Antibiotic resistance in potato cultivars to the aphid Myzus persicae

Commercially-grown potato cultivars were assessed for antibiotic resistance to Myzus persicae in a culture room at 19°C with a 2°C range and a 16 h photoperiod. The resistant rankings obtained were consistent for the following aphid metrics: maturation time, teneral adult weight, embryo complement and the production of nymphs. Intrinsic rates of increase (r_m) correlated with the above parameters of aphid growth and reproduction but this was not always the case for nymphal survival rate. The cultivars Ulster Tarn, Record and Maris Piper were relatively resistant to M. persicae and cultivars Desirée and King Edward were the most susceptible. When the aphid clone or the culture plants were changed there was no significant effect on the relative resistance of the potato cultivars under test. The relationship between the number of embryos in teneral adults and the production of nymphs by aphids was examined by grouping aphids in weight batches. There was a strong relationship between these two variables.     

Comparative analysis of Solanum stoloniferum responses to probing by the green peach aphid Myzus persicae and the potato aphid Macrosiphum euphorbiae

Abstract Plants protect themselves against aphid attacks by species‐specific defense mechanisms. Previously, we have shown that Solanum stoloniferum Schlechtd has resistance factors to Myzus persicae Sulzer (Homoptera: Aphididae) at the epidermal/mesophyll level that are not effective against Macrosiphum euphorbiae Thomas (Homoptera: Aphididae). Here, we compare the nymphal mortality, the pre‐reproductive development time, and the probing behavior of M. persicae and M. euphorbiae on S. stoloniferum and Solanum tuberosum L. Furthermore, we analyze the changes in gene expression in S. stoloniferum 96 hours post infestation by either aphid species. Although the M. euphorbiae probing behavior shows that aphids encounter more probing constrains on phloem activities–longer probing and salivation time– on S. stoloniferum than on S. tuberosum, the aphids succeeded in reaching a sustained ingestion of phloem sap on both plants. Probing by M. persicae on S. stoloniferum plants resulted in limited feeding only. Survival of M. euphorbiae and M. persicae was affected on young leaves, but not on senescent leaves of S. stoloniferum. Infestation by M. euphorbiae changed the expression of more genes than M. persicae did. At the systemic level both aphids elicited a weak response. Infestation of S. stoloniferum plants with a large number of M. persicae induced morphological changes in the leaves, leading to the development of pustules that were caused by disrupted vascular parenchyma and surrounding tissue. In contrast, an infestation by M. euphorbiae had no morphological effects. Both plant species can be regarded as good host for M. euphorbiae, whereas only S. tuberosum is a good host for M. persicae and S. stoloniferum is not. Infestation of S. stoloniferum by M. persicae or M. euphorbiae changed the expression of a set of plant genes specific for each of the aphids as well as a set of common genes.     

Positive genetic correlations among major life-history traits related to ecological success in the aphid Myzus persicae

Life-history theory is based on the assumption that evolution is constrained by trade-offs among different traits that contribute to fitness. Such trade-offs should be evident from negative genetic correlations among major life-history traits. However, this expectation is not always met. Here I report the results of a life-table experiment designed to measure the broad-sense heritabilities of life-history traits and their genetic correlations in 19 different clones of the aphid Myzus persicae from Victoria, Australia. Most individual traits, as well as fitness calculated as the finite rate of increase from the life table, exhibited highly significant heritabilities. The pattern of genetic correlations revealed absolutely no evidence for life-history trade-offs. Rather, life histories were arranged along an axis from better to worse. Clones with shorter development times tended to have larger body sizes, higher fecundities, and larger offspring. The fitness of clones estimated from the life table in the laboratory tended to be positively associated with their abundance in the field. Fitness also increased significantly with heterozygosity at the seven microsatellite loci that were used to distinguish clones and estimate their frequencies in the field. I discuss these findings in light of a recent proposition that positive genetic correlations among life-history traits for which trade-offs are expected can be explained by genetic variation for resource acquisition ability that is maintained in populations by a cost of acquisition, and I propose ways to test for such a cost in M. persicae.     

Low temperature mortality of the peach-potato aphid Myzus persicae

ABSTRACT.

