發(fā)布時間：2019-06-22 10:53

【摘要】：隨著番茄種植規(guī)模的不斷擴大,高鹽、干旱、低溫、病原菌等各種非生物和生物脅迫對番茄的生長及生產(chǎn)造成了嚴重的危害。目前為止,大量研究顯示,植物轉(zhuǎn)錄因子在植物適應和抵御各種非生物及生物脅迫的過程中起著重要的調(diào)控作用。近年來對于植物不同轉(zhuǎn)錄因子家族的研究越來越深入,HD-Zip是植物中特有的一類轉(zhuǎn)錄因子,參與植物多種生物功能,尤其對植物在非生物脅迫下的應答反應有著重要調(diào)控作用。在本研究中,對番茄HD-Zip轉(zhuǎn)錄因子基因家族進行了生物信息學分析,對番茄HDZip蛋白家族的成員進行了分類、理化特性、保守區(qū)、基因結構、染色體分布、二級和三級結構的分析。同時對番茄HD-Zip I亞家族組織特異性及非生物脅迫下的表達模式進行了分析,對應答表現(xiàn)明顯的兩個基因進行了基因沉默分析。為番茄HD-Zip基因家族在非生物脅迫應答過程中的調(diào)控功能研究提供理論依據(jù)。主要研究結果如下:鑒定出番茄HD-Zip轉(zhuǎn)錄因子家族共包含58個成員(Sl HZ1-Sl HZ58),分為SlHD-ZipⅠ-Ⅳ亞家族。這些基因不均勻的分布在12條染色體上,番茄HD-Zip基因擴增方式主要為片段復制,家族成員的二級結構主要為螺旋,三級結構具有相似性。對HD-Zip I亞族15個基因進行了組織特異性表達分析,結果顯示,這些成員在根、莖、葉、花以及果實中均有不同程度的表達。大部分基因都能在根中進行表達,而Sl HZ30基因僅在花中表達。逆境脅迫表達分析中,在鹽脅迫處理下14個基因表達上調(diào),1個基因(SlHZ14)表達下調(diào)。在干旱處理中13個基因表達上調(diào),2個基因(Sl HZ14和SlHZ28)表達下調(diào)。在低溫處理中全部15個基因表達上調(diào),其中SlHZ13基因上調(diào)最為顯著。對逆境脅迫應答分析中表達量變化顯著的基因Sl HZ11和SlHZ15進行了VIGS基因沉默分析,發(fā)現(xiàn)SlHZ11和Sl HZ15基因沉默后,植株在鹽和干旱逆境條件下的MDA指標與相同條件下的正常植株相比顯著上升,說明該基因的下調(diào)表達可以降低植株的耐鹽和抗旱性。
[Abstract]:With the continuous expansion of tomato planting scale, high salt, drought, low temperature, pathogenic bacteria and other abiotic and biological stresses have caused serious harm to the growth and production of tomato. So far, a large number of studies have shown that plant transcription factors play an important role in plant adaptation and resistance to various abiotic and biological stresses. In recent years, the study of different family of transcription factors in plants has been more and more in-depth. HD-Zip is a unique transcription factor in plants, which is involved in a variety of biological functions of plants, especially plays an important role in regulating the response of plants under abiotic stress. In this study, the HD-Zip transcription factor gene family of tomato was analyzed by bioinformatics, and the members of tomato HDZip protein family were classified, physicochemical characteristics, conserved region, gene structure, chromosome distribution, secondary and tertiary structure. At the same time, the tissue specificity of tomato HD-Zip I subfamily and the expression pattern under abiotic stress were analyzed, and the gene silencing analysis of two genes with obvious response was carried out. It provides a theoretical basis for the study of the regulatory function of tomato HD-Zip gene family in abiotic stress response. The main results are as follows: a total of 58 members (Sl HZ1-Sl HZ58) of tomato HD-Zip transcription factor family were identified, which were divided into SlHD-Zip I-IV subfamily. These genes were unevenly distributed on 12 chromosomes. The amplification mode of tomato HD-Zip gene was mainly fragment replication, the secondary structure of family members was mainly spiral, and the tertiary structure was similar. The tissue specific expression of 15 genes of HD-Zip I subfamily was analyzed. the results showed that these members were expressed in roots, stems, leaves, flowers and fruits to varying degrees. Most genes can be expressed in roots, while Sl HZ30 genes can only be expressed in flowers. In stress analysis, 14 genes were up-regulated and 1 gene (SlHZ14) was down-regulated under salt stress. In drought treatment, 13 genes were up-regulated and 2 genes (Sl HZ14 and SlHZ28) were down-regulated. The expression of all 15 genes was up-regulated in low temperature treatment, and the up-regulation of SlHZ13 gene was the most significant. VIGS gene silencing analysis of genes SlHZ11 and SlHZ15 with significant changes in stress response analysis was carried out. It was found that after SlHZ11 and SlHZ15 gene silencing, the MDA index of plants under salt and drought stress was significantly higher than that of normal plants under the same conditions, which indicated that the down-regulated expression of SlHZ11 gene could decrease the salt tolerance and drought resistance of plants.
【學位授予單位】：東北農(nóng)業(yè)大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：S641.2

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