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笃斯越橘VuLhca4基因克隆及其在非生物胁迫中的作用

Study on cloning of VuLhca4 gene from Vaccinium uliginosum L. and functional analysis under abiotic stress

  • 摘要: LHC(捕光叶绿素a/b结合蛋白)是光合系统中重要的复合体蛋白,近年发现该蛋白在植物光合作用和响应非生物胁迫中发挥重要作用。笃斯越橘(Vaccinium uliginosum L.)主要分布于我国大兴安岭、小兴安岭及长白山等地,是优良抗逆育种资源。从笃斯越橘中克隆冷响应基因VuLhca4,研究其表达模式和功能,结果表明,从笃斯越橘叶片中克隆得到的VuLhca4基因全长744 bp,编码247个氨基酸;亚细胞定位结果显示VuLhca4定位于叶绿体;VuLhca4在笃斯越橘中的表达具有组织特异性,VuLhca4在叶中的表达量显著高于根和茎,且该基因对低温和干旱胁迫敏感性较强,低温和干旱胁迫均可诱导VuLhca4基因表达;采用农杆菌介导法侵染拟南芥得到VuLhca4转基因株系,作低温和干旱两种处理后,转基因株系VuLhca4表达量及存活率高于野生型。为笃斯越橘非生物胁迫分子研究及光合作用机理方面提供理论依据。

     

    Abstract: LHC(Light harvesting chlorophyll a/b binding protein) is an important complex protein in the photosynthetic system. In recent years, it has been found that this protein plays an important role in plant photosynthesis and response to abiotic stress. Duchess blueberry is mainly distributed in the Greater Khingan Mountains, Lesser Xing’an Mountains, and Changbai Mountains of China, and is an excellent germplasm resource. However, there have been few reports on the Lhca gene in Duchess blueberry. This study cloned the VuLhca4 gene from the Duchess blueberry and investigated its expression pattern and function. The results showed that the VuLhca4 gene cloned from the leaves of Duchess blueberry had a total length of 744 bp and encoded 247 amino acids; the subcellular localization results showed that VuLhca4 was localized in chloroplasts; the expression of VuLhca4 in Duchess blueberry was tissue-specific. The expression level of VuLhca4 in leaves was significantly higher than that in roots and stems, and this gene was highly sensitive to low temperature and drought stress. Both low temperature and drought stress could induce the expression of VuLhca4 gene;Transgenic lines of VuLhca4 were obtained by infecting Arabidopsis thaliana using Agrobacterium mediated method. After two treatments of low temperature and drought, the expression level and survival rate of the transgenic strain VuLhca4 were higher than those of the wild type. This provided a theoretical basis for the molecular study of abiotic stress and the mechanism of photosynthesis in Duchess blueberry.

     

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