毛竹PheMOC1a/b基因表达与互作蛋白分析  

PheMOC1a/b Gene Expression and Interaction Protein Analysis in Moso Bamboo(Phyllostachys edulis)

作　　者：邵明侠 周敏舒 陈骄羽 刘丽[1] 郭小勤[1] SHAO Ming-Xia;ZHOU Min-Shu;CHEN Jiao-Yu;LIU Li;GUO Xiao-Qin(State Key Laboratory of Subtropical Silviculture,Zhejiang A&F University,Hangzhou 311300,China)

机构地区：[1]浙江农林大学省部共建亚热带森林培育国家重点实验室,杭州311300

出　　处：《农业生物技术学报》2023年第1期73-86,共14页Journal of Agricultural Biotechnology

基　　金：浙江省自然科学基金重点项目(LZ21C160001);国家自然科学基金(32171880)。

摘　　要：毛竹(Phyllostachys edulis)是我国重要的经济竹种,笋芽正常发育与否决定了其产量。MOC1(MONOCULM1)基因在植物腋分生组织与腋芽的形成和发育中起着重要作用。本研究通过RT-PCR从毛竹中获得2个MOC1同源基因PheMOC1a(GenBank No.ON804247)和PheMOC1b(GenBank No.ON804248),序列分析表明,PheMOC1a和PheMOC1b的ORF分别为1530和1293 bp,分别编码509和430个氨基酸,蛋白多肽链中均含有GRAS-superfamily保守结构域,属于GIBBERELLIN ACID INSENSITIVE(GAI),REPRESSOR of GA1(RGA)and SCARECROW(SCR)(GRAS)基因家族。系统进化树分析显示,PheMOC1a/b与水稻(Oryza sativa)MOC1亲缘关系较近。qRT-PCR分析表明,PheMOC1a和PheMOC1b表达呈现明显的组织特异性;PheMOC1a和PheMOC1b在不同发育阶段笋芽中呈现相反的表达特性,随着笋芽发育进程的推进,PheMOC1a表达量降低,PheMOC1b表达量上升;在不同发育阶段的分枝中,PheMOC1a和PheMOC1b的表达趋势亦相反。酵母双杂交与双分子荧光互补结果显示,PheMOC1a和PheMOC1b不仅可以自身互作,两者间也可以互作;还可与indeterminate domain(IDD)家族的2个成员发生互作,而且这些互作蛋白在细胞核中发挥生物学作用。本研究为进一步分析PheMOC1a/b的生物学功能提供参考依据,为毛竹笋芽发育的分子机制提供基础资料。Moso bamboo(Phyllostachys edulis)is an important economic bamboo species in China.The normal development of bamboo bud/shoot determines its yield.The MONOCULM1(MOC1)gene plays an important role in the formation and development of axillary meristems and axillary buds.In this study,2MOC1 homologous genes,PheMOC1a(GenBank No.ON804247)and PheMOC1b(GenBank No.ON804248),were obtained from moso bamboo by RT-PCR.The sequence analysis showed that the ORFs of PheMOC1a and PheMOC1b were 1530 and 1293 bp,encoding 509 and 430 amino acid,respectively.Both polypeptide chains contain GRAS-superfamily conserved domains and belong to the GIBBERELLIN ACID INSENSITIVE(GAI),REPRESSOR of GA1(RGA)and SCARECROW(SCR)(GRAS)gene family.Phylogenetic tree showed that PheMOC1a/b was closely related to MOC1 in rice(Oryza sativa).q RT-PCR results showed that the PheMOC1a and PheMOC1b showed obvious tissue-specific expression.PheMOC1a and PheMOC1b showed opposite expression characteristics in the bamboo bud/shoot at different developmental stages.With the development of bamboo bud/shoot,the expression of PheMOC1a decreased,while the expression of PheMOC1b increased.The trend of PheMOC1a and PheMOC1b expression was opposite in the branches at different developmental stages.The results of yeast two-hybrid and bimolecular fluorescence complementation showed that PheMOC1a and PheMOC1b not only interacted with themselves,but also interacted with each other,and interacted with 2 members of the indeterminate domain(IDD)family.These interacting proteins played biological roles in the nucleus.This study provides a reference for further analysis of the biological functions of PheMOC1a/b,and basic information for the molecular mechanism of bamboo bud/shoot development in moso bamboo.

关 键 词：毛竹 MOC1(MONOCULM1)基因 笋芽 GRAS基因家族 基因表达 蛋白互作 

分 类 号：S795.7[农业科学—林木遗传育种]