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The Cloning and Function Analysis of a Novel WRKY Transcription Factors from Pepper

Author: XiaoXiang
Tutor: HeShuiLin
School: Fujian Agriculture and Forestry University
Course: Developmental Biology
Keywords: Chili WRKY Cellular localization Expression patterns
CLC: S641.3
Type: Master's thesis
Year: 2011
Downloads: 67
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Abstract

The chili is a disease occurs more serious Solanaceae vegetables, frequent disease often leads to lower pepper yield and quality. Nurture and promote the application of resistant varieties is the fundamental technical countermeasures to solve problems of pepper diseases, the elucidation of the mechanism of resistance on pepper in the formulation of effective technology strategies conducive to the pepper disease resistance genetic improvement. A large number of studies have shown that pathogens and other biotic stresses and abiotic stress responses can activate plant resistance to adversity, and WRKY, ERF, MYB, NAC transcription factors in the induction of defense responses play an important regulatory role. Separation, identification transcription factor involved in specific Stress response and to carry out its function identification is an important way to reveal the mechanism of plant disease resistance and resilience molecular. In view of this, the uniform from exogenous SA pepper leaves isolated a cDNA library to obtain a chili WRKY gene family members of the full-length cDNA and its structure, subcellular localization, and W box binding activity, and in Different Stresses expression under a preliminary analysis, designed to lay the foundation for further analysis and to explore its value in genetic engineering of plants to environmental stresses. The main findings are as follows: 1. Cloned CaWRKY through biological software analysis found that encodes a relative molecular mass protein 37KDa the WRKY Transcription factor a typical WRKYGQK and C2HC zinc finger, and according to the sequence structure of the conserved domains Group III WRKY gene family; gene gun mediated build the good CaWRKY GFP fusion expression plasmids were introduced into onion cells, the results show CaWRKY nuclear localization of proteins; 3. effect plasmid construct CaWRKY the W-containing box reporter plasmid and total into onion cells, using plasmid transient expression analysis the CaWRKY W-box binding characteristics, The results show CaWRKY W-box binding characteristics; 4. transcriptional activation analysis show CaWRKY in its C-terminal a potential transcriptional activation domain; 5. using RT-PCR analysis the CaWRKY pepper in different periods and different tissues, results show that, CaWRKY certain expression levels in roots, stems, leaves and other tissues and organs, and in the highest expression of the maturity of the leaves; CaWRKY Real-time PCR analysis in a variety of abiotic stress and biological stress mode of expression in pepper results show, CaWRKY by the high temperature, cold, mechanical damage and pepper Ralstonia solanacearum induced After upregulated expression under drought and salt stress changes in the amount of more weak; CaWRKY by MeJA, SA and ethephon induced expression by NAA hormone-induced up-regulated expression patterns in the presence of a first down after; addition, ABA, reactive oxygen species H2O2 the induced expression against CaWRKY is not obvious; 6. the Gemini build CaWRKY Ye Chao expression vector, Agrobacterium-mediated transformation into tobacco, 30 T0 transgenic plants, of which 22 were detected by PCR positive transgenic plant. In summary, CaWRKY involved pepper for a variety of biotic and abiotic stress responses, the study of CaWRKY lay the theoretical foundation for chili improved the resilience.

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CLC: > Agricultural Sciences > Gardening > Vegetable gardening > Solanaceous > Chili
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