我国切花害虫甜菜夜蛾遗传多样性及遗传结构研究

发布时间：2018-04-22 17:22

本文选题：切花 + 甜菜夜蛾　；参考：《沈阳农业大学》2016年博士论文

【摘要】：近年来,随着全球气候变暖以及我国花卉种植面积的不断增加,危害我国切花花卉害虫种类逐渐增多、危害程度日趋严重、抗药类别及抗性水平增强,给我国切花花农造成了严重的经济损失。其中,甜菜夜蛾Spodoptera exigua是危害切花的一种重要的夜蛾科害虫,如何对其进行有效防控是切花害虫防治的一项重要工作。目前,鉴于我国危害切花的夜蛾科害虫幼虫种类难鉴定、甜菜夜蛾的种群发生动态以及我国各主要发生危害区的虫源关系不明确,使得对甜菜夜蛾尚无系统、完善和有效的防治策略的实际问题,本研究利用性信息素诱捕器对沈阳地区切花害虫甜菜夜蛾的种群动态进行为期四年的系统监测(2012-2015年)；利用DNA条形码技术对采自我国切花上的夜蛾科害虫幼虫进行分子鉴定,以明确危害我国切花的夜蛾科害虫种类；与此同时,利用不同分子标记技术,mtDNA基因(COI和Cytb)和微卫星(SSR)对我国不同地理种群甜菜夜蛾的遗传多样性及种群遗传结构进行深入研究,在分子水平上探讨甜菜夜蛾不同地理种群间的遗传进化关系,进而为深入探讨物种来源、种群发生规律及成灾遗传机理提供理论基础。最后,对我国不同地理种群甜菜夜蛾胞内共生菌Wolbachia的分布与感染特点进行研究,以明确宿主感染率及Wolbachia株系系统发育地位,为深入探讨Wolbachia与宿主物种间进化关系奠定基础,进而为害虫的生物防治提供理论依据。主要获得了以下研究结果：1.2012-2015年,沈阳地区4年全年诱集到的甜菜夜蛾数量分别为46、274、503和772头。2012年7月06日始见甜菜夜蛾发生,全年1个发生高峰期；2013年6月24日始见甜菜夜蛾发生,全年3个发生高峰期。10月末之后,甜菜夜蛾发生量锐减；2014年6月09日田间始见甜菜夜蛾发生,7月25日前,发生量较低,全年2个发生高峰期；与前3年相比较,2015年甜菜夜蛾发生量大,初春发生早,5月底始见田间甜菜夜蛾发生,全年3个发生高峰期。2.确定了危害我国切花上的5种夜蛾科害虫,即甜菜夜蛾Spodoptera exigua、棉铃虫Helicoverpa armigera、甘蓝夜蛾Mamestra brassicae、斜纹夜蛾Spodoptera litura以及粘虫Mythimna separate。基于COI基因序列的这5种夜蛾科害虫幼虫种间平均遗传距离为0.093,种内平均遗传距离为0.0018。其中,斜纹夜蛾与甜菜夜蛾之间的遗传距离最小为0.066。斜纹夜蛾与棉铃虫之间遗传距离最大为0.107。与之相比较,5种花卉夜蛾科害虫幼虫Cytb基因序列的种间平均遗传距离为0.110,种内平均遗传距离为0.002。其中,斜纹夜蛾与甜菜夜蛾间遗传距离最小为0.084。甜菜夜蛾与甘蓝夜蛾间遗传距离最大为0.138。3.利用mtDNA COI序列(578 bp)和Cytb序列(724 bp)分别对采自我国20省44县(市)和47县(市)的甜菜夜蛾遗传多样性和遗传结构进行研究。总体上,我国甜菜夜蛾遗传多样性较低(COI:Hd=0.276±0.020,Pi=0.00115±0.00014；Cytb:Hd=0.654± 0.017,Pi=0.00238±0.00020).分别确定了41和112个单倍型。各单倍型未形成明显的系统地理格局,散布在不同的地理种群中。基于COI基因的我国甜菜夜蛾不同地理种群间的平均遗传距离为0.001,怀化(HH)种群与其他48个地理种群遗传距离最远。与之相比较,基于Cytb基因的我国甜菜夜蛾不同地理种群间的平均遗传距离为0.003,总体上,TL(通辽)、NJ(南京市)和BD(保定)种群与其它地理种群遗传距离远。另外,Mantel test检验结果表明,地理距离与遗传距离存在相关性,地理隔离对甜菜夜蛾遗传分化无显著影响。分子变异分析(global AMOVA)结果表明,我国不同地理种群甜菜夜蛾遗传变异主要来自于种群内部(COI:77.14％;Cytb:84.86%),种群间的遗传变异水平较低。不同的地理条件与气候条件并不是影响甜菜夜蛾种群遗传结构与物种分布格局的主要因素。中性检验参数(Tajima's D和Fu's FS)皆为负值,且达到显著差异水平,结合单峰错配分布(Mismatch distribution)结果表明,甜菜夜蛾在我国曾经历过种群扩张。4.利用SSR对采自我国20省44县(市)甜菜夜蛾地理种群916头甜菜夜蛾的遗传多样性的研究结果表明,总体上遗传多样性偏低,即等位基因(Na)为5.909,有效等位基因数(Ne)为3.351。KM(昆明)种群具有最高遗传多样性。UPGMA聚类分析树状图结果表明,LY(凌源)、HH(怀化)、GL(桂林)、DA(海口)与其他地理种群遗传距离较远。TA(泰安)、WH(武汉)和DA(海口)种群与其他种群存在显著的遗传分化,大部分种群间遗传分化不显著。贝叶斯聚类分析表明,当K=2时,甜菜夜蛾种群明显分为两支,即ocean(S group)和inland(L group)。但是,表现出高水平的混杂、广泛的杂合和充分的基因流。Mantel Test分析表明地理距离对甜菜夜蛾遗传分化没有显著的影响。分子变异分析(global AMOVA)结果表明,95.53%遗传变异主要发生在种群内,而种群间的变异仅为4.47%。分子变异等级分析(hierarchical AMOVA)结果表明,0.16%遗传变异来源于组间,3.91%来源于此两组的种群间。5.利用wsp对我国甜菜夜蛾胞内共生菌Wolbachia进行感染率检测与系统发育分析。研究结果表明,内共生菌Wolbachia在我国不同地理种群的甜菜夜蛾体内感染率极低,在所检测的我国20省44县(市)的765头甜菜夜蛾样品中,仅仅WF种群(江苏盐城市大丰市)感染了Wolbachia,并且,确定其株系属于Supergroup B组。说明,该类内共生菌在不同地理种群的甜菜夜蛾中感染率极低。
[Abstract]:In recent years, with the increasing of global warming and the growing area of flower planting in China, the species of flower pests in China are increasing, the damage degree is becoming more and more serious, the anti drug category and the resistance level have been strengthened, which have caused serious economic losses to the flower flower farmers in China. In this case, the Spodoptera exigua of the beet armyworm is one of the harm to cut flowers. It is an important work to prevent and control the important pests of noctura noctura. At present, in view of the difficult identification of the larva species of the nocturnal insect pests in China, the population dynamics of the beet armyworm and the relationship between the main harmful regions of our country are not clear, so there is no system for the beet nocturnal moth. In this study, the population dynamics of the Spodoptera Spodoptera Spodoptera Spodoptera Spodoptera in Shenyang area was monitored for four years (2012-2015 years) by using sex pheromone trap, and the DNA barcode technique was used to identify the larvae of the nocturnal