褐飛虱achaete-scute homologue和target of rapamycin基因功能研究
發(fā)布時間：2018-08-10 19:03
【摘要】：褐飛虱(半翅目;飛虱科)是水稻上一種重要的遷飛性害蟲,具有遠距離遷飛能力。褐飛虱成蟲前翅翅脈上著生很多剛毛,其功能和發(fā)育分子機制并不清楚;褐飛虱遷飛時卵巢發(fā)育被抑制,是一種短暫的生殖滯育現(xiàn)象,生殖滯育的分子調控機制也不清楚。Achaete、scute是一類編碼b HLH結構的轉錄因子,控制果蠅鬃毛的發(fā)育。Target of Rapamycin(TOR)信號通路影響細胞的增長及分化,受外界營養(yǎng)條件的調控,Target of Rapamycin(TOR)是該通路中的關鍵基因。為弄清楚褐飛虱剛毛發(fā)育和遷飛滯育的調控關鍵基因,我們克隆褐飛虱achaete-scute homologue(ASH)、Target of Rapamycin(Nl TOR)基因,并通過RNAi干涉研究這兩個基因的功能。結果如下:一、褐飛虱achaete-scute homologue基因功能研究1.本研究發(fā)現(xiàn)Nl ASH基因的c DNA全長669個堿基,編碼233個氨基酸;并對Nl ASH進行系統(tǒng)進化樹分析,發(fā)現(xiàn)褐飛虱ASH與意大利蜜蜂和金小蜂親緣關系最近。2.Nl ASH主要在褐飛虱若蟲期高表達,成蟲期表達量較低;若蟲期不同組織部位的表達分析發(fā)現(xiàn)Nl ASH在3齡褐飛虱的翅芽中高表達。3.采用ds RNA干擾3齡褐飛虱Nl ASH的表達,發(fā)現(xiàn)Nl ASH下調對褐飛虱有顯著的致死效應并且褐飛虱羽化后,成蟲翅出現(xiàn)畸形,前翅翅脈上剛毛幾乎完全消失。二、褐飛虱Target of Rapamycin基因功能研究1.本研究發(fā)現(xiàn)Nl TOR在褐飛虱若蟲與成蟲期都表達,在成蟲期高表達,長、短翅品系Nl TOR表達存在顯著性差異,短翅品系在成蟲1、2和4天Nl TOR表達水平顯著高于長翅品系。2.沉默Nl TOR后連續(xù)飼養(yǎng)觀察15天,稻莖中沒有褐飛虱孵化出來,解剖稻莖也沒有發(fā)現(xiàn)卵。解剖注射了ds TOR的雌成蟲卵巢,發(fā)現(xiàn)褐飛虱卵巢發(fā)育被抑制,卵不能形成,卵巢小管和輸卵管都被抑制。3.沉默Nl TOR顯著延長褐飛虱若蟲發(fā)育到成蟲的時間,降低蛻皮激素合成途徑關鍵基因Nl E74B(Ecdysone-induced protein 74B)和蛻皮激素受體Nl Ec R(Ecdysteroidreceptor)基因的表達水平;Nl Fo XO(Forkhead-box O)和Nl Vg(vitellogenin)的表達量也降低。綜上所述,本研究表明Nl ASH基因對褐飛虱前翅翅脈上剛毛的發(fā)育起著決定性作用;Nl TOR基因影響褐飛虱蛻皮激素的合成及蛻皮激素信號傳遞,調控卵巢發(fā)育,可能參與調控褐飛虱遷飛生殖滯育。
[Abstract]:Brown planthopper (Hemiptera; planthopper) is an important migratory pest in rice. There are many bristles on the forewing veins of brown planthopper, and its function and developmental molecular mechanism are not clear. The ovarian development is inhibited during the migration of brown planthopper, which is a temporary phenomenon of reproductive diapause. The molecular regulatory mechanism of reproductive diapause is also unclear. AchaeteScute is a kind of transcription factor that encodes the structure of b HLH. It controls the development of mane of Drosophila mane. Target of Rapamycin (TOR) signaling pathway affects cell growth and differentiation. Target of Rapamycin (TOR) is a key gene in this pathway. In order to find out the key genes regulating the development of bristles and migration diapause of brown planthopper, we cloned the target of Rapamycin (Nl TOR) gene of achaete-scute homologue (ASH), and studied the function of these two genes by RNAi interference. The results are as follows: 1. The function of achaete-scute homologue gene in brown planthopper 1. In this study, we found that the c DNA of the NL ASH gene was 669 bases in length, encoding 233 amino acids, and analyzed the NL ASH phylogenetic tree. It was found that the relationship between ASH and Apis mellifera and A. aureus was the most recent. 2. NL ASH was mainly expressed in the nymph stage, but low in the adult stage. The expression of NL ASH was found to be highly expressed in the wing buds of the third instar brown planthopper. DS RNA was used to interfere with the expression of Nl ASH in the 3rd instar brown planthopper. It was found that the down-regulation of NL ASH had a significant lethal effect on the brown planthopper. After emergence, the adult wings were deformed and the bristles on the front fin veins almost disappeared. Second, the function of Target of Rapamycin gene of brown planthopper 1. In this study, it was found that NL TOR was expressed in both nymphs and adults of brown planthopper, but there was a significant difference in the expression of NL TOR in long and short winged lines during adult stage. The expression level of NL TOR in short-winged lines was significantly higher than that in long-winged lines at 12nd and 4th day. No brown planthopper hatched in rice stem and no eggs were found in dissected stem after silencing N _ l TOR for 15 days. The ovaries of female adults with DS TOR were dissected. It was found that the ovaries of brown planthopper were inhibited, the eggs could not be formed, and the ovarian tubules and oviducts were inhibited. Silencing Nilaparvata lugens significantly prolonged the development time of nymphs to adults, and decreased the expression level of NlE74B (Ecdysone-induced protein 74B) and Nl Ec R (Ecdysteroidreceptor) gene of ecdysone receptor (Nl Ec R (Ecdysteroidreceptor) as well as the expression of Nl Vg (vitellogenin). To sum up, this study suggests that the NL ASH gene plays a decisive role in the development of the bristles on the front fin veins of the brown planthopper. The NL TOR gene affects the synthesis of ecdysone and the signal transduction of ecdysone in the brown planthopper, and regulates the development of the ovaries. It may be involved in regulating the migration of brown planthopper to reproductive diapause.
