Establishment of the Amplified Fragment Length Polymorphism (AFLP) technique for genotyping of pollen beetle (Meligethes aeneus) - a noxious insect pest on oilseed rape (Brassica napus)

In order to conduct studies concerning genetic variability of pollen beetles (Meligethes aeneus), a genotyping protocol was established. No genome information is available for pollen beetles so the amplified fragment length polymorphism (AFLP) technique was chosen since it does not depend on any prior sequence information of the samples and also is a sensitive and robust technique. However, several modifications were needed in order to adapt the method for analysis of pollen beetles. Basic modifications included (i) alterations of DNA purification, (ii) use of two six-cutter restriction enzymes, (iii) and modified PCR conditions. This protocol resulted in a favourable number of fragments of an appropriate size range for standard gel analysis by a DNA sequencer applicable to a single insect and even body parts enabling different assays to be conducted on a single specimen. Pollen beetles from different areas of Sweden were analysed to verify the reproducibility and efficacy of the protocol as well as for phenetic analysis. The high reproducibility of the modified AFLP protocol allows it to be used as a reliable tool for genotype analysis of pollen beetles.     

Glucosinolates in oilseed rape (Brassica napus) and the incidence of pollen beetles (Meligethes aeneus)

During an early stage of migration into a field of oilseed rape in the spring of 1988, more pollen beetles were observed on the double-low variety, Ariana, than on the adjacent single-low varieties, Bienvenu and Mikado. More beetles were found on open flowers on terminal racemes than on yellow and green flower buds on lateral racemes. Five alkenyl, three indolyl and two aromatic glucosinolates were identified in the floral tissues of oilseed rape. The major compounds were progoitrin (2-OH, 3-butenyl) and glucobrassicanapin (4-pentenyl). There were no differences in the types and concentrations of glucosinolates present in the terminal and lateral racemes of any variety, and only small differences between single- and double-low varieties. Concentrations of glucosinolates in floral tissues of crops grown with a little nitrogen were twice those of well-fertilised crops. There was no association between the number and distribution of pollen beetles on plants and the amounts or types of glucosinolates present in floral tissues.     

Ecology and genetics of insecticide resistance inHelicoverpa armigera: Interactions between selection and gene flow

Pyrethroid resistance inHelicoverpa armigera provides a model system in which to study evolution in natural populations. Resistance is seen to evolve as a consequence of selection pressure that varies within and between life-stages and gene flow. Although three different mechanisms are involved, present day fluctuations in phenotype frequency can be explained by variation in only one of these, metabolic resistance, that is inherited as a single, incompletely dominant gene,mfo. Selective mortality of phenotypes occurs in both larvae and adults in the presence of the pyrethroid insecticides. Although most individuals of all three genotypes are killed in young larvae, selection in this age-class contributes significantly to evolution of resistance. While there is some evidence of reduced fitness of resistant pupae during winter diapause, most of the decline in resistance frequencies each spring occurs as a result of immigration of susceptible individuals into insecticide-treated populations.     

Assessing temporal and spatial variation in wild populations of Atlantic salmon with particular reference to Asturias (Northern Spain) rivers

The extent of genetic variation and levels of temporal and spatial heterogeneity was investigated, at six polymorphic protein‐coding loci, in wild Atlantic salmon Salmo salar populations from six rivers of Asturias (Northern Spain). Also, stocks from northern Europe that were among those introduced to repopulate Asturian Rivers, and other wild Spanish and European populations were characterized. The lack of temporal variation observed suggests that effective population sizes of Asturian populations are sufficiently large to prevent extreme levels of genetic drift and that the introduced fish had a negligible contribution to the fisheries of Asturian rivers.     

Wide variation in spatial genetic structure between natural populations of the European beech (Fagus sylvatica) and its implications for SGS comparability

Identification and quantification of spatial genetic structure (SGS) within populations remains a central element of understanding population structure at the local scale. Understanding such structure can inform on aspects of the species' biology, such as establishment patterns and gene dispersal distance, in addition to sampling design for genetic resource management and conservation. However, recent work has identified that variation in factors such as sampling methodology, population characteristics and marker system can all lead to significant variation in SGS estimates. Consequently, the extent to which estimates of SGS can be relied on to inform on the biology of a species or differentiate between experimental treatments is open to doubt. Following on from a recent report of unusually extensive SGS when assessed using amplified fragment length polymorphisms in the tree Fagus sylvatica, we explored whether this marker system led to similarly high estimates of SGS extent in other apparently similar populations of this species. In the three populations assessed, SGS extent was even stronger than this previously reported maximum, extending up to 360 m, an increase in up to 800% in comparison with the generally accepted maximum of 30-40 m based on the literature. Within this species, wide variation in SGS estimates exists, whether quantified as SGS intensity, extent or the Sp parameter. Consequently, we argue that greater standardization should be applied in sample design and SGS estimation and highlight five steps that can be taken to maximize the comparability between SGS estimates.     

