本文選題：七星瓢蟲 + 滯育�。� 參考：《中國(guó)農(nóng)業(yè)科學(xué)院》2016年碩士論文

【摘要】：隨著農(nóng)作物病蟲害綠色防控技術(shù)與應(yīng)用的發(fā)展,生物防治的重要性與日俱增。七星瓢蟲Coccinella septempunctata L.是優(yōu)良的天敵昆蟲,可捕食蚜、蚧等多種農(nóng)林害蟲,在多個(gè)國(guó)家已商品化生產(chǎn)應(yīng)用。滯育是多數(shù)天敵昆蟲具有的遺傳屬性,調(diào)控天敵昆蟲滯育可以延長(zhǎng)天敵產(chǎn)品的貨架期、保證長(zhǎng)距離運(yùn)輸、提高抗逆性,促進(jìn)天敵昆蟲產(chǎn)品的貯存、運(yùn)輸和大規(guī)模應(yīng)用。對(duì)七星瓢蟲滯育的研究涵蓋了解剖學(xué)特征、誘導(dǎo)技術(shù)、滯育后生物學(xué)、滯育機(jī)理等各方面,促進(jìn)了對(duì)七星瓢蟲滯育科學(xué)規(guī)律的深入了解,對(duì)指導(dǎo)生產(chǎn)實(shí)踐應(yīng)用具有重要意義。滯育期間營(yíng)養(yǎng)物質(zhì)的貯存和利用對(duì)昆蟲順利度過(guò)滯育期具有重要的意義。本研究以滯育七星瓢蟲解剖學(xué)為基礎(chǔ),系統(tǒng)地研究了包括含水量、脂質(zhì)、碳水化合物、蛋白質(zhì)等在內(nèi)的生化物質(zhì)含量波動(dòng)情況;歸納總結(jié)了正常發(fā)育、滯育和滯育解除三種不同發(fā)育狀態(tài)下的七星瓢蟲過(guò)冷卻點(diǎn)變化特征;測(cè)試分析了正常發(fā)育、滯育和滯育解除個(gè)體轉(zhuǎn)錄組測(cè)序結(jié)果,獲得大量的滯育差異表達(dá)基因;研究篩選出與脂質(zhì)代謝相關(guān)基因并深入分析,為探究瓢蟲滯育營(yíng)養(yǎng)物質(zhì)利用和調(diào)控提供參考。主要研究結(jié)果如下:1.明確了七星瓢蟲在正常發(fā)育、滯育和滯育解除階段體內(nèi)含水量、脂質(zhì)、碳水化合物、蛋白質(zhì)等重要生化代謝物含量有顯著差異:滯育個(gè)體含水量、總蛋白含量顯著低于正常發(fā)育和滯育解除個(gè)體,說(shuō)明滯育瓢蟲體內(nèi)代謝活動(dòng)降低;滯育個(gè)體的脂質(zhì)(173.66±19.01μg/mg)比正常發(fā)育(80.04±15.82μg/mg)、滯育解除(83.83±12.54μg/mg)個(gè)體增加近1倍,總糖和糖原含量可高出2倍和7倍,是滯育瓢蟲重要的營(yíng)養(yǎng)物質(zhì)來(lái)源;而甘油、山梨醇、海藻糖含量在三種不同發(fā)育階段的瓢蟲中差異不顯著。此外,滯育瓢蟲糖原大量積累,與正常發(fā)育和滯育解除的個(gè)體存顯著性差異;而滯育瓢蟲的海藻糖含量與正常發(fā)育和滯育解除個(gè)體之間無(wú)顯著差異,證實(shí)了七星瓢蟲的滯育屬糖原積累類型。2.證實(shí)了七星瓢蟲在滯育階段耐寒性提高:滯育七星瓢蟲過(guò)冷卻點(diǎn)為-16.53℃,低于正常發(fā)育個(gè)體(-14.07℃)和滯育解除個(gè)體(-15.29℃),且滯育馴化后七星瓢蟲的耐寒性有所提高。3.應(yīng)用轉(zhuǎn)錄組技術(shù)檢測(cè)正常發(fā)育-滯育-滯育解除瓢蟲基因轉(zhuǎn)錄情況,經(jīng)過(guò)剪接、過(guò)濾、拼接等步驟,共獲得82,820個(gè)Unigenes,滯育-正常發(fā)育差異表達(dá)基因3,501個(gè),其中滯育上調(diào)2,125個(gè)、下調(diào)1,376個(gè);滯育解除-滯育差異表達(dá)基因1,427個(gè),其中滯育上調(diào)749個(gè)、下調(diào)表達(dá)678個(gè)。差異基因在分子功能類別和脂肪酸代謝通路、藥物代謝、碳代謝等通路得到富集。4.將滯育-滯育解除差異基因和滯育解除-滯育差異基因兩兩比較,并藉助KEGG數(shù)據(jù)庫(kù),共篩選出滯育期間與脂質(zhì)合成相關(guān)的差異基因17個(gè),在滯育期間呈現(xiàn)出不同程度的上調(diào)表達(dá),涉及了脂肪酸合成、β-氧化、脂質(zhì)轉(zhuǎn)運(yùn)等代謝途徑,除脂肪酸合酶、脂鏈延伸酶、脂肪酸去飽和酶基因、脂質(zhì)轉(zhuǎn)運(yùn)蛋白等與脂質(zhì)合成相關(guān)酶類上調(diào)表達(dá)外,同時(shí)一部分與脂質(zhì)分解相關(guān)的酶,如酯酶、�；D(zhuǎn)移酶等同樣上調(diào)表達(dá),可能與提高滯育瓢蟲免疫力有關(guān);此外,與激素合成相關(guān)基因表達(dá)上調(diào),可能對(duì)滯育瓢蟲卵巢發(fā)育受抑相關(guān)。
[Abstract]:With the development of green prevention and control technology and application of crop pests and diseases, the importance of biological control is increasing. Coccinella septempunctata L. of Seven Star ladybug is a good natural enemy insect. It can catch aphids, scale insects and other agroforestry pests, and has been commercialized in many countries. Diapause is the genetic attribute of most natural enemy insects. Diapause of enemy insects can prolong the shelf life of natural enemy products, ensure long distance transportation, improve the resistance and promote the storage, transportation and large-scale application of natural enemy insect products. The study of diapause of Seven Star Ladybug covers anatomic characteristics, induction technique, biologic after diapause, and hysteretic mechanism, which promotes the diapause science of seven ladybug. The deep understanding of the law is of great significance for guiding the application of production practice. The storage and utilization of nutrients during diapause is of great significance for the insect to pass through the diapause period. Based on the diapause anatomy of the ladybug, this study systematically studies the biochemical substances, including water content, fat, carbohydrate, protein and so on. The characteristics of the changes in the subcooling point of the normal