Nuccio M L, Li Z, Hsieh T F, Thomas T L
Department of Biology, Texas A&M University, College Station 77843-3258, USA.
SAAS Bull Biochem Biotechnol. 1996;9:23-8.
Although most of the major discernible morphogenetic events in plants occur after germination, the overall architectural pattern of the mature plant is established during early events of embryogenesis. So far, few genes that are expressed specifically during embryogenesis have been identified. This is due primarily to technical difficulties associated with the mass ratios of the embryo and the surrounding maternal tissue and to the lack of molecular and cellular markers to direct screening efforts. We have developed a series of molecular approaches to study the early events of embryogenesis. These include 'virtual subtraction' of a cDNA library with high specific-activity cDNA probes generated from both seed and non-seed tissue, PCR amplification of gene family members from an immature seed cDNA library using primers specific to conserved domains, differential display analysis of mRNA populations and high throughput expressed sequence tag (EST) analysis. These techniques have led to the identification and isolation of several novel seed-specific cDNAs.
尽管植物中大多数可识别的主要形态发生事件发生在萌发之后，但成熟植物的整体架构模式却是在胚胎发生的早期事件中建立起来的。到目前为止，很少有在胚胎发生过程中特异性表达的基因被鉴定出来。这主要是由于与胚胎和周围母体组织的质量比相关的技术难题，以及缺乏用于指导筛选工作的分子和细胞标记物。我们已经开发出一系列分子方法来研究胚胎发生的早期事件。这些方法包括用从种子和非种子组织产生的高比活性cDNA探针“虚拟扣除”cDNA文库，使用保守结构域特异性引物从未成熟种子cDNA文库中PCR扩增基因家族成员，对mRNA群体进行差异显示分析以及高通量表达序列标签（EST）分析。这些技术已导致鉴定和分离出几个新的种子特异性cDNA。
