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2025年4月6日 星期日
农业生物技术学报  2022, Vol. 30 Issue (9): 1698-1712    DOI: 10.3969/j.issn.1674-7968.2022.09.005
  研究论文与报告 本期目录 | 过刊浏览 | 高级检索 |
木豆MYB-related亚家族基因鉴定及干旱胁迫下CcMYB-R48的功能解析
张修齐1, 董碧莹1, 宋治华1, 杜婷婷1, 李娜1, 薛婧怡1, 曹红燕1, 王天翼1, 宋洋波3, 孟冬1,2, 杨清1,2,*
1 北京林业大学 林学院/森林培育与保护教育部重点实验室,北京 100083;
2 北京林业大学 树木发育与基因编辑研究院,北京 100083;
3 青海大学 农牧学院,青海 810016
Identification of MYB-related Subfamily Genes in Cajanus cajan and Functional Analysis of CcMYB-R48 Under Drought Stress
ZHANG Xiu-Qi1, DONG Bi-Ying1, SONG Zhi-Hua1, DU Ting-Ting1, LI Na1, XUE Jing-Yi1, CAO Hong-Yan1, WANG Tian-Yi1, SONG Yang-Bo3, MENG Dong1,2, YANG Qing1,2,*
1 The Key Laboratory for Silviculture and Conservation of Ministry of Education, College of Forestry, Beijing Forestry University, Beijing 100083, China;
2 Institute of Tree Development and Gene Editing, Beijing Forestry University, Beijing 100083, China;
3 College of Agriculture and Animal Husbandry, Qinghai University, Xining 810016, China
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摘要 木豆(Cajanus cajan)是药食两用的木本豆科植物,含有丰富的活性物质,具有极高的经济价值。包括干旱和病害在内的非生物和生物胁迫是造成木豆减产和品质下降的重要因素,因此,挖掘木豆抗逆关键基因并分析其功能是木豆育种研究的重点。作为MYB (v-myb avian myeloblastosis viral)家族转录因子中的重要成员,MYB-related (MYB-R)亚家族基因被认为在逆境胁迫中发挥着重要作用。本研究通过生物信息学方法,利用拟南芥(Arabidopsis thaliana) MYB-R亚家族基因在木豆基因组中进行序列比对,筛选出48个CcMYB-R家族基因(CcMYB-R1~CcMYB-R48)。转录组数据分析发现,CcMYB-R48基因在脱落酸(abscisic acid, ABA)与茉莉酸甲酯(methyl jasmonate, MeJA)胁迫处理下都显著上调表达且表达量较高,说明其可能在木豆的抗逆应答和激素诱导下发挥重要功能。克隆并分析CcMYB-R48基因发现,其具有1个典型的MYB功能域,与拟南芥AT1G19000AT1G74840同源性较高。实时半定量PCR (semi-quantitative RT-PCR, SqRT-PCR)检测发现,CcMYB-R48在干旱胁迫6和12 h的条件下均显著上调表达,而在高温胁迫6和12 h的条件下则无差异,说明其可能参与木豆抵御干旱的过程。构建CcMYB-R48过表达载体,并利用木豆瞬时转化系统在木豆植株中过表达CcMYB-R48,结果显示,在干旱条件下,过表达植株具有较低的死亡率(19.44%),其电导率、丙二醛(malondialdehyde, MDA)和过氧化物酶(peroxidase, POD)含量均显著低于对照植株,表明CcMYB-R48能够增强木豆的抗旱能力。本研究对进一步探究木豆CcMYB-R亚家族基因及其抗旱机制,开展分子抗逆育种工作都具有重要意义。
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张修齐
董碧莹
宋治华
杜婷婷
李娜
薛婧怡
曹红燕
王天翼
宋洋波
孟冬
杨清
关键词 木豆MYB-related亚家族转录组分析干旱胁迫功能验证    
AbstractCajanus cajan is a medicinal and dietary woody legume with rich active substances and high economic value. Abiotic and biotic stresses, including drought and diseases, are important factors in the reduction of yield and quality of C. cajan; therefore, mining key C. cajan resistance genes and analyzing their functions are the focus of C. cajan breeding research. As an important member of the MYB (v-myb avian myeloblastosis viral) family of transcription factors, MYB-related (MYB-R) subfamily genes are thought to play an important role in adversity stress. In this study, 48 CcMYB-R (CcMYB-related) family genes (CcMYB-R1~CcMYB-R48) containing conserved structural domains of the MYB-R subfamily were screened by bioinformatic methods using Arabidopsis thaliana MYB-R subfamily genes for sequence alignment in the C. cajan genome. Analysis of transcriptome data revealed that CcMYB-R48 was significantly up-regulated and highly expressed under abscisic acid (ABA), Methyl Jasmonate (MeJA) stress treatments, suggested that it might be play an important function in stress response and hormone induction in C. cajan. Cloning and analysis of the CcMYB-R48 revealed that it had a typical MYB functional domain with high homology to Arabidopsis thaliana AT1G19000 and AT1G74840. The semi-quantitative RT-PCR (SqRT-PCR) revealed that CcMYB-R48 was significantly up-regulated in expression under both 6 and 12 h of drought stress, whereas no difference was observed under 6 and 12 h of high-temperature stress, suggested its possible involved in drought resistance in C. cajan. CcMYB-R48 overexpression vector was constructed and overexpressed CcMYB-R48 in C. cajan plants were established using the C. cajan instantaneous conversion system. The results showed that the overexpression plants had lower mortality of 19.44% under drought conditions, and their electrical conductivity, malondialdehyde (MDA) content and peroxidase (POD) content were significantly lower than those of the control plants, indicated that CcMYB-R48 could enhance the drought resistance of C. cajan. This study is important to further investigate the CcMYB-R subfamily genes of C. cajan and their drought resistance mechanism and to carry out molecular resistance breeding work.
Key wordsCajanus cajan    MYB-related subfamily    Transcriptome analysis    Drought stress    Functional verification
收稿日期: 2022-04-28     
ZTFLH:  S529  
基金资助:国家自然科学基金(31901281; 31922058); 北京林业大学杰出青年基金(2019JQ03009)
通讯作者: *yang.qing1020@163.com   
引用本文:   
张修齐, 董碧莹, 宋治华, 杜婷婷, 李娜, 薛婧怡, 曹红燕, 王天翼, 宋洋波, 孟冬, 杨清. 木豆MYB-related亚家族基因鉴定及干旱胁迫下CcMYB-R48的功能解析[J]. 农业生物技术学报, 2022, 30(9): 1698-1712.
ZHANG Xiu-Qi, DONG Bi-Ying, SONG Zhi-Hua, DU Ting-Ting, LI Na, XUE Jing-Yi, CAO Hong-Yan, WANG Tian-Yi, SONG Yang-Bo, MENG Dong, YANG Qing. Identification of MYB-related Subfamily Genes in Cajanus cajan and Functional Analysis of CcMYB-R48 Under Drought Stress. 农业生物技术学报, 2022, 30(9): 1698-1712.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/10.3969/j.issn.1674-7968.2022.09.005     或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2022/V30/I9/1698
 
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