有翅烟蚜和烟草花叶病在烟田的发生动态与不同寄主上烟蚜的生命表研究

发布时间：2018-01-19 21:19

本文关键词： 翅烟烟草花叶烟田发生动态不同寄主烟蚜生命研究　出处：《西南大学》2017年博士论文　论文类型：学位论文

【摘要】：烟草是我国重要的经济作物之一,而烟草病虫害则是严重制约烟草产量与品质的重要因素,其中烟蚜更是各烟区的重点防治对象。烟蚜一方面可以通过取食直接为害烟草,一方面还是黄瓜花叶病毒(CMV)、马铃薯Y病毒(PVY)、芜菁花叶病毒(TuMV)等多种植物病毒病原的传播媒介,这几种病毒在田间单独或复合侵染引起烟草花叶病,给烟草生产造成了更为严重的间接损失。烟蚜在烤烟的整个生长季节都有发生,具有寄主种类多,分布范围广,发育历期短,繁殖速度快,生活史复杂,世代重叠等特点。本文研究了重庆武隆烟区有翅烟蚜的种群动态和烟草花叶病的发生规律,探索了有翅烟蚜种群数量与烟田周边环境和地势条件以及有翅烟蚜种群数量与烟田烟草花叶病发生程度的关系;在田间研究的基础之上选择了烟田附近有代表性的4种蔬菜作物以及烟草作为寄主,利用年龄-龄期两性生命表技术研究了不同寄主植物对烟蚜种群生长发育和繁殖的影响,从而明确了主要蔬菜种类在有翅烟蚜迁入烟田中的作用;此外利用第二代转录组测序技术Illumina/Solexa获得了烟蚜的全转录组信息,分析了烟蚜参与寄主选择的相关基因。为进一步深入研究不同寄主植物对烟蚜生理生化影响的分子机理奠定了基础,也为制定更为行之有效的防控策略提供了新的思路。主要研究结果如下:一.武隆烟区有翅烟蚜与烟草花叶病的田间发生动态利用黄板诱集的方法研究了烤烟从团颗期到成熟期田间有翅烟蚜的种群动态。结果表明,有翅烟蚜在烟田中有两次迁飞高峰,第一个高峰是在6月中下旬,烟草正处于团颗期的阶段;而第二个高峰则是在7月上旬,烟草开始进入旺长期。第一次有翅烟蚜迁飞高峰主要受到烟田周围环境的影响,靠近菜地的烟田有翅烟蚜数量极显著地高于靠近树林烟田中的有翅烟蚜数量,迁入烟田的有翅烟蚜主要来自附近的白菜、辣椒、蚕豆和萝卜等寄主。第二次迁飞高峰则与烟田的地势条件有着密切关系,发现凹地、斜坡和坡顶三种地势烟田中诱集到的有翅烟蚜的数量从多到少依次为凹地斜坡坡顶,凹地烟田的有翅烟蚜数量显著高于斜坡和坡顶有翅烟蚜数量,而7月上中旬这种差异尤为突出。烟草花叶病在烟田中的田间症状表现较有翅烟蚜的第一个迁入高峰晚2-3周左右。烟草花叶病的发病株率与病情指数均呈S型曲线上升,7月上中旬是烟草花叶病发展最快的时期,7月下旬开始后花叶病的发生减缓乃至停止。6种生境18个不同烟田之间烟草花叶病的发生存在较大差异,靠近菜地的烟田中花叶病的病情指数显著高于靠近树林的烟田,凹地烟田的烟草花叶病发病程度也高于斜坡和坡顶两种地势条件的烟田。迁入烟田的有翅烟蚜数量与烟田烟草花叶病发生发展关系密切——靠近菜地的烟田有翅烟蚜数量极显著地高于靠近树林烟田中的有翅烟蚜数量,其烟草花叶病的病情指数显著高于靠近树林的烟田;凹地烟田的有翅烟蚜数量显著高于斜坡和坡顶烟田种的有翅烟蚜数量,其花叶病的病情指数也显著高于斜坡和坡顶烟田。二.不同寄主植物上烟蚜的生命表研究通过年龄-龄期两性生命表技术研究比较了烟草、蚕豆、萝卜、辣椒和油菜5种寄主植物上烟蚜种群的生命表参数和各个发育阶段的发育历期以及年龄-龄期特定存活率sxj,期望寿命exj,与生殖价值vxj。结果表明,5种寄主植物中烟蚜种群在蚕豆上的内禀增长力r达到0.383d-1,显著高于其他四种作物上的内禀增长力,而辣椒上的烟蚜种群内禀增长力最低,为0.3423d-1;萝卜上的烟蚜净生殖率r0为80.83头,显著高于其他四种寄主上烟蚜的净生殖率,油菜和辣椒上的烟蚜净生殖率最小,分别仅为51.93头和46.28头;烟蚜在蚕豆上的周限增长率λ最高;油菜和萝卜上的烟蚜平均世代周期t最长,烟草次之,而辣椒和蚕豆上的烟蚜平均世代周期最短。