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Molecular regulation of somatic embryogenesis in potato: an auxin led perspective

机译:马铃薯体细胞胚发生的分子调控:以生长素为主导的观点

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Potato internodal segments (INS) treated with the auxin 2,4-dichlorophenoxyacetic acid can be induced to develop somatic embryos upon their transfer to an auxin-free medium, while the continuous presence of auxin in the medium suppresses the progression of embryogenically-induced somatic cells to embryos. We have employed these contrasting pathways, in combination with potato microarrays representing circa 10,000 genes, to profile global gene expression patterns during the progression of somatic embryogenesis in potato. The induction phase, characterised by the presence of auxin, was analysed by the direct comparison of RNA isolated from freshly excised (0 days) and embryogenically induced (14 days) INS explants. RNAs from embryo-forming (withdrawal of auxin after 14 days) and embryo-inhibitory (continuous presence of auxin) conditions, isolated over a range of time-points until the emergence of somatic embryos, were compared in a loop design to identify auxin responsive genes putatively involved in the process of somatic embryogenesis. A total of 402 transcripts were found to be showing significant differential expression patterns during somatic embryogenesis ‘induction’ phase, 524 during ‘embryo-transition’ phase, while 44 transcripts were common to both phases. Functional classification of these transcripts, using Gene Ontology vocabularies (molecular and biological), revealed that a significant proportion of transcripts were involved in processes which are more relevant to somatic embryogenesis such as apoptosis, development, reproduction, stress and signal transduction. This is the first study profiling global gene expression patterns during true somatic embryogenesis initiated from mature and completely differentiated explants and has enabled the description of stage-specific expression patterns of a large number of genes during potato somatic embryogenesis (PSE). The significance of the key identified genes during critical stages of somatic embryogenesis is discussed.
机译:用生长素2,4-二氯苯氧基乙酸处理的马铃薯节间节(INS)可以在转移至无生长素的培养基中后诱导体细胞胚发育,而生长素在该培养基中的持续存在则抑制了胚性诱导体细胞的进程细胞到胚胎。我们采用了这些相反的途径,结合代表约10,000个基因的马铃薯微阵列,来分析马铃薯体细胞胚发生过程中的整体基因表达模式。通过直接比较从新鲜切下的(0天)和胚诱导的(14天)INS外植体中分离的RNA,分析了以生长素存在为特征的诱导期。在循环设计中比较了在一定时间范围内分离直至胚体胚出现的胚胎形成(生长素在14天后退出生长素)和胚胎抑制(生长素持续存在)条件下的RNA,以鉴定对生长素有反应性推测参与体细胞胚发生过程的基因。共有402个转录本在体细胞胚发生“诱导”阶段表现出显着的差异表达模式,在“胚胎过渡”阶段则有524个,而这两个阶段共有44个转录本。使用基因本体学词汇(分子和生物学)对这些转录本进行功能分类,发现相当一部分转录本参与了与体细胞胚发生更为相关的过程,例如细胞凋亡,发育,繁殖,应激和信号转导。这是首次对由成熟和完全分化的外植体引发的真实体细胞胚发生过程中的全球基因表达模式进行分析的研究,并已能够描述马铃薯体细胞胚发生(PSE)过程中大量基因的阶段特异性表达模式。讨论了关键鉴定基因在体细胞胚发生关键阶段的意义。

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