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植物研究 ›› 2017, Vol. 37 ›› Issue (3): 402-415.doi: 10.7525/j.issn.1673-5102.2017.03.011

• 研究报告 • 上一篇    下一篇

脱落酸和己糖激酶抑制剂对高表达C4-PEPC转基因稻苗耐旱性的影响

吴敏怡1,2, 李霞1, 何亚飞1,4, 张琛1,3, 严婷1,3   

  1. 1. 江苏省农业科学院粮食作物研究所, 江苏省优质水稻工程技术研究中心, 南京 210014;
    2. 南京师范大学生命科学学院, 江苏省微生物与功能基因组学重点实验室, 江苏省微生物资源产业化工程技术研究中心, 南京 210023;
    3. 南京农业大学生命科学学院, 南京 210095;
    4. 南京林业大学生物与环境学院, 南京 210037
  • 收稿日期:2016-12-28 出版日期:2017-05-15 发布日期:2017-06-03
  • 通讯作者: 李霞,E-mail:jspplx@jaas.ac.cn E-mail:jspplx@jaas.ac.cn
  • 作者简介:吴敏怡(1994-),女,本科,主要从事水稻光合生理方面的研究。
  • 基金资助:
    国家自然科学基金(31571585;31371554);江苏省自主创新基金(CX[(14)5004]);江苏省农业科学院基本科研业务专项项目(ZX(16)2002)

Drought Response in Overexpression of Maize Phosphoenolpyruvate Carboxylase Rice Seedlings Treated by Inhibitors of ABA and HXK Pathway

WU Min-Yi1,2, LI Xia1, HE Ya-Fei1,4, ZHANG Chen1,3, YAN Ting1,3   

  1. 1. Institute of Food Crops, Jiangsu High Quality Rice Research and Development Center, Nanjing Branch of China National Center Rice Improvement, Jiangsu Academy of Agricultural Sciences, Nanjing 210014;
    2. Jiangsu Key Laboratory for Microbes and Functional Genomics, Jiangsu Engineering and Technology Research Center for Industrialization of Microbial Resources, College of Life Sciences, Nanjing Normal University, Nanjing 210023;
    3. College of Life Science, Nanjing Agricultural University, Nanjing 210095;
    4. College of Biology and Environment, Nanjing Forestry University, Nanjing 210037
  • Received:2016-12-28 Online:2017-05-15 Published:2017-06-03
  • Supported by:
    National Natural Science Foundation of China(31571585,31371554);The Agricultural Science and Technology Innovation Fund of Jiangsu in China[CX(14)5004];The Basic Scientific Support Project of Jiangsu Academy of Agricultural Sciences(ZX[16]2002)

摘要: 为了研究高表达转玉米C4-磷酸烯醇式丙酮酸羧化酶(phosphoenolpyruvate carboxylase,PEPC)基因水稻(PC)的耐旱性机制,本研究以PC和未转基因野生型原种kitaake为材料,分别在光照和黑暗预处理24 h,其中光照处理中光强为600 μmol·m-2·s-1,预处理后稻苗再在15%聚乙二醇-6000模拟干旱胁迫下,同时使用药理学的方法,通过加入脱落酸和己糖激酶的专一性抑制剂100 μmol·L-1去甲二氢愈创木酸和10 mmol·L-1葡萄糖胺,观察两种水稻4~5叶期稻苗耐旱表现。结果发现,与WT水稻相比,在PEG-6000处理后,经过光预处理的PC水稻叶片相对含水量下降较少,始终比WT的高,而且丙二醛含量则较少,脯氨酸诱导增加,表现耐旱;而经过暗预处理后PC植株显著降低这个优势,表明光预处理有利于PC耐旱性的表现;黑暗预处理均显著下调了2供试材料植株叶片中可溶性糖的含量,而对可溶性蛋白的含量影响不显著;而光预处理后PC水稻叶片内可溶性糖含量比WT增加,而在黑暗预处理则PC的显著低于WT的,其中葡萄糖胺对光预处理下PC的可溶性糖含量的下调作用最显著;暗预处理逆转或消除了NO,H2O2和钙离子含量变化趋势,这些变化与暗预处理减少了两材料叶片蔗糖和葡萄糖含量变化同步;光暗预处理对两材料的PEPC酶活性的差异影响不大,表明外源玉米C4-PEPC在水稻中是组成型表达。可见PC叶片可部分通过糖组分,参与内源糖介导ABA和HXK信号途径,缓解干旱处理对叶片的伤害,稳定光合能力。

关键词: 水稻, 磷酸烯醇式丙酮酸羧化酶, 干旱, 糖, 脱落酸, 己糖激酶

Abstract: The experiment was conducted to study the mechanism of drought tolerance in C4- phosphoenolpyruvate carboxylase(PEPC) gene(C4-PEPC) transgenic rice(PC), PC and its untransformed wild-type rice lines, Kitaake(WT) as materials. Pretreated by light(light intensity:600 μmol·m-2·s-1) and dark on two rice lines with 4 leaves for 24 h, the pretreated rice lines were then conducted by 15% polyethylene glycol-6000(PEG-6000) simulated drought stress conditions for 4 h. Two inhibitors(one for Abscisic acid:100 μmol·L-1 nordihydroguaiaretic acid(NDGA) and the other for hexokinase(HXK):10 mmol·L-1 glucosamine(Gluc) were also added by root during the PEG-6000 treatment. Compared with WT rice lines, there were higher relative water content, lower malondialdehyde content and pro content in PC rice lines after drought treatment for 4 h involved in ABA and HXK pathways, conferring drought tolerance. But pre-dark treatment reduced the advantage of PC for drought tolerance. Soluble sugar content in two rice lines were increased in pre-light treatment and decreased in pre-dark treatment, which were regulated by ABA and HXK pathway. Among them, the Gluc inhibition of soluble sugar content in PC pre-light treated was the largest after the PEG treatment. There was no difference in soluble protein content of two rice lines either pretreated by light and dark during the PEG treatment. Similarly, pre-dark treatment reversed or eliminated the changes in two rice lines, which trends changed synchronously as those of sucrose, glucose and fructose contents. PEPC activities in PC lines did not change by pre-light or pre-dark treatments, showed PC belongs to constitutive expression. ABA and HXK pathways regulated negatively PEPC activity, which was synchronous with its low level of glucose in PC lines. Therefore, PC rice lines alleviated the damages of drought stress by sugar involved in ABA and HXK pathways, exhibiting stable photosynthetic ability and drought tolerance.

Key words: Oryza.sativa L., phosphoenolpyruvate carboxylase, drought, sugar, abscisic acid, hexokinase

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