非洲菊4个POD基因的克隆及表达分析
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福建省重大农业科技专项(2015NZ00021);


Cloning and Expression Analysis of Four Gerbera Peroxidase Genes
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    摘要:

    以‘玲珑’非洲菊为材料,利用RTPCR技术克隆获得4个POD基因,分别命名为GjPOD4、GjPOD5、GjPOD7和GjPOD42(GenBank登录号分别为MH708528、MH708531、MH708529和MH708530)。对这4个基因进行生物信息学分析,并分析在SA、干旱、低温胁迫和非洲菊根腐病病原菌胁迫下4个POD基因的表达模式以及POD活性的变化特征,为揭示非洲菊POD基因的功能以及抗性育种奠定基础。结果显示:(1)所克隆的4个POD基因的cDNA长度为966~1 005 bp,编码321~334个氨基酸;生物信息学分析发现,它们编码的蛋白均属于植物亚铁红素依赖Ⅲ型POD,含有保守结构域及Motif,含有信号肽、跨膜螺旋结构和磷酸化位点。(2)进化树分析结果表明,GjPOD4和GjPOD7与拟南芥AtPRX52、GjPOD5与AtPRX47、GjPOD42与AtPRX42亲缘关系较近。(3)SA、干旱、低温胁迫和非洲菊根腐病病原菌处理均可显著或极显著影响4个基因的表达以及POD活性,但GjPOD4与其他3个GjPOD基因的表达趋势存在一定的差异。(4)相关性分析显示,在不同器官中,GjPOD42的表达与POD活性均呈显著正相关关系(P<0.05);在隐地疫霉处理下, 4个POD基因表达均极显著上调且与POD活性呈正相关关系。研究表明,所克隆的 4个POD基因在非洲菊抗逆防御过程中,尤其是在响应隐地疫霉侵染过程中发挥着重要作用,但GjPOD4与其他3个GjPOD存在一定的功能差异。

    Abstract:

    In this study, we successfully cloned four POD genes, named as GjPOD4, GjPOD5, GjPOD7 and GjPOD42 (GenBank accession numbers are MH708528, MH708531, MH708529 and MH708530), from Gerbera jamesonni cv. ‘Linglong’ using RTPCR. Their characteristics were then analyzed using series of bioinformatics methods. Moreover, to provide basis for the understanding of their function and their future application in gerbera resistance breeding, we investigated their expression patterns together with POD activities under SA, PEG, cold stress and Phytophthora cryptogea treatments. Results showed that: (1) the coding sequence length of these PODs ranged from 966 bp to 1 005 bp, encoding 321-334 amino acids. Bioinformatic analysis showed that all the four PODs belong to the class Ⅲ plant hemedependent peroxidase superfamily containing conserved domains and motif, signal peptide, transmembrane structure and phosphorylation sites. (2) Phylogenetic analysis revealed that GjPOD4, GjPOD7 and AtPRX52, GjPOD5 and AtPRX47, GjPOD42 and AtPRX42 shared close relationship. (3) These gene expression and POD activities were all found to be significantly or very significantly influenced by the treatments of SA, PEG, cold stress and gerbera root rot pathogen. While the expression pattern of GjPOD4 differed from other GjPODs. (4) Correlation analysis showed that the expression of GjPOD42 was significantly positive correlated with POD activity in different organs (P<0.05). Notably, all the gene expression were significantly induced and positively correlated with the POD activity under the Phytophthora cryptogea treatment. Results obtained in this study indicated that GjPODs played an important role in stress responses of gerbera, especially during the gerberaP. cryptogea interaction. And, the function of GjPOD4 might differ from other GjPODs.

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郝向阳,孙雪丽,刘 范,等.非洲菊4个POD基因的克隆及表达分析[J].西北植物学报,2018,38(10):1777-1786

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  • 在线发布日期: 2018-11-23
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