【摘要】：沙蔥(Allium mongolicum Regel.),地方名為蒙古韭,是生長(zhǎng)在內(nèi)蒙古、甘肅、新疆等荒漠地帶、沙漠周邊和山石縫隙等干旱地的一種野生蔬菜。沙蔥作為一種潛在價(jià)值極高的野生蔬菜受到人民群眾的熱捧。為此筆者以貯藏1～10年沙蔥種子為研究對(duì)象,考查其不同貯藏年限條件下干種子、萌發(fā)3天和10天種子的萌發(fā)特性、生理關(guān)鍵酶活性、呼吸作用等方面,并通過(guò)高通量測(cè)序技術(shù)對(duì)貯藏2015年(S1)、2012年(S2)、2008年(S4)這三個(gè)年份的沙蔥種子萌發(fā)的轉(zhuǎn)錄組進(jìn)行測(cè)序,并對(duì)其進(jìn)行基因注釋和功能分類(lèi),對(duì)其S1、S2、S4的差異基因進(jìn)行了分析,為沙蔥種子在不同萌發(fā)條件下的活力及壽命的生理生化響應(yīng)機(jī)制和沙蔥分子研究提供一定的理論依據(jù)。獲得研究結(jié)果如下:1.隨著沙蔥種子貯藏陳化時(shí)間的延長(zhǎng),其發(fā)芽率、發(fā)芽勢(shì)、發(fā)芽指數(shù)和活力指數(shù)均呈現(xiàn)先升高后緩慢降低的趨勢(shì),在貯藏3年的種子活力最高,發(fā)芽率高達(dá)89.00%;含水量呈現(xiàn)逐漸降低的變化趨勢(shì),電導(dǎo)率呈現(xiàn)逐漸升高的變化趨勢(shì)。2.隨著沙蔥種子貯藏陳化時(shí)間的延長(zhǎng),呼吸速率與細(xì)胞色素C氧化酶均呈先升高后降低的變化趨勢(shì);沙蔥種子在萌發(fā)過(guò)程中的關(guān)鍵酶活性變化是不同的,其中酸性磷酸酶和蘋(píng)果酸脫氫酶活性均隨陳化時(shí)間延長(zhǎng)呈現(xiàn)先升高后降低的變化趨勢(shì),且貯藏5年酶活力最高;異檸檬酸脫氫酶活性變化規(guī)律不顯著,而過(guò)氧化物酶活性呈現(xiàn)先降低后升高再降低的變化趨勢(shì)。3.對(duì)不同貯藏年限的沙蔥種子(S1、S2、S4)萌發(fā)構(gòu)建了 RNA-seq進(jìn)行轉(zhuǎn)錄組測(cè)序文庫(kù),經(jīng)Illumina測(cè)序和組裝得到了 79,703條Unigene序列,其平均長(zhǎng)度為684bp。其中有有36,232條分別注釋到NR、Swiss-Prot、KEGG、COG數(shù)據(jù)庫(kù)中得到了基因功能注釋信息,為深入研究不同貯藏年下下沙蔥種子的相關(guān)基因提供了理論依據(jù),也為沙蔥種子生物信息學(xué)提供了數(shù)據(jù)資源。4.沙蔥種子萌發(fā)的基因表達(dá)譜研究,3個(gè)不同貯藏時(shí)期測(cè)序結(jié)果兩兩比較,分別得到差異表達(dá)基因S1-10-VS-S2-10 5,723條,有75%上調(diào);S1-10-VS-S4-10中7,347 條,有 59%上調(diào);S2-10-VS-S4-10 中 5,495 條,有 48%上調(diào)。通過(guò) GO Pathway分析發(fā)現(xiàn),S1-10-VS-S2-10主要受影響的生物功能受影響最大的是代謝過(guò)程;受影響最小的是生物功能過(guò)程的蛋白結(jié)合的轉(zhuǎn)錄因子;S1-10-VS-S4-10和S2-10-VS-S4-10參與調(diào)控最多的是生物功能。通過(guò)差異表達(dá)基因在KEGG代謝通路中的功能富集分析,發(fā)現(xiàn)S1、S2、S4三個(gè)不同貯藏年份差異表達(dá)基因參與的代謝通路差異很大,S1對(duì)應(yīng)到個(gè)類(lèi)通路的Unigene數(shù)量高于其他兩個(gè)年限,與總差異基因表達(dá)量趨勢(shì)基本一致。
[Abstract]:Allium mongolica (Allium mongolicum Regel.), is a wild vegetable growing in the desert areas of Inner Mongolia Gansu Xinjiang and other arid areas such as the desert and the gap between mountains and rocks. As a kind of wild vegetable with high potential value, Allium balsicum is popular among the people. For this reason, the seeds of Allium mongolica were stored for 1 ~ 10 years. The germination characteristics, physiological key enzyme activities and respiration of dry seeds, germination days and 10 days of germination were investigated under different storage years. The transcriptome of seed germination of Allium mongolicum in 2015 (S1), 2012 (S2) and 2008 (S4) was sequenced by high-throughput sequencing technique, and its gene annotation and functional classification were carried out, and the differential genes of S1-S2OS4 were analyzed. It provides a theoretical basis for the physiological and biochemical response mechanism of seed vigor and longevity of Allium mongolicum under different germination conditions and the molecular studies of Allium mongolicum. The results are as follows: 1. The germination rate, germination potential, germination index and vigor index of the seeds increased first and then decreased slowly with the prolonging of storage time, and the seed vigor was the highest after 3 years of storage. The germination rate was up to 89.00, the water content decreased gradually, and the electrical conductivity increased gradually. The respiration rate and cytochrome C oxidase increased firstly and then decreased with the prolongation of storage and aging time of Allium mongolica seeds, and the changes of key enzyme activities were different during germination. The activity of acid phosphatase and malate dehydrogenase increased first and then decreased with aging time, and the activity of isocitrate dehydrogenase was the highest in 5 years, but the change of isocitrate dehydrogenase activity was not significant. The activity of peroxidase decreased first and then increased and then decreased. The RNA-seq transcriptome sequencing library was constructed for the germination of the seeds of Allium mongolicum (S1, S2, S4) for different storage years. 79703 Unigene sequences were obtained by Illumina sequencing and assembly, with an average length of 684 BP. 36232 of them were annotated in the database of NRRS-Swiss-ProtKEGGG, which provided theoretical basis for the further study of the genes related to the seeds of Allium mongolicum under different storage years, and provided a data resource of .4for the bioinformatics of the seeds of Allium mongolica. The gene expression profile of seed germination of Allium mongolicum L. was studied. The sequencing results of three different storage periods showed that there were 5723 differentially expressed genes (S1-10-VS-S2-10), 7347 of S1-10-VS-S4-10, 5495 of S2-10-VS-S4-10, and 48% of S2-10-VS-S4-10, respectively, with 75% upregulation of S1-10-VS-S4-10 and 59% of up-regulation of S2-10-VS-S4-10, respectively. Go Pathway analysis showed that the major affected biological function of S1-10-VS-S2-10 was the metabolic process, and the transcriptional factor S1-10-VS-S4-10 and S2-10-VS-S4-10 were the most involved in the regulation of biological function. Based on the functional enrichment analysis of differentially expressed genes in the KEGG metabolic pathway, it was found that the number of Unigene in the three differentially expressed genes involved in the three different storage years was higher than that in the other two years. The trend of total differentially expressed genes was basically consistent with that of total differentially expressed genes.
【學(xué)位授予單位】：內(nèi)蒙古農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：S647

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