【摘要】：硫代葡萄糖苷(簡(jiǎn)稱硫苷)是十字花科植物中典型的防御代謝產(chǎn)物。在轉(zhuǎn)運(yùn)蛋白的作用下,植物體內(nèi)的硫苷形成了非均勻的分布模式,易受昆蟲取食的部位容易積累更高濃度的硫苷。芥菜中含有的特異硫苷組分是其表現(xiàn)出抗蟲性狀的主要原因,已有研究證實(shí),芥菜中主要硫苷的降解產(chǎn)物對(duì)幼蟲具有致死作用。然而,芥菜中硫苷的分布模式及硫苷介導(dǎo)的防御分子機(jī)制尚未報(bào)道。所以本試驗(yàn)中,我們檢測(cè)了葉用芥菜雪里蕻不同組織、不同生長(zhǎng)時(shí)期和不同葉位的硫苷組分及含量,討論了芥菜中硫苷分布的生理和生態(tài)意義。此外,還研究了芥菜中硫苷介導(dǎo)的響應(yīng)雜食性害蟲甜菜夜蛾取食的分子防御機(jī)制。得到如下主要研究結(jié)果:(1)芥菜的根和葉中硫苷組分和含量存在較大差異。在子葉期向十六葉期生長(zhǎng)發(fā)育的過(guò)程中,葉中的總硫苷含量逐漸減少,但八葉期之后總硫苷含量無(wú)顯著差異,且二丙烯基硫苷是葉中主要的硫苷。相反,芥菜根中的總硫苷含量在子葉期向十六葉期生長(zhǎng)發(fā)育中呈上升趨勢(shì)。我們?cè)诟羞�檢測(cè)到了葉中未檢測(cè)到的2-苯乙基硫苷,且該硫苷在八葉期成為根中含量最高的硫苷,而十二葉期和十六葉期根中二丙烯基硫苷則是含量最高的硫苷。(2)對(duì)九葉期芥菜的不同葉位之間硫苷含量進(jìn)行分析,結(jié)果表明,越幼嫩的新葉硫苷含量越高。(3)芥菜總硫苷在甜菜夜蛾取食1h后沒(méi)有明顯變化,但在取食24h后顯著增加(24.7%)(P0.05)。甜菜夜蛾取食24h后脂肪族和吲哚族硫苷分別增加了22.1%和239.4%。二丙烯基硫苷在取食24h后增加了24.0%(P0.05),其含量約占總硫苷的96.0%。3-丁烯基硫苷也在芥菜被取食后顯著增加(P0.05)。除了4OHI3M以外,所有的吲哚硫苷都在取食1h后就顯著增加,I3M增加最多。而在甜菜夜蛾取食24h后所有的吲哚族硫苷含量都顯著增加。(4)轉(zhuǎn)錄組測(cè)序分析顯示,三個(gè)樣本(CK,1 h和24 h)分別獲得20.7,20.6和20.0百萬(wàn)個(gè)clean reads。測(cè)序片段組裝后共獲得60,948個(gè)獨(dú)立基因,基于七個(gè)數(shù)據(jù)庫(kù)的相似性分析共得到40,580個(gè)功能注釋獨(dú)立基因。芥菜被取食后,硫苷合成、降解和轉(zhuǎn)運(yùn)的53個(gè)同源基因的表達(dá)量幾乎全部上調(diào)。同時(shí),植物激素JA,ET和SA基因的表達(dá)量也呈上升趨勢(shì)。JA基因的表達(dá)量在取食1小時(shí)后顯著上升,不管是取食1h還是24h后,JA基因的表達(dá)量上升幅度明顯都高于ET和SA基因。我們的試驗(yàn)結(jié)果為進(jìn)一步研究硫苷介導(dǎo)的抗蟲防御機(jī)制和芥菜中硫苷代謝關(guān)鍵基因的功能奠定了基礎(chǔ)。
[Abstract]:Glucosinolate (glucosinolate) is a typical defense metabolite in cruciferous plants. Under the action of transporter, the glucosinolates in plants form a non-uniform distribution pattern, which is easy to accumulate a higher concentration of glucosinolate in the parts vulnerable to insect feeding. The specific glucosinolates contained in mustard are the main reasons for their insect-resistance. It has been proved that the degradation products of main glucosinolates in mustard have lethal effect on larvae. However, the distribution pattern of glucosinolate in mustard and the defense molecular mechanism mediated by glucosinolate have not been reported. Therefore, in this experiment, we detected the composition and content of glucosinolate in different tissues, different growth stages and different leaf positions, and discussed the physiological and ecological significance of the distribution of glucosinolate in Brassica juncea. In addition, the molecular defense mechanism of glucosinolate mediated response to the feeding of beet Spodoptera exigua in mustard was studied. The main results were as follows: (1) there were great differences in the composition and content of glucosinolate between the roots and leaves of mustard. During the growth and development from cotyledon stage to hexadecap stage, the total glucosinolate content in leaves decreased gradually, but there was no significant difference in total glucosinolate content after eight leaf stages, and diallyl glucosine was the main glucosine in leaves. On the contrary, the total glucosinolate content in mustard root increased from cotyledon stage to hexadecyl stage. We also detected 2-phenylethyl glucosinolate, which was not detected in the leaves, and it became the highest level of glucosinolate in the root at the eight-leaf stage. However, diallyl glucosinolate was the highest content in the roots of mustard at 12 and 16 leaf stages. (2) the contents of glucosinolate in different leaf positions of mustard at nine leaf stage were analyzed, and the results showed that, The content of glucosinolate in the young leaves was higher. (3) the total glucosinolates of mustard did not change after feeding for 1 hour, but increased significantly (24.7%) after 24 hours (P0.05). After feeding for 24 hours, the glucosinolates of aliphatic group and indole group increased by 22.1% and 239.4%, respectively. The content of diallyl glucosinolate increased by 24.0% (P0.05) after ingested for 24 hours, and the content of diallyl glucosinolate increased significantly (P0.05) after ingesting mustard (P 0.05), which accounted for 96.0.3-butenyl glucosinolates of total glucosinolate (P < 0.05). Except for 4OHI3M, all indolethioside increased significantly after feeding for 1 hour. However, after 24 hours of feeding, all indolyl glucosinolates were significantly increased. (4) the three samples (CK,1 h and 24 h) obtained 20.7G 20.6 and 20.0 million clean reads., respectively. (4) the transcriptome sequencing analysis showed that the three samples (CK,1 h and 24 h) were 20.6 and 20.0 million clean reads., respectively. A total of 60948 independent genes were obtained after the sequencing fragments were assembled, and 40580 functional annotated independent genes were obtained based on similarity analysis of seven databases. After the mustard was fed, the expression of 53 homologous genes that were synthesized, degraded and transported were almost upregulated. At the same time, the expression of plant hormone JA,ET and SA genes also showed an increasing trend. The JA gene expression increased significantly after 1 hour of feeding, and the expression of JA gene was significantly higher than that of ET and SA genes after 1 hour or 24 hours of feeding. Our results provide a basis for further study of glucosinolate mediated anti-insect defense mechanism and the function of key genes involved in glucosinolate metabolism in mustard.
【學(xué)位授予單位】：浙江農(nóng)林大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：S436.344

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本文編號(hào)：2231768