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2025年8月3日 星期日
农业生物技术学报  2022, Vol. 30 Issue (12): 2290-2300    DOI: 10.3969/j.issn.1674-7968.2022.12.003
  研究论文与报告 本期目录 | 过刊浏览 | 高级检索 |
木薯ABA受体MePYL4a在拟南芥中过表达增强其耐旱性
赵东黎1,2, 鲍茹雪1,2, 李梦桃1,2, 陈新2,*, 王文泉1,*
1 海南大学 热带作物学院,海口 570228;
2 中国热带农业科学院 热带生物技术研究所,海口 571101
Overexpression of Cassava (Manihot esculenta) ABA Receptor MePYL4a in Arabidopsis thaliana Enhances Its Drought Tolerance
ZHAO Dong-Li1,2, BAO Ru-Xue1,2, LI Meng-Tao1,2, CHEN Xin2,*, WANG Wen-Quan1,*
1 College of Tropical Crops, Hainan University, Haikou 570228, China;
2 Institute of Tropical Biotechnology, Chinese Academy of Tropical Agricultural Sciences, Haikou 571101, China
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摘要 脱落酸(abscisic acid, ABA)及其受体pyrabactin resistance-like (PYL)蛋白在植物抵御干旱胁迫中发挥重要功能。为探究木薯(Manihot esculenta) MePYL4a (GenBank No. XM_021766535)基因的抗旱功能,本研究从木薯品种'SC8'中克隆MePYL4a基因,对其进行了生物信息学分析,利用农杆菌(Agrobacterium tumefaciens)介导的瞬时转化法进行亚细胞定位;通过实时荧光定量PCR检测MePYL4a基因在干旱胁迫处理下的表达情况;并构建了MePYL4a过表达载体转化拟南芥(Arabidopsis thaliana),进行拟南芥转基因植株的抗旱分析。结果表明,MePYL4a是不稳定蛋白,MePYL4a蛋白的亚细胞定位特征分析表明其定位于细胞核和细胞质内;qPCR分析显示,MePYL4a基因的表达水平显著受到干旱胁迫诱导,在1 h达到最高,随后慢慢下降。干旱处理后,观察表型发现MePYL4a能够提高干旱胁迫下拟南芥幼苗的耐旱性,过表达植株平均生物量是野生型(wild type, WT)的1.5倍,ABA合成关键限速酶基因9-顺式-环氧类胡萝卜素双加氧酶(9-cis-epoxycarotenoid dioxygenase, NCED3)和干旱胁迫诱导表达基因(responsive to desiccation 29A, RD29A)表达量显著高于WT和Atpyl4突变体植株,抗氧化酶活性也大约比WT高了2倍,表明MePYL4a的过表达植株清除活性氧的能力强于WT和Atpyl4突变体植株,在干旱胁迫下能积累更多的生物量,增强拟南芥的抗旱性。综上所述,MePYL4a过表达降低了干旱胁迫下拟南芥中活性氧的积累,诱导抗逆基因上调表达,增强拟南芥抗旱性。本研究为MePYL4a参与木薯抗逆的进一步研究提供参考依据。
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赵东黎
鲍茹雪
李梦桃
陈新
王文泉
关键词 木薯木薯脱落酸受体4a (MePYL4a)干旱胁迫基因表达量生理生化指标    
Abstract:Abscisic acid (ABA) and its receptor pyrabactin resistance like (PYL) play an important role in plant resistance to drought stress. In order to explore the drought resistance function of the MePYL4a (GenBank No. XM_021766535) gene of cassava (Manihot esculenta), the MePYL4a gene was cloned from the cassava variety 'SC8', and its bioinformatics analysis was carried out. The subcellular localization was carried out using the Agrobacterium tumefaciens mediated transient transformation method; The expression of MePYL4a gene under drought stress was detected by qPCR; MePYL4a overexpression vector was constructed and transformed into Arabidopsis thaliana for drought resistance analysis of A. thaliana transgenic plants. The results showed that MePYL4a was an unstable protein, and the subcellular localization characteristics of MePYL4a protein showed that it was located in the nucleus and cytoplasm; qPCR analysis showed that MePYL4a gene expression level was significantly induced by drought stress, reached the highest level in 1 h, and then slowly decreased. After drought treatment, the phenotype observation showed that MePYL4a could improve the drought tolerance of A. thaliana seedlings under drought stress. The average biomass of over expressed plants was 1.5 times that of wide type (WT). The expression of key rate limiting enzyme gene 9-cis-epoxycarotenoid dioxygenase (NCED3), and drought stress response index gene responsive to dehydration (RD29A) for ABA synthesis was significantly higher than that of WT and Atpyl4 mutant plant. And the activity of antioxidant enzymes was about twice higher than that of WT, indicating that the ability of over expressed plants of MePYL4a to remove active oxygen was stronger than that of WT and Atpyl4 plant. Under drought stress, it could accumulate more biomass and enhance the drought resistance of A. thaliana. In conclusion, MePYL4a overexpression reduced the accumulation of reactive oxygen species in A. thaliana under drought stress and induced higher expression of stress resistance genes, which enhanced drought resistance of A. thaliana. This study provides a reference for MePYL4a to participate in cassava stress resistance.
Key wordsCassava    M. esculenta pyrabactin resistance 1-like 4a (MePYL4a)    Drought stress    Gene expression    Physiological and biochemical indexes
收稿日期: 2022-04-27     
ZTFLH:  S533  
基金资助:国家重点研发专项(2018YFD1000500); 海南省研究创新科研课题(Hyb2020-09; Qhyb2021-25)
通讯作者: * wangwenquan@itbb.org.cn;chenxin@itbb.org.cn   
引用本文:   
赵东黎, 鲍茹雪, 李梦桃, 陈新, 王文泉. 木薯ABA受体MePYL4a在拟南芥中过表达增强其耐旱性[J]. 农业生物技术学报, 2022, 30(12): 2290-2300.
ZHAO Dong-Li, BAO Ru-Xue, LI Meng-Tao, CHEN Xin, WANG Wen-Quan. Overexpression of Cassava (Manihot esculenta) ABA Receptor MePYL4a in Arabidopsis thaliana Enhances Its Drought Tolerance. 农业生物技术学报, 2022, 30(12): 2290-2300.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/10.3969/j.issn.1674-7968.2022.12.003     或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2022/V30/I12/2290
 
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