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RcMPK3正向调控月季对灰霉病菌的响应

RcMPK3 positively regulates Rosa chinensis ‘Old Blush’ in response to Botrytis cinerea

  • 摘要: 灰霉病是月季采后和运输过程中危害最严重的真菌病害。MPK3基因参与植物对逆境的响应。基于古老月季品种‘月月粉’(Rosa chinensis‘Old Blush’ Jacq.)全基因组数据,利用RT-PCR技术获得了包含完整ORF区的RcMPK3基因,并对其进行生物信息学分析和功能检测。结果显示:RcMPK3基因ORF序列长1113 bp,编码370个氨基酸,系统进化树分析结果表明RcMPK3蛋白与FvMPK3蛋白聚为一支;对基因全长序列分析发现,RcMPK3由6个外显子和5个内含子构成,启动子序列包含10类响应激素和逆境相关的顺式元件;qPCR分析表明,RcMPK3受SA和JA诱导表达,同时,在灰霉病菌(Botrytis cinerea)侵染过程中,RcMPK3基因表达量也显著提高;VIGS分析发现,基因沉默株系病斑直径显著大于对照组株系,表明RcMPK3基因可能正向调控月季对灰霉病菌的抗性。

     

    Abstract: Gray mold is the most serious fungal disease in rose after harvest and during transportation. The MPK3 gene participates in the plant response to stress. Here, RT-PCR technology was used to isolate and clone the full length of the RcMPK3 gene based on whole-genome data of Rosa chinensis ‘Old Blush’ Jacq. Results showed that the open reading frame (ORF) sequence of RcMPK3 was 1113 bp long and encoded 370 amino acids. Phylogenetic analysis showed that the RcMPK3 protein was clustered with FvMPK3. The full-length sequence of the RcMPK3 gene was analyzed, which showed that RcMPK3 was composed of six exons and five introns. There were 10 cis-elements in RcMPK3. Quantitative real time PCR (qPCR) analysis showed that RcMPK3 was induced by salicylic acid (SA) and jasmonic acid (JA). RcMPK3 expression significantly increased under Botrytis cinerea stress. Functional analysis of RcMPK3 by VIGS showed that lesion diameter in the gene-silenced plant was significantly larger than that in the control, indicating that the RcMPK3 gene positively regulates the resistance of rose to B. cinerea.

     

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