【摘要】： 科學(xué)家近年來的研究表明,B／Q型煙粉虱成功入侵的原因主要有以下幾種：一是生殖干涉,即非對稱性交配；二是感染雙生病毒,即與病毒的互利互惠；三是煙粉虱對化學(xué)殺蟲劑的抗性。但引起這些機(jī)制的生理和分子途徑尚不明確,因此,探討煙粉虱入侵的生理和分子機(jī)制,對于日后有效控制其為害具有深遠(yuǎn)意義。本研究主要從煙粉虱的生殖發(fā)育包括：卵巢形態(tài)及發(fā)育、精巢發(fā)育及卵子發(fā)生等為切入點(diǎn),通過比較不同生物型煙粉虱卵巢內(nèi)不同發(fā)育級別卵子比例、卵子發(fā)生過程及卵黃發(fā)生動(dòng)態(tài)等的異同,分析引起不同生物型煙粉虱之間生殖差異的生殖基礎(chǔ),并通過克隆卵黃原蛋白(Vitellogenin, Vg)及其受體(Vitellogenin Receptor, VgR)基因,分析比較Vg基因序列和結(jié)構(gòu),以及檢測感染TYLCCNV病毒生物型煙粉虱轉(zhuǎn)錄水平的差異,探討入侵種煙粉虱成功入侵的生殖基礎(chǔ)與分子機(jī)制。主要結(jié)果如下： 1煙粉虱內(nèi)生殖系統(tǒng)形態(tài)和卵巢發(fā)育的比較分析 B型煙粉虱雌蟲內(nèi)生殖系統(tǒng)主要由對稱的一對卵巢、一個(gè)直徑約20μm的受精囊、側(cè)輸卵管和中輸卵管組成。B型煙粉虱卵巢管屬于端滋式發(fā)育模式,由12-22根卵巢管組成,可以根據(jù)發(fā)育時(shí)期將煙粉虱的卵巢分為四個(gè)不同階段。根據(jù)卵巢內(nèi)卵子形態(tài)和卵黃蛋白沉積的情況,可以把卵子分為四個(gè)級別,即Ⅰ、Ⅱ、Ⅲ和Ⅳ級。B煙粉虱的精巢主要由一對對稱排列的睪丸、雄性附腺和輸精管組成。比較B型和ZHJ1型煙粉虱卵巢發(fā)育顯示,兩種生物型煙粉虱卵巢形態(tài)相似,以健康棉花為寄主植物時(shí),生殖發(fā)育差異不顯著。 2煙粉虱卵子發(fā)生和精子發(fā)生 煙粉虱的卵子發(fā)生始于偽蛹期,在煙粉虱雌蟲開始形成卵巢時(shí),卵原干細(xì)胞分別分化成滋養(yǎng)細(xì)胞、卵母細(xì)胞以及濾泡細(xì)胞。紅眼期偽蛹已經(jīng)發(fā)現(xiàn)有卵巢管,成蟲羽化后12 h就觀察到有產(chǎn)卵行為,24 h后解剖出含有成熟卵子的卵巢。48 h解剖出帶有受精囊的卵巢。煙粉虱的卵子發(fā)生分為滋養(yǎng)期、卵黃發(fā)生前期、卵黃發(fā)生期和成熟期四個(gè)時(shí)期。卵母細(xì)胞通過滋養(yǎng)細(xì)胞運(yùn)輸和自身攝取兩種方式進(jìn)行Vg等營養(yǎng)物質(zhì)的轉(zhuǎn)運(yùn)。煙粉虱的精子發(fā)生同樣始于偽蛹期,精細(xì)胞先后經(jīng)歷精原細(xì)胞、初級精細(xì)胞、次級精細(xì)胞和成熟精細(xì)胞四個(gè)階段,不同階段的形態(tài)和結(jié)構(gòu)也不同。 3煙粉虱Vg單克隆抗體和多克隆抗體的制備 通過收集煙粉虱產(chǎn)的新鮮卵作為免疫抗原制備單克隆、多克隆抗體。經(jīng)小鼠脾細(xì)胞與可體外培養(yǎng)并繁殖的入骨髓瘤細(xì)胞進(jìn)行體外融合,利用ELISA篩選陽性克隆并擴(kuò)大培養(yǎng)后用上清制備腹水,收集腹水經(jīng)純化后得到Vg的單克隆抗體。經(jīng)SDS-PAGE電泳,切取雌性特異性條帶,將回收條帶電透析后濃縮制備抗原,皮下多點(diǎn)注射雄性白兔,爾后取靜脈血經(jīng)沉淀純化后獲得多克隆抗體。ELISA測定效價(jià)發(fā)現(xiàn),單抗效價(jià)為1：100,000,多抗的效價(jià)為1：68,000。Wertern-blot分析發(fā)現(xiàn)抗卵黃蛋白抗體能和幾種生物型煙粉虱卵黃蛋白及血淋巴中的雌性特異性蛋白產(chǎn)生免疫反應(yīng),但是不能與雄蟲提取物起特異性免疫反應(yīng)。 4煙粉虱卵黃發(fā)生及Vg特性的分析 利用凝膠層析和離子交換層析對煙粉虱卵黃蛋白(Vetellin, Vt)進(jìn)行了分離純化,利用非變性凝膠電泳和變性凝膠電泳分析卵黃蛋白分子組成發(fā)現(xiàn),卵黃蛋白是由兩個(gè)大小為190 kDa相似亞基組成的分子量約380 kDa的大分子蛋白。對煙粉虱Vg性質(zhì)分析顯示,煙粉虱Vg是磷酸化糖基化脂蛋白。利用雙抗夾心ELISA對取食健康棉花的煙粉虱不同發(fā)育時(shí)期Vg含量檢測發(fā)現(xiàn),煙粉虱Vg合成始于偽蛹期,羽化后血淋巴中的Vg含量先下降后逐漸上升,卵巢中的Vt基本呈逐漸上升趨勢。 5煙粉虱Vg基因的克隆及轉(zhuǎn)錄表達(dá)分析 克隆獲得B型煙粉虱Vg基因cDNA全長序列為6731 bp,開放閱讀框長度為6474bp,共編碼2158個(gè)氨基酸,預(yù)測分子量為242 kDa,等電點(diǎn)為8.9,3'端非編碼區(qū)為220bp,5'端非編碼區(qū)為37 bp。