本文關鍵詞：中間錦雞兒參與葉片衰老基因CibHLH027的克隆和功能研究　出處：《內蒙古農業(yè)大學》2017年碩士論文　論文類型：學位論文

  更多相關文章： 中間錦雞兒 CibHLH027 黑暗 干旱 葉片衰老

【摘要】：葉片衰老是植物生長發(fā)育的最后階段,也是必經的主動應答階段。當植物經歷逆境脅迫,比如黑暗、干旱和極端溫度等環(huán)境因素,會加速葉片衰老過程,嚴重影響了植物的生長發(fā)育和作物產量。運用基因工程對植物進行改良,能夠提高植物在逆境脅迫下的耐受性,增強其適應環(huán)境的能力,從而有效延緩衰老。錦雞兒屬(Caragana)植物具有抗寒、抗旱及抗熱特性,賦予其適應各種環(huán)境中不良因素的能力,是改善生態(tài)環(huán)境的理想植被,因此也成為研究抗逆機制的優(yōu)良材料。本研究從中間錦雞兒(Caragana intermedia)轉錄組數(shù)據(jù)庫篩選到一個受非生物脅迫誘導的中間錦雞兒基因CibHLH027,并對其功能進行了研究,主要結果如下:1.CibHLH027編碼區(qū)長738bp,起始密碼子為ATG,終止密碼子為TGA,編碼245個氨基酸。與擬南芥bHLH蛋白系統(tǒng)進化分析顯示,CibHLH027與聚類到同一分支上的 AtbHLH027、AtbHLH035 的一致性分別為 58%和 52%。將 CibHLH027與不同植物中一致性較高的bHLH蛋白進行多序列比對和聚類分析顯示,N-端堿性區(qū)域都包含N-E-R基序,CibHLH027與同一分支上的鷹嘴豆CabHLH035 一致性為90%。2.CibHLH027的表達受ABA、NaCl、高pH、熱、黑暗及干旱的誘導。利用擬南芥葉肉原生質體瞬時表達發(fā)現(xiàn)CibHLH027-GFP定位于細胞核中。3.過表達CibHLH027基因的擬南芥,離體葉片和整株植株在黑暗誘導下,都表現(xiàn)出葉片衰老加速,葉綠素含量降低、離子滲透率增大和細胞死亡增多的表型:并且干旱處理下,CibHLH027過表達株系與野生型相比,耐旱能力明顯降低。
[Abstract]:Leaf senescence is the last stage of plant growth and development, and it is also a required stage of active response. When plants experience environmental stresses, such as darkness, drought and extreme temperature, they will accelerate leaf senescence and seriously affect plant growth and crop yield. The use of genetic engineering to improve plants can improve the tolerance of plants under stress, enhance their ability to adapt to the environment, and effectively delay aging. Caragana (Caragana) has the characteristics of cold resistance, drought resistance and heat resistance, giving it the ability to adapt to various adverse factors in the environment, and is an ideal vegetation for improving the ecological environment. Therefore, it has also become an excellent material for studying the mechanism of stress resistance. This study from Caragana intermedia (Caragana intermedia) transcriptome database to a screening by Caragana intermedia gene CibHLH027 induced by abiotic stress, and its function is studied, the main results are as follows: 1.CibHLH027 encoding region of 738bp, ATG as initiation codon and termination codon was TGA, encoding 245 amino acids. The phylogenetic analysis of bHLH protein in Arabidopsis thaliana showed that the consistency of CibHLH027 and AtbHLH027 and AtbHLH035 on the same branch was 58% and 52% respectively. Multiple sequence alignment and cluster analysis of bHLH protein with high consistency between CibHLH027 and different plants showed that the alkaline region of N- contains N-E-R motifs, and the consistency of CibHLH027 with chickpea CabHLH035 on the same branch was 90%. The expression of 2.CibHLH027 is induced by ABA, NaCl, high pH, heat, darkness and drought. CibHLH027-GFP was located in the nucleus by transient expression of protoplasts of Arabidopsis thaliana. 3. expression of CibHLH027 gene in Arabidopsis leaves and whole plants in the dark induced, showed accelerated leaf senescence, chlorophyll content decreased, ion permeability increases and increased cell death phenotype and under drought treatment, CibHLH027 overexpression lines compared with the wild type, drought resistant ability decreased significantly.
【學位授予單位】：內蒙古農業(yè)大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：Q943.2;S793.3

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本文編號：1346137