| 王万鹏,马骊,孙柏林,郭秀娟,王世发,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.DNA去甲基化参与提高白菜型冬油菜抗寒性的生理机制[J].干旱地区农业研究,2021,39(5):29~39 |
| 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. |
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| DOI：10.7606/j.issn.1000-7601.2021.05.04 |
| 中文关键词: 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 |
| 英文关键词:Brassica rapa L. cold resistance breeding selection 5-azaC low temperature stress DNA methylation physiological mechanism |
| 基金项目: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) |
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| 中文摘要: |
| 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，1 000 μmol·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。 |
| 英文摘要: |
| 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 1 000 μmol·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 increased most significantly with 321.02%. HPLC analysis found that the methylation level of CT-2360 in materials with weak cold resistance at room temperature was 77.48%, which was higher than that of the other three materials with strong cold resistance. Significant demethylation occurred after 5-azaC treatment. This study supports that the cold resistance is regulated by DNA demethylation. |
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