insect pests on the cut flower of China, in order to harm our country clearly. At the same time, using different molecular markers, mtDNA gene (COI and Cytb) and microsatellite (SSR), the genetic diversity and population genetic structure of Spodoptera Spodoptera Spodoptera Spodoptera were studied, and the genetic relationship between different geographic populations of the Spodoptera Spodoptera was discussed at the molecular level. In order to explore the origin of species, the regularity of population generation and the genetic mechanism of disaster, the distribution of Wolbachia and the characteristics of the infection of the intracellular symbiotic bacteria of the Spodoptera Spodoptera in China were studied in order to clarify the infection rate of the host and the phylogenetic status of the Wolbachia strain, so as to explore the interrelationship between the Wolbachia and the host species. The relationship laid the foundation and provided the theoretical basis for the biological control of the pests. The following results were obtained. In 1.2012-2015, the number of beet armyworm luring in the year of 4 years in Shenyang was 46274503 and 772 of.2012 year July, the beet armyworm occurred on 06 days, 1 peaks in the whole year. In June 24, 2013, the sweet Spodoptera was sweet. The occurrence of Spodoptera Spodoptera Spodoptera Spodoptera was found at the end of the 3 peak period of.10 month, and the occurrence of beet Spodoptera Spodoptera was sharply reduced. On the 09 day of June 2014, the occurrence of beet armyworm was found in the field. Before July 25th, the occurrence of the Spodoptera Spodoptera was lower and 2 had the peak period of the year. Compared with the previous 3 years, the occurrence of beet armyworm in 2015 was large, early spring occurred early, and the field beet armyworm occurred in the end of May, and the occurrence of beet armyworm was seen in the end of May, The 3 peak.2. of the year confirmed the 5 species of nocturnal insect pests on Chinese cut flowers, namely, Spodoptera exigua of beet armyworm, Helicoverpa armigera of cotton bollworm, Mamestra brassicae of nocturnal moth, Spodoptera litura of Spodoptera Spodoptera, and Mythimna separate. based on the sequence of COI base among the 5 larvae of nocturnal pests The genetic distance was 0.093 and the intraspecific average genetic distance was 0.0018.. The genetic distance between the Spodoptera Spodoptera and the beet armyworm was the smallest between 0.066. and cotton bollworm, and the maximum genetic distance was 0.107. compared with that of the cotton bollworm. The average interspecific genetic distance of the Cytb gene sequence of the 5 species of the insect larvae was 0.110, and the intraspecific inheritance of the genetic distance was 0.110. The distance is 0.002., the genetic distance between the Spodoptera Spodoptera and the beet armyworm is the smallest, the largest genetic distance between the 0.084. beet armyworm and the Spodoptera Spodoptera is 0.138.3. using mtDNA COI sequence (578 BP) and Cytb sequence (724 BP) to study the genetic diversity and genetic structure of the Spodoptera Spodoptera in 20 provinces and 44 counties (cities) and 47 counties (cities) of China. In vivo, the genetic diversity of Spodoptera exigua was low (COI:Hd=0.276 + 0.020, Pi=0.00115 + 0.00014; Cytb:Hd=0.654 + 0.017, Pi=0.00238 + 0.00020). 