其中前16個(gè)氨基酸為信號肽,絲氨酸(S)占12.5%,天冬氨酸(N)占12.7%,谷氨酰胺(Q)8.9%。在490個(gè)氨基酸有一個(gè)RXXR酶切位點(diǎn),將Vg基因切成編碼約50 kDa和190 kDa的大小蛋白。序列相似性比對后發(fā)現(xiàn),煙粉虱Vg基因與玻璃葉蟬Homalodisca coagulata的相似性為86.7%,與葉蜂Athalia rosae的相似性為85.6%,與大鱉負(fù)蝽Lethocerus deyrollei的相似性為84.5%。利用熒光定量PCR(Q-PCR)技術(shù)檢測不同時(shí)期Vg基因轉(zhuǎn)錄水平分析發(fā)現(xiàn),羽化后煙粉虱Vg表達(dá)量呈逐漸升高趨勢。 6不同生物型煙粉虱Vg基因克隆及分析。 ZHJ2型煙粉虱Vg基因全長6721 bp,開放閱讀框長度約6549 bp,共編碼2183個(gè)氨基酸,預(yù)測分子量為245 kDa,等電點(diǎn)為8.7,3'非編碼區(qū)為109 bp,5'非編碼區(qū)為63bp。其中前22個(gè)氨基酸為信號肽,在450個(gè)氨基酸有一個(gè)RXXR酶切位點(diǎn),將Vg基因切成編碼約50 kDa和190 kDa大小蛋白。絲氨酸(S)占12.5%,天冬氨酸(N)占13.5%,谷氨酰胺(Q)7.1%,丙氨酸(A)9.2%。序列相似性比對后發(fā)現(xiàn),ZHJ-2型煙粉虱Vg基因與菜葉蜂Athalia rosae的相似性為93.9%,與大鱉負(fù)蝽Lethocerus deyrollei的相似性為93.5%,與玻璃葉蟬Homalodisca coagulate相似性為89.9%。 Q型煙粉虱Vg基因全長6871 bp,開放閱讀框長度約6651 bp,共編碼2217個(gè)氨基酸,預(yù)測分子量為246 kDa,等電點(diǎn)為8.9,3'非編碼區(qū)為107 bp,5'非編碼區(qū)為43 bp。其中前17個(gè)氨基酸為信號肽,在490和790個(gè)氨基酸各有一個(gè)RXXR酶切位點(diǎn),可能會(huì)將Vg基因切成編碼約50 kDa、90 kDa、150 kDa或者190 kDa大小蛋白。絲氨酸(S)占15%,天冬氨酸(N)占12.8%,谷氨酰胺(Q)6.2%,丙氨酸(A)9.2%。序列相似性比對后發(fā)現(xiàn),Q型煙粉虱Vg基因與菜葉蜂A. rosae的相似性較低,僅為63.4%,與大鱉負(fù)蝽L. deyrollei的相似性為57.3%,與玻璃葉蟬H. coagulata相似性為55.6%。 對三種煙粉虱Vg的核酸序列和氨基酸序列比較分析后發(fā)現(xiàn),入侵種B型／Q型與本地種ZHJ2型煙粉虱相比,在第620-640氨基酸序列間,均比ZHJ2型煙粉虱多一段長度為15個(gè)氨基酸的多聚絲氨酸片段。而入侵種Q型與B型、本地種ZHJ2煙粉虱Vg基因序列相比,Q型煙粉虱具有兩個(gè)酶切位點(diǎn),且在多聚絲氨酸區(qū)多9個(gè)重復(fù)的GHN功能結(jié)構(gòu)域。三個(gè)生物型煙粉虱Vg基因都具有卵黃蛋白典型的功能結(jié)構(gòu)域GCMG和NIIK,以及重復(fù)的多聚絲氨酸區(qū),與其它昆蟲Vg基因推導(dǎo)氨基酸序列比較,煙粉虱Vg基因不僅含有多個(gè)多聚絲氨酸區(qū)域,且存在多聚天冬氨酸和谷氨酰胺區(qū),這在昆蟲卵黃蛋白基因序列結(jié)構(gòu)上是不多見的。有意思的是在煙粉虱Vg基因序列的氮端多聚絲氨酸區(qū)之前,都會(huì)出現(xiàn)多個(gè)重復(fù)的GH/RN功能結(jié)構(gòu)域,其功能還有待進(jìn)一步研究。 7煙粉虱Vg受體基因的克隆及表達(dá)分析 克隆到B型煙粉虱Vg受體(Vg Receptor, VgR)基因全長cDNA序列,大小5774bp,編碼1919個(gè)氨基酸,5'端非編碼區(qū)201 bp,等電點(diǎn)為8.7,分子量大小約201 kDa,跨膜結(jié)構(gòu)域分析顯示,VgR跨膜結(jié)構(gòu)位于C-末端的1679-1696個(gè)氨基酸位置。煙粉虱的VgR屬于低密度脂蛋白受體(Low Density Lipoprotein Receptor, LDLR)家族,具有一些LDLR家族典型的保守結(jié)構(gòu)域,主要包括：配體結(jié)合域(Ligand-Binding Domain)、表皮生長因子前體同源域(Epidermal Growth Factor Precursor Homology Domain)、跨膜域(Transmembrane Domain)、O-聯(lián)糖功能域(O-linked Carbohydrate Domain)及胞質(zhì)尾域(Cytoplasmic Domain)。