菊花CmHsfB1基因的克隆及其功能鉴定

摘要：Hsf转录因子在植物响应多种逆境胁迫中发挥作用，目前菊花中关于Hsf的基因克隆及功能分析报道较少。本研究从菊花中克隆CmHsfB1基因，分析其表达特性，利用农杆菌介导的叶盘转化技术进行遗传转化，并对CmHsfB1基因的功能进行初步研究，主要研究结果如下：同源克隆了CmHsfB1基因，全长为1154bp，其中开放阅读框（ORF）858bp。分析发现，CmHsfB1与拟南芥中AtHsfB1同源性较高。CmHsfB1受到蚜虫取食的强烈诱导，且表现出组织特异性；对CmHsfB1超表达转基因菊花接种蚜虫进行抗蚜性鉴定发现，转基因菊花株系有较强抗蚜性。接种蚜虫后，转基因株系中的H2O2及O2∙-含量比野生型更高。28071
毕业论文关键词：菊花；CmHsfB1；抗蚜性；热激转录因子
Cloning and Functional Analysis of CmHSFB1 Gene in Chrysanthemum
Abstract：Hsf transcription factors have involved in varieties of plants stress resistance however little information about Hsfs in chrysanthemum are available. In this study, we isolated CmHsfB1 from chrysanthemum and analysed its expression feature. To identify the function of CmHsfB1, the overexpression chrysanthemum lines of CmHsfB1 were generated through Agrobacterium tumefaciens mediated leaf disc transformation. The main contents and conclusions are as follows. CmHsfB1 was cloned from chrysanthemum with the method of homologous cloning. The ORF of CmHsfB1 was 1154bp contained a 858 bp open reading frame (ORF). The amino acid sequence analysis showed that, CmHsfB1 belongs to HsfB1 Class with high homology to Arabidopsis thaliana AtHsfB1. The expression of CmHsfB1 was strongly induced by aphid stress, and tissue-specific expression analysis showed a high level of CmHsfB1 in leaf and stem. Overexpression of CmHsfB1 in chrysanthemum increased the tolerance to aphids. CmHsfB1 overexpressing chrysanthemum accelerated aphid-induced H2O2 and O2∙- production. Additionally, the overexpression of CmHsfB1 exhibited elevated expression of ROS-generating and ROS-responsive genes.
Key words: Chrysanthemum morifolium；CmHsfB1；Aphid resistance；Heat shock transcription factor
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 材料    2
1．2 CmHsfB1基因克隆、序列分析及系统进化树构建    2
1．2．2 CmHsfB1基因序列分析    3
1．2．3 系统进化树构建    3
1．3 表达特性分析    3
1．3．1 组织特异性取样    3
1．3．2 蚜虫接种处理及机械损伤处理    3
1．4 CmHsfB1在‘神马’中的遗传转化    4
1．5 CmHsfB1转基因植株抗蚜性鉴定    4
1．5．1 转基因菊花抗蚜性评价    4
1．5．2 转基因菊花接种蚜虫后活性氧（ROS）含量测定    4
1．6 数据分析    4
2 结果与分析    4
2．1 CmHsfB1基因克隆、序列分析    4
2．2 CmHsfB1表达特性    5
2．2．1 不同组织器官中CmHsfB1的表达特性    5
2．2．2 不同组蚜虫胁迫与机械伤害处理下CmHsfB1的表达特性    6
2．3 CmHsfB1转基因菊花分子鉴定    6
2．4 CmHsfB1转基因植株抗蚜性及抗蚜机理    7
2．4．1 CmHsfB1转基因植株抗蚜性评价    7
2．4．2 CmHsfB1转基因株系接种蚜虫后ROS水平    8
3 讨论    9 菊花CmHsfB1基因的克隆及其功能鉴定:http://www.youerw.com/shengwu/lunwen_22783.html