Population differentiation and genetic variation in host choice among pea aphids from eight host plant genera

Habitat choice plays a critical role in the processes of host range evolution, specialization, and ecological speciation. Pea aphid, Acyrthosiphon pisum, populations from alfalfa and red clover in eastern North America are known to be genetically differentiated and show genetic preferences for the appropriate host plant. This species feeds on many more hosts, and here we report a study of the genetic variation in host plant preference within and between pea aphid populations collected from eight genera of host plants in southeastern England. Most host-associated populations show a strong, genetically based preference for the host plant from which they were collected. Only in one case (populations from Vicia and Trifolium) was there little difference in the plant preference spectrum between populations. All populations showed a significant secondary preference for the plant on which all the aphid lines were reared: broad bean, Vicia faba, previously suggested to be a "universal host" for pea aphids. Of the total genetic variance in host preference within our sample, 61% could be attributed to preference for the collection host plant and a further 9% to systematic differences in secondary preferences with the residual representing within-population genetic variation between clones. We discuss how a combination of host plant preference and mating on the host plant may promote local adaptation and possibly ecological speciation, and whether a widely accepted host could oppose speciation by mediating gene flow between different populations.     

Resistance to a fungal pathogen and host plant specialization in the pea aphid

Herbivores that show host race formation on different plant species have proven to be valuable model systems for studying the evolution of specialization and speciation. Here, we use the pea aphid, Acyrthosiphon pisum, to investigate a possible link between specialization on two host plant species, Lotus uliginosus and Trifolium pratense, and resistance to a natural enemy, the fungal pathogen Erynia neoaphidis. Pea aphids collected on either plant species in the field showed in most cases poor survival on the alternate host plant. Furthermore, pea aphids specialized on T. pratense were very resistant to E. neoaphidis, whereas aphids specialized on L. uliginosus were susceptible. This susceptibility was not influenced by the actual food plant on which the assays were conducted. We discuss how selection from natural enemies may influence the process of specialization and race formation, and how specialization can affect the evolution of resistance.     

Functional divergence of three glutathione transferases in two biotypes of the English grain aphid,Sitobion avenae

The English grain aphid, Sitobion avenae (Fabricius) (Hemiptera: Aphididae), is a significant pest of cereal crops, but molecular factors and mechanisms that underpin its ability to develop differential biotypes on variable host plants are still not well understood. In this study, we investigated the interactions between two plant secondary metabolites (i.e., gramine and gallic acid) and three glutathione S-transferases (GSTs) of S. avenae. Using artificial diets complemented or not with one of these two plant compounds, we found that gramine had relatively stronger negative effects on fitness of S. avenae biotype 3 (adapted to barley), and gallic acid on that of biotype 4 (adapted to wheat). Gramine significantly induced overexpression of SaveGST1 and SaveGST2 in biotype 4, but not in biotype 3. Gramine also reduced SaveGST3 expression in biotype 4, but not in biotype 3, suggesting biotype-specific effect of GSTs’ regulation. In the treatments with gallic acid, the overexpression of SaveGST1, but not SaveGST2 or SaveGST3, was significantly induced in both biotypes, suggesting a critical role of SaveGST1 in detoxification of phenolic compounds such as gallic acid. The total constitutive GST activity was much higher in biotype 4 than in biotype 3. Significant increase in GST activity was obtained by the addition of both secondary metabolites in biotype 4, but not in biotype 3, showing significantly higher expression plasticity of GSTs in biotype 4. Thus, both constitutive and induced expression of GSTs could affect the adaptability of S. avenae on plants with variable secondary compounds, and thus contribute to the divergence of biotypes in this aphid species.     

Cold hardiness and overwintering of the grain aphid Sitobion avenae

ABSTRACT.

• 1 Cold hardiness as measured by supercooling ability in the active stages of the grain aphid Sitobion avenae (F.) decreased progressively with maturation.
• 2 Aphids showed no acclimation response when maintained at low temperatures.
• 3 Starvation did not improve supercooling ability.
• 4 In a single exposure, surface moisture caused inoculation above the inherent supercooling point in a small proportion of a population.
• 5 Field populations show a seasonal change in supercooling ability, which is at a maximum in summer and a minimum in late winter.
• 6 It is concluded that the act of feeding on healthy plant tissue may confer extensive supercooling ability.

