亚洲棉司笃克氏棉及其体细胞杂种纤维发育调控差异表达基因的克隆及分析

Cloning and analysis of the differentially expressed genes regulating fiber development in Gossypium arboreum, Gossypium stocksii and their somatic hybrid摘要棉花纤维是纺织工业最主要的原材料，纤维的起始和伸长是决定棉花纤维产量和品质的关键因素。尽管转录组和功能基因组学的研究已为我们提供了一些线索，但是纤维发育机制仍不清晰。利用cDNA-AFLP对三个棉种纤维发育过程进行比较转录组分析，一共分离到到598个差异表达基因；本项目拟通过生物信息学分析和表达分析进一步筛选、克隆调控纤维发育相关基因；已分离到一个果胶甲基酯酶抑制剂基因（PMEI），该基因在纤维起始和快速伸长阶段特异表达，通过生物信息学分析和超量表达转基因功能验证进一步揭示基因对纤维发育的调控作用和分子机制。76532

毕业论文关键词：棉花纤维；果胶甲基酯酶抑制剂；克隆；表达；转基因；拟南芥

Abstract Cotton fiber is the main raw material of textile industry, fiber initiation and elongation are the key determinants of fiber output and quality in cotton。 Although transcriptional profiling and functional genomics provide some data, the mechanism of fiber development is still not well understood。 The cDNA-AFLP technique was used to identify the differentially expressed genes in fiber developmental stages, with Gossypium arboreum, Gossypium stocksii and their somatic hybrid as the research materials。 And a total of 598 differentially expressed genes were isolated。 The project intends to further screen and clone the genes involved in fiber development by bioinformatic and expression analysis。 A pectin methylesterase inhibitor gene (PMEI) was isolated。 It was specifically expressed in fiber initiation and elongation stage。 Bioinformatic analysis and overexpression strategies will be applied for further revealing the functional and molecular mechanisms to fiber development。

Key Words：Cotton fiber；Pectin methylesterase inhibitor；；clone；expression；transgenic ；Arabidopsis thaliana

摘要 3

Abstract 3

1 引言 5

1。1研究的意义 5

1。2研究的历史背景 5

1。2。1 棉花纤维 5

1。2。2果胶甲基酯酶（PME）及其抑制剂（PMEI） 6

2 材料与方法 6

2。1 实验材料 6

2。2 实验试剂 7

2。2。1 栽培试剂 7

2。2。2 RT-PCR试剂 7

2。2。3 QRT-PCR试剂 7

2。3 实验仪器设备 7

2。4研究方法 7

2。4。1cDNA-AFLP分离亚洲棉、司笃克氏棉和体细胞杂种纤维发育阶段的差异表达基因 8

2。4。2 目的基因克隆及生物信息学分析 8

2。4。3目的基因的表达分析 9

2。4。4 超量表达载体构建及拟南芥转化 10

2。4。4。1 电转化方法步骤亚洲棉司笃克氏棉及其体细胞杂种纤维发育调控差异表达基因的克隆及分析:http://www.youerw.com/shengwu/lunwen_87802.html