Life cycle of the cereal aphid Sitobion avenae F.: polymorphism and comparison of life history traits associated with sexuality

1. Clones of Sitobion avenae from the west of France exhibited four types of responses in growth chambers to short days and low temperatures: in addition to permanently parthenogenetic clones (anholocyclic) and clones producing both males and parthenogenetic females (androcyclic), lineages producing both mating females and males were categorized into holocyclic and intermediate types, the latter investing into both sexual and parthenogenetic reproduction.
2. Holocyclic, intermediate and androcyclic clones were tested outdoors for sexual production in autumn and results were consistent with those obtained in growth chambers. Eggs from holocyclic and intermediate clone selfings were laid and hatched for most crosses.
3. Reciprocal crosses within and between life cycle variants showed no effect of the type of cross on the number of eggs laid per mating female, their hatching success, or the survival and fecundity of the parthenogenetic females born from eggs (fundatrices).
4. Eggs from intermediate × intermediate crosses hatched 20 days earlier than those involving at least one holocyclic parent. It is suggested that this short diapausing time gives a selective advantage to intermediate clones in oceanic climates, which could compensate for their lower sexual production in autumn.
5. Selfed offspring showed inbreeding depression, which seems to be common in non host-alternating aphids.
6. Mating with androcyclic males affects percentage (lower) and time (longer) of egg hatching, but not the other investigated traits.
7. The significance of intragenotypic strategy mixing in androcyclic and intermediate clones is discussed as an overall adaptation to the uncertainty of winter climate in oceanic zones.     

The genetics of obligate parthenogenesis in an aphid species and its consequences for the maintenance of alternative reproductive modes

Although loss of sex is widespread among metazoans, the genetic mechanisms underlying the transition to asexuality are poorly understood. Aphids are good models to address this issue because they frequently show reproductive-mode variation at the species level, involving cyclical parthenogens (CP) that reproduce sexually once a year and obligate parthenogens (OP) that reproduce asexually all year round. Here, we explore the genetic basis of OP in the cereal aphid Sitobion avenae by crossing several genotypes with contrasting reproductive modes and then characterising the reproductive phenotypes of F1 and F2 offspring. The analysis of phenotypic variation in F1 and F2 progenies suggests that at least two autosomal loci control OP in S. avenae. First, the transition to asexuality seems to depend on a single recessive locus, because the offspring from self-crossed cyclical parthenogenetic genotypes contain either 0 or 25% OP. Second, as we observed OP in the F1 progenies from crosses between CP and OP, and some CP in the offspring from outcrossed OP, a dominant ‘suppressor'' gene may also be involved, being inactive when in a recessive homozygous state in CP; this is the most parsimonious explanation for these results. This oligogenic inheritance of OP in S. avenae appears to be an efficient genetic system to generate new OP genotypes continually. It also allows asexuality-inducing alleles to be protected locally during harsh winters when extreme frost kills most OP, and then to spread very quickly after winter.     

Differences in egg hatching time between cyclical and obligate parthenogenetic lineages of aphids

Many aphid species exhibit a variation in reproductive mode which is influenced by winter climate regimes,with cyclical parthenogenetic (CP)lines dominating in cold winter areas (because they produce cold-resistant eggs)and obligate parthenogenetic (OP) ones in mild winter regions (because of their parthenogenetic overwintering).Genetic studies on several aphid species have shown that the OP trait can be transmitted during sexual events involving the 2 types of lines.This genetic system could be considered as a local safeguarding mechanism for OP alleles in case severe frost would have killed all parthenogenetically overwintering individuals.However,this strategy would only be efficient in restoring local polymorphism in breeding systems if the newly hatched OP recombinants remain competitive over their CP counterparts.In this study we compared egg hatching sequences of CP and OP F1 clones from several crosses obtained for 2 cereal aphid species,Sitobion avenae (constant 5℃,8 h of light)and Rhopalosiphum padi (winter outdoor conditions).For S.avenae,we obtained F1 offspring from 6 crosses, involving 4 clones while in R.padi F1 were obtained from 11 crosses involving 14 clones. We showed that in both species proportions of OP clones were higher in the first half of the progeny relative to the second half.In addition,F1 OP clones hatched in the mean about a week earlier than their CP sibs,which gives them a demographic advantage at the start of the growth season.We then discussed the consequences of this fitness advantage for the maintenance and spread of the OP trait in aphid populations.     