同源比對分析顯示,B型煙粉虱VgR同美洲大蠊Periplaneta americana相似度達(dá)83%,及德國小蠊Blattella germanica相似度達(dá)74%,并利用同源建模方法構(gòu)建了VgR編碼蛋白質(zhì)三維結(jié)構(gòu)。同時(shí)用熒光定量PCR (Q-PCR)技術(shù)檢測分析了B型煙粉虱Vg受體基因轉(zhuǎn)錄表達(dá)情況。 8感染TYLCCNV病毒對煙粉虱生殖力及Vg、VgR基因轉(zhuǎn)錄表達(dá)的影響 對羽化后15 d內(nèi)的B型煙粉虱雌蟲卵巢發(fā)育進(jìn)度觀察發(fā)現(xiàn),初羽化的雌蟲已經(jīng)開始卵巢發(fā)育,在卵巢內(nèi)未見成熟的卵子。羽化后2 d發(fā)現(xiàn)有卵黃沉積的卵子,發(fā)育3-4 d的卵巢內(nèi)觀察到成熟卵子,且隨著發(fā)育時(shí)間的延長,卵巢內(nèi)成熟卵子數(shù)目逐漸增多,至羽化后11-14 d卵巢內(nèi)成熟卵子數(shù)目達(dá)到最大。煙粉虱卵巢中的卵子在形態(tài)上分為四個(gè)級別,分別標(biāo)記為Ⅰ級、Ⅱ級、Ⅲ級和Ⅳ級,其中Ⅳ級為成熟卵。煙粉虱卵巢時(shí)期和卵子發(fā)育級別的劃分為評價(jià)煙粉虱生殖力奠定了基礎(chǔ)。比較B型／ZHJ1型煙粉虱取食感染中國番茄黃化曲葉病毒(TYLCCNV)煙草植株與在健康煙草植株上取食2種條件下的卵巢發(fā)育進(jìn)度結(jié)果表明,B型煙粉虱在患病植株上取食,卵子發(fā)育進(jìn)度加快,表現(xiàn)在不同發(fā)育時(shí)間Ⅱ、Ⅲ和Ⅳ級卵子數(shù)目顯著高于取食未感毒煙草的煙粉虱。然而,觀察發(fā)現(xiàn)TYLCCNV對土著ZHJ1型煙粉虱卵巢內(nèi)卵子發(fā)育進(jìn)度卻無顯著影響。提取同期煙粉虱總RNA,并分別考察了取食健康煙草和感染病毒煙草后的卵黃發(fā)生、Vg/VgR基因轉(zhuǎn)錄情況。發(fā)現(xiàn)了感染病毒后煙粉虱Vg含量、Vg/VgR基因轉(zhuǎn)錄水平顯著提高。
[Abstract]:Research in recent years has shown that the main reasons for the successful invasion of B / Q type Bemisia tabaci are as follows: one is the reproductive interference, that is, asymmetric mating; two is the infection of the double biovirus, that is, the mutual benefit of the virus, and the three is the resistance to chemical insecticides by the whitefly. Therefore, the study of the physiological and molecular mechanisms of the invasion of Bemisia tabaci is of profound significance for the effective control of its damage in the future. This study mainly from the form and development of the ovary, the development of the ovary, the development of the spermary and the oogenesis, and the comparison of the egg ratio of different developmental levels in the ovary of different biologic type of Bemisia tabaci. The reproductive basis of reproductive differences between different biotypes of Bemisia tabaci was analyzed. The sequence and structure of the Vg gene were analyzed and compared by cloning the Vitellogenin (Vg) and its receptor (Vitellogenin Receptor, VgR) gene, and the detection of the biotype of the biotype of TYLCCNV virus was detected. The differences in transcriptional level were explored to explore the reproductive basis and molecular mechanism of invasive species of Bemisia tabaci.