Spatial and temporal changes in genetic structure of greenhouse and field populations of cabbage looper,Trichoplusia ni

Trichoplusia ni is a subtropical moth that migrates annually from southern California to southern British Columbia, Canada where it invades vegetable greenhouses and field crops. The heated greenhouse environment has altered the natural extinction–recolonization dynamics of T. ni populations, and allows year‐round persistence in some locations. In addition, the extensive use of the biopesticide, Bacillus thuringiensis subspecies kurstaki (Bt) in some greenhouses has selected for resistance. Here we investigated the genetic structure of T. ni populations in British Columbia greenhouses and in field populations in California and British Columbia using amplified fragment length polymorphisms (AFLP) as related to patterns of Bt resistance. The majority of British Columbia field populations were similar to the California field populations, the potential source of migrants. However populations in two geographic areas with high concentrations of greenhouses showed local genetic differentiation. Some of these populations experienced severe bottlenecks over‐winter and following Bt sprays. Greenhouse populations showed a pattern of isolation by distance and a strong positive relationship between genetic differentiation and levels of Bt resistance. These patterns indicate that greenhouses that sometimes support year‐round populations of T. ni and the ensuing strong bottlenecking effects following winter cleanups and Bt application cause genetic differentiation of T. ni populations. Long distance migrants to field populations contribute to genetic homogeneity of these.     

Variation in spatial and temporal gradients in zooplankton spring development: the effect of climatic factors

1. We examined the temporal and spatial heterogeneity of zooplankton in lake surface waters during the spring of 3 years in Lake Washington, U.S.A., a large lake with a high production of sockeye salmon fry. 2. We show large within‐season and among‐year variation in the horizontal distribution of temperature, chlorophyll a concentration, and zooplankton in the lake. The main pattern, a delay in zooplankton population increase from the north‐ to the south‐end of the lake, recurred in each year and was persistent within each spring. 3. The delay is primarily caused by the development of a temperature gradient during spring warming, as cold mountain water enters the south end of the lake, while warm water enters the north end via a river draining a nearby lake. Climate factors, such as air temperature and precipitation during winter and spring, appear to influence the extent of the delay of zooplankton increase. 4. If the climate continues to warm, the temporal disconnection in zooplankton development between lake areas immediately influenced by cold river inflow and areas that are influenced by spring warming may increase in magnitude. Thus, the different areas of the lake may not contribute equally to fish production.     

Methodology for glutathione S-transferase purification and localization in two-dimensional gel electrophoresis performed on the pollen beetle,Meligethes aeneus (Coleoptera: Nitidulidae)

Glutathione S-transferases (GSTs) are one of the major detoxification enzymes involved in insecticide resistance. In this study, optimized methodology for GSTs purification and localization in two-dimensional gel electrophoresis is shown on the pollen beetle. Specifically, the GST proteins were purified using GSTrap 4B column, and the protein profile of the supernatant before purification was compared with the unbound fraction post-purification via two-dimensional gel electrophoresis (2D-E). The identity of these localized protein spots was confirmed by MS analysis and further analyzed by NanoLC-ESI-QUAD-TOF-MS/MS protein identification. Results indicate that 2 out of 5 protein spots were GSTs with region for class Delta and Epsilon subfamilies similar to other insects; however, the remaining 3 spots did not show any match in the current NCBInr. Both Delta and Epsilon class GSTs are specifically involved in insecticide resistance and their relatively high abundance in the 2D-E map suggests that these enzymes could play a role in the resistance of Meligethes aeneus to the most commonly used pyrethroids. The approach applied in this study for the specific localization of GSTs in 2D-E can be used for similar analyses in other organisms.     

Relative importance of temporal and spatial heterogeneity in the zooplankton community of an artificial reservoir

Zooplankton samples were collected monthly at three stations in Ogochi Reservoir during 1980, and the temporal and horizontal variations in the density of dominant taxa were evaluated by a standard 2-way ANOVA with random effects. The analysis revealed that horizontal is greater than temporal variation in the warm water season (> 18 °C), whereas temporal variation is greater in the cold water season (< 18 °C). Horizontal heterogeneity of the zooplankton community found in the warm water season seems to be associated with differences in site-specific conditions.     