Inheritance of photoperiodic response in the bird cherry aphid, Rhopalosiphum padi

Abstract. The photoperiodic induction of presexual females (gynoparae) and males of the aphid Rhopalosiphum padi (L.) was examined in single clones collected from different latitudes (N clone from Blair Atholl, 56.5N 3.1W; C clone from Leeds, 53.5N 1.4W and S clone from Exeter, 50.4N 3.3W). Critical night lengths (CNLs) for gynopara and male production (CNLs: 50% of maximum production) and inter-clonal differences, defined by the different forms of progeny produced at a single discriminating night length (12 h 30 min), were measured at 16 ± 1C. The aphid clones differed in CNL and in the morphs produced at the discriminating night length. These differences were used to investigate the inheritance of the photoperiodic response. In crosses between clones the photoperiodic response showed considerable overall variation. The variation in CNLs indicated that the photoperiodic response was unlikely to affect the overwintering success after long-distance movement of clones from one latitude to another.     

Variation in the sexual behaviour of Achnanthes longipes (Bacillariophyta). III. Progeny of crosses between monoecious and unisexual clones

The allogamous raphid diatom Achnanthes longipes C. A. Agardh possesses a complex breeding system involving interactions between three types of clone: monoecious, unisexual and bisexual. Previous studies showed that these three types can be crossed with each other, with a tendency for sexual characteristics to be inherited: inbred monoecious lineages gave rise to monoecious or, very rarely, to bisexual clones, while inbred unisexual lineages yielded unisexual and bisexual clones. The current paper reports on the progeny of crosses between monoecious and unisexual clones and their inbred offspring. All three types of clone appeared in the F₁ and F₂, although unisexual clones of opposite sex to the parental clone were not found. Inbreeding depression was observed and also a tendency for ‘normal’ auxosporulation (producing two auxospores per pair of gametangia) to be replaced by ‘reduced’ or ‘intermediate’ auxosporulation (producing one auxospore per pair). In addition, patterns of incompatibility were observed that were not seen during earlier studies of clones isolated directly from nature. These included the inability of some F₁ clones to mate with each other, in spite of compatibility with all other clones examined (unisexual, bisexual and monoecious).     

Spatial and temporal dynamics of Myzus persicae clones in fields and suction traps

1 The population of peach‐potato aphid Myzus persicae in Scotland is comprised almost entirely of long‐term asexual clones. 2 Over a ten year period, M. persicae from Scottish fields and suction traps were analysed with six microsatellite markers. 3 Out of 1497 individuals analysed, 14 clones (denoted A–N) comprised over 98% of the collection. 4 Some clones were particularly abundant but most clones had a widespread distribution on all available plants. 5 Clones E and L had distinct features in their distributions as clone L was geographically totally restricted to the north east of Scotland and clone E showed a marked preference for brassica crops. 6 Clones E and L provide direct evidence of a role for local adaptations in the distribution of M. persicae clones.     

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.     

The influence of genetic background on resistance to the cabbage aphid (Brevicoryne brassicae) in Kale (Brassica oleracea var. acephala)

Resistance to Brevicoryne brassicae has been identified in the progeny of two selected kale (B. oleracea var. acephala) plants, one from the F1 hybrid cultivar ‘Arsis RS’ and one from the landrace ‘Butzo’. These plants were crossed with susceptible B. oleracea morphotypes that have different periods to flowering. The type of susceptible plant line used had an effect on the resistance phenotypc of the progeny. Tested F2 populations derived from these crosses show that resistance is not under simple genetic control. This, in addition to variation in aphid numbers within accessions, suggests that separation of genetic components of control from environmental ‘noise’ for any accession may only be possible by the production of double haploid plant lines.     

