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[1]冯 倩 陈永富 姚银安 吴英青 张国燕 韩颖 高永峰**.烟草异源过表达胡杨PeGRF6/8a对不同逆境的响应分析*[J].应用与环境生物学报,2019,25(03):1-12.[doi:10.19675/j.cnki.1006-687x.2018.10006]
 FENG Qian,CHEN Yongfu,YAO Yinan,et al.The Response Analysis of Heterologous Overexpression of Populus euphratica PeGRF6/8a in Tobacco under Different Stresses*[J].Chinese Journal of Applied & Environmental Biology,2019,25(03):1-12.[doi:10.19675/j.cnki.1006-687x.2018.10006]
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烟草异源过表达胡杨PeGRF6/8a对不同逆境的响应分析*()
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《应用与环境生物学报》[ISSN:1006-687X/CN:51-1482/Q]

卷:
25卷
期数:
2019年03期
页码:
1-12
栏目:
研究论文
出版日期:
2019-06-25

文章信息/Info

Title:
The Response Analysis of Heterologous Overexpression of Populus euphratica PeGRF6/8a in Tobacco under Different Stresses*
文章编号:
201810006
作者:
冯 倩 陈永富 姚银安 吴英青 张国燕 韩颖 高永峰**
西南科技大学生命科学与工程学院 绵阳 621010
Author(s):
FENG Qian CHEN Yongfu YAO Yin’an WU Yingqing ZHANG Guoyang HAN Ying & GAO Yongfeng**
School of Life Science and Engineering, Southwest University of Science and Technology, Mianyang 621010, China
关键词:
PeGRF6/8a14-3-3蛋白烟草低氮高氮盐胁迫
Keywords:
PeGRF6/8a tobacco abscisic acid low nitrogen high nitrogen salt stress
DOI:
10.19675/j.cnki.1006-687x.2018.10006
摘要:
14-3-3蛋白是一种广泛存在于动植物体内的高度保守的蛋白家族,它通过与各种靶蛋白之间的互作,参与生物体内各种生理生化过程和代谢反应,且在生物与非生物胁迫响应中发挥着重要作用。为明确胡杨PeGRF6/8a在非生物胁迫中的功能,本实验克隆了胡杨14-3-3蛋白家族中PeGRF6/8a的cDNA序列,构建该基因的植物过表达载体pBI121-35S::PeGRF6/8a,并采用农杆菌介导法将其转化野生型烟草。对获得的异源过表达烟草进行以下处理:(1)浓度为1 μmol/L和2.5 μmol/L脱落酸(Abscisic acid ,ABA)处理条件下的种子萌发实验;(2)低氮(2.5 mmol/L KNO3)和高氮(150 mmol/L KNO 3)胁迫下的根长对比实验;(3)1/2霍格兰(Hongland)营养液水培实验,分别设置对照(CK)、低氮(0.2 mmol/L KNO3)、高氮(150 mmol/L KNO3)和盐胁迫(150 mmol/L NaCl)四个处理。结果表明:(1)在ABA处理下,转基因烟草种子的萌发率较野生型低,且在2.5 μmol/L处理下更加显著;(2)低氮处理下,转基因烟草的根长显著短于野生型烟草的根长,而高氮处理下的结果相反;(3)水培实验中,低氮和盐胁迫处理下,转基因烟草与野生型相比,其下部的叶片明显变黄或萎焉,所含叶绿素和类胡萝卜素含量明显降低,抗氧化酶SOD以及渗透调节物质脯氨酸和可溶性蛋白的含量也显著降低,而叶片中丙二醛(MDA)的积累量却显著升高;高氮胁迫下,野生型烟草的萎蔫速度要显著快于转基因烟草,其转基因植株的生理指标除MDA积累量外均显著高于野生型。以上结果表明,在烟草中异源过表达胡杨PeGRF6/8a能够降低植物对低氮和盐的耐受性,但可以增强植物对高氮的耐受性。
Abstract:
14-3-3 protein is a kind of highly conserved protein family widely distributed both in animals and plants. Not only i t involves in various physiological and biochemical processes and metabolic reactions in organisms by interacting with different target proteins, but also it plays important roles in response to biotic and abiotic stresses. To clarify the function of the PeGRF6/8 under abiotic stress, we cloned the PeGRF6/8a from the 14-3-3 protein family of Populus euphratica, constructed the pBI121-35S::PeGRF6/8a expression vector, and then transformed it into wild-type tobacco by Agrobacterium -mediated transformation. The obtained transgenic tobaccos were treated as follows: (1) Seed germination experiments under 1 μmol/L and 2.5 μmol/L abscisic acid (ABA) treatments; (2) Root length comparison experiments under low nitrogen (2.5 mmol/L KNO 3) and high nitrogen (150 mmol/L KNO 3) treatments; (3) 1/2 Hoagland hydroponics experiments, four treatments were performed which included control (CK), low nitrogen (0.2 mmol/L KNO 3), high nitrogen (150 mmol/L KNO 3) and salt stress (150 mmol/L NaCl). The results were as follows: (1) Under ABA treatment, the germination rate of transgenic tobacco seeds was lower than the wild type. Upon increased ABA concentration, the germination rate of transgenic tobacco seeds declined much faster than that of the wild type; (2) Under low nitrogen treatment, the root length of transgenic tobacco was distinctly shorter than wild type tobacco. However, it was in contrast with that under high nitrogen treatment. (3) Under low nitrogen and salt treatments, comparing with the wild type, the lower leaves of transgenic tobacco showed yellow or wilting, the chlorophyll and carotenoid contents of transgenic tobacco leaves were dramatically lower. Furthermore, the activity of superoxide dismutase (SOD) and osmolytes contents such as proline and soluble proteins in transgenic plants was significantly lower, but the content of malondialdehyde (MDA) was higher. However, under high nitrogen treatment, the speed of plants wilting in the wild type was markedly faster than the transgenic, and the resistance and related physiological indexes (except MDA) of transgenic plants were considerably higher than those of wild type. These results indicate that heterologous overexpression of Populus euphratica PeGRF6/8a in tobacco can weaken plant tolerance to low nitrogen and salt stresses , but it can enhance plant tolerance under high nitrogen stress.

备注/Memo

备注/Memo:
收稿日期:2018-10-08 接受日期 Accepted: 2018-12-06*国家自然基金面上项目(31770644),四川省国际合作项目(2017HH0050),四川省教育厅资助科研项目(17ZB0456),四川省青年科技基金项目(2014JQ0016),西南科技大学博士基金项目(14zx7157)资助 **通讯作者(E-mail: gaoyongfeng0263@gmail.com)点击摘要页题目后的“PDF”可下载阅读全文;本文为已录用的作者修定稿,尚未经编辑全面修改。引用本文请注明出处本刊;发表刊期和页码将以正式出版时的安排为准,但DOI确定不变。
更新日期/Last Update: 2018-12-28