DNA去甲基化参与提高白菜型冬油菜抗寒性的生理机制  

Physiological mechanism of DNA demethylation in improving the cold resistance of Brassica rapa L.

作　　者：王万鹏 马骊 孙柏林 郭秀娟[2] 王世发[3] 牛早霞 祁伟亮 蒲媛媛 路晓明 呼芳娣 许金苗 马学才 武军艳 李学才 方彦 孙万仓 刘丽君 WANG Wanpeng;MA Li;SUN Bolin;GUO Xiujuan;WANG Shifa;NIU Zaoxia;QI Weiliang;PU Yuanyuan;LU Xiaoming;HU Fangdi;XU Jinmiao;MA Xuecai;WU Junyan;LI Xuecai;FANG Yan;SUN Wancang;LIU Lijun(College of Agronomy,Gansu Agricultural University,Rapeseed Engineering Research Center of Gansu Province,Key Laboratory of Arid Land Crop Science in Gansu Province,Gansu Key Laboratory of Crop Improvement and Germplasm Enhancement,Lanzhou,Gansu 730070,China;Shanxi Academy of Agricultural Sciences Crop Research Institute of High and Cold Regions,Datong,Shanxi 037000,China;Jilin Academy of Agricultural Sciences Economic Plant Development and Research Center,Changchun,Jilin 130124,China)

机构地区：[1]甘肃农业大学农学院/甘肃省油菜工程技术研究中心/甘肃省干旱生境作物学重点实验室/甘肃省作物遗传改良与种质创新重点实验室,甘肃兰州730070 [2]山西省农业科学院高寒区作物研究所,山西大同037000 [3]吉林省农业科学院经济植物开发研究中心,吉林长春130124

出　　处：《干旱地区农业研究》2021年第5期29-39,49,共12页Agricultural Research in the Arid Areas

基　　金：国家自然科学基金(31960435;31860388);国家现代农业产业技术体系项目(CARS-12);甘肃省现代农业产业技术体系项目(GARS-TSZ-1);国家重点研发计划科技部“油菜杂交优势利用技术与强优势杂交种创制”(2016YFD0101300)。

摘　　要：利用来自不同育种环境的10份白菜型冬油菜材料,通过半致死温度的测定分析其抗寒性与环境的关系,并利用不同浓度DNA甲基化抑制剂5-azaC处理,分析DNA去甲基化对白菜型冬油菜DNA整体甲基化水平及低温胁迫下苗期生理特性的影响。结果表明,不同育种环境选育的材料,其半致死温度具有显著差异,其中材料2018-FJT、DT-7、DT-9与MXW-1的半致死温度分别为-16.04℃、-15.98℃、-15.63℃、-15.04℃;材料CT-2360、CT-2380、CT-2400、CT-2420、CT-2440、CT-2460的半致死温度分别为-11.32℃、-11.6℃、-11.42℃、-11.44℃、-12.97℃、-13.28℃。依据半致死温度,10个白菜型冬油菜抗寒性由强到弱依次为:2018-FJT>DT-7>DT-9>MXW-1>CT-2460>CT-2400>CT-2380>CT-2420>CT-2440>CT-2360。抗寒性的形成与育成环境的纬度和年平均气温呈极显著正相关。选择抗寒性及产地不同的4个材料CT-2360、MXW-1、2018-FJT、DT-7进行生理测定,结果表明,1000μmol·L^(-1)5-azaC处理能显著抑制幼苗的根生长,4个材料的根长较对照减少93.14%~95.06%;低温下5-azaC处理对抗寒性较弱材料CT-2360幼苗的相对电导率、丙二醛含量以及强抗寒性材料DT-7幼苗的脯氨酸、可溶性蛋白含量影响最显著(P<0.05);低温处理过程中,4个材料的SOD、POD、CAT活性均上升,以低温处理5d最显著(P<0.05),其中2018-FJT的SOD活性增幅最高,为45.85%,DT-7的POD活性增幅最大,为460%,CT-2360的CAT活性增幅最显著,为321.02%。HPLC分析发现常温下抗寒性较弱材料CT-2360的甲基化水平为77.48%,高于其他3个抗寒性强的材料;经过5-azaC处理后发生明显的去甲基化作用,证明抗寒能力受DNA去甲基化的调控。Ten Brassica rapa materials from different breeding environments were used to analyze the relationship between cold resistance and environment by measuring semi-lethal temperature.Treated with different concentrations of DNA methylation inhibitor 5-azaC,the effects of DNA demethylation on DNA methylation level and seedling physiological characteristics of Brassica rapa L.under low temperature stress were examined.The results showed that the semi-lethal temperatures of materials selected in different breeding environments were significantly different,and the semi-lethal temperatures of 2018-FJT,DT-7,DT-9,and MXW-1 were-16.04℃,-15.98℃,-15.63℃,and-15.04℃,respectively.The semi-lethal temperatures of CT-2360,CT-2380,CT-2400,CT-2420,CT-2440,and CT-2460 were-11.32℃,-11.6℃,-11.42℃,-11.44℃,-12.97℃and-13.28℃,respectively.According to the semi-lethal temperature,the cold resistance of 10 Brassica rapa L.was in the following order:2018-FJT>DT-7>DT-9>MXW-1>CT-2460>CT-2400>CT-2380>CT-2420>CT-2440>CT-2360.The formation of cold resistance was positively correlated with the latitude and annual average temperature of the breeding environment.Four materials CT-2360,MXW-1,2018-FJT,and DT-7 with cold resistance and different producing areas were selected for physiological determination.The results showed that 1000μmol·L^(-1)5-azaC treatment could significantly inhibited the root growth of seedlings and the root length of the four materials decreased by 93.14%and 95.06%compared with the control.Under low temperature,5-azaC treatment had the most significant effects on the REC and MDA content of CT-2360 seedlings,as well as the Pro and soluble protein content of strong cold resistant material DT-7.During the low temperature treatment,the SOD,POD,and CAT activities of the four materials all increased,and the low temperature treatment for 5 days was the most significant(P<0.05).Of which the SOD activity of 2018-FJT increased the most,which was 45.85%.The POD of DT-7 increased the most with 460%.The CAT of CT-2360 in

关 键 词：白菜型冬油菜 抗寒性 育种选择 5-azaC 低温胁迫 DNA甲基化 生理机制 

分 类 号：S565.4[农业科学—作物学] Q945.78[生物学—植物学]