     

Population differentiation and genetic variation in performance on eight hosts in the pea aphid complex

Phytophagous insects frequently use multiple host-plant species leading to the evolution of specialized host-adapted populations and sometimes eventually to speciation. Some insects are confronted with a large number of host-plant species, which may provide complex routes of gene flow between host-adapted populations. The pea aphid (Acyrthosiphon pisum) attacks a broad range of plants in the Fabaceae and it is known that populations on Trifolium pratense and Medicago sativa can be highly specialized at exploiting these species. To find out whether adaptation to a broad range of co-occurring hosts has occurred, we tested the performance of pea aphid clones collected from eight host-plant genera on all of these plants in a reciprocal transfer experiment. We provide evidence for pervasive host-plant specialization. The high performance of all aphid clones on Vicia faba suggests that this host plant could be a site of gene flow between different populations that could limit further host-associated divergence. The genetic variance in host-plant usage was partitioned into within- and among-population components, which represent different levels of host adaptation. Little evidence of within-population trade-offs in performance on different plant species was found.     

Ecological specialization correlates with genotypic differentiation in sympatric host-populations of the pea aphid

The pea aphid, Acyrthosiphon pisum, encompasses distinct host races specialized on various Fabaceae species, but the extent of genetic divergence associated with ecological specialization varies greatly depending on plant and geographic origins of aphid populations. Here, we studied the genetic structure of French sympatric pea aphid populations collected on perennial (pea and faba bean) and annual (alfalfa and red clover) hosts using 14 microsatellite loci. Classical and Bayesian population genetics analyses consistently identified genetic clusters mostly related to plant origin: the pea/faba bean cluster was highly divergent from the red clover and the alfalfa ones, indicating they represent different stages along the continuum of genetic differentiation. Some genotypes were assigned to a cluster differing from the one expected from their plant origin while others exhibited intermediate genetic characteristics. These results suggest incomplete barriers to gene flow. However, this limited gene flow seems insufficient to prevent ecological specialization and genetic differentiation in sympatry.     

Genome scans reveal candidate regions involved in the adaptation to host plant in the pea aphid complex

A major goal in evolutionary biology is to uncover the genetic basis of adaptation. Divergent selection exerted on ecological traits may result in adaptive population differentiation and reproductive isolation and affect differentially the level of genetic divergence along the genome. Genome‐wide scan of large sets of individuals from multiple populations is a powerful approach to identify loci or genomic regions under ecologically divergent selection. Here, we focused on the pea aphid, a species complex of divergent host races, to explore the organization of the genomic divergence associated with host plant adaptation and ecological speciation. We analysed 390 microsatellite markers located at variable distances from predicted genes in replicate samples of sympatric populations of the pea aphid collected on alfalfa, red clover and pea, which correspond to three common host‐adapted races reported in this species complex. Using a method that accounts for the hierarchical structure of our data set, we found a set of 11 outlier loci that show higher genetic differentiation between host races than expected under the null hypothesis of neutral evolution. Two of the outliers are close to olfactory receptor genes and three other nearby genes encoding salivary proteins. The remaining outliers are located in regions with genes of unknown functions, or which functions are unlikely to be involved in interactions with the host plant. This study reveals genetic signatures of divergent selection across the genome and provides an inventory of candidate genes responsible for plant specialization in the pea aphid, thereby setting the stage for future functional studies.     

Back to the future: genetic correlations, adaptation and speciation

Genetic correlations can affect the course of phenotypic evolution. Although genetic correlations among traits are a common feature of quantitative genetic analyses, they have played a very minor role in recent linkage-map based analyses of the genetic architecture of quantitative traits. Here, we use our work on host-associated races in pea aphids to illustrate how quantitative trait locus (QTL) mapping can be used to test specific hypotheses about how genetic correlations may facilitate ecological specialization and speciation.     