• 1 The mean supercooling points of first instar and adult Myzus persicae (Sulzer) maintained at 20°C and cooled at 1°C min^?1 were ?26.6 and ?25.0°C respectively.
• 2 The LT₅₀ (temperature) of the same age groups drawn from the same population and cooled at the same rate were ?8.1 and ?6.9°C, indicating extensive pre-freeze mortality in M.persicae under laboratory conditions.
• 3 Acclimation at 10 and 5°C did not affect supercooling but depressed the LT₅₀ of both first instars and adult aphids.
• 4 Freezing of leaves during feeding did not increase mortality above that expected from the direct effects of low temperature.
• 5 The level of cold in different winters can be expressed in terms of the total number of frost days, and the frequency of abnormally cold days. Winter temperatures differ markedly in a vertical profile from the soil to the soil or grass surface, and then to the air (and foliage) above.
• 6 The time of the first record of M.persicae in suction trap samples is correlated with January and February temperatures except in the west of England and Wales. Further north December and January temperatures are relatively more important.
• 7 Winter temperatures and the resultant aphid mortality is a primary determinant of the timing of the spring migration.

     

Location of resistance factors in the leaves of potato and wild tuber-bearing Solanum species to the aphid Myzus persicae

Analysis of electrically recorded feeding behaviour of aphids was combined with colony‐development tests to search for sources of resistance to Myzus persicae (Sulzer) (Homoptera: Aphididae) in tuber‐bearing Solanum species (Solanaceae), aiming at a reduction of potato leaf roll virus (PLRV) transmission. Twenty genotypes, originating from 14 gene bank accessions, representing 13 wild tuber‐bearing Solanum spp., three Solanum tuberosum L. (potato) cultivars, and one S. tuberosum breeding line, were selected. Colony‐development tests were carried out in no‐choice experiments by placing adult aphids on plants of each genotype and counting numbers of nymphs and adults on young plants after 8 and 15 days, and on flowering plants after 14 and 30 days. Large differences were observed among genotypes: some developed small colonies and others developed large ones. Also, in a few genotypes, resistance in mature plants was different for leaves of different ages; young leaves were resistant to aphids whereas old senescent leaves were susceptible. The electrical penetration graph (DC‐EPG system) technique was used to study aphid feeding behaviour on each Solanum genotype for 6 h. Electrical penetration graph (EPG) results also showed large differences among the genotypes, indicating resistance at the leaf surface and at three different levels of plant tissue (epidermis, mesophyll, and phloem). Therefore, it was concluded that different mechanisms of resistance to M. persicae exist among the genotypes analysed. EPGs recorded from aphids on Solanum berthaultii Hawkes and Solanum tarijense Hawkes with and without glandular trichomes showed that strong surface resistance can bias EPG parameters associated with resistance located in deeper tissues. Experimental evidence is presented that the resistance to aphids in the genotypes with glandular trichomes strongly depends on these morphological structures.     

Gustation of dietary amino acids by the aphid Myzus persicae

In simple choice-chamber tests the ability of the aphid Myzus persicae (Sulzer) to discriminate between sucrose solutions containing single or groups of amino acids and a pure sucrose solution was assessed. Each of 20 dietary amino acids was tested at a concentration of 0.1% in 20% sucrose. Each of six amino acids (asparagine, isoleucine, leucine, methionine, phenylalanine, and tryptophan) strongly enhanced the acceptability of sucrose. Methionine had a strong effect also at 0.02%. A few other amino acids had a lesser effect, while only a slight or no effect was observed with about half of the 20 dietary amino acids. A slightly deterrent effect was observed with histidine, glutamic acid, and arginine.The experimental apterous adult aphids that had been reared on radish seedlings had to be preconditioned on a pure sucrose solution for one day prior to the tests in order to show these responses within a few hours.

