毛竹PheFT6和PheFT17基因对外界环境的应答及蛋白互作分析  

Responses of PheFT6 and PheFT17 Genes in Phyllostachys pubescens to External Environment and Protein Interaction Analysis

作　　者：刘丽[1] 陈骄羽 邵明侠 柳参奎 郭小勤[1] LIU Li;CHEN Jiao-Yu;SHAO Ming-Xia;LIU Shen-Kui;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》2021年第3期506-520,共15页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金(31971735);浙江省自然科学基金重点项目(LZ21C160001);教育部创新团队发展计划(IRT-17R99);高等学校学科创新引智计划(D18008)。

摘　　要：毛竹(Phyllostachys edulis)是我国重要的经济竹种,笋芽萌发及发育是限制其产量的主要因素之一。磷脂酰乙醇胺结合蛋白(phosphatidyl ethanolamine-binding protein, PEBP)家族包含3个亚家族,分别为FLOWERING LOCUS T (FT)、TERMINAL FLOWER 1 (TFL1)和MOTHER OF FT AND TFL1 (MFT)。其中FT亚家族在植物生长发育过程中发挥着重要作用。为探究FT基因在笋芽发育过程中的功能,本研究克隆获得毛竹2个FT同源基因PheFT6 (GenBank No. MT976159)和PheFT17 (GenBank No.MT976160),进一步利用qRT-PCR分析其表达模式,利用酵母双杂交实验探究互作蛋白。结果表明,PheFT6和PheFT17编码区序列长度分别为525和528 bp,分别编码174和175个氨基酸,编码蛋白均包含PEBP结构域。PheFT6基因包含典型的4个外显子和3个内含子;而PheFT17基因有额外2个外显子插入,发生了可变剪接,形成至少4个转录本。系统进化树分析显示,PheFT6和PheFT17与水稻(Oryza sativa) Hd3a (heading date 3a)和拟南芥(Arabidopsis thaliana) FT亲缘关系较远。q RT-PCR分析表明,PheFT6和PheFT17基因表达呈现明显的昼夜节律,白天表达水平很低,夜晚表达水平较高;PheFT6基因受到低温(12℃)诱导,PheFT17基因表达受适温(28℃)诱导,28℃处理的幼苗比12℃培养的毛竹幼苗具有更高的分蘖率;PheFT6和PheFT17基因表达受干旱强烈抑制,随着干旱程度加强而表达抑制越明显。酵母双杂交结果显示,PheFT6和PheFT17蛋白均不与14-3-3 (G-box factor 14-3-3 protein)中的PheGF14b和PheGF14c互作,而与分枝过程中的Hub蛋白BRANCHED1 (BRC1)/TEOSINTE BRANCHED1 (TB1)的同源物PheTB1L-1和PheTB1L-2蛋白存在相互作用,说明PheFT6和PheFT17可能参与毛竹分枝分蘖过程。本研究为进一步分析PheFT6和PheFT17的生物学功能提供参考依据,可为毛竹笋芽发育的分子机制研究提供基础资料。Moso bamboo(Phyllostachys edulis) is an important economical bamboo species in China. Shoot bud germination and development is one of the main factors limiting its productivity. The phosphatidyl ethanolamine-binding protein(PEBP) family contains FLOWERING LOCUS T(FT) subfamily, TERMINAL FLOWER 1(TFL1) subfamily and MOTHER OF FT AND TFL1(MFT) subfamily. The FT subfamily plays an important role in the growth and development of plants. In order to explore the function of FT gene in the process of shoot bud development, 2 FT homologous genes, PheFT6 and PheFT17, were cloned in Ph. edulis,and their expression patterns were analyzed by qRT-PCR, and their interaction proteins were explored by yeast two-hybrid. The results showed that the lengths of the coding regions of PheFT6(GenBank No. MT976159)and PheFT17(GenBank No. MT976160) were 525 and 528 bp, encoding 174 and 175 amino acids,respectively, which contained the PEBP domain. The PheFT6 gene typically contains 4 exons and 3 introns,while the PheFT17 gene has additional 2 exons inserted, resulting in alternative splicing that produce at least 4 transcripts. Phylogenetic tree analysis showed that PheFT6 and PheFT17 were far from rice Heading date 3 a(Hd3 a) and Arabidopsis thaliana FT. qRT-PCR analysis showed that the expression of PheFT6 and PheFT17 genes showed obvious circadian rhythm, with low expression level in the daytime and high expression level at night. The expression of PheFT6 gene was induced by low temperature(12 ℃), while the expression of PheFT17 gene was induced by warm temperature(28 ℃). Compared with the seedlings cultured at 12 ℃, the seedlings treated at 28 ℃ had higher tillering ability. The expression of PheFT6 and PheFT17 genes was strongly inhibited by drought, and the inhibition increased with the increasing of drought degree. The yeast two-hybrid results showed that both PheFT6 and PheFT17 did not interact with PheGF14 b and PheGF14 c in14-3-3(G-box factor 14-3-3 protein), but interacted with PheTB1 L-1 and PheTB1 L-2, homologues of

关 键 词：毛竹 FT基因 分蘖 干旱 可变剪接 

分 类 号：S795.7[农业科学—林木遗传育种]