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The mechanisms underlying tolerance to B-toxicity in plants are still controversial. Our previous studies indicated that B-toxicity is mainly limited to leaves in  Citrus  and that alternations of cell-wall structure in vascular bundles are involved in tolerance to B-toxicity. Here, miRNAs and their expression patterns were first identified in B-treated  Citrus sinensis  (tolerant) and  C. grandis  (intolerant) leaves via high-throughput sequencing. Candidate miRNAs were then verified with molecular and anatomical approaches. The results showed that 51 miRNAs in  C. grandis  and 20 miRNAs in  C. sinensis  were differentially expressed after B-toxic treatment. MiR395a and miR397a were the most significantly up-regulated miRNAs in B-toxic  C. grandis  leaves, but both were down-regulated in B-toxic  C. sinensis  leaves. Four auxin response factor genes and two laccase (LAC) genes were confirmed through 5′-RACE to be real targets of miR160a and miR397a, respectively. Up-regulation of  LAC4  resulted in secondary deposition of cell-wall polysaccharides in vessel elements of  C. sinensis , whereas down-regulation of both  LAC17  and  LAC4 , led to poorly developed vessel elements in  C. grandis . Our findings demonstrated that miR397a plays a pivotal role in woody  Citrus  tolerance to B-toxicity by targeting  LAC17  and  LAC4 , both of which are responsible for secondary cell-wall synthesis.

scientific reports

Illumina microRNA profiles reveal the involvement of miR397a in Citrus adaptation to long-term boron toxicity via modulating secondary cell-wall biosynthesis