本文關鍵詞： 翅煙 煙草 花葉 煙田 發(fā)生 動態(tài) 不同 寄主 煙蚜 生命 研究　出處：《西南大學》2017年博士論文　論文類型：學位論文

【摘要】：煙草是我國重要的經(jīng)濟作物之一,而煙草病蟲害則是嚴重制約煙草產量與品質的重要因素,其中煙蚜更是各煙區(qū)的重點防治對象。煙蚜一方面可以通過取食直接為害煙草,一方面還是黃瓜花葉病毒(CMV)、馬鈴薯Y病毒(PVY)、蕪菁花葉病毒(TuMV)等多種植物病毒病原的傳播媒介,這幾種病毒在田間單獨或復合侵染引起煙草花葉病,給煙草生產造成了更為嚴重的間接損失。煙蚜在烤煙的整個生長季節(jié)都有發(fā)生,具有寄主種類多,分布范圍廣,發(fā)育歷期短,繁殖速度快,生活史復雜,世代重疊等特點。本文研究了重慶武隆煙區(qū)有翅煙蚜的種群動態(tài)和煙草花葉病的發(fā)生規(guī)律,探索了有翅煙蚜種群數(shù)量與煙田周邊環(huán)境和地勢條件以及有翅煙蚜種群數(shù)量與煙田煙草花葉病發(fā)生程度的關系;在田間研究的基礎之上選擇了煙田附近有代表性的4種蔬菜作物以及煙草作為寄主,利用年齡-齡期兩性生命表技術研究了不同寄主植物對煙蚜種群生長發(fā)育和繁殖的影響,從而明確了主要蔬菜種類在有翅煙蚜遷入煙田中的作用;此外利用第二代轉錄組測序技術Illumina/Solexa獲得了煙蚜的全轉錄組信息,分析了煙蚜參與寄主選擇的相關基因。為進一步深入研究不同寄主植物對煙蚜生理生化影響的分子機理奠定了基礎,也為制定更為行之有效的防控策略提供了新的思路。主要研究結果如下:一.武隆煙區(qū)有翅煙蚜與煙草花葉病的田間發(fā)生動態(tài)利用黃板誘集的方法研究了烤煙從團顆期到成熟期田間有翅煙蚜的種群動態(tài)。結果表明,有翅煙蚜在煙田中有兩次遷飛高峰,第一個高峰是在6月中下旬,煙草正處于團顆期的階段;而第二個高峰則是在7月上旬,煙草開始進入旺長期。第一次有翅煙蚜遷飛高峰主要受到煙田周圍環(huán)境的影響,靠近菜地的煙田有翅煙蚜數(shù)量極顯著地高于靠近樹林煙田中的有翅煙蚜數(shù)量,遷入煙田的有翅煙蚜主要來自附近的白菜、辣椒、蠶豆和蘿卜等寄主。第二次遷飛高峰則與煙田的地勢條件有著密切關系,發(fā)現(xiàn)凹地、斜坡和坡頂三種地勢煙田中誘集到的有翅煙蚜的數(shù)量從多到少依次為凹地斜坡坡頂,凹地煙田的有翅煙蚜數(shù)量顯著高于斜坡和坡頂有翅煙蚜數(shù)量,而7月上中旬這種差異尤為突出。煙草花葉病在煙田中的田間癥狀表現(xiàn)較有翅煙蚜的第一個遷入高峰晚2-3周左右。煙草花葉病的發(fā)病株率與病情指數(shù)均呈S型曲線上升,7月上中旬是煙草花葉病發(fā)展最快的時期,7月下旬開始后花葉病的發(fā)生減緩乃至停止。6種生境18個不同煙田之間煙草花葉病的發(fā)生存在較大差異,靠近菜地的煙田中花葉病的病情指數(shù)顯著高于靠近樹林的煙田,凹地煙田的煙草花葉病發(fā)病程度也高于斜坡和坡頂兩種地勢條件的煙田。遷入煙田的有翅煙蚜數(shù)量與煙田煙草花葉病發(fā)生發(fā)展關系密切——靠近菜地的煙田有翅煙蚜數(shù)量極顯著地高于靠近樹林煙田中的有翅煙蚜數(shù)量,其煙草花葉病的病情指數(shù)顯著高于靠近樹林的煙田;凹地煙田的有翅煙蚜數(shù)量顯著高于斜坡和坡頂煙田種的有翅煙蚜數(shù)量,其花葉病的病情指數(shù)也顯著高于斜坡和坡頂煙田。二.不同寄主植物上煙蚜的生命表研究通過年齡-齡期兩性生命表技術研究比較了煙草、蠶豆、蘿卜、辣椒和油菜5種寄主植物上煙蚜種群的生命表參數(shù)和各個發(fā)育階段的發(fā)育歷期以及年齡-齡期特定存活率sxj,期望壽命exj,與生殖價值vxj。結果表明,5種寄主植物中煙蚜種群在蠶豆上的內稟增長力r達到0.383d-1,顯著高于其他四種作物上的內稟增長力,而辣椒上的煙蚜種群內稟增長力最低,為0.3423d-1;蘿卜上的煙蚜凈生殖率r0為80.83頭,顯著高于其他四種寄主上煙蚜的凈生殖率,油菜和辣椒上的煙蚜凈生殖率最小,分別僅為51.93頭和46.28頭;煙蚜在蠶豆上的周限增長率λ最高;油菜和蘿卜上的煙蚜平均世代周期t最長,煙草次之,而辣椒和蠶豆上的煙蚜平均世代周期最短。煙蚜種群在蘿卜和煙草上的成蟲壽命與總壽命均最長,油菜和蠶豆次之,辣椒上的煙蚜種群最短;在蘿卜上的煙蚜種群繁殖期、繁殖力顯著高于其他4種寄主植物上的煙蚜種群。煙蚜種群在5種寄主植物上繁殖期均主要集中在第6天到第27天,而繁殖高峰期在第10天到第15天之間,煙蚜種群在蘿卜上的最高峰日繁殖量達到400頭/天,繁殖期也明顯較其他4種寄主植物上的煙蚜更長,在蠶豆上次之,煙蚜種群在辣椒、油菜上的繁殖峰值較低,繁殖期時間也最短。煙蚜的期望壽命均隨著年齡增加而縮短,而蘿卜和煙草上年齡-階段期望壽命exj要比剩余三種寄主植物上的期望壽命長;此外,5種寄主植物上煙蚜新生若蚜的存活率均在95%以上,沒有顯著性差異;煙蚜在蠶豆、辣椒、蘿卜、煙草和油菜5種寄主植物上的最高生殖價值vxj各出現(xiàn)在第9天,第7.5天,第8.5天,第10天和第9天,分別為16.93,12.32,14.88,17.19和13.76,其中煙草和蠶豆上的煙蚜種群明顯較其他三種寄主更高。根據(jù)以上研究結果表明,蘿卜、蠶豆、煙草、辣椒和油菜均是煙蚜的適宜寄主,但蠶豆和蘿卜更有利于煙蚜種群生長發(fā)育和繁殖,煙草次之,油菜和辣椒則再次之。三.煙蚜的轉錄組測序及生物信息學分析為進一步探討寄主植物對煙蚜種群發(fā)展的影響,對煙草上飼養(yǎng)了10代以上的煙蚜進行了高通量轉錄組測序。通過illuminahiseq2000平臺測序產出了51,222,318個reads,共計4,610,008,620個堿基(nt)。通過組裝獲得了50,177個contigs,平均長度為470nt,這些contigs進一步組裝最后得到了30,293條unigenes,平均長度966nt。利用nr,nt,swiss-prot,go,cog和kegg六個數(shù)據(jù)庫對這些unigenes進行了功能預測,分別有22,736條,24,498條,16,856條,10,194條,8,232條和15,512條unigenes被注釋到了相應數(shù)據(jù)庫。在利用nr數(shù)據(jù)庫注釋時發(fā)現(xiàn)絕大部分unigenes(20,896條,91.91%)與豌豆蚜(acyrthosiphonpisum)高度同源。注釋到go數(shù)據(jù)庫的10,194條unigenes則被分成了biologicalprocess,cellularcomponent和molecularfunction三大類群58個功能組,其中“cellularprocess”的unigenes最多(6,525條),緊隨其后的是“single-organismprocess”(5,122條)以及“metabolicprocess”(4,813條)兩個功能組。利用cog數(shù)據(jù)庫將基于序列同源性將總共8,232條unigene分為了25個亞類,“generalfunction”亞類最大(2,931條)。利用kegg數(shù)據(jù)庫預測了15,512條unigenes的代謝通路,結果共發(fā)現(xiàn)了255個,包含了最多unigenes的是“metabolicpathways”(id:ko01100)(2132條,13.74%)。在轉錄組測序的基礎之上,本文對與昆蟲的取食消化等過程密切相關的通路以及相關unigenes進行了發(fā)掘,包括與碳水化合物代謝相關的“starchandsucrosemetabolism”(213unigenes,1.37%),與氨基酸代謝相關的“l(fā)ysinedegradation”(188unigenes,1.21%),與脂類代謝相關的“glycerophospholipidmetabolism”(161unigenes,1.04%),另外還有參與消化系統(tǒng)代謝的“bilesecretion”(310unigenes,2.00%),“pancreaticsecretion”(231unigenes,1,49%),“proteindigestionandabsorption”(204unigenes,1.32%),“salivarysecretion”(197unigenes,1.27%),“gastricacidsecretion”(87unigenes,1.21%)以及“vitamindigestionandabsorption”(168unigenes,1.08%)。
