摘要
同源异型-亮氨酸拉链(homeodomain-leucine-zipper, HD-Zip)转录因子参与高等植物形态建成、生长发育和环境中各种逆境的调控。本研究旨在利用融合蛋白表达、RNA干扰载体(p OsHox24:GUS, pCaMV35S:OsHox24-EGFP, pCaMV35S:OsHox24-RNAi)遗传转化等分子生物学方法,解析水稻中同源异型域亮氨酸拉链转录因子OsHox24在水稻生殖生长过程中的时空表达特性及其生物学功能。生物信息学分析表明,Os-Hox24基因在水稻第2号染色体上,具有2个外显子,1个内含子,编码的蛋白质由261个氨基酸组成,在该蛋白第61～121位氨基酸残基位置处有一个典型的HB类转录因子的DNA结合区(DNA-binding domain)。系统进化分析表明该基因与OsHox22同源性很高。GUS组织染色与RT-PCR实验显示OsHox24在水稻根尖、茎节和节间、叶鞘及不同时期的幼穗中均有表达。洋葱内表皮细胞亚细胞定位显示OsHox24在细胞膜和细胞核中表达。与野生型植株相比,RNA干扰植株的结实率显著降低。本研究初步探明由OsHox24形成的转录因子参与了水稻的生殖发育的调控过程,为进一步探索水稻HD-Zip转录因子功能和育种方面的应用研究提供科学依据。
Homeodomain-leucine-zipper(HD-Zip)transcription factors participate in morphogenesis,growth and development,and regulation of various adversities in the environment of advanced plants.This study aimed to analyze the spatial and temporal expression characteristics and biological functions of the homologous leucine zipper transcription factor OsHox24 in rice during reproductive growth by using fusion protein expression and RNA interference vector(pOsHox24:GUS,pCaMV35S:OsHox24-EGFP,pCaMV35S:OsHox24-RNAi)transformation.Bioinformatic analysis showed that OsHox24 was on the second chromosome of rice,containing 2 exons and one intron.The encoded protein consisted of 261 amino acids.There was a typical DNA-binding domain of HB transcription factor at the position of 61~121 amino acid residues.Phylogenetic analysis indicated that the gene was highly homologous with OsHox22.GUS staining and RT-PCR revealed that OsHox24 was expressed in root tip,stem node and internode,leaf sheath and young panicle of rice at different stages.Subcellular localization of onion epidermis cells showed that OsHox24 had expression in cell membrane and nucleus.Compared with wild-type plants,the seed setting rate of RNA interference plants decreased significantly.This study preliminarily demonstrated that transcription factors formed by OsHox24 were involved in the regulation of reproductive development in rice,which could provide scientific basis for further exploring the function of HD-Zip transcription factors and their application in rice breeding.
作者
付亚雄
刘思琪
汪琨璧
朱骞
Sadia Nadir
熊海波
李伟
孙明姬
李梦婷
周丽
冯天
李东宣
陈丽娟
Fu Yaxiong;Liu Siqi;Wang Kunbi;Zhu Qian;Sadia Nadir;Xiong Haibo;Li Wei;Sun Mingji;Li Mengting;Zhou Li;Feng Tian;Lee Dongsun;Chen Lijuan(Rice Research Institute,Yunnan Agricultural University,Kunming,650201;State Key Laboratory for Conservation and Utilization of Bio-Resources in Yunnan,Yunnan Agricultural University,Kunming,650201;Department of Chemistry,University of Science and Technology,Bannu,28100,Pakistan)
出处
《分子植物育种》
CAS
CSCD
北大核心
2019年第4期1061-1068,共8页
Molecular Plant Breeding
基金
国家重点研发计划项目(2017YFD0100205)
国家自然科学基金NSFC-云南联合基金(U1136604)
国家自然科学基金项目(31560115)
云南省重点基金项目(2015FA023)
云南省重大专项(2018ZG005)共同资助
