Comparative transcriptional profiling analysis of the two daughter cells from tobacco zygote reveals the transcriptome differences in the apical and basal cells.
Comparative transcriptional profiling analysis of the two daughter cells from tobacco zygote reveals the transcriptome differences in the apical and basal cells.
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烟草受精卵两个子细胞的比较转录谱分析揭示了顶端细胞和基底细胞的转录组差异
DOI：
10.1186/1471-2229-10-167
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发表时间：
2010-08-11
影响因子：
5.3
通讯作者：
Zhao J
中科院分区：
文献类型：
--
作者：
Hu TX;Yu M;Zhao J
在被子植物中，合子细胞的第一次不对称分裂后，顶端和基底层的子细胞遵循不同的发育途径。对这两种细胞的整体转录组分析对于了解它们的发育差异是必不可少的。然而，由于从高等植物的胚珠和子房中分离二细胞原胚活顶层和基底层细胞的难度较大，对它们的转录组分析尚未见报道。结果本研究建立了一套烟草二细胞原胚活顶层和基底层细胞的分离程序，并结合大芯片筛选技术对两种细胞进行了比较转录组学分析。测序后，我们鉴定出797条差异表达的EST，对应于299个非基因。文库序列分析成功鉴定了烟草胚胎发育和种子发育相关基因的同源性。通过实时定量聚合酶链式反应，我们验证了40个基因的差异表达，其中6个转录本在顶端或基底细胞中特异表达。表达分析还显示，在受精卵分裂后的其中一个子细胞中，一些转录本显示了细胞特异性激活。通过原位杂交(ISH)进一步验证了这些差异表达。组织表达谱分析也揭示了这些候选基因在发育过程中的一些潜在作用。结论成功分离了两细胞原胚顶端和基底层细胞中的一些差异或特异的转录本，对这些转录本的鉴定表明，受精卵分裂后，这两个子细胞具有不同的转录图谱。对这些差异或特异表达的基因进行进一步的功能研究将有助于阐明控制早期胚胎发生的分子机制。
BackgroundIn angiosperm, after the first asymmetric zygotic cell division, the apical and basal daughter cells follow distinct development pathways. Global transcriptome analysis of these two cells is essential in understanding their developmental differences. However, because of the difficulty to isolate thein vivoapical and basal cells of two-celled proembryo from ovule and ovary in higher plants, the transcriptome analysis of them hasn't been reported.ResultsIn this study, we developed a procedure for isolating thein vivoapical and basal cells of the two-celled proembryo from tobacco (Nicotiana tabacum), and then performed a comparative transcriptome analysis of the two cells by suppression subtractive hybridization (SSH) combined with macroarray screening. After sequencing, we identified 797 differentially expressed ESTs corresponding to 299 unigenes. Library sequence analysis successfully identified tobacco homologies of genes involved in embryogenesis and seed development. By quantitative real-time PCR, we validated the differential expression of 40 genes, with 6 transcripts of them specifically expressed in the apical or basal cell. Expression analysis also revealed some transcripts displayed cell specific activation in one of the daughter cells after zygote division. These differential expressions were further validated byin situhybridization (ISH). Tissue expression pattern analysis also revealed some potential roles of these candidate genes in development.ConclusionsThe results show that some differential or specific transcripts in the apical and basal cells of two-celled proembryo were successfully isolated, and the identification of these transcripts reveals that these two daughter cells possess distinct transcriptional profiles after zygote division. Further functional work on these differentially or specifically expressed genes will promote the elucidation of molecular mechanism controlling early embryogenesis.
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影响因子：
64.8
作者：
Blilou, I;Xu, J;Scheres, B
通讯作者：
Scheres, B
影响因子：
1.1
作者：
Kim, Dong Han
通讯作者：
Kim, Dong Han
影响因子：
4.8
作者：
Kawai-Yamada, M;Saito, Y;Uchimiya, H
通讯作者：
Uchimiya, H
影响因子：
11.6
作者：
Leonhardt, N;Kwak, JM;Schroeder, JI
通讯作者：
Schroeder, JI
影响因子：
11.6
作者：
Lu, PZ;Porat, R;ONeill, SD
通讯作者：
ONeill, SD