Dissecting the function of ib-miR2111 and its targets in sweetpotato
作者单位：College of Agriculture Shanxi Agriculture University College of Life Sciences Shanxi Agriculture University
会议名称：《2019年中国作物学会学术年会》
会议日期：2019年
基 金：National Key R&D Program of China(2018YFD1000705,2018YFD1000700) Key R&D Program of Shanxi Province(201803D221008-6) NSF of Shanxi Province(201801D121238) Coal-based Key Sci-Tech Project of Shanxi Province(FT-2014-01) NSFC(31101555,31571611) Special Plan of Scientific Research for Shanxi Agriculture Valley Construction of China(SXNGJSKYZX201701-03)
关 键 词：Sweetpotato Anthocyanin biosynthesis Ib-miR2111 Pi deficiency
摘 要：【Introduction】Compared with white-fleshed sweetpotato(WFSP), purple-fleshed sweetpotato(PFSP) is a desirable resource for functional food development because of the abundant anthocyanin accumulation in its tuberous roots. Some studies have shown that the expression regulation mediated by miRNA plays an important role in anthocyanin biosynthesis in plants. However, few mi RNAs and their corresponding functions related to anthocyanin biosynthesis in tuberous roots of sweetpotato have been known. 【Materials and methods】By comprehensive analysis of transcriptome, small RNA(s RNA) and degradome data from the tuberous roots of WFSP(Xushu18) and PFSP(Xuzishu3), a set of sweetpotato miRNAs were highly associated with root-specific anthocyanin accumulation. In particular, there was a significant difference in the expression of ib-miR2111 and its target genes. Transgenic plants were generated for forward and reverse genetic investigation of ib-miR2111 and its corresponding targets, through over-expression and knockout by techniques such as STTM, RNAi, and CRISPR/Cas9. 【Results and analysis】 Two target genes of ib-miR2111, ibZFP and ibFbox were identified by degradome sequencing and RLM-5’RACE. The different expression and inverse correlation between the expression of ib-miR2111 and its targets in Xushu18 and Xuzishu3 were validated by qRT-PCR. The young seedlings of ib-miR2111 over-expressing Arabidopsis showed suppressed abundance of ibZFP and a purplish phenotype. under normal growth conditions. Transgenic sweetpotato seedlings overexpressing ib-miR2111 also appear purple. In addition, the transgenic Arabidopsis expressing p CBCDT1 T2-pHSE401-ib-miR2111 CRISPR/Cas9 vector showed decreased biomass. Overexpression and CRISPR/Cas9 vector of the two targets were constructed and transformed into sweetpotato callus cells. Interestingly, we found that ib-mi R2111 also responded to Pi deficiency in sweetpotato.【Conclusion】Ib-mi R2111 was involved in the biosynthesis of anthocyanins