本文選題：哈密瓜 + 轉錄組　； 參考：《石河子大學》2017年碩士論文

【摘要】：哈密瓜是新疆特色瓜果,采后貯藏過程中易發(fā)生冷害,造成很大經濟損失。導致冷害的主要原因是貯藏中溫度過低,因此適宜的貯藏溫度是解決這一問題的關鍵。目前哈密瓜響應冷脅迫作用機制的研究較少,通過對哈密瓜轉錄組進行研究,通過分子手段研究哈密瓜關鍵基因的調控,為哈密瓜響應冷脅迫作用機制提供理論依據(jù)。目的:本實驗研究了新疆哈密瓜早熟品種金皇后,通過研究3種貯藏溫度(21℃、3℃和0.5℃)對哈密瓜果實采后品質及生理影響,并對不同條件的哈密瓜進行轉錄組測序,篩選出差異表達基因,對差異基因進行GO分析和Pathway分析等,從分子水平上研究哈密瓜在貯藏過程中可能存在的基因種類、功能及代謝途徑等,提供哈密瓜響應冷脅迫作用機制的理論依據(jù)。方法:測定哈密瓜的生理指標,包括冷害指數(shù)、失重率、電導率、硬度和可溶性固形物,用試劑盒測定SOD、POD、CAT和CHT酶活性;利用BGISEQ-500測序平臺對哈密瓜進行轉錄組測序。結果與結論:(1)0.5℃貯藏哈密瓜果實,能有效維持果實品質;延緩冷害指數(shù)、失重率和電導率的上升以及硬度、可溶性固形物含量下降;保持較高的SOD、POD、CAT、CHT的活性,延緩果實衰老、腐爛。(2)通過BGISEQ-500測序平臺,平均得到24136070條原始reads,24105793條clean reads,clean reads與參考基因的平均比對率為64.22%,Clean reads與參考基因組平均比對率為80.67%。共鑒定到20823個基因,0.5℃的樣品中鑒定到的基因數(shù)整體均少,而在21℃鑒定到的基因數(shù)整體均較多,說明低溫抑制了哈密瓜在貯藏過程的代謝,抑制了基因表達,更有利于果實保鮮。(3)篩選出哈密瓜果實不同樣品間的所有差異基因共15741個,進行GO功能注釋,生物學過程顯著富集的差異表達的基因在23條GO-term中,主要富集在代謝、細胞、單一生物三個過程中;細胞組分顯著富集的差異表達的基因在16條GO-term中,主要富集在細胞、細胞部件和膜;分子功能顯著富集的差異表達基因在13條GO-term中,主要富集在催化活性和結合。其中免疫調節(jié)、響應刺激、抗氧化活性和酶調節(jié)活性與冷脅迫相關。(4)哈密瓜果實中共有15741個差異表達基因,富集在134-131條通路中,顯著富集的通路包括脂肪酸代謝、谷胱甘肽代謝、代謝途徑、次生代謝產物生物合成、植物激素信號轉導、植物病原菌的相互作用、淀粉蔗糖代謝、氨基酸生物合成、丙酮酸代謝、纈氨酸,亮氨酸和異亮氨酸降解等。
[Abstract]:Hami melon is a characteristic melon and fruit in Xinjiang. Cold damage is easy to occur during storage after harvest. The main cause of cold damage is low temperature in storage. Therefore, the key to solve this problem is the suitable storage temperature. At present, Hami melon response to cold stress mechanism is less, and the study of Hami melon transcriptional group is studied. Study on the regulation of Hami melon key genes by molecular means to provide a theoretical basis for Hami melon response to cold stress mechanism. Objective: this experiment studied the Xinjiang Hami melon early maturing variety gold queen, through the study of 3 storage temperatures (21, 3 and 0.5 C) on the fruit quality and physiological effects of Hami melon after harvest, and to different conditions of HA. The transcriptional group was sequenced, the differentially expressed genes were screened, the differentially genes were analyzed by GO analysis and Pathway analysis. The possible genetic types, functions and metabolic pathways of Hami Melon during storage were studied at the molecular level, and the theoretical basis of the mechanism of Hami melon in response to cold stress was provided. Indexes, including the cold damage index, weight loss rate, electrical conductivity, hardness and soluble solid, the activity of SOD, POD, CAT and CHT enzyme were measured with kits, and the transcriptional group of Hami melon was sequenced using BGISEQ-500 sequencing platform. Results and conclusions: (1) storage of Hami melon fruit at 0.5 degrees can effectively maintain fruit quality; delay the cold damage index, weight loss rate and electrical conductivity Rising and hardness, the content of soluble solids decreased; maintained high activity of SOD, POD, CAT, CHT, and delayed fruit senescence and decay. (2) the average ratio of 24136070 original reads, 24105793 clean reads, clean reads to reference basis was 64.22%, Clean reads and the average ratio of the reference genome. The total number of genes identified in the samples of the rate of 80.67%. was 20823. The number of genes identified in the samples at 0.5 C was less, and the number of genes identified at 21 C were all more, indicating that low temperature inhibited the metabolism of Hami melon in the storage process, inhibited the gene expression and was more conducive to the preservation of the fruit. (3) all the differences between the different fruits of Hami melon were screened. A total of 15741 different genes were annotated by the GO function. The differentially expressed genes in the biological processes were significantly enriched in 23 GO-term, mainly enriched in the three processes of metabolism, cells and single organisms; the differentially expressed genes in the cell components were significantly enriched in 16 GO-term, mainly enriched in cells, cell parts and membranes; the function of the molecules was significantly rich. The differentially expressed genes in the 13 GO-term were mainly concentrated in the catalytic activity and binding. The immunoregulation, response stimulation, antioxidant activity and enzyme regulation activity were related to cold stress. (4) there were 15741 differentially expressed genes in Hami melon fruits, enriched in 134-131 pathways, and the significant enrichment pathways included fatty acid metabolism and cereal caspine. Peptide metabolism, metabolic pathway, biosynthesis of secondary metabolites, plant hormone signal transduction, plant pathogen interaction, starch sucrose metabolism, amino acid biosynthesis, pyruvate metabolism, valine, leucine and isoleucine degradation, etc.

【學位授予單位】：石河子大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：S652.1

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