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    钟姗辰, 武舒, 王黎, 苏晓华, 张冰玉. 杨树种毛相关候选基因PeCFL1和PeCFL2的时空表达分析[J]. 北京林业大学学报, 2022, 44(5): 1-7. DOI: 10.12171/j.1000-1522.20200403
    引用本文: 钟姗辰, 武舒, 王黎, 苏晓华, 张冰玉. 杨树种毛相关候选基因PeCFL1和PeCFL2的时空表达分析[J]. 北京林业大学学报, 2022, 44(5): 1-7. DOI: 10.12171/j.1000-1522.20200403
    Zhong Shanchen, Wu Shu, Wang Li, Su Xiaohua, Zhang Bingyu. Spatio-temporal expression of candidate genes PeCFL1 and PeCFL2 in seed hair development of poplar[J]. Journal of Beijing Forestry University, 2022, 44(5): 1-7. DOI: 10.12171/j.1000-1522.20200403
    Citation: Zhong Shanchen, Wu Shu, Wang Li, Su Xiaohua, Zhang Bingyu. Spatio-temporal expression of candidate genes PeCFL1 and PeCFL2 in seed hair development of poplar[J]. Journal of Beijing Forestry University, 2022, 44(5): 1-7. DOI: 10.12171/j.1000-1522.20200403

    杨树种毛相关候选基因PeCFL1和PeCFL2的时空表达分析

    Spatio-temporal expression of candidate genes PeCFL1 and PeCFL2 in seed hair development of poplar

    • 摘要:
        目的  杨絮是由杨树子房胎座表皮细胞发育而来的种毛,近年来已成为我国北方城市环境问题之一。目前对杨树种毛发育基因的研究还不深入,本研究将2个在杨树子房发育过程中差异性表达、且基因注释均与棉纤维发育相关的基因(PeCFL1和PeCFL2)作为种毛发育调控的候选基因,探究二者在欧美杨‘渤丰3号’中表达的时空特异性,为深入研究该基因在杨絮发育过程中的调控作用及基因工程手段改良杨树品种奠定基础。
        方法  取‘渤丰3号’杨越冬花枝,采集水培4、5、6、7、8、12 d的雌花序,进行固定、石蜡包埋、切片,观察子房发育及种毛的形态发生过程。采用实时定量PCR检测PeCFL1和PeCFL2基因在‘渤丰3号’杨雌花序发育过程中以及在根、茎、叶等营养器官中的表达模式。利用原位杂交技术检测PeCFL1和PeCFL2基因在杨树花器官中的组织表达特异性,揭示杨絮发育调控相关候选基因PeCFL1和PeCFL2的时空表达模式。
        结果  ‘渤丰3号’杨雌花枝水培12 d后,子房胎座底部出现纤维状结构,杨絮开始形成。PeCFL1和PeCFL2在‘渤丰3号’杨根、茎、叶及雌花枝腋芽中微量表达,在水培4 ~ 7 d的雌花序中表达量较少,水培8 d后表达量开始显著升高,12 d时表达量继续大幅上升,此时的石蜡切片可见子房胎座底部纤维状结构。原位杂交结果显示,PeCFL1和PeCFL2基因在杨树子房的子房壁和胎座部位表达。
        结论  种毛发育调控相关候选基因PeCFL1和PeCFL2在‘渤丰3号’杨雌花子房胎座底部出现纤维状结构时表达显著升高,并在胎座底部纤维状结构和子房壁中特异性表达,说明其与种毛发育调控密切相关,可作为基因工程改良杨树飞絮的目标基因。

       

      Abstract:
        Objective  Poplar seed hair is produced by the development of epidermal cells of the placenta at the base of ovary and it has become one of the environmental problems in northern cities of China in recent years. At present, the research on poplar seed hair is not in-depth. In this study, two genes (PeCFL1 and PeCFL2), which are differentially expressed during the development of poplar ovary and related to cotton fiber development, were selected as candidate genes for the regulation of seed hair development. We studied the spatio-temporal specificity of PeCFL1 and PeCFL2 genes expressed in Populus × euramericana ‘Bofeng 3’ to lay a foundation for further study on the regulation of the two genes during the development of poplar seed hair and improvement of poplar varieties by genetic engineering.
        Method  After 4, 5, 6, 7, 8, 12 days of hydroponic culture, the female inflorescences of Populus × euramericana ‘Bofeng 3’ were collected, fixed, paraffin embedded and sliced to observe the ovary development and the morphogenesis of poplar seed hair. Real time quantitative PCR was used to detect the expression patterns of PeCFL1 and PeCFL2 genes during the development of female inflorescence and in vegetative organs such as roots, stems and leaves of ‘Bofeng 3’ poplar. The tissue expression specificity of PeCFL1 and PeCFL2 genes in poplar flower organs was detected by in situ hybridization, and the temporal and spatial expression patterns of candidate genes PeCFL1 and PeCFL2 related to the regulation of poplar floc development were revealed.
        Result  After 12 days of hydroponic culture, fibrous structure appeared at the bottom of ovary placentation and poplar seed hair began to form. PeCFL1 and PeCFL2 were expressed in roots, stems, leaves and axillary buds of female flower branches of ‘Bofeng 3’ poplar. For the female inflorescence of hydroponic culture, a small amount of PeCFL1 and PeCFL2 were expressed from the 4th to the 7th day, and the expression levels began to increase significantly after the 8th day. Then the expression levels continued to increase significantly on the 12th day, at the same time, the fibrous structure at the bottom of placenta could be seen in paraffin section. The results of in situ hybridization showed that PeCFL1 and PeCFL2 genes were expressed in ovary wall and placenta of poplar ovary.
        Conclusion  The expression of PeCFL1 and PeCFL2 is significantly increased when the fibrous structure appeared at the bottom of the placentation in female flowers of Populus × euramericana ‘Bofeng 3’, and they are specifically expressed in the fibrous structure at the bottom of the placenta and the ovary wall, indicating that they are closely related to the regulation of seed hair development, and can be used as target genes for genetic engineering improvement of poplar seed hair.

       

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