Life-cycle variation in the aphid Sitobion avenae: costs and benefits of male production

Abstract 1. Patterns of male production and life-cycle variation were investigated in the aphid Sitobion avenae . Seventy-seven field-collected clones were subjected to a 14 °C short day-length regime to assess their reproductive modes, and three clones were used to study life-cycle inheritance. A further analysis assessed the cost of male production.
2. In comparison with those from East Anglia, there were more holocyclic and intermediate clones found in Scottish populations, and they produced significantly more mating females. In total, 44% of clones were androcyclic.
3. The inheritance of life cycle showed a greater level of complexity than could be achieved by previously suggested monohybrid inheritance mechanisms.
4. Holocyclic and intermediate clones produced a higher proportion of males than did androcyclic clones. Inter-clonal differences were reflected in the pattern of male production in the reproductive sequence.
5. A significant cost was associated with male production in terms of a reduction in both fecundity and total offspring biomass.
6. Winged females rarely gave birth to males but the pattern of mating female production differed between holocyclic and intermediate clones.
7. Patterns of male and mating female production by the different types of clone may be related to different advantages and disadvantages of dispersal and inbreeding.
8. Recent models of aphid overwintering could be enhanced by consideration of issues raised in this study, such as the cost of male production, the inheritance of life cycle, and the patterns of sexual morph production.     

Sympatric genetic differentiation of a generalist pathogenic fungus, Botrytis cinerea, on two different host plants, grapevine and bramble

Prime candidates for sympatric ecological divergence include parasites that differentiate via host shifts, because different host species exert strong disruptive selection and because both hosts and parasites are continually co-evolving. Sympatric divergence may be fostered even more strongly in phytopathogenic fungi, in particular those where sex must occur on the host, which allows adaptation alone to restrict gene flow between populations developing on different hosts. We sampled populations of Botrytis cinerea, a generalist ascomycete fungus, on sympatric grapes and brambles in six regions in France. Microsatellite data were analyzed using standard population genetics, a population graph analysis and a Bayesian approach. In addition to confirming that B. cinerea reproduces sexually, our results showed that the fungal populations on the two hosts were significantly differentiated, indicating restricted gene flow, even in sympatry. In contrast, only weak geographical differentiation could be detected. These results support the possibility of sympatric divergence associated with host use in generalist parasites.     

麦长管蚜和禾谷缢管蚜对小麦挥发物的触角电位反应

采用活体蚜虫测定法,利用EAG技术比较分析了麦长管蚜Sitobion avenae和禾谷缢管蚜Rhopalosiphum padi有翅及无翅成蚜对小麦挥发物及麦蚜取食诱导挥发物组分的嗅觉反应,揭示了两种麦蚜的嗅觉变异特点。结果表明：麦长管蚜对水杨酸甲酯、反-2-己烯醛、反-2-己烯醇、-6-甲基-5-庚烯-2-酮和-6-甲基-5-庚烯-2醇的反应较强,禾谷缢管蚜对水杨酸甲酯、反-3-己烯乙酸酯、-6-甲基-5-庚烯-2-酮和-6-甲基-5-庚烯-2-醇的反应较强,并得到了剂量反应曲线。麦长管蚜的有翅和无翅成蚜对6-甲基-5-庚烯-2-酮、反-2-己烯醇和水杨酸甲酯的反应差异显著;禾谷缢管蚜的有翅和无翅成蚜对反-2-己烯醇、辛醛、里那醇、水杨酸甲酯和反-3-己烯乙酸酯的EAG反应差异显著,其原因与禾谷缢管蚜迁移及转主为害的生物学特性有关。     

Biochemical basis of winter wheat resistance to the grain aphid, Sitobion avenae

Six winter wheat cultivars with differing degrees of resistance to the grain aphid, Sitobion avenae (F.), were studied under field conditions. Resistance was measured in terms of non-preference and antibiosis on plants at seven growth stages. The varieties Saga and Grana were most resistant in terms of both non-preference and antibiosis to S. avenae at all growth stages examined. The varieties Liwilla and Dana were relatively susceptible to aphid attack. The number of aphids was directly proportional to the total content of free and essential amino acids. The level of resistance showed a similar, but not identical, relationship with the observed concentration of soluble proteins. A higher degree of cultivar resistance was associated with a higher value on a toxicity index, which is the ratio between the free-phenols and free-amino acids content. Obtained results suggest that the resistance of winter wheat cultivars to the grain aphid was based mainly on the mechanism of antibiosis.

