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    毛白杨PtYABBY基因家族3个成员的克隆与表达分析

    Cloning and expression analysis of three members of PtYABBYs in Populus tomentosa

    • 摘要:
        目的  YABBY是高等植物中特有的转录因子家族,在侧生器官发育和开花过程中发挥重要作用。本研究对毛白杨PtYABBY基因进行克隆和生物信息学分析,并对雌雄花芽8个关键发育时期和根茎叶等组织中PtYABBY基因表达模式进行探究,以期为阐明PtYABBY基因在毛白杨侧生器官发育过程中的功能奠定前期基础。
        方法  以毛白杨为试材,采用同源基因克隆法从毛白杨中分离FILAMENTOUS FLOWER (FIL)/YABBY3 (YAB3)同源基因PtYABBY3、PtYABBY4和PtYABBY11的编码区序列,并开展生物信息学分析。通过qRT-PCR研究这3个基因在雌雄花芽8个发育时期及根、茎、幼叶、成熟叶片中的表达规律。
        结果  PtYABBY3、PtYABBY4和PtYABBY11编码区长度分别为633、642、639 bp,分别编码210、213、212个氨基酸。所推测的氨基酸序列包含C2C2锌指结构域和YABBY结构域。系统进化分析进一步表明这3个基因属于FIL/YAB3亚家族成员。qRT-PCR结果显示这3个基因在根、茎、叶及8个时期雌雄花芽组织中均有表达,但PtYABBY基因间的表达水平存在明显差异。PtYABBY3表达水平在雌雄花芽发育初期下调,发育后期上调;花原基形成到休眠期内PtYABBY3在雌雄花芽中呈现相反的表达变化趋势。PtYABBY4和PtYABBY11基因在雌雄株成花诱导期表达水平最高,成花诱导到伸长期内呈现下降趋势,在雄花芽休眠期和小孢子发生期基因表达水平无明显变化。营养组织表达量测定结果显示这3个基因在幼叶和成熟叶片中表达水平相对较高,根系中表达量最低。
        结论  YABBY基因家族成员PtYABBY3、PtYABBY4和PtYABBY11同属FIL/YAB3亚类,在毛白杨雌雄花芽发育的8个时期中表达水平存在较大差异,且在叶片和花芽中表达量较高,表明其可能与叶片和花芽发育相关。本研究为今后深入研究YABBY家族成员在杨树器官生长发育过程中的作用奠定基础。

       

      Abstract:
        Objective  YABBY is a unique transcription factor family in higher plants, which plays an important role in the development of lateral organs and flowering. In this study, we conducted cloning and bioinformatics analysis of PtYABBY genes, and explored the expression patterns of PtYABBY genes in eight stages of male and female flower buds and vegetative tissues such as roots, stems and leaves in order to lay a foundation for clarifying the function of PtYABBY genes on the development of lateral organs of Populus tomentosa.
        Method  The coding sequences of PtYABBY3, PtYABBY4 and PtYABBY11, homologous genes of FILAMENTOUS FLOWER (FIL) and YABBY3 (YAB3), were isolated from P. tomentosa by homologous gene cloning method, and bioinformatics analysis was performed. The expression patterns of three PtYABBYs in eight stages of male and female flower buds, roots, stems, young leaves and mature leaves were studied by qRT-PCR.
        Result  The lengths of PtYABBY3, PtYABBY4 and PtYABBY11 were 633, 642, 639 bp, encoding 210, 213, 212 amino acids, respectively. The proposed amino acid sequences all contained the C2C2 zinc finger domain and the YABBY domain. Phylogenetic analysis further indicated that three PtYABBYs belong to the FIL/YAB3 subfamily. qRT-PCR results showed that three PtYABBYs were expressed in roots, stems, leaves, male and female flower buds at eight stages. However, there were significant differences in the expression levels of PtYABBY genes. The expression level of PtYABBY3 was down-regulated at the early stage and up-regulated at the late stage of male and female flower bud development. From primordia formation to dormancy stage, the expression trends of PtYABBY3 in female and male flower buds were completely different. The expression levels of PtYABBY4 and PtYABBY11 were the highest during the floral induction stage, and decreased from floral induction stage to enlargement stage. During dormancy and microsporogenesis stage of male flower buds, there was no significant change in gene expression. The fluorescence quantitative results of vegetative tissues showed that the expression levels of three PtYABBYs were relatively high in young leaves and mature leaves, and the expression levels were the lowest in roots.
        Conclusion  PtYABBY3, PtYABBY4 and PtYABBY11, members of the YABBY gene family belong to FIL/YAB3 subclass. The expression levels of PtYABBYs are obviously different during 8 stages of female and male flower bud development in P. tomentosa, and relatively high in leaves and flower buds. The expressions of PtYABBY3, PtYABBY4 and PtYABBY11 may be related to the development of buds and leaves of P. tomentosa, which lays a foundation for further study of their roles in the development of poplar.

       

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