Biocontrol Activity Research Articles

Biocontrol Stem-end Rot Disease Agent, Alternaria alternata YZU, on Pitaya by Soil Phosphate Solubilizing Bacteria

Biocontrol of stem end disease agent, Alternaria alternata YZU, on pitaya is more interested in altering the application of chemical pesticides. This study was conducted to characterize antagonistic phosphate solubilizing bacteria (PSB) from rhizosphere soil for their biocontrol activities against A. alternata YZU under laboratory and greenhouse conditions. Six PSB isolated from 31 rhizosphere soil samples were tested for inhibiting the mycelial growth of A. alternata YZU in dual cultures. Six isolates were determined to inhibit the mycelial growth of A. alternata YZU. Among them, PSB31 presented the highest level of antagonistic activity against A. alternata YZU with a mean inhibition diameter of 0.64 ± 0.02 cm, while the other strains, including PSB11, PSB21, PSB41, PSB51, and PSB61 presented a weaker inhibition. The results also showed that the strain PSB31 was identified as Bacillus sp. strain PSB31 (Accession number: ON422095) and could control the mycelial growth of A. alternata YZU by secreting antifungal metabolites. Moreover, an in vivo antagonistic experiment of PSB31 on pitaya twigs showed a significant reduction of lesions on twigs than the control. The results suggested that the isolated PSB31 is a potential biological control agent. Further studies should be done to identify the biochemical basis of their activity against A. alternata YZU.

Strain improvement of Trichoderma harzianum for enhanced biocontrol capacity: Strategies and prospects.

In the control of plant diseases, biocontrol has the advantages of being efficient and safe for human health and the environment. The filamentous fungus Trichoderma harzianum and its closely related species can inhibit the growth of many phytopathogenic fungi, and have been developed as commercial biocontrol agents for decades. In this review, we summarize studies on T. harzianum species complex from the perspective of strain improvement. To elevate the biocontrol ability, the production of extracellular proteins and compounds with antimicrobial or plant immunity-eliciting activities need to be enhanced. In addition, resistance to various environmental stressors should be strengthened. Engineering the gene regulatory system has the potential to modulate a variety of biological processes related to biocontrol. With the rapidly developing technologies for fungal genetic engineering, T. harzianum strains with increased biocontrol activities are expected to be constructed to promote the sustainable development of agriculture.

Metschnikowia pulcherrima as biocontrol agent and wine aroma enhancer in combination with a native Saccharomycescerevisiae

One of the main objectives for a sustainable winemaking process is the reduction of the use of sulfur dioxide. In this regard, non-Saccharomyces wine yeasts are proposed as biocontrol agent in different steps of wine production chain. Here, a selected strain of Metschnikowia pulcherrima (DiSVA 269) and a native Saccharomyces cerevisiae low sulfite producer strain (DiSVA 708) were investigated. After preliminary laboratory trials, winemaking process at industrial level showed an effective biocontrol action (reduction of c.a. 1 Log order of wild yeasts) of M. pulcherrima inoculated at prefermentative stage in cold clarification (48 h at 10 °C) and during the subsequent fermentation process. The combination of M. pulcherrima/S. cerevisiae led a distinctive aromatic profile of wines both in laboratory and winery trials with a significant enhancement of ethyl butyrate, ethyl hexanoate, isoamyl acetate and β-phenyl ethanol. Moreover the use of the two selected strains was the best combination to enhance volatile thiols (3-mercaptohexan-1-ol and 3-mercaptoexil acetate) that well correlate with the sensory analysis (tropical fruits). The overall results indicate that the combined use of M. pulcherrima DiSVA 269 and native S. cerevisiae DiSVA 708 led a biocontrol action and an improvement of aromatic and sensorial profile of wine with low SO2 content.