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Résumé Six cultivars de blé d'hiver, présentant différents degrés de résistance au puceron des grains, S. avenae, ont été étudiés dans la nature. La résistance a été évaluée en fonction de l'antibiose et de l'absence de préférence de plantes à 7 stades de développement. Les variétés Saga et Grana ont été les plus résistantes, suivant ces deux critères, pour tous les stades de puceron étudiés. Les variétés Liwilla et Dana ont été relativement sensibles. Les effectifs de pucerons étaient proportionnels à la teneur totale en acides aminés libres essentiels. Les concentrations en protéines solubles présentent une relation de même type, mais pas identique, avec le niveau de résistance. Un haut degré de résistance a été associé à une valeur élevée de l'indice de toxicité, rapport entre les phénols libres et la teneur en acides aminés libres. Ces résultats suggèrent une résistance des cultivars de blé d'hiver à S. avenae liée principalement aux mécanismes d'antibiose.

     

Methods of hatching the eggs and rearing the fundatrices of the English grain aphid, Sitobion avenae

Several methods for hatching the eggs and rearing individuals of the first generation (fundatrices) of Sitobion avenae were investigated. The most successful methods were incubation of the eggs on grass seedlings at 2°C and rearing the fundatrices on grass seedlings (overall survival 66%) and incubation of the eggs in plastic boxes at 2°C and rearing the fundatrices on wheat seedlings (overall survival 62%).

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Résumé L'éclosion des oeufs de S. avenae peut être induite par le transfert à 10°C ou 12°C, après une incubation de 75–120 jours à 2°C. Le pourcentage le plus élevé d'éclosions a été obtenu quand les oeufs avaient incubé pendant 100 à 110 jours à 2°C (67% at 71.5% respectivement) dans des petites boîtes de plastique, ou pendant 100 jours à 2°C sur des pousses de graminées (73.5%). Si les oeufs sont pondus sur blé, la plante ne peut pas tolérer la période d'incubation, mais cet obstacle peut être surmonté en obligeant les ovipares à pondre leurs oeufs sur de pousses de graminées, comme Poa annua, hôte convenable pour les fondatrices. Les ovipares peuvent aussi pondre sans difficultés sur autre chose que des végétaux, et des récipients peuvent ètre mis à incuber sans contenir du matériel végétal.

     

Salivation into sieve elements in relation to plant chemistry: the case of the aphid Sitobion fragariae and the wheat, Triticum aestivum

Extended sieve element salivation (E1 waveform in the electrical penetration graph) is a characteristic activity during early sieve element punctures, particularly in resistant plants. In order to explore a chemically-mediated mechanism of resistance associated with sieve element salivation, we compared the pattern of feeding behaviour of the aphid, Sitobion fragariae (Walker), on two cultivars of the wheat Triticum aestivum L., with different concentrations of hydroxamic acids (Hx). During 24 h of electronic monitoring, aphids dedicated over 50% of the total time to phloem ingestion from the sieve elements. Total time allocated to E1 in the experiment, time to first E1 within the experiment, time allocated to E1 before a sustained phloem ingestion (E2) and the contribution of sieve element salivation to the phloem phase (E1/[E1+E2]) were significantly higher in the high-Hx cultivar. The increased salivation in plants with higher contents of Hx suggests the existence, at least in this system, of a chemically-mediated sieve element constraint.     

