John A. Tsitsipis

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.

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.

A small deletion in the olive fly acetylcholinesterase gene associated with high levels of organophosphate resistance

Organophosphate resistance in the olive fly was previously shown to associate with two point mutations in the ace gene. The frequency of these mutations was monitored in Bactrocera oleae individuals of increasing resistance. In spite of the difference in resistance among the individuals, there was no correlation between mutation frequencies and resistance level, indicating that other factors may contribute to this variation. The search for additional mutations in the ace gene of highly resistant insects revealed a small deletion at the carboxyl terminal of the protein (termed Delta3Q). Significant correlation was shown between the mutation frequency and resistance level in natural populations. In addition, remaining activity of acetylcholinesterase enzyme (AChE) after dimethoate inhibition was higher in genotypes carrying the mutation. These results strongly suggest a role of Delta3Q in high levels of organophosphate (OP) resistance. Interestingly, the carboxyl terminal of AChE is normally cleaved and substituted by a glycosylphosphatidylinositol (GPI) anchor. We hypothesize that Delta3Q may improve GPI anchoring, thus increasing the amount of AChE that reaches the synaptic cleft. In this way, despite the presence of insecticide, enough enzyme would remain in the cleft for its normal role of acetylcholine hydrolysis, allowing the insect to survive. This provides a previously un-described mechanism of resistance.

Morphological discrimination of Aphis gossypii (Hemiptera: Aphididae) populations feeding on Compositae

Aphis gossypii Glover is a polyphagous aphid pest with a worldwide distribution. However, there is evidence that on a global scale the name A. gossypii is being applied to a number of forms with different life cycles and/or host-plant associations. Morphometric variation of A. gossypii samples from crops and non-cultivated plants in many parts of the world was examined, to determine whether this variation is correlated with the hosts from which the aphids originated. Samples of A. gossypii were collected from Cucurbitaceae and Malvaceae in Europe, and from Compositae in various parts of the world. Morphometric data for 13 parameters measured from 97 clonal lineages (728 specimens) and 27 field-collected samples (313 specimens) were analysed by a series of canonical variates analyses, using the field sample/clonal lineage as grouping factor. Clonal lineages were reared on a common host in controlled conditions to standardize the effect of host and environment on morphology. The analyses provided a clear morphometric separation of the aphids originating from Compositae and those collected on Cucurbitaceae and Malvaceae, regardless of the geographical origin of the aphids and the host plant on which they were reared. This indicates that within A. gossypii there are two widely distributed host races or subspecies with different plant family associations. The taxonomic implications are discussed.

Co-existence of different host-adapted forms of the Myzus persicae group (Hemiptera: Aphididae) in southern Italy

Multivariate morphometric analysis (method of canonical variates) was used to compare 38 parthenogenetic lineages and three field collections of aphids of the Myzus persicae (Sulzer) group sampled on peach and tobacco in the Caserta region of southern Italy. Comparisons were also made with the morphology of lineages collected on peach in Lehonia, in central eastern Greece away from tobacco-growing regions, and on tobacco in Naphplion, in southern Greece. The lineages were measured after parthenogenetic rearing for one to three generations on the same host (potato) under constant conditions. As in previous work, the multivariate morphometric approach separated the aphids from peach from those feeding on tobacco. The life cycle category of the lineages was also examined. Almost all the lineages from peach in Caserta were holocyclic, yet the lineages from tobacco in the same region were unable to produce sexual morphs. The results suggest that tobacco-adapted and non-tobacco-adapted forms co-exist in the same region in southern Italy, not only because they colonize different host plants, but also because they have different life cycles. This is in complete contrast to the situation in peach-growing areas of northern Greece, and shows that the ecology and population structure of M. persicae is different in neighbouring counties of the Mediterranean area, even where climatic conditions and cultivated crops are similar.