擬南芥開花調(diào)控基因EFS1的功能研究.pdf

1、河北農(nóng)業(yè)大學(xué)碩士學(xué)位論文擬南芥開花調(diào)控基因EFS1的功能研究姓名：李莉申請學(xué)位級別：碩士專業(yè)：植物學(xué)指導(dǎo)教師：董金皋；邢繼紅2009-06-12Functional Analysis of Flower-Related Gene EFS1 in Arabidopsis thaliana Author: Li Li Supervisor: ProfessorDONG Jin-gao Associate professor XING Ji

2、-hong Major: Botany Abstract Flowering is the most important result of a plant developmental process that controls the transition from vegetative maturity to the reproductive stage. It plays a key role in the phase and

3、quality of development. Investigation of flowering controlling molecular mechanism is important to regulate development and propagation, improve crop’s output and quality. In previous research, we had got a mutant from T

4、-DNA insert library named efs1(early flowering in short mutant 1), which had identified based on Genome Walking and bio-information to be a candidate gene involved with Arabidopsis thaliana flowering regulation. To inves

5、tigate the function and role in the regulation pathway, we designed the complementation and overexpression to confirm that EFS1 certainly belongs to the flowering controlling gene. Semi-quality RT-PCR and GUS histochemic

6、al staining were used to confirm the expression level and tissue localization. RT-PCR and Real-Time PCR were used to confirm the location of EFS1 in the regulating pathway. We have provided a foundation to the further re

7、gulation mechanism research. 1. EFS1 was constructed to the pCAMBIA1300 vector. complementation and overexpression were obtained via Agrobacterium-mediated transformed to efs1 mutant and Col-0. The EFS1 was certainly int

8、egrated to the efs1 mutant as multiple copies based on the PCR and Southern blotting. The RT-PCR analysis showed that in the reverse mutant, EFS1 had a higher expression than the wild type. In the complementation, the fl

9、owering time was similar to the wild type Col-0, and also the flowering time was positively correlated to the expression level of EFS1. This result showed that the phenotypic reversion was caused by the expression of exo

10、genous EFS1. 2. RT-PCR was used to analyze the expression of EFS1. The results showed that the EFS1 might be a constitutive expression in the different tissues such as rosette leaf, stem, cauline leaf and flower. It had

11、a highest expression in the flower. Binary vector pCAMBIA1300-ProEFS1::GUS-NOS was constructed to transform to Nicotiana tabacum and Arabidopsis thaliana to get the tissue localization analysis, EFS1 promoter-driven GUS

12、expression was detectable in rosette leaf, stem, cauline leaf and flower of six-week seedings, and in the whole one-week seedings，this result is consistent with RT-PCR. 3. Expressed pattern of some important genes regula