[Abstract]:Tobacco is one of the most important economic crops in China, and the tobacco diseases and insect pests is an important factor restricting the yield and quality of tobacco, which is the focus of the tobacco aphid Myzus persicae. The control objects can through a feeding or a direct damage to tobacco, cucumber mosaic virus (CMV), potato virus Y (PVY) turnip mosaic virus (TuMV), and other plant pathogenic virus media, the virus caused by tobacco mosaic virus in the field of single or multiple infection, tobacco production to cause more serious indirect losses. Aphid has occurred in flue-cured tobacco throughout the growing season, with a variety of hosts, wide distribution, developmental period short, fast propagation speed, complex life history, overlapping generations etc. This paper studied the population dynamics and occurrence regularity of tobacco mosaic virus in Chongqing tobacco growing areas of Wulong have explored the wing aphid, the number of winged aphid population With the tobacco field surrounding environment and terrain conditions and the winged aphid population and the number of tobacco mosaic disease severity; on the basis of field study chose 4 representative vegetable crops and tobacco as host tobacco field near the age -, sex life table technique to study the effects of different host plants on Myzus persicae population growth and reproduction of the use, in order to clear the main vegetable species in the tobacco fields in alate aphid; in addition to the use of second generation sequencing Illumina/Solexa obtained the whole transcriptome analysis of the population information, participate in the host selection of genes related to aphid. Laid the foundation for further study of molecular mechanism of different host plants the physiological and biochemical effects of Myzus persicae, to develop more effective prevention and control strategy provides a new way of thinking. The main results are as follows: 1. Wuhan Long winged aphid and field tobacco tobacco mosaic virus population dynamic of tobacco field from a group of winged aphid stage to mature stage by means of the yellow sticky traps. The results show that the two winged aphid migration peak in tobacco fields, the first peak is 6 months in the second half, tobacco is in group a period; while the second peak is in early July, tobacco entered the vigorous. The first winged aphid migration period is mainly affected by the tobacco field of the surrounding environment, the tobacco field near the vegetable winged aphid was significantly higher than the number of tobacco fields close to the woods winged aphid number in tobacco field the winged aphid mainly from the nearby Chinese cabbage, peppers, beans and turnips and other hosts. The second peak and migration of tobacco field terrain conditions are closely related, the slope and the slope found, three kinds of the tobacco fields were captured winged The number of aphid in decreasing order of the slope top, tobacco hollow winged number was