两种兰科植物CaM及CML基因家族全基因组分析

目的]探讨Ca M和CML基因家族在兰科植物不同组织和与菌根真菌建立共生关系过程中的潜在功能。方法]依据已发表的拟南芥Ca M和CML基因家族蛋白序列,利用生物信息学方法对小兰屿蝴蝶兰和铁皮石斛全基因组进行搜索,通过多种生物软件或在线工具对Ca M和CML基因家族蛋白序列进行筛选及确定、蛋白结构分析、系统进化分析及结构域预测;利用转录组数据绘制heatmap图,对小兰屿蝴蝶兰不同组织(花、叶、茎、根)及石斛种子与美胞胶膜菌共生条件下Ca M和CML基因家族的表达情况进行分析。结果]在小兰屿蝴蝶兰和铁皮石斛全基因组中均预测到4个Ca M蛋白和54个CML蛋白;小兰屿蝴蝶兰Ca M及CML家族基因中39个基因没有内含子,19个基因具有内含子;铁皮石斛Ca M及CML基因家族中有41个基因没有内含子,17个基因具有内含子;系统进化分析将小兰屿蝴蝶兰和铁皮石斛Ca M和CML蛋白家族各分为10个亚家族;小兰屿蝴蝶兰中9个基因在叶中的表达相对于花、茎、根为上调表达,2个基因在叶中的表达相对于花、茎、根却为下调表达; 3个基因在花中的表达相对于叶、茎、根为上调表达; 3个基因在花和叶中的表达相对于茎和根为上调表达;铁皮石斛中4个基因在与美胞胶膜菌共生萌发文库中的表达相对于非共生文库为上调表达; 4个基因在共生萌发文库中的表达相对于非共生文库为下调表达。结论]Ca M及CML家族基因在兰科植物不同组织具有重要作用,且参与铁皮石斛种子与美孢胶膜菌共生萌发的生物学调控过程。

Genome-wide Analysis of CaM/CML Gene Family in Two Orchidaceae Species

Objective] To research the CaM/CML gene family''s characteristics in different tissues and symbiosis with mycorrhizal fungi of Orchidaceae.Method] Genome-wides of Phalaenopsis equestris and Dendrobium officinale were separately predicted and determined with bioinformatics methods based on the publishing protein sequences of CaM/CML gene family of Arabidopsis. The protein structures, phylogenetic trees, exon-intron structures of CaM/CML gene family in P. equestris and D. officinale were analyzed using different biological softwares. Gene expression analysis of different tissues (flower, leave, stem and root) in P. equestris and symbiotic with Tulasnella calospora and asymbiotic germination seeds from D. officinale were conducted by using RNA-seq data.Result] Four CaMs and fifty-four CMLs were separately predicted from P. equestris and D. officinale genomes. In P. equestris, forty-one genes did not have intron, seventeen genes had intron in CaM/CML gene family; and thirty-nine genes did not have intron, nineteen genes had intron in D. officinale. Phylogenetic analysis results showed that the CaMs and CMLs could be separately divided into ten subfamilies. The results of CaMs and CMLs genes expression analysis in different tissues of P. equestris indicated that nine genes were up-regulated in the leaf relative to the flower, stem and root, two genes were down-regulated; three genes were up-regulated in the flower relative to the leaf, stem and root; three genes were up-regulated in the flower and leaf relative to the stem and root. The results of gene expression analysis in symbiotic with T. calospora and asymbiotic germinated seeds from D. officinale by using RNA-seq data indicated that four genes were up-regulated, and four genes down-regulated.Conclusion] These results will lay the basis for the further studying of potential biological function and molecular mechanism of CaMs and CMLs genes in orchid.