【學位授予單位】：華中農(nóng)業(yè)大學
【學位級別】：碩士
【學位授予年份】：2016
【分類號】：S435.112.3
本文編號：2175915
[Abstract]:Brown planthopper (Hemiptera; planthopper) is an important migratory pest in rice. There are many bristles on the forewing veins of brown planthopper, and its function and developmental molecular mechanism are not clear. The ovarian development is inhibited during the migration of brown planthopper, which is a temporary phenomenon of reproductive diapause. The molecular regulatory mechanism of reproductive diapause is also unclear. AchaeteScute is a kind of transcription factor that encodes the structure of b HLH. It controls the development of mane of Drosophila mane. Target of Rapamycin (TOR) signaling pathway affects cell growth and differentiation. Target of Rapamycin (TOR) is a key gene in this pathway. In order to find out the key genes regulating the development of bristles and migration diapause of brown planthopper, we cloned the target of Rapamycin (Nl TOR) gene of achaete-scute homologue (ASH), and studied the function of these two genes by RNAi interference. The results are as follows: 1. The function of achaete-scute homologue gene in brown planthopper 1. In this study, we found that the c DNA of the NL ASH gene was 669 bases in length, encoding 233 amino acids, and analyzed the NL ASH phylogenetic tree. It was found that the relationship between ASH and Apis mellifera and A. aureus was the most recent. 2. NL ASH was mainly expressed in the nymph stage, but low in the adult stage. The expression of NL ASH was found to be highly expressed in the wing buds of the third instar brown planthopper. DS RNA was used to interfere with the expression of Nl ASH in the 3rd instar brown planthopper. It was found that the down-regulation of NL ASH had a significant lethal effect on the brown planthopper. After emergence, the adult wings were deformed and the bristles on the front fin veins almost disappeared. Second, the function of Target of Rapamycin gene of brown planthopper 1. In this study, it was found that NL TOR was expressed in both nymphs and adults of brown planthopper, but there was a significant difference in the expression of NL TOR in long and short winged lines during adult stage. The expression level of NL TOR in short-winged lines was significantly higher than that in long-winged lines at 12nd and 4th day. No brown planthopper hatched in rice stem and no eggs were found in dissected stem after silencing N _ l TOR for 15 days. The ovaries of female adults with DS TOR were dissected. It was found that the ovaries of brown planthopper were inhibited, the eggs could not be formed, and the ovarian tubules and oviducts were inhibited. Silencing Nilaparvata lugens significantly prolonged the development time of nymphs to adults, and decreased the expression level of NlE74B (Ecdysone-induced protein 74B) and Nl Ec R (Ecdysteroidreceptor) gene of ecdysone receptor (Nl Ec R (Ecdysteroidreceptor) as well as the expression of Nl Vg (vitellogenin). To sum up, this study suggests that the NL ASH gene plays a decisive role in the development of the bristles on the front fin veins of the brown planthopper. The NL TOR gene affects the synthesis of ecdysone and the signal transduction of ecdysone in the brown planthopper, and regulates the development of the ovaries. It may be involved in regulating the migration of brown planthopper to reproductive diapause.
【學位授予單位】：華中農(nóng)業(yè)大學
【學位級別】：碩士
【學位授予年份】：2016
【分類號】：S435.112.3
【參考文獻】
相關期刊論文 前1條
1 封傳紅,翟保平,張孝羲;褐飛虱的再遷飛能力[J];中國水稻科學;2001年02期
，本文編號：2175915
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