National Engineering Laboratory for Tree Breeding, College of Biological Sciences and Technology, Beijing Forestry University, Beijing, P R China.
BMC Genomics. 2012 Jun 29;13:286. doi: 10.1186/1471-2164-13-286.
Drought is one of the main environmental factors limiting tree growth and productivity of plantation forests worldwide. Populus hopeiensis Hu et Chow is one of the most important commercial plantation tree species in China. However, the genes controlling drought tolerance in this species have not been identified or characterized. Here, we conducted differential expression analyses and identified a number of genes that were up- or downregulated in P. hopeiensis during water stress. To the best of our knowledge, this is the first comprehensive study of differentially expressed genes in water-stressed P. hopeiensis.
Using the cDNA-AFLP detection technique, we used 256 primer combinations to identify differentially expressed genes in P. hopeiensis during water stress. In total, 415 transcript derived-fragments (TDFs) were obtained from 10× deep sequencing of 473 selected TDFs. Of the 415 TDFs, 412 were annotated by BLAST searches against various databases. The majority of these genes encoded products involved in ion transport and compartmentalization, cell division, metabolism, and protein synthesis. The TDFs were clustered into 12 groups on the basis of their expression patterns. Of the 415 reliable TDFs, the sequences of 35 were homologous to genes that play roles in short or long-term resistance to drought stress. Some genes were further selected for validation of cDNA-AFLP expression patterns using real-time PCR analyses. The results confirmed the expression patterns that were detected using the cDNA-AFLP technique.
The cDNA-AFLP technique is an effective and powerful tool for identifying candidate genes that are differentially expressed under water stress. We demonstrated that 415 TDFs were differentially expressed in water-stressed poplar. The products of these genes are involved in various biological processes in the drought response of poplar. The results of this study will aid in the identification of candidate genes of future experiments aimed at understanding this response of poplar.
干旱是限制世界范围内人工林树木生长和生产力的主要环境因素之一。毛白杨是中国最重要的商业造林树种之一。然而,该物种控制耐旱性的基因尚未被鉴定或表征。在这里,我们进行了差异表达分析,鉴定出了在水分胁迫下毛白杨中上调或下调的许多基因。据我们所知,这是首次对水分胁迫下毛白杨差异表达基因进行的全面研究。
使用 cDNA-AFLP 检测技术,我们使用 256 个引物组合来鉴定水分胁迫下毛白杨中的差异表达基因。总共从 473 个选定的 TDF 中进行了 10×深度测序,获得了 415 个转录衍生片段(TDF)。在 415 个 TDF 中,412 个通过与各种数据库的 BLAST 搜索进行注释。这些基因大多数编码参与离子运输和区室化、细胞分裂、代谢和蛋白质合成的产物。根据表达模式将 TDF 聚类为 12 组。在 415 个可靠的 TDF 中,有 35 个序列与在短期或长期抗旱中起作用的基因同源。一些基因进一步选择使用实时 PCR 分析验证 cDNA-AFLP 表达模式。结果证实了使用 cDNA-AFLP 技术检测到的表达模式。
cDNA-AFLP 技术是一种有效的鉴定在水分胁迫下差异表达的候选基因的有力工具。我们证明了在水分胁迫下毛白杨中有 415 个 TDF 差异表达。这些基因的产物参与毛白杨抗旱反应中的各种生物学过程。本研究的结果将有助于鉴定未来旨在了解毛白杨这种反应的候选基因。