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Zusammenfassung In einfachen Wahlkammer-Versuchen wurde die Fähigkeit der Blattlaus Myzus persicae geprüft, Rohrzuckerlösungen, die einzelne oder Gruppen von Aminosäuren enthielten, von reiner Rohrzuckerlösung zu unterscheiden. Jede der 20 diätischen Aminosäuren wurde in einer Konzentration von 0,1% in 20% iger Rohrzuckerlösung geprüft. Jede der 6 Aminosäuren Asparagin, Isoleucin, Leucin, Methionin, Phenylalanin und Tryptophan erhöhte die Annehmbarkeit stark. Unter ihnen hatte Methionin auch bei 0,02% eine kräftige Wirkung. Einige wenige andere Aminosäuren hatten einen geringen Effekt, während bei etwa der Hälfte der 20 diätischen Aminosäuren nur ein schwacher oder kein Einfluß beobachtet wurde. Eine leicht abweisende Wirkung war bei Histidin, Glutaminsäure und Arginin festzustellen.Die im Versuch verwendeten erwachsenen apteren Blattläuse, die an Rettichsämlingen herangezogen worden waren, mußten vor den Versuchen einen Tag lang an reiner Rohrzuckerlösung gehalten werden, wenn sie diese Reaktionen innerhalb einiger weniger Stunden zeigen sollten.

     

Cytogenetic and molecular characterization of a highly repeated DNA sequence in the peach potato aphid Myzus persicae

Electrophoresis following digestion of Myzus persicae genomic DNA with HindIII showed the presence of a prominent band of approximately 200 bp whereas a faint electrophoretic band corresponding to DNA fragments of about 3000 bp was observed after digestion with ApaI. In situ digestion with restriction enzymes, followed by in situ nick translation, showed that ApaI targets are localized at the nucleolus organizer-bearing X telomeric region, whereas HindIII restriction sites are clustered in intercalary C-positive areas on the same X chromosome. Fluorescent in situ hybridization (FISH) carried out by using digoxygenin-labeled HindIII repeats as probe fully confirmed overlapping between the hybridization sites of this probe and the AT-rich intercalary heterochromatic bands on the X chromosome. These findings, together with published data, allow us to conclude that the M. persicae genome possesses three classes of C-positive heterochromatin: (i) a GC-rich argentophilic band located on one telomere of the X chromosome that contains ApaI targets; (ii) AT-rich intercalary bands located on the X chromosome containing clustered HindIII fragments; (iii) AT-rich telomeric bands, located on autosomes, consisting of HaeIII repeats. Molecular analysis has shown that the length of the HindIII repeat consensus sequence is 189 bp with an AT content of 67%. Southern blotting with HindIII monomers revealed a regular ladder of bands composed of multimers of basic length that are characteristic of satellite DNAs. The HindIII repeat displays other features typical of eukaryotic satellite arrays such as overlapping with heterochromatic bands and a high degree of sequence similarity among monomers (84%–94%). A similarity plot showed that sequences were particularly variable in the 50–100 bp region whereas they proved to be highly conservative in the first 50 bp, thus suggesting that this portion of the repeat might be functionally important. Received: 23 February 1999; in revised form: 21 July 1999 / Accepted: 28 July 1999     

Effect of latitude and acclimation on the lethal temperatures of the peach‐potato aphid Myzus persicae

• 1 Aphids, similar to all insects, are ectothermic and, consequently, are greatly affected by environmental conditions. The peach potato aphid Myzus persicae (Sulzer) has a global distribution, although it is not known whether populations display regional adaptations to distinct climatic zones along its distribution and vary in their ability to withstand and acclimate to temperature extremes. In the present study, lethal temperatures were measured in nine anholocyclic clones of M. persicae collected along a latitudinal cline of its European distribution from Sweden to Spain. The effects of collection origin and intra‐ and intergenerational acclimation on cold and heat tolerance, as determined by upper and lower lethal temperatures (ULT₅₀ and LLT₅₀, respectively), were investigated.
• 2 Lethal temperatures of M. persicae were shown to be plastic and could be altered after acclimation over just one generation. Lower lethal temperatures were significantly depressed in eight of nine clones after acclimation for one generation at 10°C (range: ?13.3 to ?16.2°C) and raised after acclimation at 25°C (range: ?10.7 to ?11.6°C) compared with constant 20°C (range: ?11.9 to ?12.9°C). Upper lethal temperatures were less plastic, although significantly increased after one generation at 25°C (range: 41.8–42.4°C) and in five of nine clones after acclimation at 10°C. There was no evidence of intergenerational acclimation over three generations.
• 3 Thermal tolerance ranges were expanded after acclimation at 10 and 25°C compared with constant 20°C, resulting in aphids reared at 10°C surviving over a temperature range that was approximately 2–6°C greater than those reared at 25°C.
• 4 There was no clear relationship between lethal temperatures and latitude. Large scale mixing of clones may occur across Europe, thus limiting local adaption in thermal tolerance. Clonal type, as identified by microsatellite analysis, did show a relationship with thermal tolerance, notably with Type O clones being the most thermal tolerant. Clonal types may respond independently to climate change, affecting the relative proportions of clones within populations, with consequent implications for biodiversity and agriculture.