Trichoderma virens exerts herbicidal effect on Arabidopsis thaliana via modulation of amino acid metabolism

Trichoderma virens is a plant beneficial fungus well-known for its biocontrol, herbicidal and growth promotion activity. Earlier, we identified HAS (HA-synthase, a terpene cyclase) and GAPDH (glyceraldehyde-3-phosphate dehydrogenase) to be involved in the production of multiple non-volatiles and non-volatile+volatile metabolites, respectively. The present study delineates the function of HAS and GAPDH in regulating herbicidal activity, using the model plant Arabidopsis thaliana. Under axenic conditions, rosette-biomass of seedlings co-cultivated with ΔHAS (HASR) and ΔGAPDH (GAPDHR) was higher than WT-Trichoderma (WTR) as well as non-colonized control (NoTR), even though the root colonization ability was reduced. However, HASR biomass was still higher than those of GAPDHR, indicating that blocking volatiles will not provide any additional contribution over non-volatile metabolites for Trichoderma-induced herbicidal activity. LC-MS analysis revealed that loss of herbicidal activity of ΔHAS/ΔGAPDH was associated with an increase in the levels of amino acids, which coincided with reduced expression levels of amino-acid catabolism and anabolism related genes in HASR/GAPDHR. RNAi-mediated suppression of an oxidoreductase gene, VDN5, specifically prevented viridin-to-viridiol conversion. Additionally, vdn5 mimics ΔHAS, in terms of amino-acid metabolism gene expression and partially abolishes the herbicidal property of WT-Trichoderma. Thus, the study provides mechanistic frame-work for better utilization of Trichoderma virens for biocontrol purposes, balancing between plant growth promotion and herbicidal activity.

New Observation in Biocontrol of Penicillium caperatum against Fusarium oxysporum on Saposhnikovia divaricata and as a Plant Growth Promoter

Fusarium oxysporum, a common fungal pathogen that infects economic crops, causes Fusarium wilt disease to Saposhnikovia divaricata at an annual incidence rate of more than 15%. This study aimed to assess the potential of rhizospheric fungi as antifungal agents against Fusarium wilt of Saposhnikovia divaricata. In this study, 104 fungi were isolated from S. divaricata rhizospheric soil. Twelve rhizospheric strains that showed antagonistic activity against F. oxysporum, MR-16, MR-32, MR-38, etc., were screened out. Biocontrol activities of the twelve strains, especially MR-16, were subsequently characterized and evaluated. Strain MR-16 as potential stock for biocontrol had good antibiotic activity against F. oxysporum in vitro experiment. Based on the analysis of morphological properties and rDNA internal transcribed spacers (ITS), we identified an isolate MR-16 as Penicillium caperatum (GenBank No. OK287146.1), a new record of this species of China. The results of the in vitro antagonistic assay indicated that the conidial germination rate was significantly decreased, and the mycelia morphology of F. oxysporum induced change via the culture filtrate of P. caperatum MR-16, such as deformation and degradation. In an outdoor pot experiment, inoculation of S. divaricata plants with F. oxysporum created severe wilting symptoms; however, in inoculation trials, MR-16 effectively suppressed disease lesions, with a strong control efficacy of 60.76%. In addition, strain MR-16 could successfully colonize and form stable populations in the soil, and it showed a continuous positive growth-promoting effect on S. divaricata plants.

Nematode predatory ability of the fungus Duddingtonia flagrans affected by in vitro sequential exposure to ovine gastrointestinal tract

Duddingtonia flagrans is a nematophagous fungus employed as a biocontrol agent of gastrointestinal nematodes in ruminants. After oral ingestion and passage through the digestive tract of animals, this microorganism captures the nematodes in the feces. The drastic conditions of ruminant digestive tract could affect fungi chlamydospores and therefore biocontrol activity. The aim of this study was to evaluate in vitro the effect of four ruminant digestive segments on the concentration and nematode predatory ability of a Colombian native strain of D. flagrans. The sequential four-step methodology proposed evaluated conditions of the oral cavity, rumen, abomasum, and small intestine such as pH (2, 6, 8), enzymes (pepsin, pancreatin), temperature (39 °C), and anaerobiosis comparing short (7 h) and long (51 h) exposure times. The results showed that the nematode predatory ability of the fungi is affected by sequential exposure to gastrointestinal segments and this effect depends on the exposure time to those conditions. After short exposure (7 h) through the four ruminant digestive segments, the fungi had a nematode predatory ability of 62%, in contrast, after long exposure (51 h) the nematode predatory ability was lost (0%). Moreover, the number of broken chlamydospores was higher in the long-exposure assay.

