非洲菊SPFH基因家族的鉴定及表达

Identification and expression analysis of SPFH gene family in Gerbera jamesonii

SPFH(Stomain,Prohibitin,Flotillin and Hflk/C)超家族蛋白是锚定在膜微区的多样化膜蛋白家族,参与调控植物的生长发育、响应生物或非生物胁迫.基于非洲菊转录组数据,利用生物信息学对SPFH(GjSPFH)家族成员进行鉴定,分析其基本理化性质、亚细胞定位和保守氨基酸序列,构建系统进化树,并进行蛋白互作预测.利用转录组FPKM值分析GjSPFHs在非洲菊健康及感病时期的表达水平;利用荧光实时定量技术检测GjSPFH家族成员在不同组织(根、茎和叶片)和不同激素(水杨酸SA和茉莉酸甲酯MeJA)处理下的表达模式.结果表明,非洲菊包含8个GjSPFH成员,其蛋白长度在285-393 aa之间,蛋白理论分子量在31.22×10^(3)-43.14×10^(3)之间,等电点为5.53-9.80,其中有4个蛋白为稳定蛋白,4个为不稳定蛋白;均定位于线粒体,含有15个保守氨基酸序列;GjSPFH可分为3个亚组;该蛋白家族不仅成员之间可以发生互作,还与其他蛋白互作.非洲菊GjSPFH基因家族能够响应根腐病胁迫,GjPHB6在感病时期表达量显著高于健康时期,可能参与了非洲菊抵御真菌侵染的过程.非洲菊GjSPFH基因的表达具有组织特异性,大部分GjSPFHs基因在叶片中表达量相对较高.本研究表明GjSPFHs能够响应SA和MeJA的胁迫并呈现多种表达调控模式,可能与非洲菊抗病性有关,但不同成员的功能有所差异.

SPFH(Stomain,Prohibitin,Flotillin,and Hflk/C)superfamily proteins are diversified membrane protein families anchored in membrane microregions that participate in the regulation of plant growth and development and respond to biological and abiotic stress.Based on the transcriptome database of Gerbera jamesonii,the SPFH gene family members(GjSPFH)of G.jamesonii were identified via bioinformatics analysis.First,we analyzed the basic physical and chemical properties,secondary structure,subcellular location,and conserved amino acid sequence of GjSPFH proteins and constructed a phylogenetic tree.Second,we predicted the protein interactions of the related genes.In addition,we used the transcriptome dataset to analyze the expression levels of the GjSPFH gene family during the healthy and susceptible periods.Finally,we also used qRT-PCR analysis to detect the expression pattern of GjSPFH family members in different tissues(including roots,petioles,and leaves)and under different hormone treatments(including salicylic acid(SA)and methyl jasmonate(MeJA)).The results showed that the GjSPFH gene family contains eight members,the length of eight GjSPFH proteins was between 285 and 393 aa,the theoretical molecular weight of the predicted proteins was between 31.22×10^(3)and 43.13×10^(3),and the isoelectric point was between 5.53 and 9.80.Among the proteins,four were stable and four were unstable.The prediction results of subcellular localization showed that the eight GjSPFH proteins were located in mitochondria and contained 15 conserved motifs.The phylogenetic tree showed that the eight GjSPFHs were divided into three groups.The protein interaction prediction analysis showed that the GjSPFH protein family not only interacted with each other but also with other proteins.The GjSPFH gene family could respond to root rot disease stress,and the expression level of GjPHB6 in the susceptible period was significantly higher than that in the healthy period,suggesting GjPHB6 involved in the resistance of G.jamesonii to fungal infection.The expression of the GjSPFH gene was tissue-specific,and most were expressed at relatively high levels in leaves.The GjSPFHs could respond to SA and MeJA stress and present a variety of expression regulation patterns,which may be related to the disease resistance of G.jamesonii,but the functions of each member were different.These results could provide support for further studies on the biological function of the SPFH gene family in G.jamesonii.