the biological-fungicide (bio-fungicide) on the microbial diversity assessed for grape biofilm and for musts was not remarkable. Neither of the tested fungicides enhanced the growth of any species or acted against the development of any microbial groups. The bio-fungicide had no significant impact on the wine microbiota whereas the chemical fungicide caused a reduction of microbial community richness and diversity. Although environmental threats might generate a detriment of the microbial species richness, in this study the tested bio-fungicide did not modify the structure of the microbial community. Indeed, some of the Bacillus applied at the grape surface, were detected at the end of the AF showing its resilience to the harsh environment of the winemaking; in contrast, its impact on wine quality during aging is yet unknown.

Isolated Pseudomonas aeruginosa strain VIH2 and antagonistic properties against Ralstonia solanacearum

The aim of this study was to isolates with antagonist activity against R. solanacearum. Thirty-two bacterial isolates were obtained from samples, and they were screened for potential antagonistic activity against R. Solanacearum. Using the agar spot method, ten out of the 21 tested bacteria showed antilisterial activity. VIH2 had the highest inhibitory effect on the growth of R. Solanacearum. Based on 16S rDNA and Biolog test analysis, the strain VIH2 was identified as Pseudomonas aeruginosa. Single-factor and Response Surface Methodology experiments were used to optimize the culture medium and conditions. This study was to explore whether the hemolysin-co-regulated protein secretion island I (HSI-I)-encoded type VI secretion system (T6SS) in Pseudomonas can be used as a biological control approach against Ralstonia solanacearum under field conditions. Bacterial competition assay showed that the HSI-I type T6SS of strain VIH2 exhibited dramatic antibacterial killing activity against R. solanacearum. The HSI-I T6SS of P. aeruginosa was regulated by the ppKA gene. We disrupted the gene ppKA in VIH2 by a single crossover to yield the VIH2 (ΔppKA) mutant. The antagonism of VIH2 was significantly decreased by ppKA gene disruption. In conclusion, our data supported the idea that HSI-I T6SS plays a crucial role in the antagonistic action of strain VIH2 against R. solanacearum. This alternative approach for antagonism against R. solanacearum might help develop attenuated strains of engineered bacteria for biological control.

Potential of some bacteria for biological control of postharvest citrus green mould caused by Penicillium digitatum

Ten bacteria isolate (4 Bacillus subtilis, 2 Bacillus pumilus, 2 Bacillus cereus, 1 Bacillus megaterium, and 1 Agrobacterium radiobacter) were tested in vitro for antagonistic properties against Penicillium digitatum, the causal agent of citrus green mould. The effect of these bacteria was also observed on mycelial growth, spore germination, and spore production of the pathogenic fungus in broth culture. Extracellular enzyme activities of the bacteria were determined. According to the results of in vitro antagonistic tests and enzymes activities, the most promising bacteria were Bacillus subtilis and Agrobacterium radiobacter. These bacteria were tested for disease suppression on lemon fruits. In addition, these bacterial isolates also showed remarkable antifungal activity against the pathogen on lemon fruits. The results of this study showed that Bacillus subtilis and Agrobacterium radiobacter showed remarkable antifungal activity against the pathogen. Chitinase and glucanase enzyme activity of all the tested bacteria was positive. Protease enzyme activity was positive in all tested bacteria with the exception of Agrobacterium radiobacter. In addition, all bacteria inhibited mycelial growth and spore germination (except Agrobacterium radiobacter) of the fungus. Bacillus subtilis, Bacillus cereus, and Agrobacterium radiobacter inhibited spore production in broth culture. Bacillus subtilis and Agrobacterium radiobacter were tested on lemon fruits significantly reduced disease severity. Consequently, these isolates can be used as new biocontrol agents in controlling the post-harvest decay of citrus fruits caused by Penicillium digitatum.