海岛棉GbNPR1基因的表达特性及功能分析-遗传学专业论文.docxVIP

万方数据 万方数据 Abstract Objective ： Sea Island cotton is one of Chinas important cultivars, Xinjiang as Chinas major cotton growing area, its favorable natural conditions provide a unique advantage for the development of Sea Island cotton. However, it is easy to be affected by fungi , bacteria and virus in the growth process, so it will suffer seriously blight, which will seriously affect the yield and quality of cotton. Therefore, we use modern genetic engineering techniques to dig resistance gene resources, and to study its function in disease resistance in Sea Island cotton, which will lay foundation for blight-resistant Sea Island cotton varieties breeding. Method: In this study, with Sea Island cotton 14, 16 and 21 in Xinjiang as the materials, we used RT-PCR technology to tap and exploit blight-resistant-related gene GbNPR1, and carried on bioinformatic analysis of the gene; we use real-time PCR technology to analyze the gene expression levels in Fusarium stress and salicylic acid and ethylene treatment; Constructed the over-expression plant vector, transfered the gene to Sea Island cotton by Agrobacterium-mediated transformation of shoot tips, for a preliminary study of its functions; We used Agrobacterium infecting stigma method and Agrobacterium buds injection method for Sea Island cotton genetic transformation in the field, to explore an effective genetic transformation method. Results: We clone the GbNPR1 gene from Sea Island cotton , which has a 1764bp full-length open reading frame , encodes 587 amino acids, its molecular weights 65393.8, and its theoretical isoelectric point is 5.99, the protein-coding gene is a hydrophilic protein; it contains a BTB / POZ domain, three ankyrin repeats (ANK) domains and the representative NPR1_like_C domains of NPR1 protein family. Random coil and α helix in the secondary structure of the protein shares a portion of 42.42% and 49.40% , respectively. qPCR analysis showed that, after fusarium induction, GbNPR1 gene expression