Niche separation in space and time between two sympatric sister species—a case of ecological pleiotropy

We investigate the niche separation in space and time between the Palearctic sister species Leptidea sinapis and L. reali (Lepidoptera, Pieridae) in central Sweden. Using field sampling, we show that L. reali is a habitat specialist confined to meadows, whereas L. sinapis is a habitat generalist also inhabiting forests. This difference in habitat utilization was corroborated by experimental release of laboratory-reared L. sinapis and L. reali in two adjacent forest and meadow habitats during their natural flight period; virtually all recaptured L. reali that were released in the forest were later caught in the meadow, whereas L. sinapis shifted equally often from meadow to forest as in the opposite direction. In the field, both species fly in May–June, but L. reali appears on average a week earlier in spring and has a substantial second generation in July, whereas L. sinapis is practically univoltine. When overwintered pupae were incubated under identical conditions in the laboratory, females did, however, not differ in phenology, and L. sinapis males actually emerged earlier than L. reali males. When larvae were reared at 23°C on the host plant Lotus corniculatus at a range of daylengths, both species produced a substantial proportion of directly developing individuals at an 18.5 h daylength or longer. When reared at 23°C and a 22 h daylength, L. reali showed an overall higher propensity to develop directly than L. sinapis on plant species originating from both the meadow and the forest habitat. Both Leptidea species showed a lower propensity to enter direct development on forest associated plants than on meadow associated plants. Hence, we suggest that the difference in phenology and voltinism between L. sinapis and L. reali is largely the result of environmentally implemented ecological pleiotropic effects caused by the between-species difference in habitat preference.     

Trade-offs of host use between generalist and specialist <Emphasis Type="Italic">Helicoverpa</Emphasis> sibling species: adult oviposition and larval performance

Much attention has been paid to the question of the relative importance of female behaviour versus larval feeding capacities in determining the host range of herbivorous insects. Host-use trade-offs displayed by generalist and specialist sister species of the genus Helicoverpa were evaluated to examine the relationship between maternal choice and offspring performance. The prediction of optimal oviposition theory, that females will choose to lay eggs on plants on which their offspring perform best as larvae, was tested by measuring oviposition preference and larval performance of Helicoverpa armigera and H. assulta on tobacco, sunflower, and hot pepper. These two measures were more highly correlated in the specialist H. assulta. Both species exhibited the same oviposition preference ranking: tobacco > sunflower > hot pepper. H. armigera larvae preferred sunflower, followed by tobacco and hot pepper; while H. assulta larvae preferred tobacco to sunflower and hot pepper, consistent with their mothers’ oviposition preference. Duration of the total period from egg to adult emergence for each species was significantly shorter on the host plant preferred by the larvae. H. assulta had shorter larval duration and higher relative growth rate than H. armigera on tobacco and hot pepper, and vice versa for sunflower, indicating species differences in host utilization. Thus, while only the specialist H. assulta displayed the predicted optimal oviposition pattern, females of both species show the least preference for the plant on which their offspring perform worst. Selection for optimal oviposition may be stronger on the specialist, which has fewer choices and lower lifetime fecundity than the generalist.     

Morphology and colonization preference of arbuscular mycorrhizal fungi in <Emphasis Type="Italic">Clethra barbinervis</Emphasis>, <Emphasis Type="Italic">Cucumis sativus</Emphasis>, and <Emphasis Type="Italic">Lycopersicon esculentum</Emphasis>

Clethra barbinervis (Ericales), Cucumis sativus, and Lycopersicon esculentum were grown in soils collected from six different vegetation sites (cedar, cypress, larch, red pine, bamboo grass, and Italian ryegrass), and morphology and colonization preference of arbuscular mycorrhizal (AM) fungi were investigated by microscopic observation and PCR detection. C. barbinervis consistently formed Paris-type AM throughout the sites. C. sativus formed both Arum- and Paris-type AM with high occurrence of Arum-type AM. L. esculentum also formed both Arum- and Paris-type AM but with high occurrence of Paris-type AM. AM diversity within the same plant species was different among the sites. Detected AM diversity from AM spores in different site soils did not consistently reflect AM fungal diversity seen in test plants. Detected families were different, depending on test plants grown even in the same soil. AM fungi belonging to Glomaceae were consistently detected from roots of all test plants throughout the sites. Almost all the families were detected from roots of C. barbinervis and L. esculentum. On the other hand, only two or three families of AM fungi (Archaeosporaceae and/or Paraglomaceae and Glomaceae) but not two other families (Acaulosporaceae and Gigasporaceae) were detected from roots of C. sativus, indicating strong colonization preference of AM fungi to C. sativus among test plants. This study demonstrated that host plant species strongly influenced the colonization preference of AM fungi in the roots.     