41 and 112 haplotypes were identified respectively. Each haplotype did not form a distinct geographic pattern and distributed in different geographic populations. The different geography of Chinese beet armyworm based on COI gene The average genetic distance between the population is 0.001, the Huaihua (HH) population is the farthest from the other 48 geographical populations. Compared with the Cytb gene, the average genetic distance between the different geographical populations of the beet armyworm is 0.003. In general, the TL (Tongliao), the NJ (Nanjing) and the BD (Baoding) population are far from the other geographical populations. The results of Mantel test test showed that there was a correlation between geographical distance and genetic distance, and geographical isolation had no significant influence on the genetic differentiation of beet armyworm. The results of molecular variation analysis (global AMOVA) showed that the genetic variation of Spodoptera Spodoptera from different geographic populations mainly came from the intra species (COI:77.14%; Cytb:84.86%), and the inheritance between populations. Different geographic conditions and climatic conditions were not the main factors affecting the genetic structure and species distribution pattern of the beet armyworm population. The neutral test parameters (Tajima's D and Fu's FS) were negative and reached significant differences. The result of single peak mismatch distribution (Mismatch distribution) showed that the beet armyworm was in our country. The genetic diversity of the 916 beet Spodoptera Spodoptera Spodoptera Spodoptera Spodoptera Spodoptera Spodoptera Spodoptera from 20 provinces and 44 counties of China was studied with population expansion.4.. The genetic diversity of the Spodoptera Spodoptera Spodoptera Spodoptera Spodoptera Spodoptera Spodoptera Spodoptera Spodoptera Spodoptera showed that the genetic diversity was low, that is, the allele (Na) was 5.909 and the number of effective alleles (Ne) was the highest genetic diversity.UPGMA cluster in 3.351.KM (Kunming) population. The dendrogram showed that the genetic distance between LY (Lingyuan), HH (Huaihua), GL (Guilin), DA (Haikou) and other geographical populations was far from.TA (Tai'an), WH (Wuhan) and DA (Haikou) population and other populations had significant genetic differentiation, and the genetic differentiation between most of the populations was not significant. The Bayesian cluster analysis showed that when K=2, the beet armyworm population was obviously divided into two groups. Two, ocean (S group) and inland (L group). However, the high level of hybrid, extensive heterozygosity and full gene flow.Mantel Test analysis showed that the geographical distance had no significant effect on the genetic differentiation of the beet armyworm. The results of molecular variation analysis (global AMOVA) showed that the 95.53% genetic variation mainly occurred in the population and between the population. The variation is only 4.47%. molecular variation grade analysis (hierarchical AMOVA). The results show that 0.16% genetic variation comes from intergroup and 3.91% from the two groups of.5. using WSP to detect the infection rate and phylogenetic analysis of the intracellular symbiotic bacteria Wolbachia of the Spodoptera exigua. The results show that the endosonde Wolbachia is not in China. The infection rate of the Spodoptera Spodoptera was very low in the same geographical population. In the samples of 765 beet armyworm in 20 provinces and 44 counties of China, only WF population (Dafeng City, Yancheng City, Jiangsu) infected Wolbachia, and it was determined that the strains belonged to group Supergroup B. It indicated that the infection rate of this kind of endophytic bacteria in the beet armyworm in different geographic populations was extremely high. Low.

【学位授予单位】：沈阳农业大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：S436.8

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