RAPD方法用于亚洲玉米螟地理种群分化的研究

玉米螟属鳞翅目(Ｌｅｐｉｄｏｐｔｅｒａ)螟蛾科(Ｐｙｒａｌｉｄａｅ)拟螟亚科(Ｐｙｒａｕｓｔｉｎａｅ)杆野螟属(Ｏｓｔｒｉｎｉａ),是世界性的重大农业害虫。70年代以来,通过形态学鉴别、遗传杂交实验和性信息素组份分析及其活性反应等方面的实验表明,在我国绝大部分地?..     

Molecular differentiation at nuclear loci in French host races of the European corn borer (Ostrinia nubilalis)

French populations of the European corn borer consist of two sympatric and genetically differentiated host races. As such, they are well suited to study processes that could be involved in sympatric speciation, but the initial conditions of host-race divergence need to be elucidated. Gene genealogies can provide insight into the processes involved in speciation. We used DNA sequences of four nuclear genes to (1) document the genetic structure of the two French host races previously delineated with allozyme markers, (2) find genes directly or indirectly involved in reproductive isolation between host races, and (3) estimate the time since divergence of the two taxa and see whether this estimate is compatible with this divergence being the result of a host shift onto maize after its introduction into Europe approximately 500 years ago. Gene genealogies revealed extensive shared polymorphism, but confirmed the previously observed genetic differentiation between the two host races. Significant departures from the predictions of neutral molecular evolution models were detected at three loci but were apparently unrelated to reproductive isolation between host races. Estimates of time since divergence between French host races varied from approximately 75,000 to approximately 150,000 years, suggesting that the two taxa diverged recently but probably long before the introduction of maize into Europe.     

Dissemination of Nosema pyrausta in feral populations of the European corn borer,Ostrinia nubilalis

Epizootiological studies of Nosema pyrausta in natural European corn borer populations show that while vertical transmission is the primary way in which N. pyrausta is transferred from one host generation to the next, it is horizontal transmission that is responsible for the annual build-up of infection in each nonoverlapping generation. During the first generation, larval corn borer migration to adjacent corn stalks is minimal and increases in the prevalence of N. pyrausta within the population result from horizontal transmission of infection among borers that inhabit the same stalk. During the second generation, corn borer larvae actively disperse to other corn plants and this results in an increased level of infection. Factors which facilitate pathogen dispersal in this generation include (1) higher host densities, (2) longer periods of larval development, (3) lower mortality among young larvae, and (4) possible mechanical transmission by the braconid parasitoid, Macrocentrus grandii.     

Insights into genome differentiation: pheromone-binding protein variation and population history in the European corn borer (Ostrinia nubilalis)

Examination of sequence variation at nuclear loci can give insights into population history and gene flow that cannot be derived from other commonly used molecular markers, such as allozymes. Here, we report on sequence variation at a single nuclear locus, the pheromone-binding protein (PBP) locus, in the European corn borer (Ostrinia nubilalis). The European corn borer has been divided into three races in New York State on the basis of differences in pheromone communication and life history. Previous allozyme data have suggested that there is a small but significant amount of genetic differentiation between these races. The PBP does not appear to be involved in the pheromone differences between these races. Examination of variation at the PBP locus in the three races reveals no fixed differences between races despite high levels of polymorphism. There also appears to have been considerable recombination in the history of the pheromone-binding protein alleles. Observation of both recombination between alleles and lack of significant nucleotide or insertion/deletion divergence between races leads us to suggest that these populations are either recently diverged or have continued to exchange genetic material subsequent to divergence in pheromone communication and life history.     

