Metagenomics of two gnotobiotically grown aromatic rice cultivars reveals genotype-dependent and tissue-specific colonization of endophytic bacterial communities attributing multiple plant growth promoting traits.

Exploration of community structures, habitations, and potential plant growth promoting (PGP) attributes of endophytic bacteria through next generation sequencing (NGS) is a prerequisite to culturing PGP endophytic bacteria for their application in sustainable agriculture. The present study unravels the taxonomic abundance and diversity of endophytic bacteria inhabiting in vitro grown root, shoot and callus tissues of two aromatic rice cultivars through 16S rRNA gene-based Illumina NGS. Wide variability in the number of bacterial operational taxonomic units (OTUs) and genera was observed between the two samples of the root (55, 14 vs. 310, 76) and shoot (26, 12 vs. 276, 73) but not between the two callus samples (251, 61 vs. 259, 51), indicating tissue-specific and genotype-dependent bacterial community distribution in rice plant, even under similar gnotobiotic growth conditions. Sizes of core bacteriomes of the selected two rice genotypes varied from 1 to 15 genera, with Sphingomonas being the only genus detected in all six samples. Functional annotation, based upon the abundance of bacterial OTUs, revealed the presence of several PGP trait-related genes having variable relative abundance in tissue-specific and genotype-dependent manners. In silico study also documented a higher abundance of certain genes in the same biochemical pathway, such as nitrogen fixation, phosphate solubilization and indole acetic acid production; implying their crucial roles in the biosynthesis of metabolites leading to PGP. New insights on utilizing callus cultures for isolation of PGP endophytes aiming to improve rice crop productivity are presented, owing to constancy in bacterial OTUs and genera in callus tissues of both the rice genotypes.

Impact of seed-transmitted endophytic bacteria on intra- and inter-cultivar plant growth promotion modulated by certain sets of metabolites in rice crop.

Exploring the beneficial interactions between plant and endophytes could be an effective strategy in the implementation of sustainable agricultural practices to enhance crop productivity. In this study, we aimed to evaluate holistically the plant growth promoting (PGP) abilities rendered by seed-transmitted endophytic bacteria isolated from in vitro grown calli of two rice cultivars. Nine bacterial endophytes, designated as PB001-PB009, were isolated and identified at the genus level through 16S rRNA gene sequence analysis. Biochemical investigations disclosed that they possess several PGP traits, such as phosphate solubilization, indole acetic acid biosynthesis, ammonia production, nitrogen fixation, amylase production and siderophore production. Results in gnotobiotic conditions revealed an increase in fresh weight, dry weight, root length and shoot length of seedlings germinated from endophyte-primed seeds than the control (uninoculated) set in a non-host and two host rice cultivars. In net house experiments, plants germinated from Micrococcus sp. PB001, Pseudomonas sp. PB002, Methylobacterium sp. PB005 and Methylorubrum sp. PB009 primed seeds showed an increase of upto 34.06 %, 38.77 %, 182.87 %, 16.59 % and 33.52 % in chlorophyll content, number of tillers/plant, number of grains/plant, grain size and grain weight, respectively than control plant sets in the non-host rice cultivar, further validating inter-cultivar PGP abilities of these endophytes. Metabolite profiling unfolded the abundance of few metabolites that are involved in pathways associated with PGP traits, in seedlings germinated from the endophyte-primed seeds. Together, the study documents the effect of seed-transmitted endophytic bacteria on intra- and inter-cultivar PGP by modulating certain sets of metabolites in rice plant, and is promising in developing bioinoculant formulations employing these selected endophytes for enhancement of rice productivity.