Evidence-based National Recovery Plan for <Emphasis Type="Italic">Leptidea sinapis</Emphasis> (wood white butterfly) in Southern Britain

We present a key part of the evidence base used to develop a National Recovery Plan for Leptidea sinapis a declining and threatened species in Britain. Knowledge of the current sites has been updated along with the macro- and micro-habitat resource requirements of the species and its autecology to set the basis for a conservation strategy for 2010 and beyond. The three main findings are that: L. sinapis uses a greater diversity of biotopes within and between sites than was previously thought; population dynamics are complex with 2–3 year peaks and troughs of annual abundance indices and two generations per year throughout its range; and dispersal ability is also greater than previously stated. This evidence has been used to develop an Action Plan for 2010 and beyond. The importance of landscape-scale management and effective collaboration between scientists, land managers and volunteers is emphasised.     

Development of <Emphasis Type="Italic">Mamestra brassicae</Emphasis> and its solitary endoparasitoid <Emphasis Type="Italic">Microplitis mediator</Emphasis> on two populations of the invasive weed <Emphasis Type="Italic">Bunias orientalis</Emphasis>

The warty cabbage Bunias orientalis is an invasive pest in much of central Europe, including much of Germany since the 1980s, whereas in other countries, such as The Netherlands, it is a less common exotic species. Here, healthy larvae of Mamestra brassicae, which has been found feeding on B. orientalis plants in Germany, and larvae parasitized by one of its major larval endoparasitoids Microplitis mediator, were reared on both herbivore-induced and noninduced leaves of B. orientalis originating from single large populations growing in The Netherlands and central Germany. Herbivore performance was less negatively affected than parasitoid performance by differences in plant quality. Development times in both M. brassicae and Mi. mediator were shorter on Dutch than German plants and also shorter on noninduced than induced plants. Moreover, survival and body size of the parasitoid was more strongly affected by plant population and induction than survival of healthy M. brassicae. Chemical analyses of defensive secondary metabolites [glucosinolates (GS)] revealed that concentrations of the major GS sinalbin were constitutively expressed in German plants whereas they were induced in Dutch plants. However, in separate feeding bioassays in which preference for induced and noninduced leaves was compared separately, L3 instars of M. brassicae preferred noninduced German plants over Dutch plants but induced Dutch plants over German plants, revealing that changes in primary metabolites or an unidentified non-GS compound mediates population-related differences in plant quality. The results reveal asymmetric effects of plant quality in exotic plants on organisms in the second and third trophic level.     

Nutritional suitability and ecological relevance of <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis> and <Emphasis Type="Italic">Brassica oleracea</Emphasis> as foodplants for the cabbage butterfly, <Emphasis Type="Italic">Pieris rapae</Emphasis>

The thale cress, Arabidopsis thaliana, is considered to be an important model species in studying a suite of evolutionary processes. However, the species has been criticized on the basis of its comparatively small size at maturity (and consequent limitations in the amount of available biomass for herbivores) and on the duration and timing of its life cycle in nature. In the laboratory, we studied interactions between A. thaliana and the cabbage butterfly, Pieris rapae, in order to determine if plants are able to support the complete development of the herbivore. Plants were grown in pots from seedlings in densities of one, two, or four per pot. In each treatment, one, two, or five newly hatched larvae of P. rapae were placed on fully developed rosettes of A. thaliana. In a separate experiment, the same densities of P. rapae larvae were reared from hatching on single mature cabbage (Brassica oleracea) plants. Pupal fresh mass and survival of P. rapae declined with larval density when reared on A. thaliana but not on B. oleracea. However, irrespective of larval density and plant number, some P. rapae were always able to complete development on A. thaliana plants. A comparison of the dry mass of plants in different treatments with controls (= no larvae) revealed that A. thaliana partially compensated for plant damage when larval densities of P. rapae were low. By contrast, single cress plants with 5 larvae generally suffered extensive damage, whereas damage to B. oleracea plants was negligible. Rosettes of plants that were monitored in spring, when A. thaliana naturally grows, were not attacked by any insect herbivores, but there was often extensive damage from pulmonates (slugs and snails). Heavily damaged plants flowered less successfully than lightly damaged plants. Small numbers of generalist plant-parasitic nematodes were also recovered in roots and root soil. By contrast, plants monitored in a sewn summer plot were heavily attacked by insect herbivores, primarily flea beetles (Phyllotreta spp.). These results reveal that, in natural populations of A. thaliana, there is a strong phenological mismatch between the plant and most of its potential specialist insect herbivores (and their natural enemies). However, as the plant is clearly susceptible to attack from non-insect generalist invertebrate herbivores early in the season, these may be much more suitable for studies on direct defense strategies in A. thaliana.     

Host-related variability in arbuscular mycorrhizal fungal structures in roots of <Emphasis Type="Italic">Hedera rhombea</Emphasis>, <Emphasis Type="Italic">Rubus parvifolius</Emphasis>, and <Emphasis Type="Italic">Rosa multiflora</Emphasis> under controlled conditions

