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

作　　者：代志国 李红梅 孟静璇 张有来 张丙秀 DAI Zhiguo;LI Hongmei;MENG Jingxuan;ZHANG Youlai;ZHANG Bingxiu(Key Laboratory of Biology and Genetic Improvement of Horticultural Crops(Northeast Region),Ministry of Agriculture and Rural Affairs,Harbin 150030,China;National-Local Joint Engineering Research Center for Development and Utilization of Small Fruits in Cold Regions,Harbin 150030,China;School of Horticulture and Landscape Architecture,Northeast Agricultural University,Harbin 150030,China)

机构地区：[1]农业农村部园艺作物生物学与遗传改良重点实验室(东北地区),哈尔滨150030 [2]寒地小浆果开发利用国家地方联合工程研究中心,哈尔滨150030 [3]东北农业大学园艺园林学院,哈尔滨150030

出　　处：《东北农业大学学报》2023年第11期34-44,共11页Journal of Northeast Agricultural University

基　　金：国家重点研发计划项目(2022YFD1600501)。

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。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.

关 键 词：笃斯越橘 VuLhca4 低温胁迫 干旱胁迫 

分 类 号：S663[农业科学—果树学] Q785[农业科学—园艺学]