Zinc solubilizing Bacillus sp (SS9) and Enterobacter sp (SS7) promote mung bean (Vigna radiata L.) growth, nutrient uptake and physiological profiles.

Zinc (Zn) is a crucial micronutrient required for optimum plant growth. Zn-solubilizing bacteria (ZSB) are potential alternatives for Zn supplementation and convert applied inorganic Zn to available forms. In this study, ZSB were isolated from the root nodules of wild legumes. From a set of 17 bacteria, the isolates SS9 and SS7 were found to be efficient in tolerating 1 g (w/v) Zn. The isolates were identified as Bacillus sp (SS9, MW642183) and Enterobacter sp (SS7, MW624528) based on morphology and 16S rRNA gene sequencing. The screening of PGP bacterial properties revealed that both isolates possessed production of indole acetic acid (50.9 and 70.8 µgmL-1), siderophore (40.2% and 28.0%), and solubilization of phosphate and potassium. The pot study experiment in the presence and absence of Zn revealed that the Bacillus sp and Enterobacter sp inoculated plants showed enhanced mung bean plant growth (45.0% to 61.0% increment in shoot length and 26.9 to 30.9% in root length) and biomass compared to the control. The isolates also enhanced photosynthetic pigments such as total chlorophyll (1.5 to 6.0-fold) and carotenoids (0.5 to 3.0-fold) and 1-2-fold increase in Zn, phosphorous (P), and nitrogen (N) uptake compared to the Zn-stressed control. The present results indicated that the inoculation of Bacillus sp (SS9) and Enterobacter sp(SS7) reduced the toxicity of Zn and, in turn, enhanced the plant growth and mobilization of Zn, N, and P to the plant parts.

Evaluation of biocontrol Bacillus species on plant growth promotion and systemic-induced resistant potential against bacterial and fungal wilt-causing pathogens.

Bacillus spp. have a wide range of activities in the biocontrol potential against various phytopathogens. This study focuses on the biocontrol potential of two species belonging to the same genera, as Bacillus subtilis (SSR2I) and Bacillus flexus (AIKDL) have contrasting activity under in vivo and in vitro conditions. In this study, two medicinal plants-associated bacteria showing antagonistic activity against wilt-causing pathogens were selected and identified as B. subtilis (SSR2I) and B. flexus (AIKDL) based on 16S rRNA gene sequencing. Crude extracts of these bacteria showed that chloroform extracts of AIKDL, and ethyl acetate extraction of SSR2I showed effective potential inhibition of both the wilt-causing pathogens in the well-diffusion method. PCR-based detection of antimicrobial peptide genes revealed the presence of five genes in B. subtilis and none in B. flexus. On the basis of in vivo analysis, the isolate SSR2I showed reduced disease incidence and enhanced biocontrol efficiency against Ralstonia solanacearum and Fusarium oxysporum compared with AIKDL and control plants. Further, the isolates SSR2I also enhanced the induced systemic resistance (ISR) against both the pathogens compared to the control. However, the isolate AIKDL showed enhanced ISR against F. oxysporum-treated plants, but not against R. solanacearum-treated plants. The results indicated that even though the isolates had strong antagonistic potential under in vitro conditions, their biocontrol efficiency differed in in vivo condition. On the basis of the overall performance, the isolate SSR2I could be formulated as biocontrol agents against both the wilt-causing pathogens tested in this study.

Characterisation of antagonistic Bacillus paralicheniformis (strain EAL) by LC-MS, antimicrobial peptide genes, and ISR determinants.