Endophytic Bacterium Lysobacter firmicutimachus Strain 5-7 Is a Promising Biocontrol Agent Against Rice Seedling Disease Caused by Pythium arrhenomanes in Nursery Trays.

Rice root rot disease caused by Pythium spp. is a highly destructive disease in rice nurseries. Biocontrol with endophytic bacteria was developed in this study to control rice seedling diseases. An in planta screening assay revealed that two bacterial endophytes, strains 5-7 and 6-4, displayed strong protection of rice seedlings from attack by Pythium arrhenomanes. Phylogenetic analysis indicated that strain 5-7 is Lysobacter firmicutimachus, while strain 6-4 belongs to the Kitasatospora genus. To quickly evaluate the disease severity of the root system damaged by Pythium spp. in nursery trays, a root surface area measurement assay was developed. By using this measurement, the control efficacy in nursery trays was evaluated, and L. firmicutimachus 5-7 showed promising biocontrol activity against Pythium disease. In a field trial, the two endophytes exhibited significant disease control efficacy on rice brown spot disease caused by Bipolaris oryzae naturally occurring in a commercial nursery field. The two endophytes exhibited multiple enzymatic activities and broad-spectrum antagonistic activities against multiple rice pathogens. The two endophytes colonized the root surface and inside of the root. L. firmicutimachus 5-7 primarily colonized the intercellular space and aerenchyma. Antibiosis is the major mechanism used by strain 5-7 to cause Bipolaris hyphal swelling and inhibit Pythium zoospore germination and sporangium formation, while a hyperparasitism-like phenomenon was found in the interaction of strain 6-4 with Pythium and Bipolaris hyphae. In conclusion, we report the promising biocontrol agent L. firmicutimachus 5-7 and the potential biocontrol agent Kitasatospora sp. 6-4 for disease control of rice seedlings in commercial nursery trays and their possible mechanisms of action.

RNA silencing proteins and small RNAs in oomycete plant pathogens and biocontrol agents.

Oomycetes cause several damaging diseases of plants and animals, and some species also act as biocontrol agents on insects, fungi, and other oomycetes. RNA silencing is increasingly being shown to play a role in the pathogenicity of Phytophthora species, either through trans-boundary movement of small RNAs (sRNAs) or through expression regulation of infection promoting effectors. To gain a wider understanding of RNA silencing in oomycete species with more diverse hosts, we mined genome assemblies for Dicer-like (DCL), Argonaute (AGO), and RNA dependent RNA polymerase (RDRP) proteins from Phytophthora plurivora, Ph. cactorum, Ph. colocasiae, Pythium oligandrum, Py. periplocum, and Lagenidium giganteum. Moreover, we sequenced small RNAs from the mycelium stage in each of these species. Each of the species possessed a single DCL protein, but they differed in the number and sequence of AGOs and RDRPs. SRNAs of 21nt, 25nt, and 26nt were prevalent in all oomycetes analyzed, but the relative abundance and 5' base preference of these classes differed markedly between genera. Most sRNAs mapped to transposons and other repeats, signifying that the major role for RNA silencing in oomycetes is to limit the expansion of these elements. We also found that sRNAs may act to regulate the expression of duplicated genes. Other sRNAs mapped to several gene families, and this number was higher in Pythium spp., suggesting a role of RNA silencing in regulating gene expression. Genes for most effector classes were the source of sRNAs of variable size, but some gene families showed a preference for specific classes of sRNAs, such as 25/26 nt sRNAs targeting RxLR effector genes in Phytophthora species. Novel miRNA-like RNAs (milRNAs) were discovered in all species, and two were predicted to target transcripts for RxLR effectors in Ph. plurivora and Ph. cactorum, indicating a putative role in regulating infection. Moreover, milRNAs from the biocontrol Pythium species had matches in the predicted transcriptome of Phytophthora infestans and Botrytis cinerea, and L. giganteum milRNAs matched candidate genes in the mosquito Aedes aegypti. This suggests that trans-boundary RNA silencing may have a role in the biocontrol action of these oomycetes.