     

Detection of potato virus Y in the aphid Myzus persicae by enzyme-linked immunosorbent assay (ELISA)

Potato virus Y was detected by enzyme linked immunosorbent assay (ELISA) in at least 50% of groups of five Myzus persicae. The mean A₄₀₅ value for groups of viruliferous aphids was 2–3 times greater than that for virus-free ones. PVY was not detected in Aphis craccivora, A. citricola or A. gossypii, three other species which transmitted the virus to peppers, and it was detected in only a small proportion of groups of Acyrthosiphon pisum. In a series of trials, success in detection of PVY by ELISA was not correlated with the ability of other aphids from the same source plant to transmit the virus to test plants. The limitation of ELISA for quantitative assay of PVY in aphids and for epidemiological work is discussed.     

Effects of potato plants expressing the nptII-gus fusion marker genes on reproduction, longevity, and host-finding of the peach-potato aphid, Myzus persicae

Transgenesis developed in the last 20 years offers new possibilities for crop protection. The transgenic process, however, requires the use of marker fusion genes to select and visualize the transformed tissues. Although the expression products of these marker genes are stably expressed in crops, little attention has been given to assess the eventual risks of these recombinant proteins on phytophage populations. Three independent transgenic potato (Solanum tuberosum) clones from the cultivar Désirée (DG5, DG18, and DG20) carrying the commonly used nptII‐gus gene construct and exhibiting different β‐glucuronidase activity (0.843 ± 0.011, 0.576 ± 0.096, and 0.002 ± 0.000 pmol min^?1.mg^?1, respectively) were evaluated to determine the impact of the encoded proteins on the behaviour, development, reproduction, and demography of the peach‐potato aphid, Myzus persicae, under laboratory‐controlled light and temperature. Our results revealed that the transgenic event can alter aphid physiology or behaviour. Experiments showed a probiotic effect of one transgenic line, the DG5, resulting in reduced prereproductive period and mortality, and enhanced daily fecundity, which was expressed in a greater population growth potential (r_m = 0.205 vs. r_m = 0.174 of the control). In contrast, aphids fed with the DG18 line exhibited reduced adult survival and reproductive period but no alteration of their demographic parameters (r_m = 0.176). Finally, no physiological alteration was induced in aphids fed on a DG20 diet (r_m = 0.170). Behavioural experiments conducted in a 4‐choice olfactometer demonstrated that insects were significantly more attracted by the odour of transgenic DG18 potato plant than that of Désirée non‐transformed plant, spending twice as much time in the DG18 plant odour. The two other transformed clones (DG5 and DG20) were as attractive as the non‐transformed cultivar. It is concluded that the β‐glucuronidase expression in potato plants might be responsible for the probiotic effect measured on the feeding aphids, whereas alteration of the foliage odour would result from a pleiotropic effect.     

