番茄sr蛋白调控致病疫霉抗性的初步研究【字数:12815】
目录
摘要Ⅲ
关键词Ⅲ
AbstractⅣ
引言
引言1
文献综述1
第一章 番茄SR家族基因的鉴定和分析 3
1 材料与方法4
1.1 番茄SR家族基因预测4
1.2 系统发育树构建4
1.3 蛋白结构域分析4
1.4 蛋白质理化性质分析 4
2 结果与分析5
2.1 番茄SR家族基因的鉴定5
2.2 番茄SR家族系统发育树分析5
2.3 番茄SR蛋白的结构域分析 6
2.4 番茄SR蛋白的理化性质分析7
3 讨论8
第二章 番茄SR30负调控植物免疫 10
1 材料与方法11
1.1 供试植物与菌株11
1.2 试剂及培养基11
1.2.1 LB液体及固体培养基11
1.2.2 RSAV8固体培养基11
1.3 大肠杆菌感受态制备和转化方法11
1.3.1 制备大肠杆菌感受态 11
1.3.2 大肠杆菌热激转化 12
1.4 农杆菌电转感受态制备和转化方法12
1.4.1 农杆菌电转感受态细胞的制备 12
1.4.2 农杆菌电激转化 13
1.5 目的基因在烟草中瞬时表达13
1.5.1 目的基因重组质粒构建13
1.5.2 农杆菌介导的瞬时表达 13
1.6 烟草叶片接种辣椒疫霉和寄生疫霉 14
1.7 RNA的提取和逆转录14
1.7.1 RNA的提取14
1.7.2 RNA的逆转录 14
1.8 荧光定量PCR检测15
1.9 番茄SR基因转录量分析15
1.10 番茄沉默载体构建15
1.11 病毒介导的基因沉默和沉默效率检测16
1.12 沉默植株感病性检测16
2 结果与分析16
2.1 SlSRs在疫霉菌侵染番茄早期差异表达16 *51今日免费论文网|www.51jrft.com +Q: @351916072@
2.2 过表达SlSR30可促进疫霉菌的侵染17
2.3 沉默SlSR30及沉默效率的检测19
2.4 番茄沉默SlSR30抑制疫霉菌侵染19
3 讨论20
参考文献22
附录24
致谢25
番茄SR蛋白调控致病疫霉抗性的初步研究
摘 要
由致病疫霉(Phytophthora infestans)侵染马铃薯、番茄等茄科作物引发的晚疫病是威胁全球粮食安全的毁灭性病害之一,曾造成19世纪的爱尔兰饥馑,每年给世界造成高达百亿美元的经济损失,因此对该病害的有效防控是我国农业发展的重要环节。由于致病疫霉的特殊的生活史,传统的病害防治无法有效防控病害,因此从致病疫霉与寄主互作角度深入解析致病疫霉的致病机理,可为寻找新的防治策略提供重要参考。
可变剪切是指一个单链的premRNA产生多个转录形式的过程,由此产生的不同的mRNA转录本可能被翻译成不同的蛋白质构体,是蛋白质功能多样性的重要来源之一。可变剪切通过影响基因的表达调控和转录多样性在生物的生长发育和环境适应中具有重要作用。SR蛋白是一类可以和剪切复合体互作的蛋白因子,在整个可变剪切早期的剪切体组装过程中起着重要的作用。本研究主要以番茄为研究材料,利用生物信息学技术鉴定到番茄SR蛋白家族,继而探究致病疫霉与番茄互作过程中,番茄SR蛋白的功能。
PHYTOPHTHORA INFESTANS RESISTANCE REGULATED BY SR PROTEIN FAMILIES IN TOMATO
ABSTRACT
Phytophthora infestans infects solanaceae crops such as potato and tomato is one of the destructive diseases threatening global food security. It caused famine in Ireland in the 19th century and caused economic losses of up to ten billion dollars every year. Due to the special life history of P. infestans, traditional disease control cannot effectively control the disease. Therefore, the analysis of the pathogenic mechanism of P. infestans between phytophthora and host plant can provide an important reference for the search for new control strategies.
Alternative splicing refers to the process of producing multiple transcriptions of a single strand of pre mRNA. Different mRNA transcripts may be translated into different protein structures, which is one of the important sources of protein functional diversity. Alternative splicing plays an important role in biological growth and environmental adaptation by influencing gene expression regulation and transcription diversity. SR protein is a kind of protein factor that can interact with splicing complex, and plays an important role in the whole assembly process of splicing body in the early stage of alternative splicing. In this study, tomato was used as the research material to find the tomato SR protein family by using bioinformatics technology, and then to explore the function of tomato SR protein in the process of interaction between Phytophthora and tomato.
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