Biocontrol efficacy of endophytic Trichoderma spp. in fresh and dry powder formulations in controlling northern corn leaf blight in sweet corn

Seventy-three endophytic Trichoderma spp. obtained from healthy corn leaves collected in the central and northeast parts of Thailand were evaluated for their biocontrol activity against northern leaf blight disease due to Exserohilum turcicum. Among them, T. asperellum KUFA0702, KUFA0703, T. hamatum KUFA0706, T. harzianum KUFA0710, and T. harzianum KUFA0713 showed promising biocontrol efficacy against the disease under laboratory and greenhouse conditions; thus, they were selected for control efficacy evaluation under field conditions using their fresh and dry powder formulations. The results found that the fresh formulation of each Trichoderma strain displayed better biocontrol effects than their dry formulation. In field trials, the application of fresh formulation of T. harzianum KUFA0710 displayed the best disease reduction at 55.70%, while its dry formulation caused 47.46% disease reduction followed by T. harzianum KUFA0713 at 49.92% and 43.84% disease reduction when applied as fresh and dry formulations, respectively. The antagonistic activities of Trichoderma spp. included fast growth with high competition for space and nutrients and production of antifungal crude extracts. Moreover, all Trichoderma strains showed compatibility fungicides at recommendation rate with azoxystrobin (1000 ppm) and difenoconazole + azoxystrobin (500 ppm). These fungi can serve as alternative fungicides or be combined with fungicides for integrated pest management.

Metschnikowia pulcherrima in Cold Clarification: Biocontrol Activity and Aroma Enhancement in Verdicchio Wine

Non-Saccharomyces wine yeasts are not only proposed to improve the sensory profile of wine but also for several distinctive promising features. Among them, biocontrol action at different steps of the wine production chain could be a suitable strategy to reduce the use of sulfur dioxide. In this work, the activity of a selected strain of Metschnikowia pulcherrima was evaluated as inoculum in cold clarification with the aim to reduce SO2 and improve the aromatic profile of the wine. Fermentation processes were carried out at the winery level for two consecutive vintages using a pied de cuve as the starter inoculum coming from indigenous Saccharomyces cerevisiae strains. M. pulcherrima revealed an effective bio-protectant action during the pre-fermentative stage even if the timely and appropriate starter inoculum in the two years permitted the effective control of wild yeasts during the fermentation also in the control trials. In general, the main oenological characters did not show differences if compared with an un-inoculated trial, while the inoculum of M. pucherrima in cold clarification determined an enhancement of ethyl hexanoate, isobutanol, acetaldehyde, and geraniol even if they are considered in different amounts for each year. Indeed, the analytical and sensory profiles of wines were also influenced by the vintage and variation pied the cuve population. Nonetheless, the overall results indicated that M. pulcherrima led to biocontrol action and an improvement of the aromatic and sensory profile of the wine.

Identification, Biocontrol Activity, and Field Application Effect of Bacillus velezensis Yb-1

