高博闻, 戎玉清, 李铁铮, 魏胜利, 王晓晖, 屠鹏飞. 白木香3个G6PDH基因的鉴定与表达分析J. 药学学报, 2020,55(9): 2216-2225. doi: 10.16438/j.0513-4870.2020-0642
引用本文: 高博闻, 戎玉清, 李铁铮, 魏胜利, 王晓晖, 屠鹏飞. 白木香3个G6PDH基因的鉴定与表达分析J. 药学学报, 2020,55(9): 2216-2225. doi: 10.16438/j.0513-4870.2020-0642
GAO Bo-wen, RONG Yu-qing, LI Tie-zheng, WEI Sheng-li, WANG Xiao-hui, TU Peng-fei. Identification and expression analysis of three G6PDH genes from Aquilaria sinensisJ. Acta Pharmaceutica Sinica, 2020,55(9): 2216-2225. doi: 10.16438/j.0513-4870.2020-0642
Citation: GAO Bo-wen, RONG Yu-qing, LI Tie-zheng, WEI Sheng-li, WANG Xiao-hui, TU Peng-fei. Identification and expression analysis of three G6PDH genes from Aquilaria sinensisJ. Acta Pharmaceutica Sinica, 2020,55(9): 2216-2225. doi: 10.16438/j.0513-4870.2020-0642

白木香3个G6PDH基因的鉴定与表达分析

Identification and expression analysis of three G6PDH genes from Aquilaria sinensis

  • 摘要: 葡萄糖-6-磷酸脱氢酶(glucose-6-phosphate dehydrogenase,G6PDH)是戊糖磷酸途径的关键酶,在植物响应胁迫方面发挥重要作用。本研究对白木香转录组数据进行分析,设计特异性引物,首次从白木香中克隆得到3条AsG6PDHs基因的cDNA序列,分别命名为AsG6PDH1AsG6PDH2AsG6PDH3,其开放阅读框(ORF)分别为1 809、1 767、1 548 bp,编码蛋白分别含有602、588、516个氨基酸,蛋白分子质量分别为68.02、67.02、59.35 kDa。3个G6PDH蛋白的氨基酸序列与其他植物G6PDH的氨基酸序列相似性比较高,都含有G6PDH蛋白的3个特征性序列。系统进化树显示AsG6PDH1和AsG6PDH2属于质体G6PDH,AsG6PDH3属于胞质G6PDH。组织特异性分析结果表明,AsG6PDH1AsG6PDH2主要在根中表达,AsG6PDH3在各组织中表达量都比较高,茎中表达量最高。实时荧光定量PCR结果显示盐、干旱、低温和重金属胁迫都能够诱导白木香中3个AsG6PDHs基因表达,其中干旱胁迫对AsG6PDH1AsG6PDH2的转录水平影响最显著,重金属胁迫对AsG6PDH3的表达水平影响最显著。同时,盐、干旱、低温和重金属胁迫都能够提高愈伤组织中G6PDH酶活性,其中干旱胁迫对白木香愈伤组织中G6PDH酶活性影响最显著。本研究为进一步阐明G6PDH基因在白木香防御反应中的作用和沉香结香机制奠定基础。

     

    Abstract: Glucose-6-phosphate dehydrogenase, a key enzyme in the pentose phosphate pathway, plays an important role in plant resistance. In this study, three full length cDNAs of G6PDH genes, namely AsG6PDH1, AsG6PDH2 and AsG6PDH3 were cloned from Aquilaria sinensis for the first time. The open reading frames (ORF) of AsG6PDH1, AsG6PDH2 and AsG6PDH3 were 1 809, 1 767 and 1 548 bp, respectively, encoding proteins of 602, 588 and 516 amino acid residues, respectively, with predicted molecular masses of 68.02, 67.02, 59.35 kDa, respectively. The three AsG6PDHs proteins shared high sequence identity with the G6PDH proteins of various plants, and possessed three conserved sequences found in G6PDH proteins. The phylogenic analysis showed that AsG6PDH1 and AsG6PDH2 were grouped in the plastidic cluster, while AsG6PDH3 was classified into the cytosolic cluster. Expression analysis indicated that AsG6PDH1 and AsG6PDH2 were primarily observed in root, while AsG6PDH3 was primarily observed in stem. The expression of AsG6PDH1, AsG6PDH2 and AsG6PDH3 was induced by salt, drought, low temperature and CdCl2 treatments, while the content of AsG6PDH1 and AsG6PDH2 was most significantly increased by drought stress, and the transcript level of AsG6PDH3 was most significantly induced by metal stress. Furthermore, G6PDH activity was stimulated under salt, drought, low temperature and CdCl2 treatments, and G6PDH activity was remarkably increased under drought stress. These results provide valuable insights into the role of AsG6PDHs in plant defense and the mechanism of agarwood formation.

     

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