Morphological, Metabolomic and Genomic Evidences on Drought Stress Protective Functioning of the Endophyte Bacillus safensis Ni7.

The metabolomic and genomic characterization of an endophytic Bacillus safensis Ni7 was carried out in this study. This strain has previously been isolated from the xerophytic plant Nerium indicum L. and reported to enhance the drought tolerance in Capsicum annuum L. seedlings. The effects of drought stress on the morphology, biofilm production, and metabolite production of B. safensis Ni7 are analyzed in the current study. From the results obtained, the organism was found to have multiple strategies such as aggregation and clumping, robust biofilm production, and increased production of surfactin homologues under the drought induced condition when compared to non-stressed condition. Further the whole genome sequencing (WGS) based analysis has demonstrated B. safensis Ni7 to have a genome size of 3,671,999 bp, N50 value of 3,527,239, and a mean G+C content of 41.58%. Interestingly the organism was observed to have the presence of various stress-responsive genes (13, 20U, 16U,160, 39, 17M, 18, 26, and ctc) and genes responsible for surfactin production (srfAA, srfAB, srfAC, and srfAD), biofilm production (epsD, epsE, epsF, epsG, epsH, epsI, epsK, epsL, epsM, epsN, and pel), chemotaxis (cheB_1, cheB_2, cheB_3, cheW_1, cheW_2 cheR, cheD, cheC, cheA, cheY, cheV, and cheB_4), flagella synthesis (flgG_1, flgG_2, flgG_3, flgC, and flgB) as supportive to the drought tolerance. Besides these, the genes responsible for plant growth promotion (PGP), including the genes for nitrogen (nasA, nasB, nasC, nasD, and nasE) and sulfur assimilation (cysL_1&L_2, cysI) and genes for phosphate solubilization (phoA, phoP_1& phoP_2, and phoR) could also be predicted. Along with the same, the genes for catalase, superoxide dismutase, protein homeostasis, cellular fitness, osmoprotectants production, and protein folding could also be predicted from its WGS data. Further pan-genome analysis with plant associated B. safensis strains available in the public databases revealed B. safensis Ni7 to have the presence of a total of 5391 gene clusters. Among these, 3207 genes were identified as core genes, 954 as shell genes and 1230 as cloud genes. This variation in gene content could be taken as an indication of evolution of strains of Bacillus safensis as per specific conditions and hence in the case of B. safensis Ni7 its role in habitat adaptation of plant is well expected. This diversity in endophytic bacterial genes may attribute its role to support the plant system to cope up with stress conditions. Overall, the study provides genomic evidence on Bacillus safensis Ni7 as a stress alleviating microbial partner in plants.

Drought-Alleviating Effects of Endophytic Bacteria Isolated from Xerophytic Plants on Capsicum annuum L. Seedlings.

In the current study, 51 endophytic bacteria were isolated from 5 different xerophytic plants. Their drought tolerance properties were screened in vitro, and from these, four endophytes with tolerance up to - 1.5 MPa water potential were further selected and identified as Acinetobacter sp. Eo3, Pseudomonas sp. Ni5, Bacillus safensis Ni7, and Stenotrophomonas sp. C3. Due to biosafety concern, Acinetobacter sp. Eo3 and Pseudomonas sp. Ni5 were excluded from further investigation, while B. safensis Ni7 and Stenotrophomonas sp. C3 were subjected to detailed study. The drought tolerance properties of these endophytes were evaluated in vivo using Capsicum annuum L. by analysing the growth parameters (leaf number, root number, shoot length, and plant fresh weight) as well as physiological and biochemical parameters (stomatal index, relative water content, chlorophyll content, and carbohydrate accumulation) of bacteria-treated and control seedlings. Here, treatment with B. safensis Ni7 and Stenotrophomonas sp. C3 was found to result in statistically significant enhancement (P ≤ 0.001) of the measured parameters of plants when compared with the control groups. In the case of fresh weight itself, Ni7 and C3 treatment was found to result in values of 157.76 and 142.8 mg, respectively, and was statistically significant enhancement as the same for nutrient broth and distilled water control were 73.3 mg and 70.5 mg only. Additionally, the endophyte-treated seedlings displayed significant improvement in other growth parameters even under induced drought stress. These findings highlight the potential of xerophytic-derived bacterial endophytes to have significant role in mitigating the drought stress effects in plants with the promises for field application.

Quinalphos Tolerant Endophytic Bacillus sp. Fcl1 and its Toxicity-Alleviating Effect in Vigna unguiculata.

In order to meet the agricultural requirement for the expanding population, pesticides have been used regularly even with their severe threat. The uncontrolled use of these pesticides can cause irreparable damage to both soil and plant-associated microbiome. Therefore, an environment friendly alternative to enhance plant productivity and yield is highly important. Here comes the importance of endophytic microorganisms with multi-plant beneficial mechanisms to protect plants from the biotic and abiotic stress factors. However, their performance can be negatively affected under pesticide exposure. Hence the present study was conducted to analyse the tolerating ability of a Bacillus sp. Fcl1 which was originally isolated from the rhizome of Curcuma longa towards the pesticide quinalphos and also its ability to reduce the quinalphos-induced toxicity in Vigna unguiculata. The results revealed that the viability of endophytic Bacillus sp. Fcl1 depended on the concentration of quinalphos used for the study. Further, Fcl1 supplementation was found to alleviate the quinalphos-induced toxicity in Vigna unguiculata seedlings. The study is environmentally significant due to the pesticide tolerating and alleviating effect of Bacillus sp. Fcl1 in quinalphos-induced plant toxicity. This could suggest the application of microbes of endophytic origin as an efficient bioinoculant for field application even in the presence of pesticide residues.