紫花苜蓿pPLA基因家族的生物信息学及表达模式分析

Bioinformatics and expression patterns analysis of pPLA gene family in alfalfa

摘要:

【目的】分析紫花苜蓿pPLA基因家族的生物信息学及表达模式,筛选诱导紫花苜蓿单倍体的候选pPLA基因。【方法】利用紫花苜蓿基因组数据库及多种生物信息学软件对MspPLA基因家族进行鉴定和分析,并利用qRT-PCR分析了其在不同器官中的表达模式,通过进化树和表达模式分析,获得诱导紫花苜蓿单倍体的候选pPLA基因。【结果】1)从紫花苜蓿基因组数据库中筛选并鉴定到12个MspPLA基因,分布于不同的染色体中;2)在12个MspPLA蛋白中,11个为稳定性亲水蛋白,亚细胞定位显示2/3的pPLA蛋白定位于叶绿体中;MspPLA家族共含有10个Motifs,除了MspPLA6外,其余蛋白均含有7个保守的Motifs,且MspPLA1/2/4/12含有特异的Motif7;MspPLA家族含有4个pPLA保守结构域;系统进化分析将其分为3个亚组,且MspPLA1/2/4/12与其他物种中能诱导单倍体的pPLA蛋白聚为一类;3)qRT-PCR分析显示,多数MspPLA基因(包括MspPLA1/2/3/4/5/6/7/9/12)在花序和荚果中有较高的相对表达量,MspPLA8/11和MspPLA10分别在根和叶片中的相对表达量最高。【结论】结合系统进化和表达模式分析,可将MspPLA1/2/4/12作为诱导紫花苜蓿单倍体的候选基因。

Abstract:

【Objective】To analyze the bioinformatic characteristics and expression patterns of the alfalfa (Medicago sativa L. ) pPLA gene family, and to screen the candidate pPLA genes for inducing alfalfa haploid. 【Method】In this study, the MspPLA gene family was identified and analyzed using alfalfa genome database and a variety of bioinformatics software, and its expression patterns in different organs were analyzed by qRT-PCR. Candidate pPLA genes for inducing alfalfa haploid were obtained through phylogenetic tree and expression pattern analyses. 【Result】1) 12 MspPLA genes were screened and identified from the genomic database of alfalfa, and they were distributed in different chromosomes. 2) Among the 12 MspPLA proteins, 11 were stable hydrophilic proteins, and subcellular localization showed that 2/3 of the pPLA proteins were located in chloroplasts. The MspPLA family contained 10 motifs; except for MspPLA6, all the other proteins contained 7 conserved motifs, and MspPLA1/2/4/12 contained a specific motif 7. The MspPLA family contains four conserved domains of pPLA. Phylogenetic analysis divided the MspPLA family into three subgroups, and MSPPLA1/2/4/12 were clustered with haploide-inducing pPLA proteins from other species. (3) qRT-PCR analysis showed that most MspPLA genes (including MspPLA1/2/3/4/5/6/7/9/12) had high relative expression levels in inflorescence and pod, whereas MspPLA8/11 and MspPLA10 had the highest relative expression levels in root and leaf, respectively. 【Conclusion】Combined with phylogenetic and expression pattern analysis, MspPLA1/2/4/12 can be used as a candidate genes for inducing alfalfa haploid. This study lays a foundation for pPLA gene-induced haploid breeding in alfalfa.