Comparative analysis of reproductive system morphology and ovarian development in 1 Bemisia tabaci
The internal reproductive system of female Bemisia tabaci B mainly consists of a symmetrical pair of ovary, a seminal vesicle with a diameter of about 20 mu m, the lateral oviduct and the middle fallopian tube composed of the.B type of the ovaries, which are composed of 12-22 ovarian tubes and can be divided into four different stages according to the development period. The condition of submorphologic and yolk protein deposition can be divided into four levels. The sperma of.B, II, III and IV is mainly composed of a pair of symmetrical arranged testis, male attached glands and vas deferens. The ovarian development of B and ZHJ1 type tobacco whitefly shows that the ovarian morphology of the two species of Bemisia tabaci is similar to that of healthy cotton. In the main plant, the difference of reproductive development is not significant.
2 oogenesis and spermatogenesis of Bemisia tabaci
The eggs of Bemisia tabaci began in the puppet stage. Oocytes were differentiated into trophoblastic cells, oocytes and follicle cells when the females began to form ovaries. The ovaries were found in the pseudochrysalis at the red eye stage. The oviposition was observed at 12 h after adult emergence. After 24 h, the ovaries containing mature ovaries were dissected and dissected by.48 H. The ovaries with the seminal vesicles. The eggs of Bemisia tabaci are divided into the nourishing period, the early stage of the yolk occurrence, the yolk occurrence period and the maturity period of four periods. Oocyte transshipment through the trophoblast transport and self uptake in two ways. The spermatogenesis of Bemisia tabaci also begins in the pseudo pupae period and spermatocytes successively experience spermatogonial cells. There are four stages of primary spermatocyte, secondary spermatocyte and mature spermatocyte, and their morphology and structure are different at different stages.