Genetic variation in natural populations of the aphid Rhopalosiphum padi as revealed by maternally inherited markers

A survey on 148 clones of the aphid Rhopalosiphum padi from 11 widespread localities has been carried out to study the genetic structure of populations of this species as revealed by mitochondrial DNA restriction site and length polymorphisms as well as by restriction site analysis of a maternally inherited plasmid carried by the aphid eubacterial endosymbiont Buchnera aphidicola. Our results support the existence in the area under study of two main aphid maternal lineages strikingly coincidental with the two main reproductive categories displayed by this species. Those aphid clones possessing an incomplete life cycle that lacks the sexual phase (anholocyclic or androcyclic clones) show mitochondrial DNA (mtDNA) haplotype I and plasmid haplotype I, whereas those clones displaying the complete life cycle (holocyclic clones) posses some other distinct mtDNA haplotypes closely related to each other and plasmid haplotype II. While restriction-site analysis of maternally inherited markers points to a relatively ancient origin of anholocycly/androcycly (between 460 000 and 1 400 000 years) followed by interrupted gene flow with respect to the ancestral holocyclic population, mtDNA size variation also suggests that historical stochastic processes have a different effect on the evolution of both main aphid lineages. Evidence of occasional nuclear gene flow between lineages and its consequences on the correspondence between maternally inherited haplotypes and life cycle are also presented and discussed.     

Fitness comparison between clones differing in their ability to produce sexuals in the aphid Thopalosiphum padi

This study aimed at evaluating the intrinsic rate of increase (r _m ) of clones of the cereal aphid Rhopalosiphum padi (L.) that differ in their ability to produce sexuals. The value of r _m was measured for wingless parthenogenetic females in experiments conducted at two temperatures (15 °C and 20 °C). We studied six holocyclic clones, five androcyclic clones and five anholocyclic clones and showed that life-cycle has no significant effect on the age at first reproduction, on fecundity nor on longevity. As a result there is no influence on (summer) fitness: on the one hand, holocyclic clones do not compensate their lower fitness in mild winters by a higher fitness during the summer. On the other hand, anholocyclic and androcyclic clones do not seem to obtain any advantage from living on a more restricted range of hosts: there appears to be no cost of polyphenism for holocyclic clones.     

Production and progeny testing of androgenetic rosy barb Puntius conchonius

Protocol for androgenetic cloning of the rosy barb, Puntius conchonius, with contrasting gray and golden strains is described. At the intensity of 4.2 W/m2, UV irradiation for 3.0 min inactivates the maternal genome in eggs of the gray barb. Following activation by the golden barb sperm, 24-min old eggs are shocked at 41 degrees C for 2 min to restore diploidy. Maternal genomic inactivation is confirmed by the (i) golden body color, (ii) karyotyping, and (iii) progeny testing of F1-F3 progenies. Estimates of stage-specific mortality of haploid and diploid androgenotes indicate no change in the time scale or developmental sequence, when sperm of related strain is used for activation, and when haploid genome regulates the development. Survival of androgenetic clones remains constant for the F1, F2, and F3 progenies and is about 15% and 7% at hatching and sexual maturity, respectively. Homozygosity of the androgenotes is shown to inflict greater mortality. Between F1 and F3 generations, the heterozygosity of the androgenetic clone is decreased, as evidenced by reduction in size hierarchy. Though the reproductive performance of the F1, F2, and F3 supermales is superior to the normal ones, the realized fecundity remains equal around 80 progenies per brood. The 92 crosses involving 16 supermales and 10 normal dams yield 75-100% male progenies, confirming the possible operation of XXfemale symbol:XYmale symbol sex determination system. The frequency of unexpected occurrence of female progenies is about 8%, the causes for which are discussed.     