Host-plant-associated genetic differentiation in Northern French populations of the European corn borer

The phytophagous insects that damage crops are often polyphagous, feeding on several types of crop and on weeds. The refuges constituted by noncrop host plants may be useful in managing the evolution in pest species of resistance to the Bacillus thuringiensis toxins produced by transgenic crops. However, the benefits of these refuges may be limited because host-plant diversity may drive genetic divergence and possibly even host-plant-mediated sympatric speciation. The European corn borer, Ostrinia nubilalis Hübner (Lepidoptera: Crambidae), is the main pest of maize in Europe and North America, where it was introduced early in the 20th century. It has a wide host range but feeds principally on mugwort (Artemisia vulgaris L.) and maize (Zea mays L.). O. nubilalis is found on mugwort only in the northern part of France, whereas it is found on maize throughout France. The extent of genetic variation at allozyme markers was investigated in populations collected from the two host plants over the entire geographical distribution of the European corn borer on mugwort in France. Allelic differentiation between pairs of populations and hierarchical analyses of pools of samples from each host plant indicate that the group of populations feeding on maize differed from the group of populations feeding on mugwort. Our results suggest (1) host-plant-related divergent selection at the genomic region surrounding the Mpi locus and (2) limited gene flow between the populations feeding on mugwort and those infesting maize fields. These data indicate that adults emerging from mugwort would not be useful for managing the evolution of resistance to the B. thuringiensis toxins in European corn borer populations.     

Peritrophic membrane structure and secretion in European corn borer larvae (Ostrinia nubilalis)

Peritrophic membrane (PM) secretion and formation occur primarily in the anterior region of the mesenteron in the European corn borer (Ostrinia nubilalis) as determined by light and electron microscopy. Nascent PM first became visible as fibrous linear chitin-containing structures stained with gold-labeled wheat germ agglutinin between and at the tips of the microvilli. No formed PM was visible at the foregut-midgut junction, but a thin single PM appeared first in the lumen between the stomodeal valves and the midgut epithelium. Just posterior to the stomodeal valves, multiple PMs were observed that became progressively thicker and more numerous in the mid and posterior regions of the mesenteron. The PM consists of an orthogonal chitin meshwork with openings slightly larger than the diameters of the microvilli. As it delaminates from the microvilli, the meshwork becomes embedded in proteinaceous matrix that greatly reduces the pore size of the PM.     

Microbial diversity in the midguts of field and lab-reared populations of the European corn borer Ostrinia nubilalis

BACKGROUND: Insects are associated with microorganisms that contribute to the digestion and processing of nutrients. The European Corn Borer (ECB) is a moth present world-wide, causing severe economical damage as a pest on corn and other crops. In the present work, we give a detailed view of the complexity of the microorganisms forming the ECB midgut microbiota with the objective of comparing the biodiversity of the midgut-associated microbiota and explore their potential as a source of genes and enzymes with biotechnological applications. METHODOLOGICAL/PRINCIPAL FINDINGS: A high-throughput sequencing approach has been used to identify bacterial species, genes and metabolic pathways, particularly those involved in plant-matter degradation, in two different ECB populations (field-collected vs. lab-reared population with artificial diet). Analysis of the resulting sequences revealed the massive presence of Staphylococcus warneri and Weissella paramesenteroides in the lab-reared sample. This enabled us to reconstruct both genomes almost completely. Despite the apparently low diversity, 208 different genera were detected in the sample, although most of them at very low frequency. By contrast, the natural population exhibited an even higher taxonomic diversity along with a wider array of cellulolytic enzyme families. However, in spite of the differences in relative abundance of major taxonomic groups, not only did both metagenomes share a similar functional profile but also a similar distribution of non-redundant genes in different functional categories. CONCLUSIONS/SIGNIFICANCE: Our results reveal a highly diverse pool of bacterial species in both O. nubilalis populations, with major differences: The lab-reared sample is rich in gram-positive species (two of which have almost fully sequenced genomes) while the field sample harbors mainly gram-negative species and has a larger set of cellulolytic enzymes. We have found a clear relationship between the diet and the midgut microbiota, which reveals the selection pressure of food on the community of intestinal bacteria.     

