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Zn是植物必需的微量元素,同时也是近年来造成环境污染的重金属元素之一.为了从基因表达水平揭示小麦Zn胁迫响应的分子机制,本研究利用抑制差减杂交(suppression subtractivehybridization,SSH)技术构建了Zn胁迫(0.5 mmol/L,1 mmol/L)下小麦的正反向SSH文库.从正反向文库中随机挑选阳性单克隆,并利用通用引物T7/Sp6对其进行验证.结果显示,正反库中分别获得307和821个EST序列,其片段长度在200～1 000 bp之间,它们反映了Zn胁迫下特异响应的基因.利用BLASTn和BLASTx将这些EST序列进行比对分析,在正、反库中分别筛选出221和641个uniESTs,其中751个uniESTs被注释(包括正库中193个和反库中558个).这些序列的功能主要涉及信号转导、抗氧化防御、转录与翻译、物质运输、核糖体结构、能量代谢,以及一些功能未知的基因.
Zn is one of the essential trace elements in plants and also one of the heavy metals that cause environmental pollution in recent years.In order to reveal the molecular mechanism of Zn stress response in wheat from the gene expression level, suppression subtractive hybridization (SSH) Positive and negative SSH libraries of wheat under Zn stress (0.5 mmol / L, 1 mmol / L) were selected.The positive clones were randomly selected from the positive and negative libraries and verified with the universal primer T7 / Sp6, 307 and 821 EST sequences were obtained in forward and reverse library, respectively, and the length of the ESTs ranged from 200 to 1 000 bp, which reflected the specific response genes under Zn stress.The EST sequences were compared using BLASTn and BLASTx, 221 and 641 uniESTs were screened out of the positive and negative pools, of which 751 uniESTs were annotated (including 193 in the positive and 558 in the reverse). The functions of these sequences are mainly involved in signal transduction, antioxidant defense, transcription And translation, material transport, ribosome structure, energy metabolism, and some unknown genes.