John T. Margaritopoulos

Micro‐evolutionary change in relation to insecticide resistance in the peach–potato aphid, Myzus persicae

1. Phenotypic diversity is the fuel that powers evolution.

Host-correlated morphological variation of Myzus persicae (Hemiptera: Aphididae) populations in Greece.

Morphological variation in nine characters of 157 clones of Myzus persicae (Sulzer) was examined by multivariate analysis. The clones were collected from peach, Prunus persica, the primary host and the secondary hosts tobacco, Nicotiana tabacum, cabbage, Brassica oleracea, sugarbeet, Beta vulgaris and pepper Capsicum annuum. The 156 clones originated from various regions of Greece, both in the north, where a large part of the population has an annual bisexual generation on peach, and in more southerly regions, where populations are predominantly unisexual. One clone was collected from tobacco in Caserta, Italy. All clones were laboratory-reared on potato. Canonical variate analysis, hierarchical cluster analysis and a non-parametric classification tree method both revealed morphological differences associated with the host-plant on which they were collected. The scores of the first two canonical variates separated the tobacco-feeding clones from those originating from other secondary host-plants. However, in tobacco-growing areas the tobacco-feeding form predominated in spring populations on peach, and was sometimes found on other secondary hosts. In addition, using cluster analysis, the clones from tobacco which were sampled in the most southeasterly region showed a relatively large phenotypic distance from those collected further north and west. Moreover, clonal phenotypes were affected both by host plant and by long-term parthenogenetic rearing. However, in spite of these effects, the tobacco form was generally distinguishable from aphids originating from other hosts, indicating that the difference must have a genetic basis. In separate analyses of the clones originating from secondary hosts no association was found between morphology and either life cycle category or colour. Discriminant analysis showed that 89% of 1723 specimens could be correctly classified into the two groups.

Life cycle variation of Myzus persicae (Hemiptera: Aphididae) in Greece.

During the years 1995-1999 the life cycle category of 2797 clones of Myzus persicae (Sulzer) was examined. The clones originated from primary and secondary hosts from different localities of North and Central Greece and the island of Crete in the south. Four different overwintering life cycle strategies were found that have also been described for M. persicae and other heteroecious species previously. A geographical variation was found in the proportion of holocyclic clones from tobacco and other secondary hosts associated with the abundance of the primary host in the sampling regions. In Central Macedonia, around the main peach-growing regions, the proportion of holocyclic clones was mostly above 50% and in some cases reached 100%. In localities of East Macedonia, holocyclic clones were also frequent. On the other hand, further south or in north-eastern Greece, where peach is not common, the proportion of holocyclic clones varied between 0 and 33%. Fifty seven percent of examined anholocyclic clones produced males under short day conditions, suggesting that androcyclic clones in Greece represent an important factor of genetic variability. Intermediate clones were sampled from all host-plants but at low frequencies (3.6% of total examined clones and 6.9% of non-holocyclic ones). Moreover, a regional variation was found in different colour forms feeding on tobacco plants. Red clones were predominant in regions where aphids overwinter parthenogenetically on weeds or winter crops. However, almost all clones from the primary host were green. The ecological aspects of life cycle variation are discussed.

Tracking the global dispersal of a cosmopolitan insect pest, the peach potato aphid

