Dissertation > Excellent graduate degree dissertation topics show

Production of AtPP2-A1 and Its Promoter Transgenic Arabidopsis and Preliminery Study on Trichome Defense Function

Author: LiangYuan
Tutor: DongHanSong
School: Nanjing Agricultural College
Course: Plant Pathology
Keywords: AtPP2-A1 Promoter Staining Fluorescence detection ParA1 Tobacco trichomes
CLC: S432.1
Type: Master's thesis
Year: 2009
Downloads: 16
Quote: 0
Read: Download Dissertation

Abstract

Arabidopsis AtPP2 phloem protein is a class with a the phloem tissue specific plant lectins, studies have shown that it may play a decisive role in the defense response of the plant phloem (plant phloem-based defense, PBD), but the specific mechanism of action is still I do not know. Transgenic Arabidopsis plants AtPP2-A1 and its promoter, obtained by vacuum the tidbits infiltration method and PCR molecular detection. Its initial positioning of the technical means by means of staining, GFP fluorescence microscopy observation. ParA1 is a small protein produced by Phytophthora parasitica elicitor can induce tobacco leaf cells hypersensitive reaction. The table fur is located in the the blade outer layer structure, plays the role of the first defensive barrier. The ParA1 whether can cause allergic reactions trichome cells What is the relationship? The reaction with blades of trichomes ParA1 other organizations reaction? Article focuses on a preliminary study of these issues. 1 Arabidopsis AtPP2-A1 gene and its promoter turn the generation and identification of the genetic material to specific promoter AtPP2-A1 gene cloned from Arabidopsis genomic DNA to construct a plant expression vector pBI121. The use of the Agrobacterium (Agrobacerium tumefaciens) EHA105-mediated transformation of Arabidopsis Col-0 ecotype plants screened positive transgenic Arabidopsis plants. PCR analysis of conclusive evidence that exogenous GUS gene has been transferred to Arabidopsis. Transgenic Arabidopsis, GUS staining results showed that the promoter and the CaMV35S promoter capable of driving expression of the GUS gene, the former only in Arabidopsis phloem-specific expression, while the CaMV35S promoter driven GUS gene expression in a constitutive expressed. Thus proving that the promoter in the phloem-specific and efficient expression characteristics, so that they can apply the Arabidopsis insect-resistant transgenic research. In order to further study the subcellular localization of the AtPP2-A1 gene, a gene from Arabidopsis AtPP2-A1 connected into a plant expression vector pBI121, labeled with a fluorescent protein, constructed plant expression plasmid pBI121, AtPP2-A1 :: GFP, and by Agrobacterium (Agrobacterium tumefaciens) the strain EHA105 mediated transformation of Arabidopsis thaliana ecotype Col-0. Kanamycin-resistant transgenic Arabidopsis plants by PCR and fluorescence microscopy to prove AtPP2-A1 gene was integrated into the Arabidopsis genome. 2.ParA1 induction of the the tobacco epidermal hair HR reactive protein to stimulate sub ParA1 injection tobacco leaves, can trigger the the plants hypersensitive cell death (hypersensitve cell death HCD). Tobacco leaves after spraying, the ParA1 can lead to plant produce micro-sensitized reaction, and accompanied by the occurrence of active oxygen burst (ROS burst), and chromatin condensation (Chromatin condensation) phenomena. Trichomes earliest contacts ParA1 as a natural barrier of the outer layer of the tobacco to elicitor and participate induced micro-HR. As an entry point to the important signaling molecules in the micro-HR during H2O2 and cell chromatin condensation, trichomes ParA1 elicitor-induced response. DCFH-DA staining and fluorescence observation, ParA1 processing trichomes within 1h after the burst of reactive oxygen species, occurs earlier than the mesophyll cells 2h, and the generation of reactive oxygen conduction to the mesophyll cells. The DAPI fluorescence analysis showed that the table than mesophyll cells as early as the occurrence of chromatin condensation within the fur 5h. Moss phenol blue and neutral red staining with plasmolysis experiments proved after in ParA1 handling 1h to 12h, the cell death to the bottom of the corresponding cells from trichomes gradually spread, trichome cell death occurred earlier than the mesophyll cells about 5h and fur for 5h each time period the number of cell death was significantly higher than the mesophyll cells. RT-PCR was used to detect the the HR reaction of marker genes PAL hin1, and hsr203 of the expression of three genes in 12h after ParAl processing trichomes have expression of gene expression in the mesophyll cells compared table fur slightly later. These results indicate that, HR signal conduction from the the table fur to the mesophyll cells.

Related Dissertations

CLC: > Agricultural Sciences > Plant Protection > Pest and Disease Control > Plant diseases and their prevention > Plant Pathology
© 2012 www.DissertationTopic.Net Mobile