Alleviation of banded leaf and sheath blight disease incidence in maize by bacterial volatile organic compounds and molecular docking of targeted inhibitors in Rhizoctonia solani.

Rhizoctonia solani (RS) is a pathogenic fungus that affects maize (Zea mays L.) plants and causes banded leaf and sheath blight (BLSB) with severe consequences leading to significant economic losses. Contrarily, rhizobacteria produce numerous volatile organic compounds (VOCs) that help in devising the environment-friendly mechanism for promoting plant growth and stress alleviation without having physical contact with plants. In the present study, 15 rhizobacterial strains were tested for their antagonism against RS. The antagonistic potential of VOCs of the tested plant growth-promoting rhizobacteria (PGPR) strains ranged from 50% to 80% as compared to the control (without PGPR). Among these 15 strains, the maximum (80%) antagonistic activity was exhibited by Pseudomonas pseudoalcaligenes SRM-16. Thus, the potential of VOCs produced by P. pseudoalcaligenes SRM-16 to alleviate the BLSB disease in maize was evaluated. A pot experiment was conducted under greenhouse conditions to observe the effect of VOCs on disease resistance of BLSB-infected seedlings. Overall, maize seedlings exposed to VOCs showed a significant increase in disease resistance as indicated by a reduced disease score than that of unexposed infected plants. The VOCs-exposed maize exhibited lower (11.6%) disease incidence compared to the non-inoculated maize (14.1%). Moreover, plants exposed to VOCs displayed visible improvements in biomass, photosynthetic pigments, osmoregulation, and plant antioxidant and defense enzyme activities compared to the healthy but unexposed seedlings. Simultaneous application of RS and VOCs enhanced superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), phenylalanine ammonia lyase (PAL), ascorbate peroxidase (APX), and polyphenol oxidase (PPO) activities by 96.7%, 266.6%, 313.7%, 246.6%, 307%, and 149.7%, respectively, in the roots and by 81.6%, 246.4%, 269.5%, 269.6%, 329%, and 137.6%, respectively, in the shoots, relative to those of the control plants. The binding affinity of the VOCs (2-pentylfuran, 2,3-butanediol, and dimethyl disulfide) with CRZ1 and S9 protein receptors of RS was assessed by deploying in silico methods. Overall, 2-pentylfuran exhibited a binding affinity with both the selected receptors of RS, while 2,3-butanediol and dimethyl disulfide were able to bind S9 protein only. Hence, it can be deduced that S9 protein receptors are more likely the target RS receptors of bacterial VOCs to inhibit the proliferation of RS.

Drought-tolerant Bacillus megaterium isolated from semi-arid conditions induces systemic tolerance of wheat under drought conditions.

KEY MESSAGE: A detailed study of the response of wheat plants, inoculated with drought-tolerant PGPR is studied which would be beneficial to achieve genetic improvement of wheat for drought tolerance. Drought stress, a major challenge under current climatic conditions, adversely affects wheat productivity. In the current study, we observed the response of wheat plants, inoculated with drought-tolerant plant growth-promoting rhizobacteria (PGPR) Bacillus megaterium (MU2) and Bacillus licheniformis (MU8) under induced drought stress. In vitro study of 90 rhizobacteria exhibited 38 isolates showed one or more plant growth-promoting properties, such as solubilization of phosphorus, potassium, and exopolysaccharide production. Four strains revealing the best activities were tested for their drought-tolerance ability by growing them on varying water potentials (- 0.05 to - 0.73 MPa). Among them, two bacterial strains Bacillus megaterium and Bacillus licheniformis showed the best drought-tolerance potential, ACC deaminase activities, IAA production, and antagonistic activities against plant pathogens. Additionally, these strains when exposed to drought stress (- 0.73 MPa) revealed the induction of three new polypeptides (18 kDa, 35 kDa, 30 kDa) in Bacillus megaterium. We determined that 106 cells/mL of Bacillus megaterium and Bacillus licheniformis were enough to induce drought tolerance in wheat under drought stress. These drought-tolerant strains increased the germination index (11-46%), promptness index (16-50%), seedling vigor index (11-151%), fresh weight (35-192%), and dry weight (58-226%) of wheat under irrigated and drought stress. Moreover, these strains efficiently colonized the wheat roots and increased plant biomass, relative water content, photosynthetic pigments, and osmolytes. Upon exposure to drought stress, Bacillus megaterium inoculated wheat plants exhibited improved tolerance by enhancing 59% relative water content, 260, 174 and 70% chlorophyll a, b and carotenoid, 136% protein content, 117% proline content and 57% decline in MDA content. Further, activities of defense-related antioxidant enzymes were also upregulated. Our results revealed that drought tolerance was more evident in Bacillus megaterium as compared to Bacillus licheniformis. These strains could be effective bioenhancer and biofertilizer for wheat cultivation in arid and semi-arid regions. However, a detailed study at the molecular level to deduce the mechanism by which these strains alleviate drought stress in wheat plants needs to be explored.

Volatile organic compounds produced by Pseudomonas pseudoalcaligenes alleviated drought stress by modulating defense system in maize (Zea mays L.).

Research on plant growth-promoting bacteria (PGPR) revealed an effective role of bacterial volatile organic compounds (VOCs) in stress alleviation. Out of 15 PGPR strains, infection with VOCs from Pseudomonas pseudoalcaligenes' resulted in maximum germination, growth promotion, and drought tolerance in maize plants. The VOCs of P. pseudoalcaligenes caused induced systemic tolerance in maize plants during 7 days of drought stress. The VOCs exposed plants displayed resistance to drought stress by reducing electrolyte leakage and malondialdehyde content and increasing the synthesis of photosynthetic pigments, proline, and phytohormones contents. Maize plants revealed enhanced resistance by showing higher activities of antioxidant defense enzymes both in shoots and roots under drought stress. Activities of antioxidant enzymes were more pronounced in shoots than roots. Gas chromatography and mass spectrophotometric (GC-MS) analysis comparing VOCs produced by the most efficient P. pseudoalcaligenes strain and inefficient strains of Pseudomonas sp. grown in culture media revealed nine compounds that they had in common. However, dimethyl disulfide, 2,3-butanediol, and 2-pentylfuran were detected only in P. pseudoalcaligenes, indicating these compounds are potential candidates for drought stress induction. Further studies are needed to unravel the molecular mechanisms of VOCs-mediated systemic drought tolerance in plants related to each identified VOC.