development, diapause and diapause in three different developmental states were summarized, and the results of normal development, diapause and diapause were analyzed, and a large number of diapause differential tables were obtained, and the genes related to lipid metabolism were screened. The main research results are as follows: 1. the water content in the normal development, diapause and diapause release stage of the ladybug is clear. The content of the important biochemical metabolites, such as lipid, carbohydrate, protein, and other important biochemical metabolites in the stage of normal development, diapause and diapause are clearly different: the water content and total protein content of diapause individual The metabolic activity of diapause Ladybug was lower than normal development and diapause, and the lipid (173.66 + 19.01 g/mg) of diapause individual was more than normal (80.04 + 15.82 mu g/mg), and the diapause (83.83 + 12.54 g/mg) individual increased nearly 1 times, the total sugar and glycogen content could be higher than 2 times and 7 times, and it was an important nutrient of diapause ladybug. The contents of glycerol, sorbitol and trehalose were not significantly different among the ladybug of three different developmental stages. In addition, the accumulation of glycogen in diapause Ladybug was significantly different from those of normal development and diapause, while the trehalose content of diapause Ladybug was not significantly different from that of normal development and diapause. The accumulation type.2. of the diapause of Ladybug confirmed that the cold tolerance of the ladybug was improved at the diapause stage: the cross cooling point of diapause was -16.53 C, lower than the normal developing individuals (-14.07 C) and diapause individual (-15.29 C), and the cold tolerance of the ladybug was improved after the diapause and domestication, and the.3. application transcriptional group technique was used to detect normal development. Diapause - diapause relieved the gene transcription of ladybug. Through the steps of splicing, filtering and splicing, there were 82820 Unigenes, diapause and normal development differentially expressed genes, including 2125 diapause, 1376 down regulation and 1427 diapause diapause differentially expressed genes, including 749 diapause and 678 down regulated genes. In the molecular functional category and fatty acid metabolism pathway, drug metabolism, carbon metabolism and other pathways,.4. was enriched to compare diapause diapause release differential genes and diapause diapause differential genes 22, and 17 differential genes related to lipid synthesis during diapause were screened by KEGG database, and different degrees were presented during diapause. The up-regulated expression involves fatty acid synthesis, beta oxidation, lipid transport and other metabolic pathways. Besides fatty acid synthase, fat chain elongation enzyme, fatty acid desaturase gene, lipid transporter protein and lipid synthesis related enzymes up regulation, some enzymes, such as esterase and acyl transferase, are also up-regulated, which are related to lipid decomposition. It may be related to the enhancement of the immunity of diapause ladybug. In addition, the up regulation of genes related to hormone synthesis may be related to the inhibition of ovarian development in diapause ladybirds.
【學(xué)位授予單位】：中國(guó)農(nóng)業(yè)科學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：S476.2

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