【摘要】：杏是薔薇科(Rosaceae)李亞科(Prunoideae)杏屬(Armeniaca)植物,果肉、果仁均可食用。原產(chǎn)中國(guó),野生種和栽培品種資源都非常豐富。在人民生活和園藝產(chǎn)業(yè)中占據(jù)非常重要地位�？扇苄蕴堑姆e累是杏果實(shí)的重要品質(zhì)之一,如何提高杏果實(shí)的糖含量非常重要。蔗糖是果樹(shù)體內(nèi)長(zhǎng)距離運(yùn)輸?shù)闹饕镔|(zhì),不僅可以通過(guò)韌皮部長(zhǎng)距離運(yùn)輸?shù)焦麑?shí)中積累糖分,提高果實(shí)糖含量,還可以長(zhǎng)距離運(yùn)輸?shù)焦麡?shù)其他部位,為營(yíng)養(yǎng)生長(zhǎng)提供所需的能量。而蔗糖的主要運(yùn)輸途徑是質(zhì)外體途徑,在完成蔗糖韌皮部裝載和卸載的跨膜運(yùn)輸過(guò)程中,需要一種載體蛋白(Sucrose carrier,SUC)的作用。這種載體蛋白叫做蔗糖-質(zhì)子共轉(zhuǎn)運(yùn)蛋白,也叫做蔗糖轉(zhuǎn)運(yùn)蛋白(Sucrose transporter,SUT)。本研究運(yùn)用RT-PCR方法,從“吉農(nóng)紅杏”中獲得了蔗糖轉(zhuǎn)運(yùn)蛋白基因PaSUC4,所得序列長(zhǎng)度為2480bp,其中完整開(kāi)放閱讀框1500 bp,預(yù)測(cè)其編碼499個(gè)氨基酸,Genbank登錄號(hào)為KT223003;3’非翻譯區(qū)長(zhǎng)度為945bp。預(yù)測(cè)編碼的蛋白質(zhì)分子量為53.58KD,等電點(diǎn)為9.31。與張啟祥等在杏屬梅(Prunus mume)中的PmSUC4氨基酸序列的結(jié)果存在98%的相似性。相似性及系統(tǒng)進(jìn)化分析發(fā)現(xiàn)所得蔗糖轉(zhuǎn)運(yùn)蛋白與薔薇科其他李屬植物中的同類蛋白序列相似性很高,推測(cè)其功能也具有保守性。為進(jìn)一步研究PaSUC4的生物學(xué)功能,構(gòu)建了植物表達(dá)載體pCAM-UPN-PaSUC4并進(jìn)行了煙草遺傳轉(zhuǎn)化,獲得陽(yáng)性植株4株,為后續(xù)基因的生物功能鑒定工作提供重要試驗(yàn)材料。采集杏不同時(shí)期的7個(gè)部位進(jìn)行熒光定量PCR分析。熒光定量PCR顯示PaSUC4在杏樹(shù)的幼果、幼葉、成齡葉、花瓣和新梢器官中均有表達(dá),雌蕊中表達(dá)量很少,說(shuō)明表達(dá)水平具有組織差異性。PaSUC4不僅在成齡葉(源組織)中表達(dá),在花瓣、新梢、幼果、幼葉、雌蕊等庫(kù)組織中也有表達(dá),因此推測(cè)PaSUC4可能具有雙重功能。
[Abstract]:Apricot is the (Armeniaca) plant of (Prunoideae), (Rosaceae) of Rosaceae. Its flesh and kernel are edible. Native to China, wild species and cultivated varieties are rich in resources. It plays a very important role in people's life and horticultural industry. The accumulation of soluble sugar is one of the important qualities of apricot fruit, and how to improve the sugar content of apricot fruit is very important. Sucrose is the main substance of long distance transportation in fruit trees. It can not only accumulate sugar through phloem and increase sugar content in fruit, but also transport it to other parts of fruit trees. Provide the energy needed for nutritional growth. The main transport pathway of sucrose is the ectoplast pathway, and a carrier protein (Sucrose carrier,SUC) is needed to complete the transmembrane transport of sucrose phloem. This carrier protein is called sucrose-proton co-transport protein, also known as sucrose transport protein (Sucrose transporter,SUT). In this study, the length of sucrose transporter gene PaSUC4, obtained from Jinong Red apricot by RT-PCR method was 2480bp. in which the complete open reading frame 1500 bp, predicted its encoding 499 amino acids, and the Genbank accession number was KT223003;. The length of 3 'untranslated area is 945 BP. The predicted protein molecular weight is 53.58 KD and the isoelectric point is 9.31. There was a 98% similarity between the result of PmSUC4 amino acid sequence and that of Zhang Qi-xiang in (Prunus mume) of Apricot. Similarity and phylogenetic analysis showed that the sucrose transporter had high similarity with the similar proteins in other plants of Rosaceae, and its function was conserved. In order to further study the biological function of PaSUC4, a plant expression vector pCAM-UPN-PaSUC4 was constructed, and 4 positive plants were obtained by genetic transformation of tobacco. Seven parts of apricot in different periods were collected for fluorescence quantitative PCR analysis. Fluorescence quantitative PCR showed that PaSUC4 was expressed in young fruit, young leaf, adult leaf, petal and shoot organs, but not in pistil, indicating that the expression level was different. PaSUC4 was not only expressed in adult leaf (source tissue). It was also expressed in petals, shoots, young fruits, young leaves, pistil and so on. Therefore, we speculated that PaSUC4 might have dual function.
【學(xué)位授予單位】：吉林農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：S662.2

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