Colletotrichum capsici is one of the most important pathogens on chili peppers. Unreasonable application of chemical fungicides will lead to threats to human and animal health, environmental damage, and increased fungicide resistance to pathogens. As an alternative strategy, biological control has been paid more and more attention by academics. In this study, the Yb-1 strain was isolated from healthy cucumber leaves, which could significantly restrict the mycelium growth of C. capsici and Botrytis cinerea, with inhibition rates of 93.49% and 74.03%, respectively. Strain Yb-1 was identified as Bacillus velezensis by morphological features and 16S rRNA gene, gyrA gene, and gyrB gene sequence analysis. When pepper seeds were treated with different concentrations of bacterial suspension, we found that a medium-concentration treatment (104 CFU/mL and 105 CFU/mL) could promote seed germination and growth, while high-concentration treatments (108 CFU/mL) limited seed germination and growth. In addition, the root-irrigation method, acupuncture-leaf method, and leaf-cutting method were used to evaluate the safety of strain Yb-1 on peppers. The results indicated that Yb-1 did not affect the normal growth of peppers. The results of further field trials showed that the control effect of strain Yb-1 on pepper anthracnose was 59.45%. Thus, the strain B. velezensis Yb-1 has great biocontrol potential for pepper anthracnose and promotes plant growth.

Evaluation of the application of <i>Wickerhamomyces anomalus</i> supernatant for the control of relevant spoilage yeasts in wines

SO2 is traditionally used to limit or nullify the proliferation of spoilage yeasts in must/wine, but its use has become controversial due to its negative effects on human health. An alternative strategy for the control of spoilage yeasts is the use of biocontrol yeasts. Wickerhamomyces anomalus is an outstanding biocontroller yeast that has been examined by our group and that is a good biocontrol agent against Brettanomyces bruxellensis and Zygosaccharomyces rouxii. The challenge regarding the application of the results of microbial biocontrol studies lies in exploring technologically simple and economical ways to make use of microbial biocontrol activity. The introduction of foreign microbiota adds complexity to the fermentation medium and can cause technological difficulties and have oenological consequences inherent to the implementation of mixed cultures. Therefore, the objective of this study was to characterise the concentrated culture supernatant of W. anomalus and evaluate its biocontrol action in liquid medium on two main wine spoilage yeasts. W. anomalus BWa156 supernatant demonstrated inhibitory killer protein characteristics. The production of the inhibitory supernatant by the biocontroller yeast was independent of the presence or absence of the spoilage yeast. Supernatant can be produced faster under aerobic conditions than in traditional fermentation (i.e., with around 24 h), thus increasing its potential for technological development. The treatment with the supernatant of BWa156 was effective against the two spoilage populations Zygosaccharomyces rouxii and Brettanomyces bruxellensis, which are considered problematic for the wine industry. The supernatant of the biocontrol yeast can be considered to be a relevant additional stress factor for the spoilage population, which together with other factors such as ethanol, competition for nutrients, oxygen and pH, contributes to the elimination of the polluting population. This technology would allow a simple future application, through its production in a bioreactor parallel to the fermentation and subsequent inoculation in the must/wine.

Optimization of the growth conditions through response surface methodology and metabolomics for maximizing the auxin production by Pantoea agglomerans C1.

The fermentative production of auxin/indole 3-acetate (IAA) using selected Pantoea agglomerans strains can be a promising approach to developing novel plant biostimulants for agriculture use. By integrating metabolomics and fermentation technologies, this study aimed to define the optimal culture conditions to obtain auxin/IAA-enriched plant postbiotics using P. agglomerans strain C1. Metabolomics analysis allowed us to demonstrate that the production of a selected. Array of compounds with plant growth-promoting- (IAA and hypoxanthine) and biocontrol activity (NS-5, cyclohexanone, homo-L-arginine, methyl hexadecenoic acid, and indole-3-carbinol) can be stimulated by cultivating this strain on minimal saline medium amended with sucrose as a carbon source. We applied a three-level-two-factor central composite design (CCD) based response surface methodology (RSM) to explore the impact of the independent variables (rotation speed and medium liquid-to-flask volume ratio) on the production of IAA and IAA precursors. The ANOVA component of the CCD indicated that all the process-independent variables investigated significantly impacted the auxin/IAA production by P. agglomerans strain C1. The optimum values of variables were a rotation speed of 180 rpm and a medium liquid-to-flask volume ratio of 1:10. Using the CCD-RSM method, we obtained a maximum indole auxin production of 208.3 ± 0.4 mg IAAequ/L, which was a 40% increase compared to the growth conditions used in previous studies. Targeted metabolomics allowed us to demonstrate that the IAA product selectivity and the accumulation of the IAA precursor indole-3-pyruvic acid were significantly affected by the increase in the rotation speed and the aeration efficiency.