The nature and reality of the aphid clone: genetic variation, adaptation and evolution

1 When aphid clones and clonality are discussed, it is still often said that they are ‘genetically identical’, a statement for which there is no direct evidence, and certainly not for the entire genome. By contrast, there is a growing body of empirical data from the application of high resolution molecular (DNA) markers that aphid asexual lineages rapidly mutate and that, in some documented cases, this variation is selectable, either positively or negatively. 2 Although it is true that, in enclosed conditions (e.g. laboratory or field cage), a so‐called clone as defined as the asexual progeny of a single foundress may be traceable, this is rarely if ever possible in the field without the use of genetic markers, and even then, usually only at a relatively few loci (multilocus genotypes, ‘MLGs’). 3 The continued use of the term clone without qualification of its true nature and the reality of its interesting biology is likely to hamper a proper understanding of the ecology and evolution of these insects (which are interesting in their own right because of their complex life histories, but also because they are important as major pests globally, both by causing direct feeding damage and by transmitting pathogenic plant viruses and thereby leading to huge economic losses in the agricultural, horticultural and forestry industries). 4 In this short review, I provide evidence of what is now known about aphid clonality after the widespread use of molecular markers, comprising information mainly gained within the last 15 years or so. 5 The data demonstrate widespread adaptation and evolution, sometimes involving introgression and hybridization. Because of this new knowledge, our ideas of what constitutes a clone are in need of serious re‐evaluation.     

Responses of holocyclic and anholocyclic Rhopalosiphum padi populations to low‐temperature and short‐photoperiod induction

The different life cycles of aphid species make these organisms good models for studying the short‐term consequences of sex. The bird cherry‐oat aphid Rhopalosiphum padi has a wide geographic distribution and correspondingly different life cycles. In this study, the life cycles of R. padi collected from six different regions in China were characterized experimentally by comparing the responses of holocyclic and anholocyclic populations to low‐temperature and short‐photoperiod induction. Clones collected from Chuzhou, Taian, and Taigu consistently reproduced via obligate parthenogenesis, whereas clones from Hami and Baicheng were holocyclic in their response, and those from Lanzhou were both holocyclic and anholocyclic. Prolonged exposure to low temperature and a short photoperiod (LS) had negative effects on the offspring of anholocyclic aphids with regard to adult lifespan, total longevity, and fecundity compared with aphids maintained at a normal temperature and a long photoperiod (NL). Holocyclic LS R. padi had longer developmental times at all nymph stages, a shorter adult lifespan, shorter total longevity, and a lower fecundity than NL counterparts. The adult prereproduction period of gynoparae was significantly longer than that of virginoparae, and the total longevity of gynoparae was significantly shorter than that of virginoparae. Moreover, the net reproductive and gross reproduction rates, as well as the total fecundity, were roughly fivefold higher in virginoparae than in gynoparae, indicating that there is the short‐term cost of sex. When maintained on their secondary host (Triticum aestivum), gynoparae, males, and oviparae produced by holocyclic populations could survive, and gynoparae produced oviparae. However, under NL conditions, oviparae could not produce overwintering eggs on the secondary host, whereas a few overwintering eggs were generated by oviparae under LS conditions. Taken together, these results illuminate the complexity of insect responses and contribute to a complete understanding of the aphid life cycle and its evolution.     

The ploidy composition of offspring from Solanum tuberosum (4×) ×S. phureja (2×) crosses with special reference to triploid frequencies

Data on the production of offspring of 4×× 2× crosses in potato are presented. The ploidy composition of seedlings varied from year to year and parent to parent. Products of dihaploid induction crosses were chiefly 2× or 3× whereas crosses between S. tuberosum cultivars and unreduced gamete-producing S. phureja clones were mainly 3× or 4×. Substantial percentages of 3× seedlings were produced by some crosses in some years. It was concluded that the so-called ‘triploid block’ in potatoes is a variable phenomenon and that factors which increase seed production may suppress the proportion of triploids produced.     

