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利用来自 Solanum chacoense 胚珠的 7.7 K cDNA 微阵列对茄科物种的受精和早期种子发育事件进行转录谱分析。

Transcription profiling of fertilization and early seed development events in a solanaceous species using a 7.7 K cDNA microarray from Solanum chacoense ovules.

机构信息

Département de Sciences Biologiques, Institut de Recherche en Biologie Végétale, Université de Montréal, 4101 rue Sherbrooke est, Montréal, Québec, H1X 2B2, Canada.

出版信息

BMC Plant Biol. 2010 Aug 12;10:174. doi: 10.1186/1471-2229-10-174.

Abstract

BACKGROUND

To provide a broad analysis of gene expression changes in developing embryos from a solanaceous species, we produced amplicon-derived microarrays with 7741 ESTs isolated from Solanum chacoense ovules bearing embryos from all developmental stages. Our aims were to: 1) identify genes expressed in a tissue-specific and temporal-specific manner; 2) define clusters of genes showing similar patterns of spatial and temporal expression; and 3) identify stage-specific or transition-specific candidate genes for further functional genomic analyses.

RESULTS

We analyzed gene expression during S. chacoense embryogenesis in a series of experiments with probes derived from ovules isolated before and after fertilization (from 0 to 22 days after pollination), and from leaves, anthers, and styles. From the 6374 unigenes present in our array, 1024 genes were differentially expressed (>or= +/- 2 fold change, p value <or= 0.01) in fertilized ovules compared to unfertilized ovules and only limited expression overlap was observed between these genes and the genes expressed in the other tissues tested, with the vast majority of the fertilization-regulated genes specifically or predominantly expressed in ovules (955 genes). During embryogenesis three major expression profiles corresponding to early, middle and late stages of embryo development were identified. From the early and middle stages, a large number of genes corresponding to cell cycle, DNA processing, signal transduction, and transcriptional regulation were found. Defense and stress response-related genes were found in all stages of embryo development. Protein biosynthesis genes, genes coding for ribosomal proteins and other components of the translation machinery were highly expressed in embryos during the early stage. Genes for protein degradation were overrepresented later in the middle and late stages of embryo development. As expected, storage protein transcripts accumulated predominantly in the late stage of embryo development.

CONCLUSION

Our analysis provides the first study in a solanaceous species of the transcriptional program that takes place during the early phases of plant reproductive development, including all embryogenesis steps during a comprehensive time-course. Our comparative expression profiling strategy between fertilized and unfertilized ovules identified a subset of genes specifically or predominantly expressed in ovules while a closer analysis between each consecutive time point allowed the identification of a subset of stage-specific and transition-specific genes.

摘要

背景

为了对茄科植物胚胎发育过程中的基因表达变化进行广泛分析,我们利用从 Solanum chacoense 胚珠中分离的 7741 个 EST 制作了扩增子衍生的微阵列,这些胚珠带有来自所有发育阶段的胚胎。我们的目标是:1)鉴定组织特异性和时间特异性表达的基因;2)定义具有相似时空表达模式的基因簇;3)鉴定用于进一步功能基因组分析的阶段特异性或过渡特异性候选基因。

结果

我们通过一系列实验分析了 S. chacoense 胚胎发生过程中的基因表达,这些实验使用的探针来自授粉后 0 至 22 天(授粉后)的未受精和受精胚珠以及叶片、花药和花柱中分离得到。在我们的微阵列中,6374 个基因中,有 1024 个基因(差异表达倍数>或=2 倍,p 值<或=0.01)在受精胚珠中与未受精胚珠相比差异表达,并且这些基因与在其他测试组织中表达的基因之间观察到的表达重叠有限,绝大多数受精调节基因在胚珠中特异性或主要表达(955 个基因)。在胚胎发生过程中,确定了三个与胚胎发育早期、中期和晚期相对应的主要表达谱。从早期和中期开始,发现了大量与细胞周期、DNA 处理、信号转导和转录调控相关的基因。在胚胎发育的所有阶段都发现了与防御和应激反应相关的基因。在胚胎发生的早期阶段,蛋白质生物合成基因、核糖体蛋白和翻译机制的其他成分的编码基因高度表达。在胚胎发生的中晚期,降解蛋白的基因表达增加。正如预期的那样,贮藏蛋白转录本在胚胎发育的晚期阶段主要积累。

结论

我们的分析首次在茄科植物中研究了植物生殖发育早期阶段发生的转录程序,包括在全面时间过程中发生的所有胚胎发生步骤。我们在受精和未受精胚珠之间进行的比较表达谱分析鉴定了一组在胚珠中特异性或主要表达的基因,而对每个连续时间点之间的更密切分析则鉴定了一组阶段特异性和过渡特异性基因。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a38d/3095305/bc6b388bf35d/1471-2229-10-174-1.jpg

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