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,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,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1μmol/L和2.5μmol/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 KNO3)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)4个处理.结果显示:(1)在ABA处理下,转基因烟草种子的萌发率较野生型低,且在2.5μmol/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.上述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.
14-3-3 Protein is a type of highly conserved protein family widely distributed both in animals and plants. It is not only involved in various physiological and biochemical processes and metabolic reactions in organisms by interacting with different target proteins but also plays important roles in response to biotic and abiotic stresses. To clarify the function of the PeGRF6/8 a under abiotic stress, we cloned the PeGRF6/8 a from the 14-3-3 protein family of Populus euphratica, constructed pBI121-35 S::PeGRF6/8 a, an 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 KNO3) and high nitrogen(150 mmol/L KNO3) treatments;(3) 1/2 Hoagland hydroponics experiments-four treatments that included control(CK), low nitrogen(0.2 mmol/L KNO3), high nitrogen(150 mmol/L KNO3), 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 that in 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 that of wild type tobacco. However, it was in contrast with the root length under high nitrogen treatment.(3) Under low nitrogen and salt treatments, compared to the wild type, in transgenic tobacco, the lower leaves showed yellowing or wilting and the chlorophyll and carotenoid content in the leaves were markedly lower. Furthermore, the activity of superoxide dismutase and osmolyte content such as proline and soluble proteins was significantly lower in transgenic than in wild-type plants but the malondialdehyde(MDA) content was higher. However, under high nitrogen treatment, the speed
作者
冯倩
陈永富
姚银安
吴英青
张国燕
韩颖
高永峰
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)
出处
《应用与环境生物学报》
CAS
CSCD
北大核心
2019年第3期665-671,共7页
Chinese Journal of Applied and Environmental Biology
基金
国家自然科学基金面上项目(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~~
