白羊草BiMYB52基因的克隆及转基因拟南芥抗旱性表达分析

doi:10.11733/j.issn.1007-0435.2023.01.004

草地学报 ›› 2023, Vol. 31 ›› Issue (1): 29-39.DOI: 10.11733/j.issn.1007-0435.2023.01.004

白羊草BiMYB52基因的克隆及转基因拟南芥抗旱性表达分析

李春艳^1,2, 钟华¹, 杜利霞^1,2, 侯向阳^1,2

1. 山西农业大学草业学院, 山西太谷 030801;
2. 农业农村部饲草高效生产模式创新重点实验室, 山西太谷 030801

收稿日期:2022-06-13 修回日期:2022-08-27 出版日期:2023-01-15 发布日期:2023-02-07
通讯作者: 侯向阳, E-mail:houxy16@vip.126.com
作者简介:李春艳(1978-)，女，山西岚县人，博士，副研究员，主要从事牧草遗传育种研究，E-mail:licyjzgjkx@163.com；
基金资助:
山西省农业科学院博士基金项目(YBSJJ2008)；山西省重点研发计划(202102140601006)共同资助

The Cloning BiMYB52 Gene from Bothriochloa ischaemum and Analysis of Drought Resistance Expression of Transgenic Arabidopsis thaliana of that clone

LI Chun-yan^1,2, ZHONG Hua¹, DU Li-xia^1,2, HOU Xiang-yang^1,2

1. College of Grassland Science, Shanxi Agricultural University, Taigu, Shanxi Province 030801, China;
2. Key Laboratory for Model Innovation in Forage Production Efficiency, Ministry of Agriculture and Rural Affairs, P. R. China, Taigu, Shanxi Province 030801, China

Received:2022-06-13 Revised:2022-08-27 Online:2023-01-15 Published:2023-02-07

摘要/Abstract

摘要： 白羊草(Bothriochloa ischaemum)是一种耐旱的优良乡土草种。为了探索白羊草转录因子MYB的结构和功能，本研究以白羊草转录组测序中筛选出的一段对干旱胁迫响应表达量变化显著的MYB序列为基础，采用基于拓扑异构酶快速克隆法从白羊草中克隆了一个R2R3-MYB基因BiMYB52，该基因编码237个氨基酸。为了深入研究BiMYB52基因的抗旱性，利用染色体步移技术扩增出了BiMYB52启动子序列，构建了pCAMBIA1301-BiMYB52过表达载体，采用农杆菌介导的花序侵染法对拟南芥(Arabidopsis thaliana)进行遗传转化，然后对野生拟南芥和转基因拟南芥进行干旱胁迫处理。结果表明，干旱胁迫复水后，转基因拟南芥的存活率显著高于野生拟南芥，丙二醛(Malondialdehyde,MDA)含量是干旱胁迫后转基因拟南芥的显著低于对照，脯氨酸(Proline,Pro)含量则显著高于对照。由此推测BiMYB52基因可能增加了拟南芥对干旱胁迫的抵抗能力。这为白羊草抗旱育种提供候选基因和理论基础。

关键词: 白羊草, MYB转录因子, 干旱胁迫, 表达分析, 转基因

Abstract: Bothriochloa ischaemum is an excellent native grass species with drought tolerance. In order to explore the structure and function of the transcription factor MYB of Bothriochloa ischaemum,a R2R3-MYB gene,BiMYB52 encoding 237 amino acids,was cloned from Bothriochloa ischaemum by topoisomerase rapid cloning method based on a segment of MYB sequence with significant changes in expression in response to drought stress. To further study the drought resistance of BiMYB52,the BiMYB52 promoter sequence was amplified by genome walking technique,and the pCAMBIA1301-BiMYB52 expression vector was constructed.A wild Arabidopsis thaliana was transformed by Agrobacterium mediated inflorescence infection to form a transgenic Arabidopsis,then drought stress was applied to the transgenic plants. The results showed that the survival rate of the transgenic Arabidopsis was significantly higher t，the MDA content significantly lower,and the content of proline was significantly higher than those of the control,non-transgenic Arabidopsis. It is speculated that BiMYB52 may increase the resistance of Arabidopsis to drought stress. This provides a candidate gene and theoretical basis for drought resistance breeding of Bothriochloa ischaemum.

Key words: Bothriochloa ischaemum, MYB transcription factor, Drought stress, Expression analysis, Transgenic

中图分类号:

Q945

李春艳, 钟华, 杜利霞, 侯向阳. 白羊草BiMYB52基因的克隆及转基因拟南芥抗旱性表达分析[J]. 草地学报, 2023, 31(1): 29-39.

LI Chun-yan, ZHONG Hua, DU Li-xia, HOU Xiang-yang. The Cloning BiMYB52 Gene from Bothriochloa ischaemum and Analysis of Drought Resistance Expression of Transgenic Arabidopsis thaliana of that clone[J]. Acta Agrestia Sinica, 2023, 31(1): 29-39.

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白羊草BiMYB52基因的克隆及转基因拟南芥抗旱性表达分析

The Cloning BiMYB52 Gene from Bothriochloa ischaemum and Analysis of Drought Resistance Expression of Transgenic Arabidopsis thaliana of that clone

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