小麦中MIR160基因家族的生物信息学分析及靶基因鉴定

Bioinformatics Analysis and Target Gene Identification of MIR160 Gene Family in Wheat

MicroRNA(MIRNA)在植物的生长、发育和逆境胁迫应答等生物学过程中发挥着重要的作用。为了研究小麦中MIR160家族(MIR160s)的进化规律、表达模式及其靶基因,以小麦中11个MIR160s成员的前体和成熟体序列为基础,进行生物信息学分析,利用RT-qPCR结合RNA-seq数据明确MIR160s在不同小麦组织的表达模式,利用降解组测序和5'RLM-RACE技术验证小麦中MIR160s与靶基因的靶向关系。结果表明,MIR160s成熟体的序列在32个植物物种中高度保守,仅在第15和21位碱基存在多样性;小麦中MIR160s成员分别定位于1、5、6和7号染色体上;Tae-MIR160s成熟体在成熟叶片中表达量较高,其中Tae-MIR160c表达量最低;Tae-MIR160s分别可以在第1716、1562、1305和1931位碱基靶向切割TaARF11、TaARF25、TaARF34、TaARF54生长素响应因子(Auxinresponse factor)。本研究通过揭示Tae-MIR160s的进化规律、表达模式和靶基因,为小麦中MIR160s应答响应机制的解析提供了参考。

MicroRNA(MIRNA) plays an important role in plant growth, development, and response to adversity stress. To study the evolutionary laws, expression patterns and target genes of the MIR160 family(MIR160s) in wheat, bioinformatics analysis was conducted based on the precursor and mature body sequences of 11 MIR160s members in wheat. RNA-seq dates combined with RT-qPCR was used to clarify the expression patterns of MIR160s in different wheat tissues. Degradome sequencing and 5’RACE was used to verify the targeting relationship between MIR160s and target genes in wheat. The results showed that the sequence of MIR160s mature body was highly conserved among 32 plant species, with diversity only in the 15^(th) and 21^(st) bases. The members of Tae-MIR160s in wheat are located on chromosomes Chr.1, Chr.5, Chr.6, and Chr.7 respectively. Mature Tae-MIR160s have a higher expression level in mature leaves, and Tae-MIR160c has the lowest expression level. Tae-MIR160s can target and cleave TaARF11, Ta ARF25, TaARF34, TaARF54(Auxin Response Factor) at bases 1 716, 1 562, 1 305,and 1 931, respectively. This study provides a reference for the analysis of MIR160s response mechanism in wheat by revealing the evolution, expression pattern and target genes of Tae-MIR160s.