本文選題：WRKY + DoWRKY6�。� 參考：《西南交通大學》2017年碩士論文

【摘要】：WRKY是植物轉錄因子家族中較為重要的一員,在植物的生長發(fā)育過程中起著極為重要的轉錄調控作用。本論文基于鐵皮石斛原球莖轉錄組數據,獲得了 20個DoWRKY轉錄因子家族成員的保守區(qū)片段,并進一步研究了DoWRKY 基因在不同齡級、莖不同部位及實驗室組培幼苗的鐵皮石斛材料中的表達情況。通過對20個DoWRKY基因的保守區(qū)序列進行系統(tǒng)發(fā)育與序列保守性分析,發(fā)現這些成員都,有至少一個WRKYGQK構成的氨基酸殘基保守結構域,并且通過系統(tǒng)發(fā)育樹的構建,發(fā)現這20個基因親緣性有較大差異。通過生物信息學分析獲得了鐵皮石斛DoWRKY6基因的蛋白序列特征,鐵皮石斛DoWRKY6基因具有長度為1,017bp的完整ORF,編碼339個氨基酸,該蛋白只有一個WRKY結構域和一個結構為CX5C23HX1H的鋅指結構,屬于II類WRKY轉錄因子,且WRKY 結構域與鋅指結構會和DNA的W盒進行特異性結合。通過Protparam在線預測分析DoWRKY6蛋白的分子式為C1641H2556N454O530S13,相對分子質量為37,541.8,理論等電點為 4.86,總平均親水性(Grand average of hydropathicity,GRAVY)為-0.756,判定DoWRKY6灰轉錄因子為疏水性蛋白。利用熒光實時定量PCR技術獲得了DoWRKY6基因在不同齡級及莖不同部位的鐵皮石斛中的表達特點和在組培幼苗的不同器官中的相對表達量,發(fā)現DoWRKY6基因主要在莖中表達,且隨著生長年限增長其表達量會大大降低;而在莖中又主要是在莖的芽端部分表達,其次是在莖的基部表達,從而表明DoWRKY6對鐵皮石斛莖的生長發(fā)育有重要的調控作用。
[Abstract]:WRKY is an important member of the family of plant transcription factors, which plays an important role in the process of plant growth and development. Based on the transcriptome data of protocorm of Dendrobium candidum, the conservative regions of 20 members of DoWRKY transcription factor family were obtained, and the DoWRKY gene was further studied at different age levels. Expression of Dendrobium candidum in different parts of stem and tissue culture seedlings in laboratory. Based on the phylogenetic and conserved analysis of 20 conserved regions of DoWRKY gene, it was found that all of these members had at least one amino acid residues conserved domain of WRKYGQK, and the phylogenetic tree was constructed. It was found that there were significant differences in the genetic relationship of the 20 genes. The protein sequence of Dendrobium candidum DoWRKY6 gene was obtained by bioinformatics analysis. Dendrobium candidum DoWRKY6 gene has a complete ORFwhose length is 1017bp and encodes 339 amino acids. The protein has only one WRKY domain and a zinc finger structure with CX5C23HX1H structure. It belongs to class II WRKY transcription factors, and the WRKY domain binds specifically to the zinc finger structure and the W box of DNA. The molecular formula of DoWRKY6 protein was C1641H2556N454O530S13, the relative molecular weight was 37541.8, the theoretical isoelectric point was 4.86, and the total average hydrophobic average of hydropathicity GRAVYwas -0.756. The grey transcription factor of DoWRKY6 was determined as hydrophobic protein. The expression characteristics of DoWRKY6 gene in Dendrobium candidum at different age levels and different parts of stem were obtained by real-time quantitative PCR. The results showed that DoWRKY6 gene was mainly expressed in stems of Dendrobium candidum in different organs of tissue culture seedlings. The expression of DoWRKY6 was mainly expressed at the end of shoot and then at the base of stem, which indicated that DoWRKY6 played an important role in the growth and development of Dendrobium candidum stem.
【學位授予單位】：西南交通大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：Q943.2;S567.239

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本文編號：1997285