Spatial and temporal variation in distribution of Gelidium canariensis (Rhodophyta) from natural populations of the Canary Islands

This study was designed to investigate spatial and temporal variation in Gelidium canariensis populations at two shores in northern Gran Canaria during two years. Spatial scales ranged from some hundred meters (distance between shores), 10 to 30 m (distance between plots) to less than 3 m (distance between quadrats). Gelidium individuals were defined as distinct Gelidium clumps. The results show a significant difference in size of clumps between shores, but not on the smaller spatial scales. No significant temporal variation was found. There was no significant temporal or spatial variation in standing crop or density (counts made in quadrats where Gelidium was present, rather than counts for the total shore). Sporophytic and gametophytic clumps were also distinguished by identifying reproductive structures in the field. The total proportion of sporophytes was larger than the proportion of gametophytes, but at a smaller scale there could be a shift in dominance. The survival rate of clumps was similar between shores with a mean survival rate of 85%, but there was a significant difference in recruitment between shores. The results indicate a stable population structure. This revised version was published online in June 2006 with corrections to the Cover Date.     

Spatial and temporal occurrence of invertebrates in a chalk stream,Berkshire, England

The invertebrate assemblages of the River Lambourn and its tributary, the Winterbourne stream, were investigated as part of a broad ecological study prior to water resource development in the catchment. The longitudinal distribution of the invertebrate fauna within the Winterbourne stream was examined in detail and supporting data for the R. Lambourn revealed that the upstream intermittent section of each water course had an invertebrate assemblage which was distinct from the fauna downstream where flow was permanent. Within the perennial zone taxon richness tended to increase downstream.A one-year programme of monthly quantitative sampling on five distinct biotopes (Ranunculus, Berula, Callitriche, gravel and silt) at two contrasted sites (shaded and unshaded) was undertaken in the lower perennial of the R. Lambourn below the confluence with the Winterbourne. On each study site, the taxon richness, numerical abundance and biomass of invertebrates were significantly higher on the three macrophytes than on gravel and silt. There were also statistically significant seasonal changes in the biomass of invertebrates on each site. Estimates of total biomass of invertebrates per site revealed a late spring peak (May/June) and a late autumn (November/December) peak, neither of which coincided with the summer peak biomass of macrophytes. This suggested that, in the lower perennial, the level of food resources (epiphytic algae in late spring and fallen tree leaves plus decaying macrophytes in autumn) rather than available habitat, were a major influence on population levels for most of the year.Quantitative sampling of each biotope on each site continued in June and December only for a period of 8 years (1971–78), during which time the river experienced a minor drought (1973) and a severe drought (1976). The Chironomidae showed a strong positive response to conditions of low flow in June of 1973 and 1976, when exceptionally high densities of larvae were recorded. Although further between-year differences were recorded in several other major components of the invertebrate fauna, which may have been related to conditions of low discharge, they did not have long-term consequences for the fauna in the lower perennial of the R. Lambourn.In contrast, in the upper perennial, the prolonged drought of 1976 resulted in the loss of wetted perimeter and macrophytes, coupled with deposition of silt, all of which had deleterious effects on the invertebrate assemblages. The following year, despite a normal discharge regime and the removal of silt, some submerged macrophytes were slow to return to their pre-drought condition. In the intermittent section, the drought of 1976 resulted in the total loss of flow for a 16 month period instead of a more typical four month period of water loss.Thus, chalk streams encompass within a single system both stable and extreme hydrological regimes which offer unique opportunities for investigating the processes of community formation and maintenance.     

