摘要
[目的]OsGRF6隶属于GRF家族,参与调控植物的生长发育和植物对非生物胁迫的耐受性.试验旨在探究OsGRF6参与调控水稻初生根发育的分子机制,为深入研究OsGRF6参与调控水稻根的发育提供新线索.[方法]通过PCR从水稻品种'ZH11'中克隆得到转录因子基因OsGRF6,对其进行进化树构建、启动子分析;同时对OsGRF6-OE(过表达材料)和osgrf6(突变体)进行基因型分析、qRT-PCR鉴定和水稻表型观察,采用酵母单杂交技术对OsGRF6的结合基序进行检测,并通过ChIP-seq和RNA-seq数据对OsGRF6下游的直接靶基因进行分析.[结果](1)OsGRF6含有1个QLQ和1个WRC保守的功能结构域,与拟南芥AtGRF1和AtGRF2以及水稻OsGRF7亲缘关系较近,且OsGRF6启动子区域含有多个非生物胁迫和激素响应顺式作用元件;(2)OsGRF6在水稻种子、幼穗和根中表达量较高,而在叶中表达量较低;OsGRF6定位于细胞核,并且具有转录激活活性;(3)与野生型相比,OsGRF6-OE转基因材料初生根长度较长,而osgrf6突变体材料表现出初生根变短的表型;(4)OsGRF6能与CGGCA基序结合;(5)ChIP-seqs和RNA-seqs分析发现,OsGRF6靶基因中包含OsARF7、OsARF4等多个参与调控水稻根发育的基因.[结论]OsGRF6除了参与调控水稻株型和种子发育外,还能通过直接调控生长素途径中基因(如OsARF7、OsARF4等)的表达调控水稻根的发育.
Abstract
[Objective]OsGRF6 belongs to the GRF family and is involved in the regulation of plant growth and development as well as tolerance to abiotic stress.The purpose of this study was to explore the molec-ular mechanism of OsGRF6 in regulating the primary root development of rice,and provide clues for the in-depth understanding of OsGRF6 function in regulating root development in rice.[Methods]The tran-scription factor gene OsGRF6 was cloned from the rice variety'ZH11'by PCR.The evolutionary tree construction and promoter analysis were carried out.The genotype analysis,qRT-PCR identification,and phenotype observation of OsGRF6-OE(overexpression line)and osgrf6(mutant)were carried out.The binding sequence of OsGRF6 was detected through yeast one-hybrid assays,and the target genes down-stream of OsGRF6 were analyzed through ChIP-seqs and RNA-seqs data.[Results](1)OsGRF6 contained the conservative domains of QLQ and WRC,which is closely related to AtGRF1,AtGRF2,and OsGRF7.The promoter region of OsGRF6 contained multiple cis-acting elements responding to abiotic stress and hormones.(2)The expression of OsGRF6 was high in seeds,young spikes,and roots,but was low in leaves.Subcellular localization analysis showed that OsGRF6 was located in the nucleus,and OsGRF6 had transcription activation activity.(3)Compared with the wild type,overexpression of OsGRF6 showed a longer primary root,while loss of function mutant osgrf6 showed a shorter primary root.(4)OsGRF6 bond to the CGGCA sequence.(5)Using the ChIP-seq and RNA-seq assays,it was found that many Os-GRF6 target genes were involved in regulating rice root development,such as OsARF7 and OsARF4,etc.[Conclusion]In addition to regulating rice architecture and seed development,OsGRF6 also regulates rice root development by regulating the expression of genes involved in the auxin pathway,such as OsARF7 and OsARF4,etc.
基金项目
甘肃省科技厅自然科学基金面上项目(21JR7RA484)
厅市共建甘薯及特色豆科作物种质创新与利用四川省重点实验室开放课题基金资助项目(2021CGIHL04)
河南大学作物逆境适应与改良国家重点实验室开放课题项目(2022KF04)
国家科技期刊平台
NSTL
万方数据