BackgroundGlobal commerce and human transportation are responsible for the range expansion of various insect pests such as the plant sucking aphids. High resolution DNA markers provide the opportunity to examine the genetic structure of aphid populations, identify aphid genotypes and infer their evolutionary history and routes of expansion which is of value in developing management strategies. One of the most widespread aphid species is the peach-potato aphid Myzus persicae, which is considered as a serious pest on various crops in many parts of the world. The present study examined the genetic variation of this aphid at a world scale and then related this to distribution patterns. In particular, 197 aphid parthenogenetic lineages from around the world were analysed with six microsatellite loci.ResultsBayesian clustering and admixture analysis split the aphid genotypes into three genetic clusters: European M. persicae persicae, New Zealand M. persicae persicae and Global M. persicae nicotianae. This partition was supported by FST and genetic distance analyses. The results showed two further points, a possible connection between genotypes found in the UK and New Zealand and globalization of nicotianae associated with colonisation of regions where tobacco is not cultivated. In addition, we report the presence of geographically widespread clones and for the first time the presence of a nicotianae genotype in the Old and New World. Lastly, heterozygote deficiency was detected in some sexual and asexual populations.ConclusionThe study revealed important genetic variation among the aphid populations we examined and this was partitioned according to region and host-plant. Clonal selection and gene flow between sexual and asexual lineages are important factors shaping the genetic structure of the aphid populations. In addition, the results reflected the globalization of two subspecies of M. persicae with successful clones being spread at various scales throughout the world. A subspecies appears to result from direct selection on tobacco plants. This information highlights the ultimate ability of a polyphagous aphid species to generate and maintain ecologically successful gene combinations through clonal propagation and the role of human transportation and global commerce for expanding their range.

Worldwide variability of insecticide resistance mechanisms in the codling moth, Cydia pomonella L. (Lepidoptera: Tortricidae)

The activity of detoxifying enzymes (glutathione-S-transferases (GST), mixed-function oxidases (MFO), and esterases (EST)) and the presence of insensitive variants of target proteins (sodium channel and acetylcholinesterase) were examined in individual male and female codling moths. Twenty-nine populations from 11 countries and two laboratory strains were examined. Populations were classified as either unsprayed or sprayed. The ranges of enzyme activities across field populations varied 15-fold, 485-fold and fourfold for GST, MFO and EST, respectively. MFO was the only enzyme whose activity differed in a binomial classification of orchards based on their spray history. Few differences in enzyme activities were found due to sex among populations; and, in these cases, males had higher GST and lower MFO and EST activities than females. Activities of the three enzymatic systems across all populations were positively correlated. Populations from Greece, Argentina and Uruguay had significant percentages of moths with elevated GST and MFO activities. The co-occurrence of moths expressing both elevated MFO and low EST activities was found in conventional orchards from the Czech Republic and France. Chile was the only country where populations from treated orchards did not include a significant proportion of individuals with enhanced enzyme activity. The kdr mutation was found at significant levels in ten populations from five countries, including all French and Argentinean populations. The mutation in AChE was only detected in the Spanish population.

Host selection by winged colonisers within the Myzus persicae group: a contribution towards understanding host specialisation

Abstract.  1. Myzus persicae sensu lato demonstrates considerable genetic variation in respect to adaptation to host plants. The subspecies M. persicae nicotianae shows a preference for tobacco, while M. persicae sensu stricto (s. str.) for other herbaceous plants. Given that winged colonisers of several aphid species play an important role in selecting host plants, here their role in the host specialisation observed in M. persicae was examined in choice and no‐choice tests conducted outdoors, in performance studies, and in DC Electrical Penetration Graph (DC‐EPG) studies.

Biological characteristics of the mirids Macrolophus costalis and Macrolophus pygmaeus preying on the tobacco form of Myzus persicae (Hemiptera: Aphididae).

Biological and population parameters of the predatory bugs Macrolophus costalis Fieber and Macrolophus pygmaeus Rambur preying on the tobacco aphid Myzus persicae (Sulzer) were examined. Tobacco was used as host plant and all experiments were carried out at 23 degrees C and L16:D8. In M. costalis, the developmental time for eggs and the total duration of nymphal instars was 13.1 and 22.2 days and in M. pygmaeus 13.0 and 20.2 days, respectively. Both species completed their nymphal development feeding only on plant juices although they required significantly more time to attain adulthood than when feeding on prey. The longevity of M. costalis females and the preoviposition period were 49.9 and 5.6 days and those of M. pygmaeus were 50.3 and 6.3 days, respectively. Adult males showed a higher longevity than females in both species. The intrinsic rate of increase and the mean total number of eggs laid were 0.0644 and 121 in M. costalis and 0.0615 and 104 in M. pygmaeus, respectively. Mean total aphid consumption by nymphs of M. costalis was 61 aphids, about twice that of M. pygmaeus (37 aphids). Total aphid consumption by a male and female pair of M. costalis and M. pygmaeus until the death of the female was 244 and 285 aphids, respectively. The results of the study are discussed in relation to the impact of the two predatory bugs on tobacco aphid populations.

