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园艺学报 ›› 2017, Vol. 44 ›› Issue (2): 268-278.doi: 10.16420/j.issn.0513-353x.2016-0507

• 研究论文 • 上一篇    下一篇

枇杷脱水素基因EjDHN5抗低温胁迫功能研究

徐红霞1,李晓颖1,徐昌杰2,陈俊伟1,*   

  1. 1浙江省农业科学院园艺研究所,杭州 310021;2浙江大学农业与生物技术学院,农业部园艺作物生长发育和品质生物学实验室,杭州 310058
  • 出版日期:2017-02-25 发布日期:2017-02-25
  • 基金资助:

    国家自然科学基金项目(31101530);浙江省农业新品种选育重大科技专项(2016C02052-3);浙江省农业科学院地方科技合作项目(LS20140003);浙江省农科院青年人才培养项目

A Study on Cold Resistance Function of Dehydrin Gene EjDHN5 from Loquat

XU Hongxia1,LI Xiaoying1,XU Changjie2,and CHEN Junwei1,*   

  1. 1Institute of HorticultureZhejiang Academy of Agricultural SciencesHangzhou 310021China2 College of Agriculture and BiotechnologyThe State Agriculture Ministry Laboratory of Horticultural Plant GrowthDevelopment and Quality ImprovementZhejiang UniversityHangzhou 310058China
  • Online:2017-02-25 Published:2017-02-25

摘要:

为研究枇杷(Eriobotrya japonica Lindl.)脱水素基因(EjDHN5)在枇杷抗低温胁迫中的功能,将其在烟草中进行过量表达,其中表达水平比较高的两个株系L24和L26被用来进行功能研究。野生型和转基因烟草苗分别用0、2、4和8 μmol · L-1的甲基紫精(Methyl viologen,自由基发生剂)处理并进行光照培养,结果显示,4和8 μmol · L-1甲基紫精处理使野生型烟草苗存活率分别下降到73.6%和56.9%,光系统Ⅱ(PSⅡ)活性显著降低,而转基因苗的存活率仍维持在100%,PSⅡ活性明显高于野生型。对六叶期烟草的叶盘进行甲基紫精处理,野生型烟草叶盘的叶绿素含量显著低于转基因株系,而活性氧含量和MDA含量明显高于转基因株系。对野生型和转基因烟草苗进行低温处理,转基因株系生长状况明显优于野生型,活性氧含量低于野生型,膜质过氧化程度轻于野生型。这些结果表明,过量表达EjDHN5能提高烟草的抗氧化胁迫和抗低温胁迫能力,推测EjDHN5在提高枇杷抗低温能力方面起着重要作用可能与其能提高抗氧化胁迫能力有关。

关键词: 枇杷, EjDHN5, 转基因, 烟草, 氧化胁迫, 甲基紫精, 低温胁迫

Abstract:

In order to examine the cold resistance function of dehydrin gene EjDHN5 from loquat (Eriobotrya japonica),it was overexpressed in tobacco plants. Two lines with relatively higher transcripts levels of the target gene,L24 and L26 were chosen for function characterization. Both of wide type(WT)and transgenic lines(L24 and L26)were treated with 0,2,4 and 8 μmol · L-1 methyl viologen(MV). After being exposed to 0,2,4 and 8 μmol · L-1 MV under light,the survival rate of WT plants decreased to 73.6% and 56.9%,respectively,and the PSⅡ activity reduced dramatically,while the transgenic lines still remained a survival rate of 100% as well as a much higher PSⅡ activity. MV treatment in leaf discs from 6-leaf-stage tobaccos confirmed that transgenic lines maintained higher  chlorophyll content,accumulated less reactive oxygen species(ROS)and MDA. Moreover,compared to the WT plants,the EjDHN5- overexpressing plants exhibited a better growth status,had less ROS and suffered lighter membrane damage in leaves after cold stress. In summary,these results indicated that overexpressing EjDHN5 improved oxidative and cold tolerance in tobacco. It also provides evidence that dehydrins contribute to cold tolerance in loquat,which may be partly ascribed to its oxidative stress defense in plant.

Key words: loquat, EjDHN5, transgenic, tobacco, oxidative stress, methyl viologen, cold stress

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