significantly higher than that of slope and slope, the winged aphid number, and in mid July this difference is particularly prominent. In the field of tobacco mosaic virus in tobacco fields with symptoms of winged aphid first immigration peak late 2-3 weeks. The incidence of strain rate and disease index of tobacco mosaic virus are increasing type S curve in mid July is the fastest growing period of tobacco mosaic, beginning in late July after the occurrence of mosaic slow or stop.6 between 18 different habitats of tobacco mosaic disease has great differences, the disease index of tobacco field near the vegetable mosaic was significantly higher than that in tobacco field close to the woods, the tobacco field incidence of tobacco mosaic virus in tobacco fields. The slope and slope is higher than that of two kinds of terrain conditions. In tobacco field and tobacco aphid number winged The relationship between the occurrence and development of tobacco mosaic virus in tobacco field near the close of vegetable alate Myzus persicae number was significantly higher than the tobacco field close to the woods winged aphid number, the disease index of tobacco mosaic virus in tobacco field was significantly higher than that in the tobacco field close to the woods; hollow winged aphid number was significantly higher than that of the slope and the slope of the winged tobacco aphid the number, the disease index of its mosaic is significantly higher than that of slope and slope in tobacco fields. Two. Different host plants on the life table of Myzus persicae by age - sex life table of comparison of tobacco, beans, radish, 5 host plants of pepper and rape aphid population life table parameters and various developmental stages the developmental duration and age - age specific survival rate sxj, life expectancy and reproductive value of vxj. exj, the results showed that 5 species of aphid host plant population intrinsic in faba bean growth reached 0.3 R 83d-1 was significantly higher than that of the other four crops on the intrinsic growth capacity, and pepper on the aphid population intrinsic growth capacity of the lowest, 0.3423d-1; turnip aphids on the net reproductive rate of R0 was 80.83, significantly higher than the other four host on the net reproductive rate of Myzus persicae on pepper, rape and tobacco aphid net reproduction the minimum rate were only 51.93 and 46.28; the week limit on broad bean aphid growth rate is the highest; the average generation time t aphid rape and radish on the longest time, tobacco, and tobacco mean generation time of pepper and bean on the short. Life expectancy of adult population of Myzus persicae in radish and tobacco. And the total life are the longest, followed by rape and beans, pepper on the aphid population shortest; on the radish aphid population during the breeding period, fecundity was significantly higher than the other 4 kinds of host plants on the aphid population. Breeding population of Myzus persicae on 5 host plants were mainly concentrated in the 6 to twenty-seventh days, and the breeding peak in tenth to fifteenth days, breeding on the radish aphid population peak day reached 400 head / day, breeding period was also compared to the other 4 kinds of host plants on the aphid in broad bean last longer, the aphid population in pepper breeding, the peak of rape