Bna-miR43-FBXL调控模块参与甘蓝型油菜铝胁迫的功能分析  

Functional analysis of Bna-miR43-FBXL regulatory module involved in aluminum stress in Brassica napus

作　　者：张盈川 吴晓明玉 陶保龙 陈丽[1,2] 鲁海琴 赵伦 文静 易斌[1] 涂金星[1] 傅廷栋[1] 沈金雄[1] ZHANG Ying-Chuan;WU Xiao-Ming-Yu;TAO Bao-Long;CHEN Li;LU Hai-Qin;ZHAO Lun;WEN Jing;YI Bin;TU Jing-Xing;FU Ting-Dong;SHEN Jin-Xiong(National Key Laboratory of Crop Genetic Improvement/National Engineering Research Center of Rapeseed,Huazhong Agricultural University,Wuhan 430070,Hubei,China;School of Advanced Agriculture and Bioengineering,Yangtze Normal University,Chongqing 408100,China)

机构地区：[1]华中农业大学作物遗传改良全国重点实验室/国家油菜工程技术研究中心,湖北武汉430070 [2]长江师范学院现代农业与生物工程学院,重庆408100

出　　处：《作物学报》2023年第5期1211-1221,共11页Acta Agronomica Sinica

基　　金：国家自然科学基金项目(31871654)资助。

摘　　要：在酸性土壤中,铝是制约作物生长和产量的一个重要因素,如何利用酸性土地意义重大。本研究的对象是前期本课题鉴定到的一个未被报道的miRNA——Bna-miR43。系统地对甘蓝型油菜中Bna-miR43及其靶基因进行生物信息学分析以及表达模式鉴定,并构建Bna-miR43过表达载体探讨了Bna-miR43-FBXL模块在甘蓝型油菜响应铝胁迫的分子机制。5’-RACE结果表明Bna-miR43在甘蓝型油菜中真实切割BnaA09g03940D、BnaCnng24950D。生物信息学分析表明,BnaA09g03940D、BnaCnng24950D在拟南芥中的同源基因为AT5G27950,编码含F-box的E3泛素连接酶。qRT-PCR结果显示,Bna-miR43及其靶基因在甘蓝型油菜不同组织以及铝胁迫瞬时处理下的表达水平存在此消彼长的现象。油菜转基因试验表明,Bna-miR43的过表达株系地上部分长势良好,体内积累了较少的MDA和H2O2,同时转基因植株根系的铝积累量较对照少。本研究结果为甘蓝型油菜中铝胁迫响应提供参考。Aluminum is one of the main factors that limited crop growth and yield in acid soil and how to utilize acid soil is of great significance.This study is based on Bna-miR43,an unreported miRNA identified in the previous study.We analyzed the molecular characteristics,conserved structure,phylogenetic analysis,and the expression level of Bna-miR43 and its target genes in Brassica napus.The Bna-miR43 over expression vector was constructed to explore the molecular mechanism of Bna-miR43-FBXL module in Brassica napus in aluminum stress.5’-RACE showed that Bna-miR43 actually cleaved BnaA09g03940D and BnaCnng24950D.Bioinformatics analysis revealed that the homologous gene of BnaA09g03940D and BnaCnng24950D in Arabidopsis was AT5G27950,encoding E3 ubiquitin ligase contained F-box.The qRT-PCR indicated that the relative expression levels of Bna-miR43 and its target genes had a phenomenon of trade-off in different tissues of Brassica napus and under transient treatment of aluminum stress.Aluminum treatment showed that the entire aerial part of the transgenic plant grew much better than control and accumulated less MDA and H2O2.Meanwhile,the aluminum accumulation in the root of transgenic plants was less than the control.In this study,these results may provide reference for the response of aluminum stress in Brassica napus.

关 键 词：甘蓝型油菜 Bna-miR43 F-BOX 铝胁迫 

分 类 号：S565.4[农业科学—作物学]