發(fā)布時間：2018-05-19 03:43

  本文選題：苜蓿 + 秋眠性��； 參考：《中國農(nóng)業(yè)科學院》2016年博士論文

【摘要】：秋眠性是紫花苜蓿(Medicago sativa L.)(以下簡稱“苜�！�)對晚秋生長環(huán)境變化的一種適應性反應,是實現(xiàn)越冬、返青等過程的關鍵基礎,秋眠性已被作為草地栽培與種植區(qū)劃的主要依據(jù)之一。本研究以國外引進和國內(nèi)審定登記的苜蓿品種為試驗材料,以11個國際標準秋眠級苜蓿品種為對照材料,按照Teuber等方法對所有供試材料進行秋眠級和抗寒指數(shù)的評價,進而闡述秋眠級對建植次年苜蓿生產(chǎn)性能、越冬階段根系性狀及相關基因表達的影響,利用二代測序平臺對不同秋眠型苜蓿根系進行轉錄組測序分析,篩選不同秋眠級苜蓿的差異表達基因,并研究差異基因的表達模式及可能參與的代謝通路,旨在為揭示苜蓿秋眠性的分子機理提供理論依據(jù)。主要結論如下:1.我國審定的苜蓿品種基本上屬于秋眠型,秋眠性較強,其中公農(nóng)2號、公農(nóng)3號、海拉爾苜蓿及龍牧系列苜蓿品種的秋眠級為1級,甘農(nóng)1號的秋眠級為2級。秋眠級對苜蓿建植次年第一茬產(chǎn)量等生產(chǎn)性能無顯著影響,盡管夏季生長階段株高與秋眠級之間沒有關系,但不同秋眠級苜蓿的莖稈伸長策略顯著不同,節(jié)間長和節(jié)間數(shù)與秋眠級或秋眠類型顯著相關,高秋眠級(弱秋眠性)苜蓿的節(jié)間長較長,但節(jié)間數(shù)卻隨著秋眠級的增加呈遞減趨勢。2.通過對46個苜蓿品種(材料)秋眠級與抗寒指數(shù)、越冬率的關系進行分析發(fā)現(xiàn),苜蓿秋眠級與抗寒指數(shù)呈極顯著的正相關關系,與越冬率呈極顯著的負相關關系,即秋眠性越強,苜蓿的越冬能力越強。不同秋眠級苜蓿的根系性狀表現(xiàn)出對越冬低溫脅迫的差異化響應特征,苜蓿主要通過增加根頸直徑和側根數(shù)來提高其抗寒性,并不取決于根系其他表型性狀的絕對大小。3.在低溫馴化階段,隨著秋眠級的增加,標準秋眠級苜蓿葉綠素含量、凈光合速率呈現(xiàn)先增加后降低的趨勢,葉片可溶性糖含量呈現(xiàn)下降的趨勢。越冬期標準品種苜蓿根頸、主根、側根氨基酸含量隨秋眠級表現(xiàn)一致的變化規(guī)律,隨著苜蓿秋眠性增強,各種氨基酸呈現(xiàn)顯著的增加態(tài)勢。隨著秋眠性的增強,越冬期標準品種苜蓿根頸、主根、側根抗寒基因cas18、vsp、corF的表達量呈現(xiàn)顯著的增加趨勢。4.應用Illumina NextSeq 500測序平臺,對秋眠1級和9級的苜蓿根系樣品進行轉錄組測序,共獲得23,470個Unigene。其中16,190個Unigene被注釋到一個或多個GO條目中,7,696個Unigene被成功注釋到KEGG蛋白數(shù)據(jù)庫的5條KEGG生化途徑和321個代謝通路中,18,617個Unigene被注釋到eggNOG蛋白數(shù)據(jù)庫的25個功能類別中。經(jīng)差異表達分析,共檢測到2,326個差異表達基因,其中包括1,308個上調(diào)表達基因和1,018個下調(diào)表達基因。1,721個差異表達基因獲得了GO功能注釋。差異表達基因顯著富集在能量代謝過程、次級代謝物的生物合成過程、碳水化合物代謝過程及氨基酸代謝過程等。此外,從苜蓿根系轉錄組中鑒定出1,497個轉錄因子和7,005個SSR。
[Abstract]:Autumn dormancy is Medicago sativa L. (hereinafter referred to as "alfalfa") an adaptive response to the late autumn growth environment is the key basis for the process of overwintering and returning to green. Autumn dormancy has been regarded as one of the main bases of grassland cultivation and planting regionalization. In this study, 11 international standard alfalfa varieties were used as experimental materials and 11 international standard alfalfa varieties were used as control materials. All the tested alfalfa varieties were evaluated for fall dormancy grade and cold resistance index according to Teuber and other methods. Furthermore, the effects of autumn dormancy on alfalfa production performance, root traits and related gene expression in winter stage were discussed, and transcriptional sequence analysis of different autumn dormant alfalfa roots was carried out by using the second generation sequencing platform. The differentially expressed genes of alfalfa with different fall dormancy levels were screened, and the expression patterns of differential genes and the metabolic pathways involved were studied in order to provide theoretical basis for revealing the molecular mechanism of autumn dormancy of alfalfa. The main conclusions are as follows: 1. The alfalfa varieties approved in our country basically belong to autumn dormancy type and have strong autumn dormancy. Among them, Gongnong 2, Gong Nong 3, Hailaer alfalfa and Longmu series alfalfa varieties have autumn dormancy grade 1 and Gannong 1 fall dormancy grade 2. Autumn dormancy had no