Set of miRNAs Involved in Sulfur Uptake and the Assimilation Pathway of Indian Mustard (B. juncea) in Response to Sulfur Treatments

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Abstract
MicroRNAs (miRNAs) play an important role in the regulation of gene expression. They play a regulatory role in various nutrient assimilatory pathways of plants; however, their role in the regulation of sulfur uptake and assimilatory pathways in mustard cultivars under high/low sulfur conditions is not elucidated. Sulfur is essential for plant growth and development, and its deficiency can cause a decline in oil seed content and thus lower the economic yield in Brassica juncea. In this study, different miRNAs involved in the regulation of sulfur uptake and assimilation pathways in B. juncea were identified using a psRNA target analyzer and miRanda database tools. The predicted miRNAs that belong to 10 highly conserved families were validated using stem-loop RT-PCR. The B. juncea cultivars Pusa Jaikisan, Pusa Bold, and Varuna were kept in sulfur-excessive (high) and -deficient (insufficient) conditions, and expression studies of miRNAs and their target mRNAs were carried out using qRT-PCR. The correlation between the expression pattern of miRNAs and their target genes showed their potential role in sulfur uptake and assimilation. Analysis with 5′ RACE revealed the authentic target of miRNAs. The influence of S treatments on metabolites and sulfur content was also studied using GC-MS and a CHNS analyzer. Our study showed the potential role of miRNAs in the regulation of sulfur uptake and assimilation and put forward the implications of these molecules to enhance the sulfur content of B. juncea.

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