low breeding time is the shortest. The life expectancy of Myzus persicae was shortened with increasing age, and the carrot and tobacco age stage life expectancy of exj than the remaining three kinds of host plants on the long life expectancy; in addition, 5 kinds of host plants on neonatal survival if aphid Myzus persicae rates are above 95%., no significant difference; Myzus persicae in faba bean, pepper, radish, 5 kinds of host plants on tobacco and rape the highest reproductive value of vxj appeared at the ninth day, the 7.5 day, 8.5 day, tenth days and ninth days, 16.93,12.32,14.88,17.19 and 13.76 respectively, the tobacco and the bean Aphid population obviously than the other three kinds of host more. According to the above research results show that radish, bean, tobacco, pepper and rape are suitable host aphid, but more beans and turnips to aphid population growth and reproduction of tobacco, rape, and chili again. Three. Analysis of myzuspersicae transcription sequencing and bioinformatics to further explore the effects of host plants on the development of tobacco aphid population, feeding on more than 10 generations of Myzus persicae were high-throughput transcriptome sequencing. Through illuminahiseq2000 sequencing platform output 51222318 reads, a total of 4610008620 base pairs (NT). The assembly obtained 50177 contigs, the average length of for 470nt, the contigs assembly further finally got 30293 unigenes, the average length of 966nt. by NR, NT, SWISS-PROT, go, COG and KEGG six database for the work of these unigenes Can predict, respectively 22736, 24498, 16856, 10194, 8232 and 15512 unigenes were annotated to the corresponding database. We find that most of unigenes in the NR database using the annotation (20896, 91.91%) and pea aphid (acyrthosiphonpisum) is highly homologous to 10194 annotated go database is unigenes is divided into biologicalprocess, cellularcomponent and molecularfunction three groups of 58 functional groups, including "cellularprocess" unigenes (6525), followed by "single-organismprocess" (5122) and metabolicprocess (4813) of two functional groups. The use of COG database sequence will be a total of 8232 UniGene divided into 25 subgroups based on the "generalfunction" sub class maximum (2931). The prediction using the KEGG database of 15512 unigenes pathways, the results found a total of 2 55, contains most of the unigenes is "metabolicpathways" (id:ko01100) (2132, 13.74%). On the basis of transcriptome sequencing, this pathway is closely related to the process of digestion and insect feeding and related unigenes were discovered, including water and carbon starchandsucrosemetabolism compounds related to metabolism "(" 213unigenes, 1.37%), associated with the metabolism of amino acids in "lysinedegradation" (188unigenes, 1.21%), associated with lipid metabolism in "glycerophospholipidmetabolism" (161unigenes, 1.04%), in addition to participate in the metabolism of digestive system of "bilesecretion" (310unigenes, 2%), "pancreaticsecretion" (231unigenes, 1,49%), "proteindigestionandabsorption" (204unigenes, 1.32%), "salivarysecretion" (197unigenes, 1.27%), "gastricacidsecretion" (87unigenes, 1.21%) and "vitamindigestio Nandabsorption "(168unigenes, 1.08%).

【學位授予單位】：西南大學
【學位級別】：博士
【學位授予年份】：2017
【分類號】：S435.72

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