Genetic mapping in sugarcane, a high polyploid, using bi-parental progeny: identification of a gene controlling stalk colour and a new rust resistance gene

Modern sugarcane cultivars (Saccharum spp) are highly polyploïd and aneuploid interspecific hybrids (2n=100–130). Two genetic maps were constructed using a population of 198 progeny from a cross between R570, a modern cultivar, and MQ76-53, an old Australian clone derived from a cross between Trojan (a modern cultivar) and SES528 (a wild Saccharum spontaneum clone). A total of 1,666 polymorphic markers were produced using 37 AFLP primer combinations, 46 SSRs and 9 RFLP probes. Linkage analysis led to the construction of 86 cosegregation groups for R570 and 105 cosegregation groups for MQ76-53 encompassing 424 and 536 single dose markers, respectively. The cumulative length of the R570 map was 3,144 cM, while that of the MQ76-53 map was 4,329 cM. Here, we integrated mapping information obtained on R570 in this study with that derived from a previous map based on a selfed R570 population. Two new genes controlling Mendelian traits were localized on the MQ76-53 map: a gene controlling the red stalk colour was linked at 6.5 cM to an AFLP marker and a new brown rust resistance gene was linked at 23 cM to an AFLP marker. Besides another previously identified brown rust resistance gene (Bru1), these two genes are the only other major genes to be identified in sugarcane so far.     

Electrophysiological and behavioural responses of the pollen beetle, Meligethes aeneus, to volatiles from a non-host plant, lavender, Lavandula angustifolia (Lamiaceae)

A semiochemical based push-pull strategy for control of oilseed rape pests is being developed at Rothamsted Research. This strategy uses insect and plant derived semiochemicals to manipulate pests and their natural enemies. An important element within this strategy is an understanding of the importance of non-host plant cues for pest insects and how such signals could be used to manipulate their behaviour. Previous studies using a range of non-host plants have shown that, for the pollen beetle Meligethes aeneus (Coleoptera: Nitidulidae), the essential oil of lavender, Lavandula angustifolia (Lamiaceae), was the most repellent. The aim of this study was to identify the active components in L. angustifolia oil, and to investigate the behaviour of M. aeneus to these chemicals, to establish the most effective use of repellent stimuli to disrupt colonisation of oilseed rape crops. Coupled gas chromatography-electroantennography (GC-EAG) and gas chromatography-mass spectrometry (GC-MS) resulted in the identification of seven active compounds which were tested for behavioural activity using a 4-way olfactometer. Repellent responses were observed with (±)-linalool and (±)-linalyl acetate. The use of these chemicals within a push-pull pest control strategy is discussed. Handling Editor: Yvan Rahbe.     

Genetic diversity within and among 11 juvenile populations of green alder (Alnus crispa) in Canada

Germinated seeds from 11 populations of green alder [ Alnus crispa (Ait.) Pursh] sampled in four Canadian provinces were analysed for electrophoretically demonstrable diversity of 10 enzymes encoded by 15 structural loci. Of these, nine were polymorphic, and on average, 52% of the loci per population were polymorphic. Assuming a diploid model of expression, average level of expected heterozygosity was 0.11 with nearly all populations in Hardy-Weinberg equilibrium for the set of polymorphic loci analysed. No significant inbreeding and associated subpopulation structuring were noted. Rates of gene flow appeared high within and among populations. Although little divergence was observed among populations, genetic and geographical distances between populations were related. Discriminant and cluster analyses revealed a pattern of genetic variation associated with geography. Populations from northern Quebec were poorly differentiated, whereas western populations from Alberta exhibited a larger degree of genetic differentiation. Introgresive hybridization with the sympatric species Alnus sinuata (Regel) Rydberg and partial isolation in the West are suggested as an explanation for this larger differentiation. The occurrence and significance of rare alleles is discussed in relation to the importance of geographical distance in the process of population differentiation in this species.     

Genetic diversity of wild Cymbidium goeringii (Orchidaceae) populations from Hubei based on Inter-simple sequence repeats analysis

Cymbidium goeringii is a diploid and nonrewarding,bumblebee-pollinated species,which is distributed in China,Japan and Korea Peninsula.This species is now highly endangered due to the mass collection and forest clearance in China.In the present study,we investigated the distribution of genetic variation within and between eleven populations of Cymbidium goeringii in central China by using Inter-simple sequence repeats (ISSR) markers.Eleven primers produced a total of 127 clear and reproducible bands of which 112 were polymorphic.High genetic diversity was detected in Cymbidium goeringii for both population level (P = 63.1%;He = 0.194 5) and species level (P = 88.2%;He = 0.262 8).A higher level of genetic differentiation was detected among populations (GST = 0.244 0,FST = 0.220 7)with Nei's Gsr analysis and analysis of molecular variance (AMOVA),and no correlation was found between geographical and genetic distance.Genetic drift rather than gene flow played an important role in forming the present population structure of Cymbidium goeringii.Limited gene flow among populations and gene drift increase the extinction risk of local populations.Some conservation concerns are therefore discussed together with possible strategies for implementing in situ and ex situ conservation.     

