高温胁迫对葡萄高温相关基因和蛋白表达的影响

doi:10.3864/j.issn.0578-1752.2017.09.013

摘要/Abstract

摘要： 【目的】适宜的温度是葡萄生长发育的必要条件，南方地区葡萄在成熟期常常经历连续高温环境，叶片容易失水干枯，果实会发生明显的日灼，严重地影响了其经济效益。选择不同树龄葡萄树进行高温胁迫的相关研究，以解释高温对葡萄的伤害，为生产中高温逆境的抵御措施提供理论依据。【方法】选择南方地区常见的鲜食葡萄品种‘巨峰’‘巨玫瑰’‘醉金香’‘夏黑’‘申玉’‘申丰’‘申华’和‘沪培1号’为试验材料，其中葡萄幼苗选择长势基本一致的1年生扦插苗移至人工气候培养箱，在可控条件下培养7 d后于每天10: 00—16: 00(模拟夏季高温发生时段)进行不同温度处理(25℃、35℃、45℃)，观察处理后0、3、6和150 h高温应答反应;葡萄成年树选择6年生树为试验材料，在2015年夏季观测到2个处理点分别为：“37℃”处理(2015年7月23日13: 00 h最高气温为37℃)和“42℃”处理(2015年7月31日13: 00 h最高气温为42℃)，观察以上处理的8个不同葡萄品种在高温胁迫下叶片表型变化，对抗逆基因(HSFA2、GLOS1、HSP70、HSP17.9)和HSP21蛋白的表达量进行分析。【结果】 ‘申玉’‘夏黑’和‘申华’在高温发生初期，顶部新梢和老叶有失水，并在150 h时有所恢复;‘巨峰’和‘巨玫瑰’叶片在高温发生3 h时轻微失水，6 h时大面积失水，‘申丰’‘沪培1号’‘醉金香’叶片在高温发生3 h时大面积失水，6 h时整株干枯。同时对葡萄幼苗和成年树的抗逆基因的表达进行检测发现：不同温度处理(35℃/45℃/“42℃”)下不同葡萄品种的抗逆基因均被诱导表达，但是表达丰度有所不同，表现为温度水平越高，基因的上调倍数越大。另外，‘申玉’‘申华’‘醉金香’‘巨峰’和‘巨玫瑰’的GLOS1、HSFA2和‘申玉’‘申华’‘夏黑’和‘巨峰’HSP70表达在长期高温处理后恢复至原初水平;并观察到HSP17.9在高温发生后上调表达，但并未因为时间的延长而减弱表达倍数。高温处理(45℃/“42℃”)会诱导葡萄幼苗和成年树的HSP21蛋白水平，但是在一些品种(‘醉金香’和‘申丰’)中HSP21蛋白表达量有所减少。【结论】不同葡萄品种对高温的耐性有差异，其中‘申玉’‘申华’和‘夏黑’抗性最强，而‘申丰’‘沪培1号’和‘醉金香’属于抗性较弱的葡萄品种。树龄和高温发生方式是影响葡萄幼苗和成年树的抗逆基因和蛋白表达差异的主要因素。

关键词: 葡萄, 高温胁迫, 抗逆基因, HSP21

Abstract: 【Objective】The suitable air temperature is necessary for growth of grapes. Heat stress often happens in south area of China during grape mature period. The industrial economic benefits of grape are affected by the water-lost leaves and sunburn fruits. So study on the responsive mechanism of grapevines under high temperature stress is particularly important. The present research is mainly focused on the high temperature stress response of young grapes under control condition. As studies on response to high temperature stress in adult grapes are limited, so detailed explanation of high temperature damage in grapevines at different ages could provide a theoretical foundation for prevention of high temperature adversity. 【Method】 This study used 8 table grape varieties in south area of China as experimental materials, including Zuijinxiang, Summer Black, Kyoho, Jumeigui, Shenyu, Shenhua, Shenfeng, and Hupei 1#. Young grapevines (1-year-old) at the same growth stage were acclimated for 7 days in a controlled environment room, with a photoperiod of 14 h light under cool-white fluorescent light at 100 μmol·m-2·s-1. The grape plants were divided into 3 groups and were acclimated for 7 days at normal temperature (25℃). On the following day, 1 group of grapevines was kept at 25℃ in this controlled environment room. The other 2 groups were treated at 35℃or 45℃, respectively, from 10：00 h to 16：00 h (which was stimulated the high temperature levels in the day). The heat response was tested at 0, 3 h, 6 h and 150 h. In addition, the 6-year-old adult grapevines were selected to study heat stress response under field conditions in the present experiment, July 23, 2015 (denoted as “37℃”; highest temperature <40℃) stood for pre-temperature stress and July 31, 2015 (denoted as “42℃”, highest temperature >40℃) stood for post-temperature stress. 【Result】 The phenotype of young grapevines under high temperature stress was observed. The young sprout and aging leaves of Shenyu, Summer Black and Shenhua showed lightly water-loss at the beginning of high temperature stress, and recovered at 150 h. The leaves of Kyoho and Jumeigui showed slightly water-loss at 3 h and seriously water-loss at 6 h under high temperature stress. The leaves of Shenfeng, Hupei 1# and Zuijinxiang showed a seriously water-loss at 3 h, and the leaves of whole plant dry rot at 6 h under high temperature stress. And different temperature treatments (35℃/45℃/42℃) induced the expression of resistance genes in young and adult grapevines. However, the expression of genes in levels were different. The higher temperature level induced the greater gene expression. In addition, the expression of GLOS1 and HSFA2 in Shenyu, Shenhua, Zuijinxiang, Kyoho, and Jumeigui and the expression of HSP70 in Shenyu, Shenhua, Summer Black, and Kyoho recovered to the original levels at 6 h/150 h after high temperature treatments. The expression of HSP17.9 was increased by high temperature stress and the expression was not decreased with the prolongation of time. High temperature treatments (45℃/42℃) induced the expression levels of HSP21 protein in young and adult grapevines, however, the expression levels of HSP21 protein decreased in some varieties (Shenfeng and Zuijinxiang). 【Conclusion】 It was found that there are differences between different grape cultivars in the tolerance to high temperature stress. Firstly, Shenyu, Shenhua, and Summer Black are more tolerant to high temperature than others. Secondly, the system of heat-tolerance in Kyoho and Jumeigui collapsed. And then, Shenfeng, Hupei 1# and Zuijinxiang belong to the heat-sensitive grape cultivars. In addition, the differences of expression of genes and proteins between the grape seedling and adult trees were due to the tree age and the happening way of high temperature stress.

Key words: grapevine, high temperature stress, resistance genes, HSP21

查倩，奚晓军，蒋爱丽，田益华. 高温胁迫对葡萄高温相关基因和蛋白表达的影响[J]. 中国农业科学, 2017, 50(9): 1674-1683.

ZHA Qian, XI XiaoJun, JIANG AiLi, TIAN YiHua. Influence of Heat Stress on the Expression of Related Genes and Proteins in Grapevines[J]. Scientia Agricultura Sinica, 2017, 50(9): 1674-1683.

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