Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, 100193, Beijing, People's Republic of China.
Mol Biol Rep. 2010 Feb;37(2):745-53. doi: 10.1007/s11033-009-9590-7. Epub 2009 Jul 2.
An alfalfa cDNA library induced by salt stress was constructed by suppression subtraction hybridization (SSH) technology. Total RNA from 10-day-old seedlings was used as a "driver," and total RNA from seedlings induced by salt was used as a "tester". One hundred and nineteen clones identified as positive clones by reverse Northern dot-blotting resulted in 82 uni-ESTs comprised of 16 contigs and 66 singletons. Blast analysis of deduced protein sequences revealed that 51 ESTs had identity similar to proteins with known function, while 24 could not be annotated at all. Most of the annotated sequences were homologous to genes involved in abiotic or biotic stress in plants. Among these proteins, beta-amylase, fructose-1,6-bisphosphate, aldolase, and sucrose synthase are related to osmolyte synthesis; a CCCH-type zinc finger protein, DNA binding protein, His-Asp phosphotransfer protein, and the RelA/SpoT protein partake in transcription regulation and signal transduction; and ribulose-l,5-bisphosphate carboxylase/oxygenase, chlorophyll a/b binding proteins, and an early light-inducible proteins are related to photosynthesis. In addition, several ESTs, similar to genes from other plant species, closely involved in salt stress were isolated from alfalfa, such as an aquaporin protein, a late embryogenesis-abundant protein, and glutathione peroxidase.
采用抑制性消减杂交(SSH)技术构建了盐胁迫诱导的紫花苜蓿 cDNA 文库。以 10 日龄幼苗总 RNA 为“驱动子”,盐诱导的幼苗总 RNA 为“测试子”。经反向 Northern dot-blotting 鉴定为阳性克隆的 119 个克隆导致 82 个单 ESTs,包括 16 个连续序列和 66 个单序列。推测蛋白序列的 Blast 分析表明,51 个 EST 与具有已知功能的蛋白具有相似的身份,而 24 个 EST 根本无法注释。大多数注释序列与植物中涉及非生物或生物胁迫的基因同源。在这些蛋白质中,β-淀粉酶、果糖-1,6-二磷酸、醛缩酶和蔗糖合酶与渗透剂合成有关;CCCH 型锌指蛋白、DNA 结合蛋白、His-Asp 磷酸转移蛋白和 RelA/SpoT 蛋白参与转录调控和信号转导;核酮糖-1,5-二磷酸羧化酶/加氧酶、叶绿素 a/b 结合蛋白和早期光诱导蛋白与光合作用有关。此外,还从紫花苜蓿中分离出了几种与其他植物物种密切相关的 ESTs,如水通道蛋白、晚期胚胎丰富蛋白和谷胱甘肽过氧化物酶等,这些 ESTs与盐胁迫密切相关。