significant effect on the first crop yield of alfalfa in the following year. Although there was no relationship between plant height and autumn dormancy in summer, the culm elongation strategies of alfalfa of different autumn dormancy levels were significantly different. Internode length and internode number were significantly correlated with autumn dormancy or fall dormancy type. The internode length of alfalfa with high autumn dormancy (weak autumn dormancy) was longer, but the internode number decreased with the increase of autumn dormancy. Based on the analysis of the relationship between fall dormancy grade and winter resistance index and winter resistance rate of 46 alfalfa varieties (materials), it was found that there was a very significant positive correlation between autumn hibernation level and winter resistance index, and a very significant negative correlation between fall hibernation level and wintering rate of alfalfa. That is, the stronger the autumn sleep, the stronger the winter ability of alfalfa. The root characters of different autumn dormancy alfalfa showed differential response to overwintering low temperature stress. Alfalfa improved its cold resistance mainly by increasing the diameter of root neck and the number of lateral roots, and did not depend on the absolute size of other phenotypic characters of root. In the low temperature acclimation stage, the chlorophyll content and net photosynthetic rate of standard autumn dormancy alfalfa increased first and then decreased, and the soluble sugar content of leaf decreased with the increase of autumn dormancy level. The content of amino acids in root neck, main root and lateral root of alfalfa during wintering period showed the same change rule with autumn dormancy grade. With the increase of autumn dormancy of alfalfa, all kinds of amino acids showed a significant increase trend. With the increase of autumn dormancy, the expression of root neck, main root and lateral root resistance gene cas18vspspcorF of standard varieties of alfalfa in overwintering stage showed a significant increasing trend .4. Using Illumina NextSeq 500 sequencing platform, 23470 Unigenes were obtained from alfalfa root samples of autumn sleep level 1 and 9 by transcriptome sequencing. 16190 Unigene were annotated into one or more go items, 7696 Unigene were successfully annotated into 5 KEGG biochemical pathways and 321 metabolic pathways in KEGG protein database, and 1 617 Unigene were annotated into 25 functional categories of eggNOG protein database. A total of 2326 differentially expressed genes were detected by differential expression analysis, including 1308 up-regulated genes and 1018 down-regulated genes. Differentially expressed genes were significantly enriched in energy metabolism, biosynthesis of secondary metabolites, carbohydrate metabolism and amino acid metabolism. In addition, 1497 transcription factors and 7005 SSRs were identified from alfalfa root transcriptome.
【學位授予單位】：中國農(nóng)業(yè)科學院
【學位級別】：博士
【學位授予年份】：2016
【分類號】：S541.9

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