烟蚜种群在萝卜和烟草上的成虫寿命与总寿命均最长,油菜和蚕豆次之,辣椒上的烟蚜种群最短;在萝卜上的烟蚜种群繁殖期、繁殖力显著高于其他4种寄主植物上的烟蚜种群。烟蚜种群在5种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天之间,烟蚜种群在萝卜上的最高峰日繁殖量达到400头/天,繁殖期也明显较其他4种寄主植物上的烟蚜更长,在蚕豆上次之,烟蚜种群在辣椒、油菜上的繁殖峰值较低,繁殖期时间也最短。烟蚜的期望寿命均随着年龄增加而缩短,而萝卜和烟草上年龄-阶段期望寿命exj要比剩余三种寄主植物上的期望寿命长;此外,5种寄主植物上烟蚜新生若蚜的存活率均在95%以上,没有显著性差异;烟蚜在蚕豆、辣椒、萝卜、烟草和油菜5种寄主植物上的最高生殖价值vxj各出现在第9天,第7.5天,第8.5天,第10天和第9天,分别为16.93,12.32,14.88,17.19和13.76,其中烟草和蚕豆上的烟蚜种群明显较其他三种寄主更高。根据以上研究结果表明,萝卜、蚕豆、烟草、辣椒和油菜均是烟蚜的适宜寄主,但蚕豆和萝卜更有利于烟蚜种群生长发育和繁殖,烟草次之,油菜和辣椒则再次之。三.烟蚜的转录组测序及生物信息学分析为进一步探讨寄主植物对烟蚜种群发展的影响,对烟草上饲养了10代以上的烟蚜进行了高通量转录组测序。通过illuminahiseq2000平台测序产出了51,222,318个reads,共计4,610,008,620个碱基(nt)。通过组装获得了50,177个contigs,平均长度为470nt,这些contigs进一步组装最后得到了30,293条unigenes,平均长度966nt。利用nr,nt,swiss-prot,go,cog和kegg六个数据库对这些unigenes进行了功能预测,分别有22,736条,24,498条,16,856条,10,194条,8,232条和15,512条unigenes被注释到了相应数据库。在利用nr数据库注释时发现绝大部分unigenes(20,896条,91.91%)与豌豆蚜(acyrthosiphonpisum)高度同源。注释到go数据库的10,194条unigenes则被分成了biologicalprocess,cellularcomponent和molecularfunction三大类群58个功能组,其中“cellularprocess”的unigenes最多(6,525条),紧随其后的是“single-organismprocess”(5,122条)以及“metabolicprocess”(4,813条)两个功能组。利用cog数据库将基于序列同源性将总共8,232条unigene分为了25个亚类,“generalfunction”亚类最大(2,931条)。利用kegg数据库预测了15,512条unigenes的代谢通路,结果共发现了255个,包含了最多unigenes的是“metabolicpathways”(id:ko01100)(2132条,13.74%)。在转录组测序的基础之上,本文对与昆虫的取食消化等过程密切相关的通路以及相关unigenes进行了发掘,包括与碳水化合物代谢相关的“starchandsucrosemetabolism”(213unigenes,1.37%),与氨基酸代谢相关的“lysinedegradation”(188unigenes,1.21%),与脂类代谢相关的“glycerophospholipidmetabolism”(161unigenes,1.04%),另外还有参与消化系统代谢的“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%).

【学位授予单位】：西南大学
【学位级别】：博士
【学位授予年份】：2017
【分类号】：S435.72

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