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微切割拟南芥线虫诱导巨型细胞中的早期转录组事件。

Early transcriptomic events in microdissected Arabidopsis nematode-induced giant cells.

机构信息

Facultad de Ciencias del Medio Ambiente, Universidad de Castilla-La Mancha, Avenida de Carlos III s/n, 45071 Toledo, Spain.

出版信息

Plant J. 2010 Feb;61(4):698-712. doi: 10.1111/j.1365-313X.2009.04098.x. Epub 2009 Dec 9.

DOI:10.1111/j.1365-313X.2009.04098.x
PMID:20003167
Abstract

Root-knot nematodes differentiate highly specialized feeding cells in roots (giant cells, GCs), through poorly characterized mechanisms that include extensive transcriptional changes. While global transcriptome analyses have used galls, which are complex root structures that include GCs and surrounding tissues, no global gene expression changes specific to GCs have been described. We report on the differential transcriptome of GCs versus root vascular cells, induced in Arabidopsis by Meloidogyne javanica at a very early stage of their development, 3 days after infection (d.p.i.). Laser microdissection was used to capture GCs and root vascular cells for microarray analysis, which was validated through qPCR and by a promoter-GUS fusion study. Results show that by 3 d.p.i., GCs exhibit major gene repression. Although some genes showed similar regulation in both galls and GCs, the majority had different expression patterns, confirming the molecular distinctiveness of the GCs within the gall. Most of the differentially regulated genes in GCs have no previously assigned function. Comparisons with other transcriptome analyses revealed similarities between GCs and cell suspensions differentiating into xylem cells. This suggests a molecular link between GCs and developing vascular cells, which represent putative GC stem cells. Gene expression in GCs at 3 d.p.i. was also found to be similar to crown galls induced by Agrobacterium tumefaciens, a specialized root biotroph.

摘要

根结线虫通过特征描述不清的机制在根系(巨型细胞,GCs)中分化出高度特化的取食细胞,这些机制包括广泛的转录变化。虽然全转录组分析使用了根瘤,根瘤是包括 GCs 和周围组织的复杂根结构,但尚未描述专门针对 GCs 的全局基因表达变化。我们报告了在拟南芥中,由爪哇根结线虫诱导的 GC 与根维管束细胞的差异转录组,在感染后非常早期的 3 天(dpi)。激光微切割用于捕获 GC 和根维管束细胞进行微阵列分析,通过 qPCR 和启动子-GUS 融合研究进行了验证。结果表明,在 3 dpi 时,GC 表现出主要基因抑制。尽管一些基因在根瘤和 GCs 中表现出相似的调控,但大多数基因表现出不同的表达模式,这证实了 GCs 在根瘤中的分子独特性。GCs 中差异调节的大多数基因以前没有赋予功能。与其他转录组分析的比较表明,GCs 与分化成木质部细胞的细胞悬浮液之间存在相似性。这表明 GCs 与发育中的维管束细胞之间存在分子联系,维管束细胞代表潜在的 GC 干细胞。在 3 dpi 时,GC 中的基因表达也与根癌农杆菌诱导的冠瘿瘤相似,根癌农杆菌是一种专门的根生物。

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