Biodiversity of vespid wasps in spatial and temporal dimensions in northern Zanjan Province of Iran

Vespidae is one of the major families of Hymenoptera with more than 115000 described species. In the present paper, Simpson and Shannon-Wiener diversity indices, Simpson,Camargo, Smith and Wilson, and modified Nee evenness indices, and richness index based on rarefaction method were adopted to study the biodiversity of vespid wasps in spatial and temporal dimensions in northern Zanjan Province of Iran. In spatial dimension, Zanjan and Gilvan showed the highest and the lowest species diversity, evenness, and richness, respectively; while in tem-poral dimension, 16 July-6 August showed the highest species diversity and richness, and 18 May -27 May and 23 August -2 September showed the highest and the lowest species evenness,respectively. Significant differences in species abundance between localities of study area were observed by Kruskal-Wallis nonparametric test.     

Genetic variability and large scale differentiation in two species of littorinid gastropods with planktotrophic development, Littorina littorea (L.) and Melarhaphe (Littorina) neritoides (L.) (Prosobranchia: Littorinacea), with notes on a mass occurrence of M. neritoides in Sweden

A planktotrophic larval development suggests a relatively high gene flow over long distances. Both Littorina littorea and Melarhaphe neritoides have an egg capsule and planktotrophic larvae which taken together are pelagic over a period of 4–8 weeks. In the absence of differential selection one would expect low levels of genetic differentiation over large distances in these two species. In this study of allozyme variation, low levels of differentiation were found over thousands of kilometres (Norway to Spain for L. littorea and Sweden to Greece for M. neritoides). This supports the hypothesis of two species with a high dispersal potential. A second expectation from neutral theory is that effective population size is positively correlated with average levels of genetic variation within species. In light of the generally high densities of local populations of these species and the high interpopulation migration rate, both L. littorea and M. neritoides may be considered as having high N_e s. Contrary to neutral expectation, L. littorea revealed very low levels of heterozygosity over its whole European range (mean H_exp= 0.015, 15 loci), while average heterozygosity of M. neritoides (H_exp= 0.084, 11 loci) was no more than in other littorinid species. This paper also reports the occurrence of M. neritoides on the Swedish west coast in 1988, 89 and 90, and two factors which may have promoted this unusually large invasion are discussed.     

鄱阳湖生态经济区成立前后NDVI时空动态变化及其驱动因素

鄱阳湖生态经济区是中部地区正在加速形成的增长极之一,研究其植被变化特征可以为鄱阳湖生态经济区更合理的调控经济发展与生态保护提供支持。本研究基于2000—2017年MODIS NDVI,采用最小二乘法斜率探讨生态经济区成立前后NDVI时空变化特征,并借助相关性分析定量化研究气候因素和人为因素对NDVI变化的驱动作用。结果表明:生态经济区成立前(2000—2008年)和成立后(2009—2017年),区域植被覆盖整体均呈现增长趋势。生态经济区成立前NDVI变化趋势为0.052 10 a^-1(P<0.05),成立后NDVI变化趋势为0.025 10 a^-1(P<0.05),成立后NDVI增长出现放缓现象;空间上,成立前植被发生退化的区域主要分布在地级市和省会周围;成立后,省会和中心城市周围植被退化现象进一步加剧,同时湖区周围、区域北部以及南部农耕区植被也出现退化现象;在影响因素方面,在全区域尺度上,气候因素是NDVI变化的主导因素,气温对NDVI影响大于降水;在NDVI显著变化的区域,人类活动则是影响NDVI变化的主要因素;生态经济区成...     

On the worldwide spread of an insecticide resistance gene: a role for local selection

Adaptation occurs by gene replacement (or transient balanced polymorphism). Replacement may be caused by selection (local or global) and/or genetic drift among alleles. In addition, historical events may blur the respective effects of selection and drift during the course of replacement. We address the relative importance of these processes in the evolution of insecticide resistance genes in the mosquito Culex pipiens. The resistance allele, Ester2, has a broad geographic distribution compared to the other resistance alleles. To distinguish between the different processes explaining this distribution, we reviewed the literature and analysed updated data from the Montpellier area of southern France. Overall, our data indicate that Ester2 prevails over other Ester resistance alleles in moderately treated areas. Such conditions are common and favour the hypothesis of selection acting at a local level. This places an emphasis on the importance of ecological conditions during the evolution of resistance. Finally, we highlight that historical events have contributed to its spread in some areas.