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The Ramie Auxin-binding Proteins (BnABP1) Gene cDNA Transformation and Over Expressed in Elongate Cells

Author: PengZhenZi
Tutor: ZhangXueWen
School: Hunan Agricultural University
Course: Cell Biology
Keywords: Auxin binding protein cDNA Cotton E6 gene promoter Genetic transformation Cell elongation
CLC: Q943
Type: Master's thesis
Year: 2011
Downloads: 17
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Abstract


Auxin-binding protein (Auxin Binding Protein, ABP1) response to the plant the auxin signal cell elongation growth play an important regulatory role. Early, we have cloned the ramie auxin combination of the the protein genes cDNA (BnABP1). Ramie function and role of auxin binding protein ABP1 cloned in order to further understand the in elongation growth characteristics of cells were highly expressed genes - cotton fiber cells highly expressed the E6 gene promoters, the E6 gene promoter and the reorganization of BnABP1 cDNA transformed Arabidopsis and cotton, preliminary study of BnABP1 plant cell elongation. In order to verify the effect of cotton E6 gene promoter expression, we first after the reorganization of the reporter gene GUS (encoding β-glucuronidase) into Arabidopsis. Transformed seedlings by GUS chromogenic transgenic Arabidopsis table have a small amount of blue fur, flower, stem and radicle presented, but the most obvious blue leaf trichomes, E6 promoter control genes in Arabidopsis mustard trichomes advantage of expression. Use of Ti plasmid pBIl21 for the original vector was constructed by the E6 promoter regulate BnABP1 gene the plant table fur the overexpression vector pBI121-E6-BnABP1. Transferred to Agrobacterium tumefaciens GV3101 transformed after dip inflorescence type Arabidopsis of Columbia, three positive transgenic plants. Microscopic observation and statistical analysis showed that the length of the transgenic Arabidopsis leaf trichomes (trichome) elongation of about 10% on average compared with the control. The driven by E6 promoter, BnABP1 Arabidopsis trichome cells can express a certain extent, promote the growth of cell elongation. The amount of plant growth, including growth in the size of the leaf discs, stem height and bolting number. Overexpression can also affect the the trichome start and development, performance fur density increases, more uniform and dense leaf table. For ABP1 auxin regulates plant growth and development of key protein provides new evidence. As a parallel study, the use of the constructed vector cotton genetic transformation. Agrobacterium tumefaciens ovary injection of the the engineered Agrobacterium injected directly before and after pollination cotton ovary, of the eight genotypes cotton genetic transformation, and the first to establish a more efficient compared to the previous method cotton plants in vivo genetic transformation technology roadmap. Using Coomassie blue mark the injections showed that, 48 h (-2DPA) to 24 H (1DPA) injection of bacteria solution before the cotton pollination Du can be entering beads embryo, taking into account the injections on the pollination of the impact, a living body transformed pollination 24 H when injected 5μL about 104 bacteria, in eight genotypes materials are screened positive transgenic plants via pollen tube pathway. However, due to time constraints, the quality and development of genetically modified cotton fibers still further study.

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