Spatial variation in local pollen flow and mating success in a Picea abies clone archive and their implications for a novel ��breeding without breeding�� strategy

Currently, Norway spruce (Picea abies) breeding in Sweden is based on crosses between the best clones followed by clonal testing of the progenies to select for the long-term breeding population. An alternative breeding strategy called “Breeding without Breeding” (BwB) is proposed, which, in principle, relies on the DNA marker-based pedigree reconstruction from wind-pollinated progenies instead of controlled crosses. To test whether the pedigree structure could be established from progenies of clonal trials, we investigated the spatial pattern of local pollen flow and paternity assignment in a clone archive of Norway spruce. The results showed that 42% of the progeny can be assigned to fathers within 30-m distance with high confidence. Effective pollen dispersal decreased rapidly with distance and followed exponential distribution on local scale. The extent of close-neighbor (within 6 m) mating ranged from 0% to 48% among grafts with an average of 13%. Distance explained 25% deviance in mating success, and other factors such as phenology and spatial configuration of the clones should have contributed the rest. The success of parentage assignment in clone archive opens up the possibility to apply BwB in clonal trials of species that are easy to propagate vegetatively. This procedure could substantially shorten the breeding cycle and still give similar gain per year as the conventional breeding.     

Host selection and performance of the giant willow aphid, Tuberolachnus salignus Gmelin – implications for pest management

1 The recent increase in planting of selected willow clones as energy crops for biomass production has resulted in a need to understand the relationship between commonly grown, clonally propagated genotypes and their pests. 2 For the first time, we present a study of the interactions of six willow clones and a previously unconsidered pest, the giant willow aphid Tuberolachnus salignus. 3 Tuberolachnus salignus alatae displayed no preference between the clones, but there was genetic variation in resistance between the clones; Q83 was the most resistant and led to the lowest reproductive performance in the aphid 4 Maternal effects buffered changes in aphid performance. On four tested willow clones fecundity of first generation aphids on the new host clone was intermediate to that of the second generation and that of the clone used to maintain the aphids in culture. 5 In the field, patterns of aphid infestation were highly variable between years, with the duration of attack being up to four times longer in 1999. In both years there was a significant effect of willow clone on the intensity of infestation. However, whereas Orm had the lowest intensity of infestation in the first year, Dasyclados supported a lower population level than other monitored clones in the second year.     

Mode of serotype inheritance in exoconjugant progeny of the ciliate Dileptus anser

Two clones of Dileptus anser, originally isolated from natural reservoirs and referred to below as B and D clones, were found to display different serotypes, when cultured under identical laboratory conditions. On being tested with two different polyclonal rabbit immune sera against each particular clone (the classic immobilization test) these clones showed no cross-reaction. At a standard dilution (1:50) and at a standard exposure time (4 h), either of the two immune sera immobilized 100% or commonly 0% of homologous and heterologous clone cells, respectively. In addition, the difference in serotypes was confirmed by the immunofluorescence analysis. By crossing (conjugation) between B (mating type I) and D (mating type III) cells, exconjugant F1 clones were obtained. Their serotypes were then tested (the same immobilization test) with antisera against both the "parental" clones: some clones were tested before their sexual maturation in ca. one month after conjugation, while others were examined in approximately 4 months after conjugation, i.e. after reaching maturity. Each of the F1 clones could react with both immune sera, which means that they possessed the intermediate, "hybrid" phenotype. Five different F1 clones were selected, and each of them was back-crossed to both "parental" clones, B and D. We succeeded in raising 25 exconjugant F2 (B1, to be more exact) clones from F1 x B crosses and 26 clones from F1 x D crosses. The conventional testing of these clones in 5-10 weeks after conjugation provided quite unexpected results, since among them no segregation for "parental" serotypes was observed. Each of the 51 tested clones demonstrated the "hybrid" serotype--seemingly the same as that of F1 clones. Such a non-Mendelian inheritance of the character is hardly to explain from the standard, canonical assumptions on the genetic control of serotype difference between original "parental" clones (different alleles in one locus? different loci?). Also it does not seem likely that the absence of segregation could result from differential survival of various phenotypes in F2 (although the total viability of exconjugant clones appeared rather low). The above data obviously need further confirmations and experimental analyses. We attempt to discuss the obtained results in terms of the epigene hypothesis (Tchuraev, 1975) and in relation to the epigenetic control of serotype expression in species of the Paramecium aurelia complex and in Tetrahymena thermophila, which are "the chosen few" subjects in ciliate genetics.     