Factors affecting the cold hardiness of the peach-potato aphid Myzus persicae

Supercooling point studies were used to investigate the factors influencing the cold hardiness of the peach-potato aphid Myzus persicae, a freezing-susceptible insect. Overwintering adults lost cold hardiness as winter progressed, with a variable proportion showing a marked reduction in supercooling ability. Cold hardiness increased in spring so that all individuals demonstrated extensive supercooling ability typical of aphids reared in the laboratory at 20°C with a long photoperiod; these levels of cold hardiness were maintained in the field during summer and early autumn. First instar nymphs demonstrated considerable cold hardiness all year. Surface moisture caused inoculative freezing in some first instar nymphs and adults when supercooled, but the majority were unaffected. In the laboratory, adults starved for 7 days at 5°C showed distinct losses of supercooling potential equivalent to those observed in the field during mid to late winter. No loss of cold hardiness was found in first instar nymphs starved under the same conditions. The results demonstrate that the cold hardiness characteristics of M. persicae are atypical of those observed in other freezing-susceptible insects and it is suggested that continued feeding during mild winter conditions allows maintenance of cold hardiness particularly in adult aphids, and provides a possible explanation for the successful anholocyclic overwintering of M. persicae during such winters.     

Redox responses of Arabidopsis thaliana to the green peach aphid,Myzus persicae

The green peach aphid (Myzus persicae) is a phloem-feeding insect that causes economic damage on a wide array of crops. Using a luminol-based assay, a superoxide-responsive reporter gene (Zat12::luciferase), and a probe specific to hydrogen peroxide (HyPer), we demonstrated that this aphid induces accumulation of reactive oxygen species (ROS) in Arabidopsis thaliana. Similar to the apoplastic oxidative burst induced by pathogens, this response to aphids was rapid and transient, with two peaks occurring within 1 and 4 hr after infestation. Aphid infestation also induced an oxidative response in the cytosol and peroxisomes, as measured using a redox-sensitive variant of green fluorescent protein (roGFP2). This intracellular response began within minutes of infestation but persisted 20 hr or more after inoculation, and the response of the peroxisomes appeared stronger than the response in the cytosol. Our results suggest that the oxidative response to aphids involves both apoplastic and intracellular sources of ROS, including ROS generation in the peroxisomes, and these different sources of ROS may potentially differ in their impacts on host suitability for aphids.     

Glutathione S-transferases in the adaptation to plant secondary metabolites in the Myzus persicae aphid

Glutathione S-transferases (GST) in insects play an important role in the detoxification of many substances including allelochemicals from plants. Induction of GST activity in Myzus persicae in response to secondary metabolites from Brassica plants was determined using different host plant species and confirmed using artificial diet with pure allelochemicals added. The 2,4-dinitro-1-iodobenzene (DNIB) was found to be a useful substrate for identifying particular GSTs in insects. GSTs from M. persicae were purified using different affinity chromatography columns and related kinetic parameters were calculated. GST isoenzymes were characterised using electrophoretic methods. Although SDS-PAGE results indicated similarity among the purified enzymes from each affinity column, biochemical studies indicated significant differences in kinetic parameters. Finally, the GST pattern of M. persicae was discussed in terms of insect adaptation to the presence of plant secondary substances such as the glucosinolates and the isothiocyanates, from Brassicaceae host plants.     

Rapid method to screen resistance of potato plants against Myzus persicae (Homoptera: Aphididae) in the laboratory

With the objective to develop a potato, Solanum tuberosum L., resistance program against aphids, we propose a rapid screening method with Myzus persicae (Sulzer) in the laboratory. We aimed to optimize the duration of the whole procedure and to decrease the frequency of measurements. In a first experiment, intrinsic rate of natural increase (r(m)) values were compared between adult aphids reared throughout their entire life and adults reared only during a period equivalent to their prereproductive period. No significant differences were observed. In a second experiment, four groups of aphids were distinguished according to the sampling frequency, i.e., those whose biological parameters were evaluated every single, second, third, and fourth day. Except for the fourth-day experiment, the r(m) values estimated on aphids reared on the three potato lines were not significantly different whatever sampling frequency of single, second, or third day used to check aphids. Thus, screening efforts in laboratory can be largely optimized by evaluating adult aphids only during a period equivalent to their prereproductive period and assessing M. persicae populations every third day. Our method is reliable and adapted to screen a large number of potato plants against M. persicae because it allows an average 70% reduction in the time required for the whole experimental process.     

