锌指蛋白基因ZAT12提高转基因高油酸花生抗寒性  

Enhancing Cold Tolerance of Transgenic High Oleic Peanut by Zinc Finger Protein Gene ZAT12

作　　者：王旭达[1] 张高华[1] 王鹤[1] 于树涛 李怀梅[1] 王晓燕[1] 范琦[1] Wang Xuda;Zhang Gaohua;Wang He;Yu Shutao;Li Huaimei;Wang Xiaoyan;Fan Qi(Liaoning Ocean and Fisheries Science Research Institute,Liaoning Academy of Agricultural Sciences,Dalian,116023;Sandy Land Amelioration and Utilization Research Institute of Liaoning,Fuxin,123000)

机构地区：[1]辽宁省农业科学院海洋水产科学研究院,大连116023 [2]辽宁省沙地治理与利用研究所,阜新123000

出　　处：《分子植物育种》2020年第16期5351-5360,共10页Molecular Plant Breeding

基　　金：辽宁省自然科学基金指导项目(20180551263)资助。

摘　　要：低温和冻害严重影响高油酸花生的产量和品质。为了提高高油酸花生的抗寒性,本研究利用农杆菌介导的转化方式将可以激活低温响应基因的锌指蛋白基因ZAT12导入高油酸花生。同时对影响高油酸花生转化的几种因素(植物生长素ZT和6-BA的使用浓度,侵染时间,烟草提取物添加量)进行改良优化。通过PCR和实时荧光定量PCR检测证明PnZAT12已经整合到高油酸花生基因组中。对转基因植株进行低温胁迫处理,分别测定其相对电导率(relative electrolytic leakage, REL)、丙二醛(malondialdehyde, MDA)含量及超氧化物岐化酶(superoxide dismutase, SOD)活性。结果表明,转基因花生的MDA含量和REL显著低于非转基因花生,而SOD活性明显高于非转基因花生。在表型比对方面,转基因花生的叶片黄化程度和植株枯萎程度远低于非转基因花生。结果证实,转基因花生比非转基因花生有更高的抗寒性。研究结果对抗逆基因的改良研究及高油酸花生的分子育种工作都具有重要的参考价值。Cold and freezing exert dramatic influences on yield and quality of high oleic peanut.To improve the cold tolerance of peanut,a low temperature-responsive zinc finger protein gene ZAT12 was transferred into high oleic acid peanut through Agrobacterium transformation in this study.Factors affectingthe transformation efficiency(auxin concentrations of ZT and 6-BA,infection time and dosages of tobacco extracts)were optimized.The results suggested the PnZAT12 gene was integrated into the genome of high oleic acid peanut identified by PCR and Real-time quantitative PCR.The transgenic peanuts were treated by chilling stress for measuring relative electrolyte leakage(REL),malondialdehyde(MDA)content and activities ofsuperoxide dismutase(SOD).The results indicated the MDA content and REL of the transgenic peanuts were significantly lower than that of the non-transgenic peanuts,and the activity of SOD was higher than that of the non-transgenic peanuts.The degrees of leaf yellowing and wilting of transgenic peanuts were far below the non-transgenic peanuts in phenotype.This study illuminated that the transgenic peanuts had higher cold tolerance ability than non-transgenic peanuts.The study took great references to modification of the stress-tolerance genes and high oleic acid peanut molecular breeding.

关 键 词：高油酸花生 农杆菌转化 PnZAT12基因 抗寒性 

分 类 号：S565.2[农业科学—作物学]