茶树中CsHSF-A1和CsHSF-A8基因的克隆与表达研究
毕业论文关键词：茶树；温度胁迫；HSF转录因子；表达调控
Clone and Expression on CsHSF-A1 and CsHSF-A8 in Camellia sinensis
Abstract:The heat shock transcription factors (HSFs) widely exist in the whole biology world, HSFs regulate heat stress response by regulating the expression of heat shock proteins (Hsps). The CsHSF-A1 and CsHSF-A8 gene sequences, which encoding two HSF transcription factors, were isolated based on transcriptome and genome sequence data of tea plant (Camellia sinensis). The full-length of the open reading frame of CsHSF-A1 was 1032 bp, which encoded 343 amino acids. The full-length of the open reading frame of CsHSF-A8 was 837 bp, which encoded 278 amino acids. Quantitative real-time PCR analysis of the expression profiles showed that the CsHSF-A1 and CsHSF-A8 genes could be induced by low (4 ℃)or high (38 ℃)temperature treatments. The expression levels of CsHSF-A1 and CsHSF-A8 genes exhibited difference in tea plant cultivar of ‘Anjibaicha’. CsHSF-A1 only to high stress, CsHSF-A8 to low or high treatments. This study provided a relatively detailed summary of HSFs in tea plant and may serve as a reference for studies on the mechanism of temperature stress regulation by CsHSFs.
Key words: Camellia sinensis；temperature stress；HSF transcription factor；expression profiles
目 录
摘要 1
关键词 1
Abstract 1
Key words 1
引言 2
1 材料与方法 2
1．1 实验材料 2
1．1．1 植物材料，生长环境 2
1．1．2 菌株与质粒 3
1．1．3 逆境处理 3
1．2 实验方法 3
1．2．1 RNA的提取以及cDNA的合成 3
1．2．2 茶树CsHSF-A1和CsHSF-A8转录因子基因的检索及引物设计 3
1．2．3 转录因子基因的克隆 3
1．2．4 cDNA的回收 3
1．2．5 连接与转化 4
1．2．6 实时定量PCR分析 4
2 结果与分析 5
2．1 茶树CsHSF-A1和CsHSF-A8基因的克隆 5
2．2 茶树CsHSF-A1和CsHSF-A8基因在不同温度下胁迫下的表达情况 6
3 讨论 7
致谢 8
参考文献 9
茶树中CsHSF-A1和CsHSF-A8基因的克隆与表达研究
引言
茶是世界上具有非常高经济价值的一种非酒精类饮品[1]，近期由于茶的健康价值以及药用性能茶树引起了人们广泛的关注。茶树是一种多年生常绿木本植物，起源于我国西南部亚热带地区的湿润多雨的原始森林中[2-4]。高盐、重金属、干旱、低温、高温等非生物胁迫会影响到茶树的生长，这些非生物胁迫影响了茶叶的颜色、道以及香气[5-8]。在茶树的进化过程之中，由于茶树所处的地理环境使得其形成了喜温怕寒的生活习性[9]，茶产区的众多环境条件之中，温度对茶树生长和茶叶生产具有非常大的影响。正是因为如此，研究茶树中对非生物胁迫有重要调控作用的转录因子非常重要。
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