Preparation of 3 monoclonal antibodies and polyclonal antibodies against Bemisia tabaci Vg
By collecting the fresh eggs produced by Bemisia tabaci as immunogen to prepare monoclonal and polyclonal antibodies, the mouse splenocytes were fused with the myeloma cells which could be cultured and propagated in vitro. The positive clones were screened by ELISA and the ascites were prepared with the supernatant, and the monoclonal antibodies of the Vg were obtained after the purification of the abdominal water. The SDS-P The female specific bands were cut by AGE electrophoresis. The antigen was concentrated by electrified dialysis, and the male rabbits were injected subcutaneously. Then the venous blood was purified and purified to obtain the titer of the polyclonal antibody.ELISA. The titer of the monoclonal antibody was 1:100000. The titer of the polyclonal antibody was 1: 68000.Wertern-blot analysis to find the yolk antibody. It can react with the yolk protein of several biotype Bemisia tabaci and the female specific protein in the hemolymph, but it can not react with the male insect extract specific immune response.
Analysis of the yolk occurrence and Vg characteristics of 4 whitefly
Vetellin (Vt) was purified by gel chromatography and ion exchange chromatography. The analysis of the molecular composition of yolk protein was found by non denaturing gel electrophoresis and denaturing gel electrophoresis. The yolk protein was a large molecular weight of about 380 kDa with two molecular weights of 190 kDa similar subunits. To Vg of Bemisia tabaci. The properties analysis showed that the Vg of whitefly was phosphorylated glycosylated lipoprotein. Using the double anti sandwich ELISA to detect the Vg content in the different developmental stages of the healthy cotton Bemisia tabaci, the Vg synthesis began in the pseudo pupa period. The Vg content in the hemolymph was decreased first and then gradually increased, and the Vt in the ovary increased gradually.
Cloning and transcriptional expression analysis of the Vg gene of 5 whitefly
The full-length sequence of Vg gene cDNA of the B type Bemisia tabaci was 6731 BP, the length of the open reading frame was 6474bp, a total of 2158 amino acids were encoded, the predicted molecular weight was 242 kDa, the isoelectric point was 220bp in the non coding region of the 8.9,3'terminal, the non coding region of the 5' terminal was 37 bp., the first 16 amino acids were signal peptides, the serine (S) was 12.5%, the aspartic acid (N) was 12.7%, and the valley ammonia was 12.7%. Amide (Q) 8.9%. has a RXXR enzyme cut site at 490 amino acids, and the Vg gene is cut into the size protein of about 50 kDa and 190 kDa. Sequence similarity comparison shows that the similarity between the Vg gene of the whitefly and the Homalodisca coagulata of the leafhopper is 86.7%, and the similarity between the Athalia Rosae of the leaf bee and the Athalia Rosae is 85.6%. The similarity of 84.5%. using fluorescence quantitative PCR (Q-PCR) technique to detect the transcriptional level of Vg gene in different periods showed that the Vg expression of whitefly was gradually increasing.
6 cloning and analysis of Vg gene of different biotype Bemisia tabaci.
The Vg gene of Bemisia tabaci ZHJ2 is 6721 BP in length, the length of the open reading frame is about 6549 BP, which encodes 2183 amino acids, the predicted molecular weight is 245 kDa, the isoelectric point is 109 BP in 8.7,3'non coding region, the non coding region of 5' is 63bp. and the first 22 amino acids are signal peptides, and there is a RXXR enzyme cut site in the 450 amino acids, and the Vg gene is cut into about 50 kDa and the Vg gene is cut into about 50 kDa and 190 kDa serine (S) was 12.5%, aspartic acid (N) accounted for 13.5%, glutamine (Q) 7.1%, and alanine (A) 9.2%. sequence similarity comparison found that the similarity between the Vg gene of ZHJ-2 type tobacco whitefly and the Athalia Rosae of the leaf wasp was 93.9%. The similarity between the ZHJ-2 and the Lethocerus deyrollei was 93.5%. The similarity is 89.9%.
The Vg gene of Bemisia tabaci Q has a full length of 6871 BP, the length of the open reading frame is about 6651 BP, encoding a total of 2217 amino acids, the predicted molecular weight is 246 kDa, the isoelectric point is 107 BP in the non coding region of 8.9,3', the non coding region of 5' is 43 bp. and the first 17 amino acids are signal peptides, and there is a RXXR enzyme cutting site in the 490 and 790 amino acids each, and the Vg gene may be cut into the Vg gene. Coding about 50 kDa, 90 kDa, 150 kDa or 190 kDa protein. Serine (S) accounted for 15%, aspartic acid (N) accounted for 12.8%, glutamine (Q) 6.2%, and alanine (A) 9.2%. sequence similarity comparison found that the Vg gene of Q type tobacco whitefly was less similar to 63.4%, and 57.3%, and vitreous leaf. The H. coagulata similarity of cicadas is 55.6%.