棉花型和黄瓜型棉蚜(Aphis gossypii Glover)的寄主适应性及转移通道

采用寄主转接建立生命表的方法研究了棉花型和黄瓜型棉蚜对不同寄主植物的适应性,以及两寄主型棉蚜是否可通过中间桥梁寄主实现寄主互换的问题。结果表明,两寄主型棉蚜直接互换寄主后,其存活和繁殖力显著下降,表现为棉花型和黄瓜型棉蚜的净增殖率比在原寄主上分别下降980倍和12倍,平均世代寿命缩短5～12d。两寄主型棉蚜均能利用木槿植物,并且适应性没有显著差异。但是两寄主型棉蚜均不能在车前草和大叶黄杨上存活和繁殖后代。西葫芦作物对棉蚜在木槿、棉花和黄瓜寄主上的相互转移起到了重要的桥梁寄主作用。冬寄主木槿上棉蚜可通过甜瓜或西葫芦转移到黄瓜寄主上,棉花和黄瓜上棉蚜也可通西葫芦作物分别转移到黄瓜和棉花作物上,从而形成棉蚜在不同寄主植物间的相互转移通道,造成为害和病毒病的扩张。     

Effects of extreme,fluctuating temperature events on life history traits of the grain aphid,Sitobion avenae

Altered temperatures affect insects’ life history traits, such as development period and fecundity, which ultimately determine population growth rates. Understanding insects’ thermal biology is therefore integral to population forecasting and pest management decision‐making such as when to utilise crop spraying or biological control. Aphids are important crop pests in temperate regions, causing considerable yield losses. The aphid thermal‐biology literature is, however, heavily biased towards the effects of rising mean temperatures, whereas the effects of fluctuating, extreme climatic events (e.g., heat waves and sub‐zero cold periods) are largely overlooked. This study assessed the effects of laboratory‐simulated heat waves and sub‐zero cold periods on the survival, development period, and fecundity of the grain aphid, Sitobion avenae (Fabricius) (Hemiptera: Aphididae: Microsiphini), in addition to assessing maternal effects on the birth weight and development period of the offspring of exposed individuals. Exposure to heat stress periods (total of 16 h at 30 °C) significantly reduced aphid fecundity and increased physiological development period (in day‐degrees) resulting in a reduced population growth rate. Cold exposure (total of 1.33 h at ?15 °C) reduced population growth rate due to an elongated development period (in days), but did not affect fecundity or physiological development period (in day‐degrees). Both cold and heat stress significantly reduced aphid survival. Maternal experience of heat stress reduced nymphal birth weight although nymphal development period was not affected by either cold or heat stress. The results suggest that including the effects of fluctuating, extreme temperature events on aphid life history in population forecast models is likely to be of great importance to pest management decision‐making. The demonstration of maternal effects on birth weight also suggests that cross‐generational effects of heat waves on population growth rates could occur.     

Extremely divergent host plant acceptance with gene flow between sympatric host races of the phytophagous ladybird beetle Henosepilachna diekei

Ecological divergence alone can prevent the majority of gene flow in the absence of other forms of reproductive isolation. Although the importance of ecological divergence in promoting reproductive isolation has been broadly recognized, its net impact on speciation has rarely been estimated in the wild. The phytophagous ladybird beetle Henosepilachna diekei Jadwiszczak & Wegrzynowicz includes two sympatric host races that are reproductively isolated solely by extreme specialization to either of the host plants Mikania micrantha Kunth (Asteraceae) or Leucas lavandulifolia Sm. (Lamiaceae) in West Java, Indonesia. To investigate the impact of differential host use as an isolating barrier, we carried out adult host acceptance tests and molecular population genetic analyses based on mitochondrial ND2 and nucleic ITS2 gene sequences using 13 wild populations of the host races, including four sympatric population pairs. Almost all individuals of these host races persistently accepted only the original host plant. We detected restricted but a degree of gene flow between these host races. A migration event occurred only in very recent time compared to their divergence time, indicating recent secondary contact of these host races in the surveyed area. These results reveal the remarkably large impact of host‐plant shift over almost the entire process of speciation and illustrate that ecological divergence has been maintained even under the presence of a certain degree of gene flow.