褐飞虱achaete-scute homologue和target of rapamycin基因功能研究
发布时间：2018-08-10 19:03
【摘要】：褐飞虱(半翅目;飞虱科)是水稻上一种重要的迁飞性害虫,具有远距离迁飞能力。褐飞虱成虫前翅翅脉上着生很多刚毛,其功能和发育分子机制并不清楚;褐飞虱迁飞时卵巢发育被抑制,是一种短暂的生殖滞育现象,生殖滞育的分子调控机制也不清楚。Achaete、scute是一类编码b HLH结构的转录因子,控制果蝇鬃毛的发育。Target of Rapamycin(TOR)信号通路影响细胞的增长及分化,受外界营养条件的调控,Target of Rapamycin(TOR)是该通路中的关键基因。为弄清楚褐飞虱刚毛发育和迁飞滞育的调控关键基因,我们克隆褐飞虱achaete-scute homologue(ASH)、Target of Rapamycin(Nl TOR)基因,并通过RNAi干涉研究这两个基因的功能。结果如下:一、褐飞虱achaete-scute homologue基因功能研究1.本研究发现Nl ASH基因的c DNA全长669个碱基,编码233个氨基酸;并对Nl ASH进行系统进化树分析,发现褐飞虱ASH与意大利蜜蜂和金小蜂亲缘关系最近。2.Nl ASH主要在褐飞虱若虫期高表达,成虫期表达量较低;若虫期不同组织部位的表达分析发现Nl ASH在3龄褐飞虱的翅芽中高表达。3.采用ds RNA干扰3龄褐飞虱Nl ASH的表达,发现Nl ASH下调对褐飞虱有显著的致死效应并且褐飞虱羽化后,成虫翅出现畸形,前翅翅脉上刚毛几乎完全消失。二、褐飞虱Target of Rapamycin基因功能研究1.本研究发现Nl TOR在褐飞虱若虫与成虫期都表达,在成虫期高表达,长、短翅品系Nl TOR表达存在显著性差异,短翅品系在成虫1、2和4天Nl TOR表达水平显著高于长翅品系。2.沉默Nl TOR后连续饲养观察15天,稻茎中没有褐飞虱孵化出来,解剖稻茎也没有发现卵。解剖注射了ds TOR的雌成虫卵巢,发现褐飞虱卵巢发育被抑制,卵不能形成,卵巢小管和输卵管都被抑制。3.沉默Nl TOR显著延长褐飞虱若虫发育到成虫的时间,降低蜕皮激素合成途径关键基因Nl E74B(Ecdysone-induced protein 74B)和蜕皮激素受体Nl Ec R(Ecdysteroidreceptor)基因的表达水平;Nl Fo XO(Forkhead-box O)和Nl Vg(vitellogenin)的表达量也降低。综上所述,本研究表明Nl ASH基因对褐飞虱前翅翅脉上刚毛的发育起着决定性作用;Nl TOR基因影响褐飞虱蜕皮激素的合成及蜕皮激素信号传递,调控卵巢发育,可能参与调控褐飞虱迁飞生殖滞育。
[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.
【学位授予单位】：华中农业大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：S435.112.3
本文编号：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.
【学位授予单位】：华中农业大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：S435.112.3
【参考文献】
相关期刊论文 前1条
1 封传红,翟保平,张孝羲;褐飞虱的再迁飞能力[J];中国水稻科学;2001年02期
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