溴氰菊酯对亚洲玉米螟雄蛾感受雌性信息素的影响

为了探索溴氰菊醋对亚洲玉米螟雄蛾感受雌性信息素的影响,本文运用触角电位仪和风洞装置对亚洲玉米螟雄蛾进行了电生理和行为反应的测试．结果表明,处理雄蛾对不同比例雌性信息素的触角电位反应分别在E／Z;1／9、5／5和9／1处出现峰值,与对照相比无显著改变,但反应幅度为0．25—0．45mV,比对照下降0．15—0．35mV．对不同剂量的性信息素,引起可见触角电位反应的低限阈值为1ng,饱和反应剂量为10000ng,但反应幅度亦下降0．1—0．5mV,说明溴氰菊酯主要是影响雄蛾周缘感受系统的敏感性．但在风洞测试中,雄蛾除在反应率上比对照明显降低外,其峰值感受比例从E／Z＝1／1漂移到7／3以上．雄蛾对雌性信息素的低限反应剂量也从10ng漂移到100ng,并且高限反应剂量也从100000ng降低到50000ng,使有效作用剂量范围缩小．这说明溴氰菊酯对雄蛾的行为反应有较大影响,不仅使雄蛾对性信息素感受的敏感性降低,也影响到其感受的专化性。     

Intercrop effects on the distribution of populations of the European corn borer, Ostrinia nubilalis, in maize

Field investigations conducted from 1982 to 1984 to assess intercrop effects on the distribution of Ostrinia nubilalis (Hübner), Lepidoptera, Pyralidae, in maize revealed that the incidence of damage by O. nubilalis was influenced significantly by the proximity of maize to potato. Maize fields adjacent to potato fields had a lower incidence of early season damage by O. nubilalis and a higher incidence of late-season damage than maize isolated from potato.     

亚洲玉米螟不同地理种群基于COⅠ基因序列初步分析

本研究通过对我国亚洲玉米螟Ostrinia furnacalis(Guenée)6个地理种群的线粒体DNACOⅠ基因进行测序,运用分子进化遗传分析软件MEGA5对序列进行对比和同源性分析,并建立系统发育关系。结果表明,COⅠ基因在6个地理种群中进化具有一定的差异,平均遗传距离为0.013;进化树显示,亚洲玉米螟的三亚种群、广州种群与南昌种群聚为一支,然后与泰安种群聚为一支,再与哈尔滨种群聚为一支;而廊坊种群另聚成一支。遗传分歧度与系统发育进化关系均表明不同地理种群的遗传分化与地理距离之间具有一定的相关性。     

亚洲玉米螟两个不同地理种群对暗期干扰的滞育反应

为了探明亚洲玉米螟Ostrinia furnacalis (Guenée)不同地理种群对暗期干扰的滞育反应, 本文在研究了亚洲玉米螟南昌种群（28°41′N, 115°53′E）和哈尔滨种群（44°56′N, 127°10′E）光周期反应的基础上, 在室内分别测试了25℃和28℃温度下1 h的光脉冲干扰光周期L9∶D15和L12∶D12对这两个不同地理种群幼虫滞育抑制的影响。结果表明： 在25℃和28℃下, 哈尔滨种群的临界日长分别比南昌种群延长了1 h 40 min和2 h; 在所有测试的暗期干扰实验中, 除了极少数光脉冲干扰点外, 南昌种群幼虫滞育的发生率显著低于哈尔滨种群; 1 h光脉冲干扰光周期L9∶D15的滞育发生率明显低于干扰光周期L12∶D12, 且前者表现了暗期的中间对光脉冲最敏感, 而后者显示了暗期的初期对光脉冲最敏感; 28℃下光脉冲对滞育的抑制效果强于25℃。这些结果进一步提示, 即使在同种昆虫中, 如果地理种群和实验条件不同, 暗期干扰对滞育抑制的效果也可能不同。     

亚洲玉米螟滞育诱导外源性因子研究

研究了食料、温度、光周期等外源性因子对亚洲螟滞育的影响,试验证明,食料性质对其滞育诱导影响显著,取食玉米雌的幼虫以滞育诱导光周反应的敏感性强于取食棉铃的幼虫,在一定范围内,食料中可溶性糖含量增加可促进幼虫的滞育诱导,而粗蛋白含量的增加则具有抑制滞育诱导的作用,两者的互作效应不显著。高温抑制滞育诱导,你 促进滞育诱导。短光周对幼虫的滞育诱导无特定的敏感龄期,但以３龄与２龄对短光周刺激敏感性较强,包光     