On critical night lengths and temperature compensation in the photoperiodic response of two geographical clones of the black bean aphid, Aphis fabae

Photoperiodic response curves were determined for two clones of the black bean aphid, Aphis fabae Scopoli, at three temperatures, 12.5, 15 and 17.5°C. Critical night lengths for the induction of winged females in an English clone (52° N) were 10.5, 11 and 11.5 h, respectively, and 10, 10.5 and 11 h in a Scottish clone (57° N). Critical night lengths for male induction were 10.5, 11 and 11 h at 12.5, 15 and 17.5°C in the English clone, and 10, 10.5 and 10.5 h, respectively, in the Scottish clone. High incidences of winged females and males were observed at all scotophases longer than the critical night length in both clones. In addition, in the English clone, the incidences of winged female and male producers in continuous darkness were 0% at 15 and 17.5°C, and 6% at 12.5°C. In the Scottish clone, however, continuous darkness resulted in high incidences of both winged female and male producers at 12.5 and 15°C, but 0% winged female producers and 6% male producers at 17.5°C. In scotophases shorter than the critical night length, including continuous light, no males or winged females were observed in either clone under the non‐crowded rearing conditions used. The results are discussed in terms of the ‘double circadian oscillator model’ for photoperiodic induction.     

Intraspecific variability in response to stimuli for male and ephippia formation in Daphnia pulex

Natural clones and clones from laboratory crossings of Daphnia pulex have been tested in their response to stimuli for male and ephippia formation. The clones are from ponds in Illinois, USA, an area with both obligate and cyclical clones of D. pulex. The progeny from two types of crosses were studied: 1) crosses between an obligate clone producing males and a cyclical clone producing only females, and 2) self-fertilization within a cyclical clone producing both sexes. Both the natural and the artificial clones showed great variation in response to environmental stimuli for males. Ephippia response seems more equal among the clones, but two of the artificial clones could hardly be stimulated to produce ephippia in the induction experiments. In crowded cultures, however, they produced a few. Crossings between cyclicals and obligates yielded mostly cyclical progeny. Progeny from the selfed clone had low survival and fecundity.     

Weak spatial and temporal population genetic structure in the rosy apple aphid, Dysaphis plantaginea, in French apple orchards

We used eight microsatellite loci and a set of 20 aphid samples to investigate the spatial and temporal genetic structure of rosy apple aphid populations from 13 apple orchards situated in four different regions in France. Genetic variability was very similar between orchard populations and between winged populations collected before sexual reproduction in the fall and populations collected from colonies in the spring. A very small proportion of individuals (～2%) had identical multilocus genotypes. Genetic differentiation between orchards was low (F(ST)<0.026), with significant differentiation observed only between orchards from different regions, but no isolation by distance was detected. These results are consistent with high levels of genetic mixing in holocyclic Dysaphis plantaginae populations (host alternation through migration and sexual reproduction). These findings concerning the adaptation of the rosy apple aphid have potential consequences for pest management.