桂花OfACOs基因家族鉴定及表达分析

Identification and expression of OfACOs gene family in Osmanthus fragrans

【目的】对桂花Osmanthus fragrans乙烯合成路径氨基环丙烷羧酸氧化酶(1-aminocyclopropane-1-carbox-ylate oxidase, ACO)基因家族进行了全基因组鉴定及表达分析,以探索参与桂花花瓣乙烯合成的关键ACO家族成员。【方法】以桂花品种‘柳叶金桂’O. fragrans‘Liuye Jingui’为参考基因组,对桂花OfACOs家族进行鉴定、进化分析、基因结构分析以及根、茎、叶、芽和不同开花阶段的表达模式分析。【结果】通过蛋白保守结构域(protein families,pfam)分析,从桂花基因组中共鉴定出了122个OfACOs成员,分布于22条染色体上;保守基序分析表明:大多数OfACOs基因同时包含Motif 1、 Motif 2、Motif 3、Motif 4和Motif 7这5个保守基序,它们共同组成OfACOs保守结构域;共线性分析结果表明:与拟南芥Arabidopsis thaliana基因为共线性对的基因可能具有相似的功能。结合swiss-prot同源序列对比分析,筛选出27个成员作为候选研究对象,进行不同组织部位根、茎、叶、芽和不同开花阶段的转录组测序分析,结果发现:12个成员在花瓣中显著表达,其中LYG006223、LYG007706、LYG007045、LYG035696等4个成员在开花后期极显著差异上调表达。进一步对该4个成员进行荧光定量PCR验证分析,结果与转录组测序结果一致。【结论】通过OfACOs基因家族的全基因组鉴定及基因克隆分析,筛选到在开花后期显著差异上调的4个ACO家族成员可能参与桂花花瓣衰老的调控。图6参34。

[Objective]The objective of this study is to identify and analyze the gene family of OfACOs in ethylene biosynthesis pathway,so as to explore the key 1-aminocyclopropane-1-carbox-ylate oxidase(ACO)family members involved in the ethylene synthesis of Osmanthus fragrans petals.[Method]The identification,evolutionary analysis,gene structure analysis,and expression pattern analysis of OfACOs gene family at root,stem,leaf,bud and different flowering stages were carried out taking O.fragrans‘Liuye Jingui’as the reference genome.[Result]Through the analysis of protein conserved domain(protein families,pfam),a total of 122 OfACOs members were identified from O.fragrans genome,which were distributed on 22 chromosomes.The conserved motif analysis showed that most OfACOs genes contained Motif 1,Motif 2,Motif 3,Motif 4,and Motif 7,which together formed the OfACOs conserved domain.Collinearity analysis showed that OfACOs genes that were colinear with Arabidopsis thaliana genes might have similar functions.Combined with comparative analysis of switch-prot homologous sequence,27 OfACOs members were selected as candidate research objects for transcriptome sequencing analysis of different tissue parts,including root,stem,leaf,bud and different flowering stages,and the results showed that 12 OfACOs members were significantly expressed in petals,among which LYG006223,LYG007706,LYG007045,and LYG035696 were significantly up-regulated in the late flowering stage.The four members were further analyzed by RT-qPCR analysis,and the results were consistent with those of transcriptome sequencing.[Conclusion]Through the whole genome identification and gene cloning analysis of OfACOs gene family,four members of OfACOs which are significantly up-regulated in the late flowering stage are screened,which may participate in O.fragrans petal senescence.[Ch,6 fig.34 ref.]