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Quantitative expression analysis of selected transcription factors in pavement, basal and trichome cells of mature leaves from Arabidopsis thaliana

机译:拟南芥成熟叶片的路面,基底细胞和毛状体细胞中所选转录因子的定量表达分析

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Gene expression levels of several transcription factors from Arabidopsis thaliana that were described previously to be involved in leaf development and trichome formation were analysed in trichome, basal and pavement cells of mature leaves. Single cell samples of these three cells types were collected by glass micro-capillaries. Real-time reverse transcription (RT)-PCR was used to analyse expression patterns of the following transcription factors: MYB23, MYB55, AtHB1, FILAMENTOUS FLOWER (FIL)/YABBY1 (YAB1), TRIPTYCHON (TRY) and CAPRICE (CPC). A difference in the expression patterns of TRY and CPC was revealed. Contrary to the CPC expression pattern, no transcripts of TRY could be detected in pavement cells. FIL/YAB1 was exclusively expressed in trichome cells. AtHB1 was highly expressed throughout all three cell types. MYB55 was higher expressed in basal cells than in trichome and pavement cells. MYB23 showed a pattern of low expression in pavement cells, medium in basal cells and high expression in trichomes. Expression patterns obtained by single cell sampling and real-time RT-PCR were compared to promoter GUS fusions of the selected transcription factors. Therefore, we regenerated two transgenic Arabidopsis lines that expressed the GUS reporter gene under control of the promoters of MYB55 and YAB1. In conclusion, despite their function in leaf morphogenesis, all six transcription factors were detected in mature leaves. Furthermore, single cell sampling and promoter GUS staining patterns demonstrated the predominant presence of MYB55 in basal cells as compared to pavement cells and trichomes.
机译:在成熟叶片的毛状体,基部和铺面细胞中分析了拟南芥中几种转录因子的基因表达水平,这些转录因子先前被描述与叶片发育和毛状体形成有关。通过玻璃微毛细管收集这三种细胞类型的单细胞样品。实时逆转录(RT)-PCR用于分析以下转录因子的表达模式:MYB23,MYB55,AtHB1,丝状花丝(FIL)/ YABBY1(YAB1),TRIPTYCHON(TRY)和CAPRICE(CPC)。揭示了TRY和CPC的表达方式的差异。与CPC表达方式相反,在路面细胞中未检测到TRY的转录本。 FIL / YAB1仅在毛状体细胞中表达。 AtHB1在所有三种细胞类型中都高度表达。 MYB55在基底细胞中的表达高于在毛状体和路面细胞中的表达。 MYB23在路面细胞中低表达,在基底细胞中中等,在毛状体中表达高。通过单细胞采样和实时RT-PCR获得的表达模式与所选转录因子的GUS融合启动子进行了比较。因此,我们再生了两个在MYB55和YAB1启动子控制下表达GUS报告基因的转基因拟南芥系。总之,尽管它们在叶片形态发生中起作用,但在成熟叶片中检测到所有六个转录因子。此外,单细胞采样和启动子GUS染色模式表明,与路面细胞和毛线虫相比,基底细胞中主要存在MYB55。

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