张岗,赵明明,李标,宋超,张大为,郭顺星. 一个受菌根真菌诱导的铁皮石斛钙依赖蛋白激酶基因的克隆及表达分析J. 药学学报, 2012,47(11): 1548-1554.
引用本文: 张岗,赵明明,李标,宋超,张大为,郭顺星. 一个受菌根真菌诱导的铁皮石斛钙依赖蛋白激酶基因的克隆及表达分析J. 药学学报, 2012,47(11): 1548-1554.
ZHANG Gang, ZHAO Ming-ming, LI Biao, SONG Chao, ZHANG Da-wei, GUO Shun-xing. Cloning and expression analysis of a calcium-dependent protein kinase gene in Dendrobium officinale in response to mycorrhizal fungal infectionJ. 药学学报, 2012,47(11): 1548-1554.
Citation: ZHANG Gang, ZHAO Ming-ming, LI Biao, SONG Chao, ZHANG Da-wei, GUO Shun-xing. Cloning and expression analysis of a calcium-dependent protein kinase gene in Dendrobium officinale in response to mycorrhizal fungal infectionJ. 药学学报, 2012,47(11): 1548-1554.

一个受菌根真菌诱导的铁皮石斛钙依赖蛋白激酶基因的克隆及表达分析

Cloning and expression analysis of a calcium-dependent protein kinase gene in Dendrobium officinale in response to mycorrhizal fungal infection

  • 摘要:

    钙依赖蛋白激酶 (calcium-dependent protein kinases, CDPKs) 在从枝菌根、根瘤菌-宿主植物共生体系中起重要调控作用。然而, CDPKs在兰科菌根共生体系中的作用机制尚不清楚。本研究利用RT-PCRRACE方法, 首次从小菇真菌 (Mycena sp.) 侵染的铁皮石斛根中克隆到1钙依赖蛋白激酶基因, 命名为DoCPK1 (GenBank注册号JX193703)DoCPK1全长2 137 bp, 编码一条由534个氨基酸组成的肽链, 分子量59.61 kD, 等电点6.03DoCPK1蛋白包含CDPKs蛋白家族保守的丝氨酸/苏氨酸蛋白激酶催化结构域4Ca2+ 结合的EF-hand基元。DoCPK1与人参PgCPK1 (ACY78680) 同源性最高 (85%), 小麦、水稻、拟南芥CDPKs基因 (ABD98803, ADM14342, Q9ZSA2) 等次之, 与单子叶植物CDPKs基因亲缘关系最近。DoCPK1为组成型表达, 其转录本在各器官中相对表达量依次为根 > > 种子 > 叶。在小菇真菌侵染30 d的石斛根中, DoCPK1显著上调, 为对照根中的5.16, 表明DoCPK1参与小菇真菌-铁皮石斛菌根早期互作, 可能在该共生体系中起作用。本研究为进一步阐明DoCPK1在小菇真菌-铁皮石斛菌根共生中的潜在功能奠定基础。

     

    Abstract:

    Calcium-dependent protein kinases (CDPKs) play an important regulatory role in the plant- arbuscular mycorrhiza/rhizobium nodule symbiosis.  However, the biological action of CDPKs in orchid mycorrhiza (OM) symbiosis remains unclear.  In the present study, a CDPK encoding gene, designated as DoCPK1 (GenBank accession No. JX193703), was identified from D. officinale roots infected by an OM fungus-Mycena sp. using the reverse transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE) methods, for the first time.  The full length cDNA of DoCPK1 was 2 137 bp in length and encoded a 534 aa protein with a molecular weight of 59.61 kD and an isoelectric point (pI) of 6.03.  The deduced DoCPK1 protein contained the conserved serine/threonine-protein kinase catalytic domain and four Ca2+ binding EF hand motifs.  Multiple sequence alignment demonstrated that DoCPK1 was highly homologous (85%) to the Panax ginseng PgCPK1 (ACY78680), followed by CDPKs genes from wheat, rice, and Arabidopsis (ABD98803, ADM14342, Q9ZSA2, respectively).  Phylogenetic analysis showed that DoCPK1 was closely related to CDPKs genes from monocots, such as wheat, maize and rice.  Real time quantitative PCR (qPCR) analysis revealed that DoCPK1 was constitutively expressed in the included tissues and the transcript levels were in the order of roots > stems > seeds > leaves.  Furthermore, DoCPK1 transcripts were significantly accumulated in roots 30 d after fungal infection, with 5.16 fold compared to that of the mock roots, indicating involvement of DoCPK1 during the early interaction between D. officinale and Mycena sp., and a possible role in the symbiosis process.  This study firstly provided important clues of a CDPK gene associated with OM symbiosis, and will be useful for further functional determination of the gene involving in D. officinale and Mycena sp. symbiosis.

     

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