發(fā)布時(shí)間：2018-03-23 14:24

  本文選題：甜菜夜蛾　切入點(diǎn)：突變體　出處：《華中農(nóng)業(yè)大學(xué)》2015年碩士論文

【摘要】：昆蟲表皮的主要成分是幾丁質(zhì)和表皮蛋白。一直以來,表皮蛋白種類和數(shù)量的變化被認(rèn)為是影響昆蟲表皮結(jié)構(gòu)和功能的主要因素。甜菜夜蛾是一種世界性的雜食性害蟲,在我國(guó)分布范圍廣,且寄主植物種類多樣。甜菜夜蛾蛹黑突變體由本實(shí)驗(yàn)室正常體甜菜夜蛾于室內(nèi)正常飼養(yǎng)條件下突變而來,蛹黑性狀能穩(wěn)定遺傳且未發(fā)生性狀分離。本研究以蛹黑突變和正常品系甜菜夜蛾為研究對(duì)象,第二代高通量測(cè)序?yàn)槠脚_(tái),對(duì)2個(gè)品系甜菜夜蛾的5齡幼蟲、預(yù)蛹、0小時(shí)蛹的表皮組織進(jìn)行數(shù)字基因表達(dá)譜升級(jí)版測(cè)序。主要研究結(jié)果如下:1通過高通量測(cè)序由正常體甜菜夜蛾5齡、預(yù)蛹、蛹的表皮組織樣本中分別得到了350734013bp,346909171bp,345310840bp的核苷酸序列,reads數(shù)目分別為7157837,7079779,7079779。在蛹黑突變體甜菜夜蛾5齡、預(yù)蛹、蛹的表皮組織樣本中分別得到了7397772,7188944,7305754個(gè)reads,核苷酸序列長(zhǎng)度分別為362490828bp,352258256bp,357981946bp。以甜菜夜蛾轉(zhuǎn)錄組數(shù)據(jù)為參考基因進(jìn)行注釋,6個(gè)表達(dá)譜測(cè)序數(shù)據(jù)中的reads的比對(duì)率基本維持在75%,其中蛹黑突變品系的5齡幼蟲的reads比對(duì)率偏低,為65.35%。2在差異基因篩選中,5齡幼蟲蛹黑突變體較正常體甜菜夜蛾有7257個(gè)基因表達(dá)上調(diào),664個(gè)基因表達(dá)下調(diào)。預(yù)蛹中460個(gè)基因表達(dá)上調(diào),439個(gè)基因表達(dá)下調(diào);0小時(shí)蛹中162個(gè)基因表達(dá)上調(diào);293個(gè)基因表達(dá)下調(diào)。3在甜菜夜蛾3個(gè)時(shí)期的表皮組織中,均篩選到了多個(gè)表皮蛋白基因的表達(dá)有顯著性差異。其中,在預(yù)蛹時(shí)期有9個(gè)表皮蛋白基因顯著性上調(diào),上調(diào)倍數(shù)最大為23.6,僅有一個(gè)表皮蛋白基因表達(dá)量下調(diào),下調(diào)倍數(shù)為21.2;在蛹期5個(gè)表皮蛋白基因表達(dá)顯著性上調(diào),上調(diào)倍數(shù)最大為210.3倍。4通過熒光定量PCR分析目的基因CPR97在2個(gè)品系甜菜夜蛾預(yù)蛹晚期、化蛹初期中的表達(dá)量。在預(yù)蛹晚期和化蛹初期,蛹黑突變體中CPR97基因的表達(dá)量均高于正常體甜菜夜蛾。CPR63在化蛹初期蛹黑突變體中表達(dá)量明顯較高。
[Abstract]:The main components of insect epidermis are chitin and epidermal protein. The variety and quantity of epidermal protein are considered to be the main factors that affect the structure and function of insect epidermis, and Spodoptera exigua is a worldwide omnivorous pest. The black mutant of Spodoptera exigua chrysalis was derived from a normal beet moth in our laboratory under normal rearing conditions. The pupa black traits can be inherited stably and no trait segregation occurred. In this study, the fifth instar larvae of two sugarbeet Spodoptera exigua were studied by using pupa black mutation and normal sugarbeet Spodoptera exigua on the platform of the second generation high-throughput sequencing. The epidermal tissues of prepupae were sequenced with the digital gene expression profile. The main results were as follows: 1. The results were as follows: 1: 1 by high-throughput sequencing from the 5th instar of normal beet exiguous moth, prepupae, prepupae, The number of nucleotide sequences of 35073404013bpnb, 346909171bptio 345310840bp was 7157837, 70797797079779 779 779 779 9. In epidermal tissue samples of pupae, 7397772n 7188944and 7305754 readswere obtained respectively, the nucleotide sequence length was 362490828bpn352258256bpn357981946bp. the reads alignment rate of the six expression profiling data was maintained at 755.The nucleotide sequence of the pupae was annotated using the transcriptome data of Spodoptera exigua as the reference gene. The reads ratio of the 5th instar larva of the medium pupa black mutant strain was lower than that of the middle pupa black mutant strain. 65.35.2 in differential gene screening, 7257 genes were up-regulated in black mutants of 5th instar larvae compared with normal beet Spodoptera exigua, while 460 genes were up-regulated in prepupae, 439 genes were down-regulated, 162 genes were down-regulated in 0-hour pupae. The expression of 293 genes was down-regulated in the epidermis of Spodoptera exigua in three stages. Among them, 9 epidermal protein genes were significantly upregulated in prepupae stage, the maximum up-regulation was 23.6, and only one epidermal protein gene was down-regulated. The expression of 5 epidermal protein genes was significantly up-regulated in pupal stage, and the maximum up-regulation was 210.3 times. The target gene CPR97 was analyzed by fluorescence quantitative PCR in the late stage of prepupae of two strains of Spodoptera exigua. The expression of CPR97 gene in the pupae black mutant was significantly higher than that in the normal beet Spodoptera exigua in the early pupae stage and in the early pupae stage.
【學(xué)位授予單位】：華中農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：S433.4

【參考文獻(xiàn)】

相關(guān)碩士學(xué)位論文 前1條

1 劉向陽;甜菜夜蛾Spodoptera exigua （Hübner）黑化機(jī)理的研究[D];華中農(nóng)業(yè)大學(xué);2007年

，

本文編號(hào)：1653845