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2025年4月4日 星期五
  2018, Vol. 26 Issue (8): 1288-1295    
  研究论文与报告 本期目录 | 过刊浏览 | 高级检索 |
巨桉锌指结构蛋白基因EgrZFP7在低温胁迫中的功能研究
倪晓详,王晓荣,陆军,从青,程龙军
浙江农林大学
Study on the Function of Zinc Finger Protein Gene EgrZFP7 in Cold Stress Response of Eucalyptus grandis
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摘要 摘 要 锌指结构蛋白(zinc finger protein, ZFP)是一类在植物生长、发育和逆境胁迫响应中发挥重要作用的转录因子。为了挖掘桉树(Eucalyptus)抗寒分子响应中的重要基因资源,为桉树抗低温分子辅助育种提供理论依据,本实验对巨桉(Eucalyptus grandis)在低温处理下受到强烈诱导表达的锌指结构蛋白基因EgrZFP7(Eucalyptus grandis zinc finger protein 7)编码蛋白结构及其功能进行了研究。该基因是典型的C2H2型锌指结构蛋白基因,编码蛋白序列中含有2个锌指结构基序、1个L-Box基序和1个乙烯响应元件结合因子(ethylene response factor, ERF)相关双性抑制子(ERF-associated amphiphilic repression, EAR)基序。通过构建EgrZFP7∷GFP表达载体,用基因枪轰击洋葱表皮的方法,进行基因表达蛋白的亚细胞定位,结果表明,EgrZFP7表达蛋白定位于细胞核。构建35S∷EgrZFP7超表达载体,转化拟南芥(Arabidopsis thaliana),得到超表达转基因纯合株系EgrZFP7-OX1和EgrZFP7-OX2。与野生型相比,其正常生长条件下侧根的数目和长度增加。以-8 ℃低温处理3 d、再缓苗恢复3 d,结果显示,超表达转基因株系恢复速度慢,同时幼苗因受低温胁迫的死亡率远高于野生型,说明该基因超表达增加了拟南芥转基因植株的低温敏感性。酵母双杂交文库中筛选到1个与EgrZFP7互作的乙烯响应因子蛋白EgrERF4(Eucalyptus grandis ethylene response factor 4)。上述研究结果表明,EgrZFP7可能通过与乙烯响应因子的相互作用,在巨桉低温胁迫响应中发挥负调控。本研究为进一步了解桉树抗寒分子机理提供了基础资料。
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倪晓详
王晓荣
陆军
从青
程龙军
关键词 锌指蛋白巨桉锌指结构蛋白基因7(EgrZFP7)基因功能巨桉低温胁迫    
Abstract:Abstract Zinc finger proteins (ZFPs) are a class of transcriptional factors that play important roles in growth, development and stress responses in plants. Finding more genes which have key function in cold stress response can give support for cold tolerance breeding in Eucalyptus trees with molecular assisted selection. A zinc finger protein gene induced under treatment at low temperature was screened in Eucalyptus grandis. The gene was named EgrZFP7. EgrZFP7 protein structure and its function were analyzed in the research.This gene is a classical C2H2 type zinc finger protein gene. The protein sequence it encoding contains 2 zinc finger motifs, 1 L-Box motif and 1 EAR motif. EgrZFP7∷GFP vector was constructed and subcellular localization of EgrZFP7 was verified by bombardment of onion epidermis with a gene gun.The result showed that it localized in nuclear. The vector of 35S∷EgrZFP7 was also constructed and transformed into Arabidopsis thaliana. Two homozygoteslines of EgrZFP7-OX1 and EgrZFP7-OX2 were obtained. Under normal growth condition, the 2 EgrZFP7 over-expression lines showed more lateral roots and the length of lateral roots were also longer than that of the wide type. However, when EgrZFP7-OX1 and EgrZFP7-OX2 were treated at -8 ℃ for 3 d, then recovered for 3 d under normal temperature, the speed of growth recovery of over-expression lines was slower than that of the wild type. Finally, death percentage of total seedlings EgrZFP7-OX1 and EgrZFP7-OX2 lines were obviously higher than that of the wild type, implying that over-expression of EgrZFP7 could increase sensitivity to low temperature in seedlings. A protein, EgrERF4 (Eucalyptus grandis ethylene response factor 4), which can interact with EgrZFP7 was screened and verified from Yeast Two Hybrid Library. It could be concluded that EgrZFP7 possibly play a role in gene regulation in cold stress response through interact with EgrERF4 in Eucalyputs grandis. The present study gives more information to learn the mechanism of cold stress response in Eucalyptus.
Key wordsZinc finger protein    Eucalyptus grandis zinc finger protein 7 gene (EgrZFP7)    Gene function    Eucalyptus grandis    Cold stress response
收稿日期: 2017-12-05      出版日期: 2018-07-20
基金资助:国家自然科学基金;浙江省科技厅林木新品种选育重大科技专项
通讯作者: 程龙军     E-mail: ljcheng@zju.edu.cn
引用本文:   
倪晓详 王晓荣 陆军 从青 程龙军. 巨桉锌指结构蛋白基因EgrZFP7在低温胁迫中的功能研究[J]. , 2018, 26(8): 1288-1295.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/     或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2018/V26/I8/1288
 
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