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丝裂原活化蛋白激酶激酶9(mitogen-activated protein kinase kinase 9, MKK9)作为丝裂原活化蛋白激酶(mitogenactivated protein kinase, MAPK)级联通路成员,在调控植物抗逆胁迫中具有重要作用。为研究山金柑(Fortunella hindsii)FhMKK9基因在柑橘溃疡病抗性中的功能,本研究以山金柑叶片c DNA为模板,克隆获得Fh MKK9基因CDS(coding sequence)全长序列,其开放阅读框(open reading frame, ORF)为972 bp,编码323个氨基酸残基。保守结构域分析表明, Fh MKK9具有MAPKK家族特有的PKc＿MAPKK＿Plant-like蛋白激酶结构域序列。系统发育分析显示,山金柑Fh MKK9蛋白与芸香科(Rutaceae)其他物种同源蛋白亲缘关系较近。启动子顺式作用元件预测显示,其启动子区含多个光信号及激素信号响应元件。亚细胞定位结果表明, Fh MKK9蛋白定位于细胞膜和细胞核中。组织特异性RT-qPCR结果显示, Fh MKK9基因在山金柑叶片中表达量最高,在根和茎中也有较高表达,表明其可能在山金柑叶片中发挥更为重要的作用。柑橘溃疡菌(Xanthomonas axonopodis pv. citri, Xac)侵染可诱导山金柑叶片中Fh MKK9基因显著上调表达。外源乙烯利处理浓度低于50 mg~/L时, FhMKK9基因的表达与乙烯利浓度呈正相关。瞬时表达Fh MKK9基因可增强感病种质‘纽荷尔’脐橙(Citrus sinensis Osbeck′Newhall′)叶片对溃疡病的抗性。本研究表明Fh MKK9基因可能通过乙烯信号通路正向调控山金柑对柑橘溃疡病的抗性,为柑橘抗病育种提供了候选基因。
Abstract：Mitogen-activated protein kinase kinase 9(MKK9), a member of the mitogen-activated protein kinase(MAPK) cascade pathway, plays an important role in regulating plant stress resistance. In order to study the function of the Fh MKK9 gene from Fortunella hindsii in resistance to citrus canker, the full-length CDS of Fh MKK9 was amplified from the c DNA of F. hindsii leaves. The ORF of Fh MKK9 is 972 bp in length, encoding 323 amino acids. Conserved domain analysis revealed that Fh MKK9 contains the specific PKc＿MAPKK＿Plant-like protein kinase domain of the MAPKK family. Phylogenetic analysis showed that the Fh MKK9 protein of Fortunella hindsii is closely related to homologous MKK9 proteins from other Rutaceae species. Prediction of promoter cis-acting elements indicated that its promoter region contains multiple light signal and hormone signal response elements. Subcellular localization results indicated that the Fh MKK9 protein is located in the cell membrane and nucleus. Tissue-specific RT-qPCR results revealed that the expression of the Fh MKK9 gene is highest in Fortunella hindsii leaves, with relatively high expression also in roots and stems, indicating that it may play a more important role in Fortunella hindsii leaves. Infection by the citrus canker pathogen can significantly induce the up-regulated expression of the Fh MKK9 gene in Fortunella hindsii leaves. When the concentration of ethephon applied was lower than 50 mg/L, the expression of the Fh MKK9 gene is positively correlated with ethylene concentration. Transient expression of the Fh MKK9gene demonstrated that it enhances the resistance of leaves from the susceptible germplasm ′Newhall′ navel orange to citrus canker.This study indicates that the Fh MKK9 gene may positively regulate the resistance of Fortunella hindsii to citrus canker through the ethylene signaling pathway, providing a candidate gene for citrus disease-resistant breeding.
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基本信息:
DOI：10.13417/j.gab.045.000586
中图分类号:S666
引用信息:
[1]代文婧,肖璇.山金柑FhMKK9基因克隆、表达及功能的初步鉴定[J].基因组学与应用生物学,2026,45(03):586-599.DOI:10.13417/j.gab.045.000586.
基金信息:
江西省教育厅项目(GJJ2201234)资助
2026-06-25
2026-06-25