Bacillus subtilis SPB1 lipopeptide biosurfactant: antibacterial efficiency against the phytopathogenic bacteria Agrobacterium tumefaciens and compared production in submerged and solid state fermentation systems.

Bacillus subtilis SPB1 derived biosurfactants (BioS) proved its bio-control activity against Agrobacterium tumefaciens using tomato plant. Almost 83% of disease symptoms triggered by Agrobacterium tumefaciens were reduced. Aiming potential application, we studied lipopeptide cost-effective production in both fermentations systems, namely the submerged fermentation (SmF) and the solid-state fermentation (SSF) as well as the use of Aleppo pine waste and confectionery effluent as cheap substrates. Optimization studies using Box-Behnken (BB) design followed by the analysis with response surface methodology were applied. When using an effluent/sea water ratio of 1, Aleppo pine waste of 14.08g/L and an inoculum size of 0.2, a best production yield of 17.16 ± 0.91mg/g was obtained for the SmF. While for the SSF, the best production yield of 27.59 ± 1.63mg/g was achieved when the value of Aleppo pine waste, moisture, and inoculum size were, respectively, equal to 25g, 75%, and 0.2. Hence, this work demonstrated the superiority of SSF over SmF.

Deciphering the antimicrobial activity of multifaceted rhizospheric biocontrol agents of solanaceous crops viz., Trichoderma harzianum MC2, and Trichoderma harzianum NBG.

The Solanaceae family is generally known to be the third most economically important plant taxon, but also harbors a host of plant pathogens. Diseases like wilt and fruit rot of solanaceous crops cause huge yield losses in the field as well as in storage. In the present study, eight isolates of Trichoderma spp. were obtained from rhizospheric micro-flora of three solanaceous crops: tomato, brinjal, and chili plants, and were subsequently screened for pre-eminent biocontrol activity against three fungal (Fusarium oxysporum f. sp. lycopersicum, Colletotrichum gloeosporioides, and Rhizoctonia solani) and one bacterial (Ralstonia solanacearum) pathogen. Morphological, ITS, and tef1α marker-based molecular identification revealed eight isolates were different strains of Trichoderma. Seven isolates were distinguished as T. harzianum while one was identified as T. asperellum. In vitro antagonistic and biochemical assays indicated significant biocontrol activity governed by all eight isolates. Two fungal isolates, T. harzianum MC2 and T. harzianum NBG were further evaluated to decipher their best biological control activity. Preliminary insights into the secondary metabolic profile of both isolates were retrieved by liquid chromatography-mass spectrometry (LC-MS). Further, a field experiment was conducted with the isolates T. harzianum MC2 and T. harzianum NBG which successfully resulted in suppression of bacterial wilt disease in tomato. Which possibly confer biocontrol properties to the identified isolates. The efficacy of these two strains in suppressing bacterial wilt and promoting plant growth in the tomato crop was also tested in the field. The disease incidence was significantly reduced by 47.50% and yield incremented by 54.49% in plants treated in combination with both the bioagents. The results of scanning electron microscopy were also in consensus with the in planta results. The results altogether prove that T. harzianum MC2 and T. harzianum NBG are promising microbes for their prospective use in agricultural biopesticide formulations.

The posttranscriptional regulator CsrA affects multidrug resistance and biocontrol activity in Lysobacter enzymogenes.

