Ptcorp gene induced by cold stress was identified by proteomic analysis in leaves of Poncirus trifoliata (L.) Raf.

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• 作者：Guiyou Long (12) longgy63@163.com
  Jinyu Song (2) 301105649@qq.com
  Ziniu Deng (2) deng7009@163.com
  Jie Liu (1) oicqliuliu@163.com
  Liqun Rao (1) raoliqun@163.com
• 关键词：Poncirus trifoliata – Mass spectrometry – Cold stress – Resistance protein – 2 ; D IEF/SDS ; PAGE – Ptcorp
• 刊名：Molecular Biology Reports
• 出版年：2012
• 出版时间：May 2012
• 年：2012
• 卷：39
• 期：5
• 页码：5859-5866
• 全文大小：367.6 KB
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• 作者单位：1. College of Bioscience and Biotechnology, Hunan Agricultural University, Changsha, 410128 Hunan, China2. National Center for Citrus Improvement, Changsha, 410128 Hunan, China
• 刊物类别：Biomedical and Life Sciences
• 刊物主题：Life Sciences
  Animal Anatomy, Morphology and Histology
  Animal Biochemistry
  
• 出版者：Springer Netherlands
• ISSN：1573-4978

文摘

A proteomic approach was employed to investigate the cold stress-responsive proteins in trifoliate orange (Poncirus trifoliata (L.) Raf.), which is a well-known cold tolerant citrus relative and widely used as rootstock in China. Two-year-old potted seedlings were exposed to freezing temperature (?6°C) for 50 min (nonlethal) and 80 min (lethal), and the total proteins were isolated from leaves of the treated plants. Nine differentially accumulated proteins over 2-fold changes in abundance were identified by two-dimensional gel electrophoresis and mass spectrometry. Among these proteins, a resistance protein induced by the nonlethal cold treatment (protein spot #2 from P. trifoliata) was selected as target sequence for degenerated primer design. By using the designed primers, a PCR product of about 700 bp size was amplified from P. trifoliata genomic DNA, which was further cloned and sequenced. A nucleotide sequence of 676 bp was obtained and named Ptcorp. Blast retrieval showed that Ptcorp shared 88% homology with an EST of cold acclimated Bluecrop (Vaccinium corymbosum) library (Accession number: CF811080), indicating that Ptcorp had association with cold acclimation. Semiquantitative RT-PCR analysis demonstrated that Ptcorp gene was up-regulated by cold stress which was consistent with the former result of protein expression profile. As the resistance protein (NBS-LRR disease resistance protein family) gene was up-regulated by cold stress in trifoliate orange and satsuma mandarin, it may imply that NBS-LRR genes might be associated with cold resistance in citrus.