Genes related to fuzz initiation and development in gossypium arboretum identified by cDNA microarray | cDNA芯片筛选亚洲棉短绒分化发育相关基因

Chinese. 【目的】筛选亚洲棉短绒分化发育相关基因。【方法】利用cDNA芯片技术在棉纤维发育前期开花前3天(-3DPA)、开花当天(0DPA)、开花后3天(+3DPA)、5天(+5DPA)、7天(+7DPA)筛选野生型DPL971与其短绒突变体DPL972的差异基因。通过与模式植物拟南芥数据库相结合,找到与棉纤维起始发育相关基因,并通过RT-PCR和Real-Time PCR（QRT-PCR）验证部分侯选基因。【结果】筛选出与亚洲棉纤维起始分化发育相关基因8个,其中相对应的与拟南芥同源性较高的蛋白有7个,分别为KAK（DR461366）、MYB5（ES812048）、TTG1（ES811600）、MYB23（DR453866）、CSLD3（DR459646）、RHD2（DR461821）和ZWI（ES791383）,这些基因在拟南芥的表皮毛起始发育过程中都起着重要的作用。RT-PCR和QRT-PCR结果表明MYB23、MYB5、TTG1、CSLD3、RHD2这5个基因在短绒突变体DPL972与其野生型DPL971的+3DPA胚珠中表达差异都有差异。【结论】上述结果表明这5个基因可能与短绒分化发育相关,第1个基因可能抑制短绒分化和发育,后4个则促进短绒分化和发育。

English. 【Objective】 The objective of this study is to isolate genes related to fuzz initiation and development in Gossypium arboretum. 【Method】 The filter arrays of cDNAs were performed to identify the differentiation expressed genes between the wild-type DPL971 and its fuzz mutant line DPL972 in the early developing stages of -3DPA, 0DPA, +3DPA, +5DPA, and +7DPA. Compared with the database of model plant Arabodopsis thaliana, some genes related to the initiation of the cotton fiber were isolated, and part of them have been verified by RT-PCR and QRT-PCR. 【Result】 Eight genes related to fiber initiation and development in Gossypium arboretum were identified, and seven of them, KAK(DR461366), MYB5(ES812048), TTG1(ES811600), MYB23(DR453866), CSLD3(DR459646), RHD2(DR461821), ZWI(ES791383), were highly homologous to the Arabodopsis proteins which played important roles in the initiation of the trichomes development in Arabidopsis. The result of RT-PCR and QRT-PCR indicated that the 5 genes of MYB23, MYB5, TTG1, CSLD3, and RHD2 had significantly different expressions in +3DPA ovules between the fuzz mutant line DPL972 and its wild line DPL971. 【Conclusion】 It implied that these genes may be related to the initiation of the fuzz fiber, the first gene may inhibit the fuzz differentiation and the latter four may promote the fuzz development.