Cervigni Gerardo D L, Paniego Norma, Pessino Silvina, Selva Juan P, Díaz Marina, Spangenberg Germán, Echenique Viviana
Centro de Recursos Naturales Renovables de la Zona Semiárida- CONICET, Camino de La Carrindanga Km 7,0, Bahia Blanca, Argentina.
Plant Mol Biol. 2008 May;67(1-2):11-23. doi: 10.1007/s11103-008-9305-9. Epub 2008 Mar 3.
The molecular nature of gene expression during the initiation and progress of diplosporous apomixis is still unknown. Moreover, the basis of the close correlation between diplospory and polyploidy is not clarified yet. A comparative expression analysis was performed based on expressed sequence tags (ESTs) sequencing and differential display in an Eragrostis curvula diplosporous tetraploid genotype (T, 4x apo), a sexual diploid derivative obtained from tissue culture (D, 2x sex) and an artificial sexual tetraploid obtained from the diploid seeds after colchicine treatment (C, 4x sex). From a total of 8,884 unigenes sequenced from inflorescence-derived libraries, 112 (1.26%) showed significant differential expression in individuals with different ploidy level and/or variable reproductive mode. Independent comparisons between plants with different reproductive mode (same ploidy) or different ploidy level (same reproductive mode) allowed the identification of genes modulated in response to diplosporous development or polyploidization, respectively. Surprisingly, a group of genes (Group 3) were differentially expressed or silenced only in the 4x sex plant, presenting similar levels of expression in the 4x apo and the 2x sex genotypes. A group of randomly selected differential genes was validated by QR-PCR. Differential display analysis showed that in general the 4x apo and 4x sex expression profiles were more related and different from the 2x sex one, but confirmed the existence of Group 3-type genes, in both inflorescences and leaves. The possible biological significance for the occurrence of this particular group of genes is discussed. In silico mapping onto the rice genome was used to identify candidates mapping to the region syntenic to the diplospory locus.
二倍体无融合生殖起始和进程中基因表达的分子本质仍然未知。此外,二倍体孢子生殖与多倍体之间紧密关联的基础也尚未阐明。基于表达序列标签(EST)测序和差异显示,对弯叶画眉草的二倍体孢子生殖四倍体基因型(T,4x apo)、从组织培养获得的有性二倍体衍生物(D,2x sex)以及用秋水仙碱处理二倍体种子后获得的人工有性四倍体(C,4x sex)进行了比较表达分析。从花序来源文库测序的总共8884个单基因中,112个(1.26%)在不同倍性水平和/或不同生殖模式的个体中表现出显著差异表达。对具有不同生殖模式(相同倍性)或不同倍性水平(相同生殖模式)的植株进行独立比较,分别鉴定出响应二倍体孢子发育或多倍化而被调控的基因。令人惊讶的是,一组基因(第3组)仅在4x sex植株中差异表达或沉默,在4x apo和2x sex基因型中表现出相似的表达水平。通过定量实时聚合酶链反应(QR-PCR)对一组随机选择的差异基因进行了验证。差异显示分析表明,总体而言,4x apo和4x sex的表达谱更相关,与2x sex的表达谱不同,但在花序和叶片中均证实了第3组类型基因的存在。讨论了这一特定基因组出现的可能生物学意义。利用水稻基因组的电子定位来鉴定映射到与二倍体孢子位点同线区域的候选基因。