菠萝R2R3-MYB基因家族S20亚族的鉴定

Identification of S20 subgroup of R2R3-MYB superfamily of Ananas comosus L.Merr.

【目的】鉴定菠萝R2R3-MYB基因家族S20亚族成员,分析其进化、结构与表达模式,为菠萝S20亚族的生物学功能研究奠定基础。【方法】以菠萝基因组数据库为基础,利用生物信息学方法对菠萝S20亚族基因进行鉴定,并对其系统进化、结构、保守基序和表达模式进行系统分析。同时,克隆并原核表达代表性基因AcMYB108a,利用Western blot进行杂交验证。【结果】在菠萝基因组上共鉴定到5个S20亚族基因,分布于5条染色体上。系统进化与保守基序分析发现,不同植物的S20蛋白具有较高的保守性且均具有motif3保守基序。顺式作用元件分析表明,菠萝S20亚族基因与植物逆境胁迫响应过程有关,且受乙烯诱导。基因表达分析结果显示,S20亚族基因的表达存在组织特异性,其中AcMYB108a在果实中的表达水平最高。利用pET原核表达系统成功表达了AcMYB108a重组蛋白,并通过了Western杂交验证。【结论】菠萝S20亚族基因与逆境胁迫响应相关,且受到乙烯诱导。

【Objective】This research identified the S20 subgroup genes of R2R3-MYB superfamily from pineapple genome and analyzed their evolution,structures and expression patterns to help understand the biological functions of pineapple S20 subgroup.【Method】Based on pineapple genome database,S20 subgroup genes of pineapple were identified by bioinformatic means,and their phylogenetic evolution,structures,conserved motifs and expression patterns were systematically analyzed.Meanwhile,the representative gene AcMYB108a was cloned and expressed in E.coli for further verification by Western blot.【Result】Five S20 subgroup genes distributed on five individual chromosomes were identified in pineapple genome.Phylogenetic evolution and conserved motif analysis showed that S20 proteins were highly conserved and all equipped with motif3.Cis-acting regulatory element analysis revealed that pineapple S20 subgroup genes were related to plant stress response and could be induced by ethylene.According to the transciptome data,the expressions of S20 subgroup genes were tissue specific,and AcMYB108a had the highest expression level in fruit.Recombinant AcMYB108a protein was successfully expressed by pET system and verified by Western blot.【Conclusion】The genes of pineapple S20 subfamily related to stress response and could be induced by ethylene.