Plants have their own defense mechanisms such as induced systemic resistance (ISR) and systemic-acquired resistance. Bacillus spp. are familiar biocontrol agents that trigger ISR against various phytopathogens by eliciting various metabolites and producing defense enzyme in the host plant. In this study, B. paralicheniformis (strain EAL) was isolated from the medicinal plant Enicostema axillare. Butanol extract of B. paralicheniformis showed potential antagonism against Fusarium oxysporum compared to control well (sterile distilled water) A liquid chromatography mass spectrometry analysis showed 80 different compounds. Among the 80 compounds, we selected citrulline, carnitine, and indole-3-ethanol based on mass-to-charge ratio, database difference, and resolution of mass spectrum. The synthetic form of the above compounds showed biocontrol activity against F. oxysporum under in vitro condition in combination, not as individual compounds. However, the PCR amplification of 11 antimicrobial peptide genes showed that none of the genes amplified in the strain. B. paralicheniformis inoculation challenged with F. oxysporum on tomato plants enhanced production of defense enzymes such as peroxidase (POD), superoxide dismutase (SOD), phenylalanine ammonia lyase (PAL), polyphenol oxidase (PPO), and proline compared to control plants (without inoculation of B. paralicheniformis) at significant level (p < 0.005). Stem of tomato plants expressed higher POD (2.2-fold), SOD (2.2-fold), PPO (1.9-fold), and PAL (1.3-fold) contents followed by the leaf and root. Elevated proline accumulation was observed in the leaf (1.8-fold) of tomato plants. Thus, results clearly showed potentiality of B. paralicheniformis (EAL) in activation of antioxidant defense enzyme against F. oxysporum-infected tomato plants and prevention of oxidative damage though hydroxyl radicals scavenging activities that suppress the occurrence of wilt diseases.

Amelioration effect of chromium-tolerant bacteria on growth, physiological properties and chromium mobilization in chickpea (Cicer arietinum) under chromium stress.

In this study, chromium (Cr)-tolerant bacteria were test for their efficiency in alleviating Cr stress in Cicer arietinum plants. On the basis of 16S rRNA gene analysis, the isolates were identified belonging to genus Stenotrophomonas maltophilia, Bacillus thuringiensis B. cereus, and B. subtilis. The strains produced a considerable amount of indole-3-acetic acid in a medium supplemented with tryptophan. The strains also showed siderophore production (S2VWR5 and S3VKR17), phosphorus production (S1VKR11, S3VKR2, S3VKR16, and S2VWR5), and potassium solubilization (S3VKR2, S2VWR5, and S3VKR17). Furthermore, the strains were evaluated in pot experiments to assess the growth promotion of C. arietinum in the presence of chromium salts. Bacterization improved higher root and shoot length considerably to 6.25%-60.41% and 11.3%-59.6% over the control. The plants also showed increase in their fresh weight and dry weight in response to inoculation with Cr-tolerant strains. The accumulation of Cr was higher in roots compared to shoots in both control and inoculated plants, indicating phytostabilization of Cr by C. arietinum. However, phytostabilization was found to be improved manifold in inoculated plants. Apart from the plant attributes, the amendment of soil with Cr and Cr-tolerant bacteria significantly increased the content of total chlorophyll and carotenoids, suggesting the inoculant's role in protecting plants from deleterious effects. This work suggests that the combined activity of Cr-tolerant and plant growth-promoting (PGP) properties of the tested strains could be exploited for bioremediation of Cr and to enhance the C. arietinum cultivation in Cr-contaminated soils.

Characterization of medicinal plant-associated biocontrol Bacillus subtilis (SSL2) by liquid chromatography-mass spectrometry and evaluation of compounds by in silico and in vitro methods.

This study explores the antimicrobial properties of bioactive secondary metabolites extracted from the medicinal plant (Solanum surattense)-associated Bacillus subtilis strain SSL2. The secondary metabolites were extracted from B. subtilis (SSL2) using ethyl acetate, acetone, butanol, chloroform and methanol solvents. The crude extract was tested against two wilt causing pathogens: Ralstonia solanacearum and Fusarium oxysporum. The results revealed that the ethyl acetate extract has maximum inhibition against both the pathogens tested in this study. Furthermore, liquid chromatography-mass spectrometry (LC-MS) analysis of ethyl acetate extract identified 80 different compounds based on mass-to-charge ratio, database difference, resolution of mass spectrum and so on. Among the 80 compounds, citrulline (m/z = 158.0917), chloramphenicol (m/z = 195.075) and carnitine (m/z 162.11) were further selected based on m/z ratio for in silico and in vitro analyses. The in silico analysis revealed that citrulline, chloramphenicol and carnitine inhibited the virulent genes phcA (R. solanacearum) and ste12 (F. oxysporum). Further, under in vitro condition, citrulline and chloramphenicol were found to inhibit the growth of R. solanacearum and F. oxysporum. On the basis of the biocontrol activity of B. subtilis (SSL2) in in silico and in vitro conditions, the bacteria could be used as a biocontrol agent against both bacterial and fungal wilt-causing pathogens. However, this needs to be tested in pot studies or field conditions before being used as biocontrol agents.Communicated by Ramaswamy H. Sarma.