The posttranscriptional regulator CsrA regulates many cellular processes, including stress responses in diverse bacteria. However, the role of CsrA in multidrug resistance (MDR) and biocontrol activity in Lysobacter enzymogenes strain C3 (LeC3) remains unknown. In this study, we demonstrated that deletion of the csrA gene resulted in the initial slow growth of LeC3 and reduced its resistance to multiple antibiotics, including nalidixic acid (NAL), rifampicin (RIF), kanamycin (Km), and nitrofurantoin (NIT). Loss of the csrA gene also reduced its ability in inhibiting hypha growth of Sclerotium sclerotiorum and influenced its extracellular cellulase and protease activities. Two putative small noncoding regulatory RNAs (sRNAs), referred to as csrB and csrC, were also revealed in the genome of LeC3. Double deletion of csrB and csrC in LeC3 led to increased resistance to NAL, RIF, Km, and NIT. However, no difference was observed between LeC3 and the csrB/csrC double mutant in their suppression of S. sclerotiorum hypha growth and production of extracellular enzymes. These results suggest that CsrA in LeC3 not only conferred its intrinsic MDR, but also contributed to its biocontrol activity.

Tomato Production under Synergistic Application of Phosphate Solubilizing Bacteria and Phosphate Amendments

Phosphate solubilizing bacteria have multi-dimensional benefits in broad host range interaction, accessing nutrients, phytohormone induction, stress alleviation, biocontrol activity, and eco-friend approach. This study aimed to evaluate the efficacy of PSB isolates coinoculated with compost, bone meal, and DAP fertilizer on tomato growth response. Tomato seeds were treated with 10 selected PSB isolates separately and grown for 20 days on seedbed, then transplanted to field that was treated with external P-sources and enriched by PSB inoculum. PSB isolates showed positive interaction and achieved significant plant assays including plant height, leaves, branches, flowers, and fruit development. Isolate K-10-41 significantly promoted tomato plant height, floral development, and fruit yield, Mk-20-7 enhanced height and fruit weight whereas K-10-27 induced tomato fruit numbers. Compost application promoted tomato-PSB interaction and induced tomato vegetative growth whereas bone meal was least promotor for most of tomato plant assays. Bone meal was however, one of the top fruit development inducers (harvested 20.94 fruits/plant weighing 881.97 gm). Mixing 50% of recommended compost and DAP fertilizer application enhanced tomato vegetative growth and fruit yield (21 fruits/plant harvested that weighed 872.46 gm). Based on the overwhelming performance, K-10-41 and Mk-20-7 application together with compost and fertilizer mixture were found effective. Therefore, the results of this study imply that application of competent PSB isolates together with nutrient supplements improved symbiotic effectiveness, sustainable production, and environmental health. Consequently, these promising isolates would be recommended for tomato production of higher yield and sustainability after verifying their efficacy at different agroecology and taxonomic identification. Screening potential strains and evaluating their competence under different conditions would be the future perspectives to develop efficient inoculants. Moreover, synergetic application of organic supplements (compost, farmyard, bone meal, or other biowastes), bioinoculants, and proper agrochemicals maximize production and environmental health and is feasible for the economic, social, and ecological sense of balance.

Bacillus cereus B8W8 an effective bacterial antagonist against major postharvest fungal pathogens of fruit

The antifungal activity of Bacillus cereus (B8W8) was investigated against major post-harvest fungal pathogens affecting citrus and apples fruits namely Penicillium digitatum, Penicillium italicum, Geotrichum citri-aurantii, Penicillium expansum, Botrytis cinerea, Monilinia laxa and Monilinia fructigena. The in vitro dual culture of B8W8 and fungal pathogens showed high inhibition rates of the mycelial growth of Monilinia laxa and Penicillium digitatum (71.4% and 72.5% respectively). Subsequently, the volatile compounds (VOCs) have a high antagonistic effect against Monilinia laxa (86.4%) and Monilinia fructigena (73.4%). In addition, the bacterial-free cell filtrate of B8W8 had a significantly important effect on the inhibition of spore’s germination and mycelial growth of Penicillium expansum, Botrytis cinerea, Penicillium italicum and Geotrichum citri-aurantii. The in vivo bioassays revealed a significant impact of B8W8 on fruit decays either by reducing the disease severity of causative agents or by completely preventing their occurrence, particularly brown rot of apple caused by M. laxa and green mold of citrus caused by P. digitatum (DS = 0%). However, the severity of sour rot of citrus caused by G. citri-aurantii was only reduced by 50%. The semi-commercial trials showed a high potential of B. cereus B8W8 to reduce the incidence of gray mold of apple and green mold of citrus to very low values during at storage. In addition, the biological treatment with B8W8 was shown generally more effective than imazalil, the chemical treatment reference. Furthermore, the rapid colonization of fruit wounds by B8W8 could promote its antagonistic ability. Moreover, biochemical tests showed the ability of B8W8 to produce protease, amylase, and hydrocyanic acid (HCN). Interestingly, the molecular analysis demonstrated the ability of this bacterium to produce the fengycin. The secretion of this lipopeptide and lytic enzymes is more likely the major mechanism of action involved in the biocontrol activity of this bacterium. Results indicated that quality parameters of fruit, when challenged post-harvest pathogens and treated with B8W8, were mostly very close to those of the untreated control, suggesting that B8W8 did not affect the fruit quality. Accordingly, it was concluded that B. cereus strain B8W8 could be an effective biological agent to control the main fungal pathogens of apples and citrus fruits during post-harvest period.