After comparison and analysis of nucleic acid sequence and amino acid sequence of three species of whitefly Vg, it was found that the invading species B / Q type was more than the local ZHJ2 type of Bemisia tabaci and was more than 15 amino acid polyserine fragments in the 620-640 amino acid sequence compared to the ZHJ2 type tobacco whitefly. The invasive species Q and B, and the Vg gene sequence of native ZHJ2 In comparison, the Q type of Bemisia tabaci has two enzyme cutting sites and 9 repetitive GHN functional domains in the polyserine region. Three biotype Vg genes of Bemisia tabaci have typical functional domains GCMG and NIIK, and repeated polyserine regions. Compared with other insect Vg genes, the Vg gene of the Bemisia tabaci Vg gene is derived. It not only contains multiple polyserine regions, but also has polyaspartic acid and glutamine region, which is not much in the sequence structure of the insect yolk protein gene. It is interesting that many repetitive GH/RN functional domains appear before the N-terminal polyserine region of the Vg gene sequence of the Bemisia tabaci, and its function remains to be further studied. Research.
Cloning and expression analysis of Vg receptor gene of 7 Bemisia tabaci
The full-length cDNA sequence of the Vg Receptor (Vg Receptor, VgR) gene was cloned, the size 5774bp, the encoding 1919 amino acids, the non coding region of 5'end 201 BP, the isoelectric point 8.7 and the molecular weight about 201 kDa, the transmembrane domain analysis showed that the VgR transmembrane structure was located at the 1679-1696 amino acid position at the end of the C-. The Low Density Lipoprotein Receptor (LDLR) family has some typical conservative domains of the LDLR family, including the ligand binding domain (Ligand-Binding Domain), the epidermal growth factor precursor domain (Epidermal Growth Factor Precursor), the trans membrane domain, and the carbohydrate domain. D Carbohydrate Domain) and cytoplasmic tail region (Cytoplasmic Domain). Homologous analysis showed that the Americana similarity between B type VgR and cockroach of cockroach of American cockroach was 83%, and the Blattella germanica similarity of German cockroach was 74%, and the three-dimensional structure of the VgR coding protein was constructed by using the homologous modeling method. The transcriptional expression of Vg receptor gene in Bemisia tabaci B was detected by technology.
8 effects of TYLCCNV infection on fecundity and transcription of Vg and VgR genes in Bemisia tabaci
The ovarian development of female Bemisia tabaci (Bemisia tabaci) in 15 d after eclosion was observed. It was found that the ovaries were developed and no mature eggs were found in the ovaries. The egg yolk deposited in the 2 D was found, and the mature ovaries were observed in the ovary of 3-4 D, and the number of mature ovaries in the ovary increased with the development time. The number of mature ovaries in the ovary of 11-14 d after eclosion reached the maximum. The ovaries in the ovaries of the Bemisia tabaci were divided into four grades, which were marked as grade I, grade II, grade III and IV respectively, of which the grade IV was mature. The division of the ovary period and the egg development level of the whitefly laid a foundation for evaluating the fertility of the whitefly. The results of ovarian development of type / ZHJ1 type / ZHJ1 Bemisia tabaci (Bemisia tabaci) infected with Chinese tomato yellow leaf curl virus (TYLCCNV) tobacco plants and feeding on healthy tobacco plants showed that the B type of Bemisia tabaci was fed on the sick plants, and the development of eggs was accelerated. The number of eggs of grade III and IV was significantly higher than that of the number of grade III and IV. However, it was observed that TYLCCNV had no significant effect on the development of ovaries in the aboriginal ZHJ1 - type tobacco whitefly. The total RNA of the tobacco whitefly was extracted at the same time, and the yolk occurrence and the Vg/VgR gene transfer in the healthy tobacco and infected tobacco were investigated. The Vg content of the infected whitefly was found. The transcriptional level of Vg/VgR gene was significantly increased.
【學(xué)位授予單位】：浙江大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2010
【分類號】：R384.3

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1 劉佳;含有Bph15基因的水稻對褐飛虱卵巢發(fā)育相關(guān)基因表達(dá)的調(diào)控[D];華中農(nóng)業(yè)大學(xué);2012年

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