Dynamics of Nosema pyrausta in natural populations of the European corn borer, Ostrinia nubilalis: A six-year study

Nosema pyrausta (Paillot) (Microsporida: Nosematidae) is an obligatory intracellular parasite of the European corn borer, Ostrinia nubilalis (Hübner) (Lepidoptera: Crambidae). This pathogen is maintained in natural populations of O. nubilalis by both horizontal and vertical transmission. The impact of N. pyrausta on fecundity of adults and survival of larvae has been well documented in laboratory and field research. In an extensive study covering a 6 year period at one site, we described the effect of N. pyrausta within O. nubilalis populations in a continuous corn following corn ecosystem. We documented the presence of the pathogen through all life stages of O. nubilalis (egg, larva, pupa, adult), by collecting throughout the crop season and examining each insect stage in the laboratory for the frequency of infection with N. pyrausta. The percentage of infection with N. pyrausta and magnitude of the O. nubilalis population fluctuated throughout generation 1 and generation 2. Both horizontal and vertical transmission played a role in maintaining N. pyrausta in the population in both generations. There were strong correlations between percentage adults with N. pyrausta and percentage larvae with N. pyrausta, and between percentage eggs with N. pyrausta and percentage larvae with N. pyrausta. There was a weak correlation between percentage adults with N. pyrausta and percentage eggs with N. pyrausta. The percentage of insects infected with N. pyrausta was always lowest in the egg.     

Frequency of alleles conferring resistance to Bt maize in French and US corn belt populations of the European corn borer,Ostrinia nubilalis

Farmers, industry, governments and environmental groups agree that it would be useful to manage transgenic crops producing insecticidal proteins to delay the evolution of resistance in target pests. The main strategy proposed for delaying resistance to Bacillus thuringiensis ( Bt) toxins in transgenic crops is the high-dose/refuge strategy. This strategy is based on the unverified assumption that resistance alleles are initially rare (<10(-3)). We used an F(2) screen on >1,200 isofemale lines of Ostrinia nubilalis Hübner (Lepidoptera: Crambidae) collected in France and the US corn belt during 1999-2001. In none of the isofemale lines did we detect alleles conferring resistance to Bt maize producing the Cry1Ab toxin. A Bayesian analysis of the data indicates that the frequency of resistance alleles in France was <9.20 x 10(-4) with 95% probability, and a detection probability of >80%. In the northern US corn belt, the frequency of resistance to Bt maize was <4.23 x 10(-4) with 95% probability, and a detection probability of >90%. Only 95 lines have been screened from the southern US corn belt, so these data are still inconclusive. These results suggest that resistance is probably rare enough in France and the northern US corn belt for the high-dose plus refuge strategy to delay resistance to Bt maize.     

Age- and density-related oviposition behavior of the European corn borer,Ostrinia nubilalis (Lepidoptera: Pyralidae)

Age-related oviposition patterns ofOstrinia nubilalis were studied at three population densities in the laboratory by releasing newly eclosed adults in wire-screened cages and analyzing their oviposition throughout the adult stage with digital analysis. Oviposition sequences of individual females depositing egg masses were documented on the third and seventh nights after eclosion with a video camcorder. During a sequence, a female produced an egg in an average time of 15 or 26 s on the third and seventh nights, respectively, and completed depositing an egg mass the size of 20–39 eggs in an average time of 316 and 525 s, respectively. Females were not easily disturbed during egg mass deposition and pulsated their abdomen before deposition of each egg. Females produced few egg masses the first night after eclosion. Oviposition increased on nights 2 and 3 but declined steadily thereafter as females matured. Females older than 6 nights produced fewer egg masses; the proportion of egg masses with fewer than 20 eggs increased gradually. By the end of the adults' lifetime, nearly 100% of the egg masses had fewer than 20 eggs. The data are fundamental to our research to define the role of phytochemicals in modifying oviposition behavior of the European corn borer.