花生AhTFL1基因的克隆与功能分析  

Cloning and functional analysis of AhTFL1 of cultivated peanuts(Arachis hypogaea L.)

作　　者：齐飞艳[1] 石磊[1] 孙子淇[1] 郑峥[1] 苗丽娟[1] 田梦迪 张忠信[1] 黄冰艳[1] 董文召[1] 张新友[1] QI Fei-yan;SHI Lei;SUN Zi-qi;ZHENG Zheng;MIAO Li-juan;TIAN Meng-di;ZHANG Zhong-xin;HUANG Bing-yan;DONG Wen-zhao;ZHANG Xin-you(Henan Academy of Crops Molecular Breeding/Key Laboratory of Oil Crops in Huanghuaihai Plains,Ministry of Agri-culture and Rural Affairs,China/Henan Provincial Key Laboratory for Oil Crops Improvement/Henan Biological Breeding Center CO.,Ltd.,Zhengzhou 450002,China)

机构地区：[1]河南省农业科学院河南省作物分子育种研究院/农业部黄淮海油料作物重点实验室/河南省油料作物遗传改良重点实验室/河南生物育种中心有限公司,河南郑州450002

出　　处：《中国油料作物学报》2023年第4期728-734,共7页Chinese Journal of Oil Crop Sciences

基　　金：河南省科技攻关计划项目(212102110254);国家现代农业产业技术体系资助(CARS-13);河南省花生产业技术体系(S2012-5);河南省农业科学院自主创新项目(2021ZC81);河南省农业科学院科技创新创意项目(2020CX07);河南省重大科技专项(201300111000);河南省科技攻关计划项目(212102110256)。

摘　　要：栽培花生(Arachis hypogaea L.)连续开花亚种和交替开花亚种的开花时间、分枝模式和分枝数量均不同,这与PEBP蛋白家族基因时空表达差异有关。本研究从花生品种豫花25号的幼嫩腋芽组织中克隆到AhTFL1基因,属PEBP基因家族,比对参考基因组显示其位于18号染色体。AhTFL1基因开放阅读框为543 bp,编码180个氨基酸,拥有PEBP蛋白家族的保守的D-P-D-x-P和G-x-H-R结构域,91、115、145、147和159位的氨基酸与拟南芥TFL1保守的His88、Glu112、Ser142、Asp144和Asp156残基相同,系统进化树显示其属于TFL1-like亚家族。基于AhTFL1基因与拟南芥TFL1基因序列同源,所以利用拟南芥tfl1突变体进行功能互补试验验证其功能。35S::AhT⁃FL1互补拟南芥tfl1突变体表现为主茎增高、侧枝变长、荚果数增加、主茎侧枝数增多、莲座侧枝数减少,基本恢复至野生型性状。转化拟南芥野生型和tfl1突变体的35S::AhTFL1阳性株系都表现开花延迟,说明AhTFL1基因功能为促进营养生长,延迟营养生长向生殖生长的转变。Cultivated peanuts(Arachis hypogaea L.)are divided into two subspecies(hypogaea and fastigiata),according to the branching patterns of vegetable shoot and inflorescence.In these two subspecies,the flowering time,branching pattern and the numbers of branches are all different,which possibly due to the disparate spatiotemporal expressions of PEBP family genes.In this study,AhTFL1 was isolated from young axillary buds of Yuhua 25,and was alignned to chromosome 18.The 543-bp open reading frame of AhTFL1 was predicted to encode a pro-tein with 180 amino acids.The encoded protein contained the conservative D-P-D-x-P and G-x-H-R motifs,be-longing to PEBP family members.Moreover,five conservative amino acid residues in 91,115,145,147,and 159 of AhTFL1 were identical to His88,Glu112,Ser142,Asp144,and Asp156 in AtTFL1.The result of phylogenetic tree analysis suggested that AhTFL1 belongs to TFL1-like subfamily.Subsequently,AhTFL1 gene function was studied using functional complementation assay through Arabidopsis thaliana tfl1 mutant,based on the homology of AhTFL1 gene and Arabidopsis TFL1 gene.The transformation of 35S::AhTFL1 into the Arabidopsis thaliana tfl1 mutant can mainly recover the mutant phenotypes into wild type,such as resulting in the mutant with heightened main stem,lengthened lateral branches,increased main stem lateral branches and pod number,and decreased rosette lateral branches.Furtermore,over-expressing AhTFL1 in both wild type Arabidopsis and tfl1 mutant delayed flowering,im-plying AhTFL1 functions in the promotion of vegetative growth and the delay of transition from vegetative to repro-ductive growth.

关 键 词：栽培花生 PEBP基因家族 TFL1基因 基因功能 营养生长 

分 类 号：S565.2[农业科学—作物学]