‘凤丹’牡丹PoKAS基因的克隆及表达分析
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云南省重大基础专项生物资源数字化开发应用项目(202002AA10007);云南省千人计划“高层次人才”专项(2019);云南省万人计划“云岭产业技术领军人才”专项(2018-212)


Cloning and Expression Analysis of PoKAS Gene of Paeonia ostii ‘Feng Dan’
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    摘要:

    为探究‘凤丹’牡丹(Paeonia ostii ‘Feng Dan’)PoKAS基因在脂肪酸合成中的功能,从转录组数据中获得3个PoKAS基因,克隆基因全长并进行生物信息学分析,通过qRT-PCR检测它们在牡丹落花后第23、45、75、100和125 天时的表达。结果显示:(1)克隆得到的3个基因序列全长分别为1 401、1 692和1 215 bp,分别编码466、563和404 aa;保守结构域分析发现,它们都含有KAS保守结构域,属于cond-enzymes超蛋白家族。(2)系统进化树将三者分为三大类,表明其在进化上相对独立,分别命名为PoKASⅠPoKASⅡPoKASⅢ(GenBank登录号分别为OP056413、OP056412和OP056414)。(3)qRT-PCR分析发现,在牡丹落花后种子发育的5个时期中,PoKASⅠPoKASⅡ基因在落花后75 d和45 d时的表达量分别显著高于其他发育时期;PoKASⅢ基因在落花后45~125 d时的表达量均显著高于落花后23 d,说明PoKASⅢ基因在牡丹种子脂肪酸合成的整个过程中发挥着重要作用,而PoKASⅡ基因主要在种子油脂快速累积前期发挥作用。研究推测,该基因家族在‘凤丹’牡丹种子脂肪酸形成过程中的不同发育时期发挥不同的作用。

    Abstract:

    In order to investigate the function of PoKAS gene in the fatty acid synthesis of P. ostii ‘Feng Dan’, we obtained three PoKAS genes from the transcriptome data, cloned the genes and conducted the bioinformatics analysis. Their expressions were examined by qRT-PCR at 23, 45, 75, 100 and 125 days after flowering. The results showed that: (1) the full sequence length three cloned genes were 1 401, 1 692 and 1 215 bp, encoding 466, 563 and 404 aa, respectively. The analysis of the conserved domains revealed that they all contained the conserved domain of KAS and belonged to the superprotein family of cond-enzymes. (2) The phylogenetic tree analysis showed that they were clearly divided into three categories, indicating that they were relatively evolutionarily independent, named PoKASⅠ, PoKASⅡ and PoKASⅢ (GenBank accession numbers OP056413, OP056412, and OP056414, respectively). (3) The qRT-PCR analysis revealed, among the five periods of peony seed development after the flower drop, the expressions of PoKASⅠ and PoKASⅡ were significantly higher at 75 d and 45 d after flowering than those at other development periods; the expression of PoKASⅢ wassignificantly higher at 45-125 d after anthesis than that at 23 d after anthesis, indicating that PoKASⅢ could play an important role in the whole process of fatty acid synthesis in peony seeds, while PoKASⅡ mainly could play a role in the early stage of rapid accumulation of seed oil. It is hypothesized that this gene family from P. ostiis ‘Feng Dan’ could play different roles in different developmental periods during the formation of seed fatty acids.

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刀 梅,刘 琅,杨自云,等.‘凤丹’牡丹PoKAS基因的克隆及表达分析[J].西北植物学报,2023,43(2):211-219

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  • 在线发布日期: 2023-03-09
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