内源H_(2)S对不结球白菜硒耐受的影响及相关基因克隆与分析  

Effect of Endogenous H_(2)S on Selenium Tolerance in Brassica rapa ssp.chinensis and Cloning and Analysis of Related Genes

作　　者：辛爱景 杨会敏 薛延丰[2] 刘晓丽 夏芊蔚 王永竹 石志琦[2] 陈健[2] 杨立飞[1,3] XIN Ai-Jing;YANG Hui-Min;XUE Yan-Feng;LIU Xiao-Li;XIA Qian-Wei;WANG Yong-Zhu;SHI Zhi-Qi;CHEN Jian;YANG Li-Fei(College of Horticulture,Nanjing Agricultural University,Nanjing 210095,China;Institute of Food Quality and Detection,Jiangsu Academy of Agriculture Science,Nanjing 210014,China;Hexian New Rural Development Research Institute,Nanjing Agricultural University,Hexian 238200,China)

机构地区：[1]南京农业大学园艺学院,南京210095 [2]江苏省农业科学院农产品质量安全与营养研究所,和县238200 [3]南京农业大学和县新农村发展研究院,南京210014

出　　处：《农业生物技术学报》2022年第2期260-271,共12页Journal of Agricultural Biotechnology

基　　金：江苏省农业科技资助创新资金(CX(20)3166);现代农业产业技术体系专项资金(CARS-23-B16)。

摘　　要：工业污染和过量硒肥施用导致局部地区土壤环境硒污染,进而胁迫植物生长。硫化氢(hydrogen sulfide,H_(2)S)是植物体内的一种气体信号分子,参与调控植物生长发育和逆境响应。为探索H_(2)S合成相关基因L/D半胱氨酸脱巯基酶(L/D-cysteine desulfhydrase,LCDs/DCDs)的功能,本研究以不结球白菜(Brassica rapa ssp.chinensis)为材料研究了内源H_(2)S及其相关基因对硒胁迫的响应方式。通过对不同硒浓度下幼苗根长和株高进行测定发现,硒胁迫显著抑制幼苗生长,并呈现浓度和时间效应;脂质过氧化和细胞膜损伤染色以及WSP-1原位荧光标记技术分析H_(2)S对硒胁迫诱导氧化损伤的影响,结果表明提高内源H_(2)S含量能够显著缓解硒胁迫诱导的氧化损伤效应。qRT-PCR分析显示硒胁迫前期诱导12个H_(2)S合成家族基因(BrLCD1~BrLCD10,BrDCD1~BrDCD2)表达上调,有助于内源H_(2)S的应激产生。最后,克隆了表达量上调较高的基因全长cDNA序列。序列分析显示这些基因编码的氨基酸序列均包含保守的半胱氨酸脱氢酶典型特征。本研究结果为揭示植物响应硒胁迫的作用机制提供了新证据。The industrial pollution and the overuse of selenium fertilizer result in selenium pollution in some area,which further inhibit plant growth.Hydrogen sulfide(H_(2)S),a kind of gaseous signaling molecule in plants,regulates plant development and stress responses.In order to explore the function of H_(2)S synthesis related genes LCDs/DCDs,In this study,Brassica campestris ssp.chinensis was used to study response of the endogenous H_(2)S and its related genes to selenium stress.The root length and plant height of seedlings at different selenium concentrations were measured,it was found that selenium stress significantly inhibited the growth of seedlings,and showed concentration and time effects.The effect of H_(2)S on oxidative damage induced by selenium stress was analyzed by lipid peroxidation and loss of membrane integrity and WSP-1 fluorescent probe technology,the results showed that enhancing endogenous H_(2)S alleviated selenium-induced oxidative injury in seedlings.qRT-PCR analysis showed that selenium stress induced up-regulation of 12 H_(2)S producing family genes(BrLCD1~BrLCD10,BrDCD1~BrDCD2)in the early stage,which might result in the rapid production of endogenous H_(2)S in the early period of selenium stress.Finally,the full-length cDNA of H_(2)S-producing genes with relative high expression level in response selenium stress were cloned.Sequence analysis showed that the amino acid sequences encoded by these genes contained the typical characteristics of conserved cysteine dehydrogenase.The results of this study provides new evidences for the revealing the mechanisms of plant response to selenium stress.

关 键 词：不结球白菜 硒胁迫 H_(2)S L/D半胱氨酸脱巯基酶(LCDs/DCDs)基因 

分 类 号：S634.3[农业科学—蔬菜学]