Selection Of Lactic Acid Bacteria as Starter Culture for Cocoa Fermentation (Theobroma Cacao L.)

This research aims to select the potential lactic acid bacteria (LAB) for starter culture for cocoa beans fermentation. The bio-control activity against mycotoxigenic fungi were also evaluated. The indigenous LAB was analized from Sukabumi cocoa beans fermentation by spontaneous method for four days (96 hours). The preliminary screening of LAB were observed by clear zone forming, morphology characterization and catalase activity. The LAB were screened for their ability to resist acidity, ethanol, and heat, and their ability to produce high acid and antifungal. The antifungal capability was antagonistic assays to Aspergillus ochraceus IPBCC 88.033 on Potato Dextrose Agar (PDA) medium. The result showed that the selected acid bacteria isolate, H 2.34, were antifungal-producing strains, and resist acidity, ethanol and heat, which was suitable property for starter culture and biopreservation agent. The study demonstrated the selection of local Indonesian LAB from Sukabumi fermented cocoa bean to observe the potential of isolate as a starter culture and biopreservation agent

Biopriming of seed with plant growth-promoting bacteria for improved germination and seedling growth.

Several seed priming methods can be used to improve seed germination, seedling vigor, and to overcome abiotic stress. In addition to these benefits, only the biopriming method provides the additional benefit of biotic stress management, earning it special attention. Seed biopriming is useful in almost all crops around the world and is an environmentally friendly alternative to chemical fungicides. Biopriming usually refers to use of beneficial microorganisms, in particular plant growth-promoting bacteria (PGPB) able to survive under various harsh environmental conditions. In this study, various bacterial strains were isolated from samples of different origins, i.e., rhizospheric soil, desert sand, and sea mud. Preliminary screening of 156 bacterial isolates was conducted on the basis of their potassium (K), phosphorus (P) solubilization ability, and production of plant growth hormone, i.e., indole acetic acid (IAA). The most efficient bacteria were identified by 16S rRNA gene nucleotide sequences and further examined for their ACC deaminase activity, ammonia production, and biocontrol activity (defined via chitinolytic activity, HCN, and siderophores production). Finally, carrot seed germination assay was conducted with 10 shortlisted most potent isolates. 68.6, 58.3, and 66.7% of tested bacterial isolates were capable of P, K, and Zn solubilization, respectively. Klebsiella aerogenes AF3II1 showed the highest P and K solubilization, while isolate AF4II5, AF7II3, and PC3 showed the highest IAA synthesis ability. Serratia plymuthica EDC15 and Pseudomonas putida AF1I1 showed the strongest chitinolytic and siderophore production activity, respectively. Seven isolates demonstrated strong HCN production ability. Five isolates improved carrot seed germination. Only selected isolates with plant growth-promoting properties can improve carrot germination. The results of this study demonstrate that mainly auxins are involved in seed germination. Furthermore, the data suggest that phosphate solubilization ability may play an additional role in seed germination.