Concanavalin A inhibits development of tomato moth (Lacanobia oleracea) and peach-potato aphid (Myzus persicae) when expressed in transgenic potato plants

The effects of concanavalin A (ConA), a glucose/mannose-specific lectin from jackbean (Canavalia ensiformis), on insect crop pests from two different orders, Lepidoptera and Homoptera, were investigated. When fed to larvae of tomato moth (Lacanobia oleracea) at a range of concentrations (0.02–2.0% of total protein) in artificial diet, ConA decreased survival, with up to 90% mortality observed at the highest dose level, and retarded development, but had only a small effect on larval weight. When fed to peach-potato aphids (Myzus persicae) at a range of concentrations (1–9μM) in liquid artificial diet, ConA reduced aphid size by up to 30%, retarded development to maturity, and reduced fecundity (production of offspring) by >35%, but had little effect on survival. With both insects, there was a poor correlation between lectin dose and the quantitative effect. Constitutive expression of ConA in transgenic potatoes driven by the CaMV 35S promoter resulted in the protein accumulating to levels lower than predicted, possibly due to potato not being able to adequately reproduce the post-translational processing of this lectin which occurs in jackbean. However, the expressed lectin was functionally active as a haemagglutinin. Bioassay of L. oleracea larvae on ConA-expressing potato plants showed that the lectin retarded larval development, and decreased larval weights by >45%, but had no significant effect on survival. It also decreased consumption of plant tissue by the larvae. In agreement with the diet bioassay results, ConA-expressing potatoes decreased the fecundity of M. persicae by up to 45%. ConA thus has potential as a protective agent against insect pests in transgenic crops. This revised version was published online in June 2006 with corrections to the Cover Date.     

The evolution of insecticide resistance in the peach-potato aphid,Myzus persicae

The peach-potato aphid Myzus persicae (Sulzer) can resist a wide range of insecticides, but until recently (1990) the only mechanism identified was the increased production of carboxylesterases (E4 or FE4), which cause enhanced degradation and sequestration of insecticidal esters. We have now identified two forms of target-site resistance involving changes in the acetylcholinesterase (AChE) and sodium channel (kdr) genes. Biochemical and DNA diagnostic methods can be used to identify all three mechanisms in individual aphids, and thereby establish their spatial distributions and temporal dynamics. Amplified genes underlie the increased production of esterases but their expression is modulated by DNA methylation. Amplification of the E4 gene is in strong linkage disequilibrium with the kdr mechanism. This may reflect strong insecticidal selection favouring aphids with multiple mechanisms, tight chromosomal linkage and/or the prominence of parthenogenesis in many M. persicae populations. The decreased fitness of resistant aphids under winter conditions may be a consequence of the altered sodium-channel gene affecting behaviour and/or the perception of external stimuli.     

An amino acid substitution on the second acetylcholinesterase in the pirimicarb-resistant strains of the peach potato aphid,Myzus persicae

cDNAs encoding two acetylcholinesterases (AChEs) were isolated from the peach potato aphid, Myzus persicae. MpAChE1 was orthologous and MpAChE2 was paralogous with the ace of Drosophila melanogaster. The deduced amino acid sequence of MpAChE1 cDNA was identical between the pirimicarb susceptible and resistant strains. However, a single amino acid substitution of Ser431Phe on MpAchE2 was found in the pirimicarb resistant strains. This substitution was located in the acyl pocket of the enzyme and was thought to alter the ligand specificity.     

Characterization of a new densovirus infecting the green peach aphid Myzus persicae

A new icosahedral DNA virus was isolated from aphids (Myzus persicae) that showed abnormal growth and development. The purified virus particles have a diameter of 20 nm and contain a single-stranded DNA molecule of approximately 5.7 kb. The viral particles are composed of five structural proteins (92, 85, 68, 64, and 57 kDa). As the main biophysical properties of this virus are similar to those of the members of the genus Densovirus it was tentatively named Myzus persicae densovirus (MpDNV). A PCR-based detection method and a polyclonal antiserum raised against MpDNV allowed the detection of the virus in a single-infected aphid. MpDNV is immunologically related to Junonia coenia densovirus, but not to other members of the subfamily Densovirinae. Biological assays showed that MpDNV could be both transmitted transovarially and horizontally via honeydew and saliva. MpDNV was able to infect whiteflies but not other aphid species tested.