Huanca-Mamani Wilson, Garcia-Aguilar Marcelina, León-Martínez Gloria, Grossniklaus Ueli, Vielle-Calzada Jean-Philippe
Laboratory of Reproductive Development and Apomixis, Department of Genetic Engineering, Centro de Investigación y de Estudios Avanzados-Unidad Irapuato, Apartado Postal 629, CP 36500, Irapuato, Guanajuato, Mexico.
Proc Natl Acad Sci U S A. 2005 Nov 22;102(47):17231-6. doi: 10.1073/pnas.0508186102. Epub 2005 Nov 14.
Chromatin-remodeling factors regulate the establishment of transcriptional programs during plant development. Although 42 genes encoding members of the SWI2/SNF2 family have been identified in Arabidopsis thaliana, <10 have been assigned a precise function on the basis of a mutant phenotype, and none have been shown to play a specific role during the gametophytic phase of the plant life cycle. A. thaliana chromatin-remodeling protein 11 (CHR11) encodes an imitation of switch (ISWI)-like chromatin-remodeling protein abundantly expressed during female gametogenesis and embryogenesis in Arabidopsis. To determine the function of CHR11 in wild-type plants, we introduced a hairpin construct leading to the production of double-stranded RNA, which specifically degraded the endogenous CHR11 mRNA by RNA interference (RNAi). Transcription of the RNAi-inducing hairpin RNA was driven by either a constitutive cauliflower mosaic virus 35S promoter (CaMV35S) acting at most stages of the sporophytic phase or a newly identified specific promoter acting at the onset of the female gametophytic phase (pFM1). All adult transformants that constitutively lacked sporophytic CHR11 activity showed reduced plant height and small cotyledonary embryos with limited cell expansion. In contrast, RNAi lines in which CHR11 was specifically silenced at the onset of female gametogenesis (megagametogenesis) had normal height and embryo size but had defective female gametophytes arrested before the completion of the mitotic haploid nuclear divisions. These results show that CHR11 is essential for haploid nuclear proliferation during megagametogenesis and cell expansion during the sporophytic phase, demonstrating the functional versatility of SWI2/SNF2 chromatin-remodeling factors during both generations of the plant life cycle.
染色质重塑因子在植物发育过程中调控转录程序的建立。尽管在拟南芥中已鉴定出42个编码SWI2/SNF2家族成员的基因,但基于突变体表型,只有不到10个基因被赋予了确切功能,而且尚无基因被证明在植物生命周期的配子体阶段发挥特定作用。拟南芥染色质重塑蛋白11(CHR11)编码一种类似模仿开关(ISWI)的染色质重塑蛋白,在拟南芥雌配子发生和胚胎发生过程中大量表达。为了确定CHR11在野生型植物中的功能,我们引入了一个发夹结构,导致产生双链RNA,该双链RNA通过RNA干扰(RNAi)特异性降解内源性CHR11 mRNA。RNA干扰诱导发夹RNA的转录由在孢子体阶段大多数时期起作用的组成型花椰菜花叶病毒35S启动子(CaMV35S)或在雌配子体阶段开始时起作用的新鉴定的特异性启动子(pFM1)驱动。所有组成型缺乏孢子体CHR11活性的成年转化体植株高度降低,子叶胚较小,细胞扩展受限。相比之下,在雌配子发生(大配子发生)开始时CHR11被特异性沉默的RNAi株系具有正常的高度和胚胎大小,但雌配子体有缺陷,在有丝分裂单倍体核分裂完成之前就停止发育。这些结果表明,CHR11对于大配子发生过程中的单倍体核增殖以及孢子体阶段的细胞扩展至关重要,证明了SWI2/SNF2染色质重塑因子在植物生命周期两代中的功能多样性。