Xinjiang Key Laboratory of Biological Resources and Genetic Engineering, College of Life Science and Technology, Xinjiang University, Urumqi 830046, China.
Mol Biol Rep. 2011 Nov;38(8):5285-95. doi: 10.1007/s11033-011-0678-5. Epub 2011 Jan 19.
To identify genes expression in Chenopodium album exposed to NaCl stress and screen ESTs related to salt stress, a subtractive suppression hybridization (SSH) library of C. album under salt stress was constructed in the present study. Random EST sequencing produced 825 high-quality ESTs with GenBank ID GE746311-GE747007, which had 301 bp of average size and were clustered into 88 contigs and 550 singletons. They were classified into 12 categories according to their function annotations. 635 ESTs (76.97%) showed similarities to gene sequences in the non-redundancy database, while 190 ESTs (23.03%) showed low or no similarities. The transcriptional profiles of 56 ESTs randomly selected from 347 unknown or novel ESTs of SSH library under varying NaCl concentration and at different time points were analyzed. The results indicated that a high proportion of tested ESTs were activated by salt stress. Four in 56 ESTs responded to NaCl were also enhanced in expression level when exposed to ABA and PEG stresses. The above four ESTs were validated by northern blotting which was consistent with the results of RT-PCR. The results suggested that genes corresponded to these ESTs might be involved in stress response or regulation. The complete sequences and detailed function of these ESTs need to be further studied.
为了鉴定盐胁迫下藜（Chenopodium album）中的基因表达并筛选与盐胁迫相关的 ESTs，本研究构建了藜盐胁迫消减杂交文库。随机 EST 测序产生了 825 条高质量 EST，GenBank ID 为 GE746311-GE747007，平均大小为 301bp，聚类为 88 个 contigs 和 550 个 singletons。根据功能注释，它们被分为 12 类。635 条 EST（76.97%）与非冗余数据库中的基因序列具有相似性，而 190 条 EST（23.03%）具有较低或无相似性。从 SSH 文库的 347 个未知或新的 EST 中随机选择的 56 条 EST 的转录谱在不同 NaCl 浓度和不同时间点进行了分析。结果表明，测试的 ESTs 中有很大一部分被盐胁迫激活。在 56 条 EST 中，有 4 条对 NaCl 有响应，在暴露于 ABA 和 PEG 胁迫时其表达水平也增强。通过 northern blot 验证了这 4 个 EST，与 RT-PCR 的结果一致。结果表明，这些 EST 对应的基因可能参与了应激反应或调节。这些 ESTs 的完整序列和详细功能需要进一步研究。