Performance of the tobacco aphid Myzus persicae (Hemiptera: Aphididae) on Oriental and Virginia tobacco varieties

Abstract  1 The life parameters of one red and one green clone of Myzus persicae reared on 11 Oriental tobacco type varieties (KP7, K63, KZ10/z, MA13/b, BX2/a, N34/4, KP14/a, Kolindros, S79, TA21 and BX81) and on five Virginia ones (VE9, VE3, SPG28‐VE2, Kleio‐KN1 and Niki‐NK3) at 20 °C and LD 16 : 8 h were examined.

Discrimination of the closely related biocontrol agents Macrolophus melanotoma (Hemiptera: Miridae) and M. pygmaeus using mitochondrial DNA analysis

The separation of the closely related predatory species Macrolophus melanotoma Costa (= M. caliginosus Wagner) and Macrolophus pygmaeus (Rambur) based exclusively on the different colour pattern of the first antennal segment (white central band in M. melanotoma and entirely black in M. pygmaeus) is rather precarious and their taxonomic status is still in doubt. In the present study their taxonomic status was evaluated by DNA confirmatory analysis and hybridization experiments between M. pygmaeus and a Macrolophus taxon, resembling M. melanotoma, with a first antennal segment entirely black or with a white central band collected from Dittrichia viscosa (L.) W. Greuter (Asteraceae) in southern Greece. Adult females from Dittrichia plants hybridized with males of M. pygmaeus and vice versa did not produce viable eggs. The Macrolophus species from Dittrichia irrespective of the first antennal segment coloration differed from M. pygmaeusin digestive patterns generated by AseI, XbaI, and MseI on 16S rRNA and in RAPD profiles produced by the primers OPA-18 and OPA-20. These results demonstrate that on Dittrichia plants there is a distinct dimorphic taxon, M. melanotoma, as it is the only species of the genus Macrolophus bearing a first antennal segment with a central white band. Given the limitation of the coloration pattern, the mtDNA genetic markers are the appropriate method for the identification of M. melanotomaand M. pygmaeus.

Efficacy of the pyrethroid alpha-cypermethrin against Bactrocera oleae populations from Greece, and improved diagnostic for an iAChE mutation

BACKGROUND The most important pest of olive orchards worldwide is the olive fruit fly Bactrocera oleae (Gmelin). Its control in Greece has been based on organophosphates (OPs), but their intense use has led to the development of resistance. A test previously developed to monitor the trait may not be as robust as originally thought. The pyrethroid alpha-cypermethrin has recently been registered for bait sprays, as an alternative to OPs. RESULTS The susceptibility of 20 B. oleae populations to alpha-cypermethrin was examined. Variation was observed in their response, with LD(50) ranging from 0.14 to 3.28 ng insect(-1) and resistance factors from 2.3 to 54.7. Resistance mechanisms were investigated. Cytochrome P450 monoxygenase activities showed an association with resistance. Sequences in the domain IIS4-IIS6 of the B. oleae para-type sodium channel were also analysed, but no resistance-associated mutations were identified. Finally, a novel diagnostic assay able to reliably monitor the frequency of the iAChE G488S resistance mutation was developed. CONCLUSION This is the first attempt to evaluate the efficacy of alpha-cypermethrin against B. oleae from Greece. Data showed that it can be used effectively, but also highlighted the importance of continuous monitoring. The IIS4-IIS6 sodium channel region is the default area in which to look for resistance mutations if target-site resistance to pyrethroids arises. The application of the novel iAChE molecular diagnostic may facilitate the introduction of pyrethroids alongside OPs currently in use.