王顺才,梁东,师守国,马锋旺,束怀瑞.苹果中新抗旱相关基因GR-RBPs的克隆与表达分析[J].干旱地区农业研究,2011,29(5):75~81
苹果中新抗旱相关基因GR-RBPs的克隆与表达分析
Cloning and expression analysis of two novel drought tolerance genes coding glycine-rich RNA-binding proteins in Malus plants
  
DOI：10.7606/j.issn.1000-7601.2011.05.14
中文关键词:  苹果  富含甘氨酸RNA结合蛋白  干旱  基因表达
英文关键词:Malus  glycine-rich RNA-binding protein  drought stress  gene expression
基金项目:国家科技部重大转基因“863”项目（2008AA10Z157）
作者单位
王顺才 天水师范学院生命科学与化学学院， 甘肃 天水 748100
西北农林科技大学园艺学院， 陕西 杨凌 712100 
梁东 西北农林科技大学园艺学院， 陕西 杨凌 712100 
师守国 西北农林科技大学园艺学院， 陕西 杨凌 712100 
马锋旺 西北农林科技大学园艺学院， 陕西 杨凌 712100 
束怀瑞 西北农林科技大学园艺学院， 陕西 杨凌 712100 
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中文摘要:
      在构建楸子叶片SSH cDNA文库及EST分析的基础上，通过电子克隆（in silico cloning）和RT-PCR方法分别从楸子和平邑甜茶叶片中各克隆得到了1个富含甘氨酸RNA结合蛋白（GR-RBP）基因，分别命名为MpGR-RBP1（HM042682）和MhGR-RBP1（HQ380209）。它们的cDNA全长分别为781和558 bp，ORF分别编码171和164个氨基酸的多肽。MpGR-RBP1和MhGR-RBP1基因编码蛋白均含有一个RNA识别结构域（RRM）和一个富含甘氨酸区（glycine-rich region）。生物信息学分析表明，MpGR-RBP1和MhGR-RBP1蛋白属于植物GR-RBPs蛋白家族。定量PCR分析表明，MpGR-RBP1和MhGR-RBP1基因在根、茎和叶中均有表达；在干旱胁迫下，它们的表达量增加，而楸子MpGRRBP1表达水平明显高于平邑甜茶MhGR-RBP1。结果表明，MpGR-RBP1和MhGR-RBP1可能参与植物对干旱胁迫的响应反应，并推测抗旱蛋白结构的差异可能对植物的抗旱性有影响。
英文摘要:
      Two Malus (apple) drought-related genes, MpGR-RBP1 (HM042682) and MhGR-RBP1 (HQ380209), were successfully identified from the drought stress-treated Malus prunifolia and M. hupehensis, respectively, using library screening, in silico cloning and RT-PCR. The proteins of two homologous genes, MpGR -RBP1 and MhGR-RBP1, were consisted of 171 and 164 amino acids, respectively. Their deduced amino acids contain a N-terminal RNA recognition motif (RRM ) and a C-terminal glycine-rich domain, a structure which was found in stress-induced GR-RBP protein family in other plants. Bioinformatics analysis confirmed that both MpGR-RBP1 and MhGR-RBP1 proteins belong to the plant GR-RBP family, members of which play important roles in post transcriptional regulation of gene expression under various stress conditions. The expression profiles of the two apple GR-RBP transcripts were detected by quantitative real-time RT-PCR. MpGR-RBP1 and MhGR-RBP1 were expressed in various plant tissues including roots, shoots, and leaves. Both MpGR-RBP1 and MhGR-RBP1 were up-regulated under drought stress, with expression levels in the former being higher than in the latter. These results indicated that MpGR-RBP1 and MhGR-RBP1 might be involved in apple plants response to drought stress, and it suggested that differential configuration of drought-tolerant proteins may be contributed to capability of drought resistance in apple plants.
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