桃PpILR1基因调控分枝角度的功能解析  

Function analysis of PpILR1 gene regulating branch angle in peach

作　　者：张杰 沈芮先 裴弯 谢贺芳 王小贝 郑先波[1,2] 连晓东 张海朋[1,2] 谭彬 冯建灿 ZHANG Jie;SHEN Ruixian;PEI Wan;XIE Hefang;WANG Xiaobei;ZHENG Xianbo;LIAN Xiaodong;ZHANG Haipeng;TAN Bin;FENG Jiancan(College of Horticulture,Henan Agricultural University,Zhengzhou 450046,China;Henan Engineering and Technology Center for Peach Germplasm Innovation and Utilization,Zhengzhou 450046,China)

机构地区：[1]河南农业大学园艺学院,河南郑州450046 [2]河南省桃种质资源创新及利用工程技术研究中心,河南郑州450046

出　　处：《河南农业大学学报》2023年第6期958-966,共9页Journal of Henan Agricultural University

基　　金：国家自然科学基金青年基金项目(32202412);河南省重大公益科技专项项目(201300110500);河南农业大学科技创新基金项目(KJCX2020A15);河南省现代农业产业技术体系建设项目(HARS-22-09-G1)。

摘　　要：【目的】明确桃生长素酰胺水解酶PpILR1基因功能,探讨生长素响应因子PpARF4和PpARF8A,通过结合并抑制PpILR1启动子活性调控分枝角度的分子机制,为桃树型遗传改良提供理论依据。【方法】构建PpILR1基因过表达载体稳定转化micro-Tom番茄;克隆PpILR1基因启动子序列及PpARF4、PpARF8A基因编码序列,通过酵母单杂交试验和烟草瞬时试验解析他们之间的调控关系。【结果】与野生型番茄相比,2个PpILR1转基因番茄株系均呈现分枝角度显著减少的表型;酵母单杂交结果表明,转录因子PpARF4和PpARF8A均能结合PpILR1启动子;烟草瞬时结果显示,PpARF4和PpARF8A为转录抑制子,能显著降低PpILR1启动子活性。【结论】PpILR1基因过表达导致分枝角度显著变小,表明PpILR1基因参与调控植物分枝角度形成。【Object】In order to provide theoretical basis for genetic improvement of peach tree architecture,the function of PpILR1 was verified,and the molecular mechanism of auxin response factor PpARF4 and PpARF8A to regulate branching angle by binding and inhibiting PpILR1 promoter activity was explored.【Method】The overexpression vector of PpILR1 was constructed,and was used for stable transformation in micro-Tom tomato.The promoter sequences of PpILR1 gene and the CDS sequence of PpARF4 and PpARF8A genes were cloned,and the regulation relationship between them was further verified via yeast single hybridization test and tobacco transient expression test.【Result】Compared with the wild type tomato,the two transgenic lines of the overexpression PpILR1 significantly reduced branch angle.Y1H results suggested that PpARF4 and PpARF8A transcription factors could bind to the promoter of PpILR1,and the transient expression assay in tobacco leaves indicated that PpARF4 and PpARF8A were transcriptional suppressor,which could obviously inhibit the activity of PpILR1.【Conclusion】The overexpression of PpILR1 gene resulted in the significant reduction of branch angle,which indicated that PpILR1 may participate in regulating the branch angle.

关 键 词：桃 分枝角度 生长素响应因子 生长素酰胺水解酶 

分 类 号：S662.1[农业科学—果树学]