The arbuscular mycorrhizal (AM) morphology of three host plant species inoculated with single and mixed fungal culture and the distribution of AM fungal species in roots of the hosts treated with a mixed culture of AM fungi were determined. The aim was to investigate the effect of host plants and AM fungi on AM morphology of coexisting plant species. Noncolonized rooted cuttings of Hedera rhombea (Miq) Bean, Rubus parvifolius L., and Rosa multiflora Thunb. were inoculated with five fungal species as single and mixed culture inocula. The fungal species used were Gigaspora rosea and Scutellospora erythropa, previously isolated from H. rhombea; Acaulospora longula and Glomus etunicatum from R. parvifolius; and Glomus claroideum from both plant species. A few hyphal and arbusculate coils were seen in the mixed culture-inoculated roots of R. parvifolius; all fungal treatments produced this Paris-type AM in H. rhombea and Arum-type AM in R. parvifolius, and R. multiflora indicates that AM morphology is strongly controlled by the identity of the host plants used in this study. AM fungal rDNA was extracted separately from roots of each replicate plant species inoculated with the mixed fungal culture, amplified, cloned, sequenced, and analyzed to determine the AM fungal species and their respective proportions in roots of each plant species. Glomus etunicatum and G. claroideum of the family Glomaceae generally occurred more frequently in R. parvifolius and R. multiflora, which form Arum-types, whereas S. erythropa, of the family Gigasporaceae, was the most frequently detected species in H. rhombea, which produced Paris-type AM. Although the genotype of the plant species used appears to determine the AM morphologies formed, there was preferential association between the hosts and AM fungal inoculants.     

<Emphasis Type="Italic">Piriformospora indica</Emphasis> and <Emphasis Type="Italic">Sebacina vermifera</Emphasis> increase growth performance at the expense of herbivore resistance in <Emphasis Type="Italic">Nicotiana attenuata</Emphasis>

A Sebacinales species was recovered from a clone library made from a pooled rhizosphere sample of Nicotiana attenuata plants from 14 native populations. Axenic cultures of the related species, Piriformospora indica and Sebacina vermifera, were used to examine their effects on plant performance. Inoculation of N. attenuata seeds with either fungus species stimulated seed germination and increased growth and stalk elongation. S. vermifera inoculated plants flowered earlier, produced more flowers and matured more seed capsules than did non-inoculated plants. Jasmonate treatment during rosette-stage growth, which slows growth and elicits herbivore resistance traits, erased differences in vegetative, but not reproductive performance resulting from S. vermifera inoculation. Total nitrogen and phosphorous contents did not differ between inoculated and control plants, suggesting that the performance benefits of fungal inoculation did not result from improvements in nutritional status. Since the expression of trypsin proteinase inhibitors (TPI), defensive proteins which confer resistance to attack from Manduca sexta larvae, incur significant growth and fitness costs for the plant, we examined the effect of S. vermifera inoculation on herbivore resistance and TPI activity. After 10 days of feeding on S. vermifera-inoculated plants, larval mass was 46% higher and TPI activity was 48% lower than that on non-inoculated plants. These results suggest that Sebacina spp. may interfere with defense signaling and allow plants to increase growth rates at the expense of herbivore resistance mediated by TPIs.     

<Emphasis Type="Italic">In vitro</Emphasis> propagation of <Emphasis Type="Italic">Zingiber petiolatum</Emphasis> (Holttum)

Summary We describe an in vitro propagation protocol for Zingiber petiolatum (Holttum), I. Theilade, a rare species from the southern part of Thailand. Fruits were surface-sterilized and seeds germinated on Murashige and Skoog medium (MS) medium supplemented with 3% sucrose. Three-month-old seedlings were used as initial plant material for in vitro propagation. Terminal buds of the plants were inoculated on MS medium containing 6-benzylaminopurine (BA; 2.2–35.5 μM) alone or in combination with 1-naphthaleneacetic acid (0.5 μM). Eight weeks after inoculation, the cultures were transferred to MS medium without plant growth regulators for 4wk. The cultures transferred from MS medium with 17.8 μM BA revealed the highest shoot induction rate of 6.1±0.7 shoots per explant. Rooting was spontaneously achieved in MS medium without plant growth regulators. Rooted plants were successfully transplanted to soil.     

Specialist <Emphasis Type="Italic">Osmia</Emphasis> bees forage indiscriminately among hybridizing <Emphasis Type="Italic">Balsamorhiza</Emphasis> floral hosts

Pollinators, even floral generalists (=polyleges), typically specialize during individual foraging bouts, infrequently switching between floral hosts. Such transient floral constancy restricts pollen flow, and thereby gene flow, to conspecific flowers in mixed plant communities. Where incipient flowering species meet, however, weak cross-fertility and often similar floral traits can yield mixed reproductive outcomes among pollinator-dependent species. In these cases, floral constancy by polyleges sometimes serves as an ethological mating barrier. More often, their foraging infidelities instead facilitate host introgression and hybridization. Many other bee species are oligolectic (taxonomic specialists for pollen). Oligoleges could be more discriminating connoisseurs than polyleges when foraging among their limited set of related floral hosts. If true, greater foraging constancy might ensue, contributing to positive assortative mating and disruptive selection, thereby facilitating speciation among their interfertile floral hosts. To test this Connoisseur Hypothesis, nesting females of two species of oligolectic Osmia bees were presented with randomized mixed arrays of flowers of two sympatric species of their pollen host, Balsamorhiza, a genus known for hybridization. In a closely spaced grid, the females of both species preferred the larger flowered B. macrophylla, evidence for discrimination. However, both species’ females showed no floral constancy whatsoever during their individual foraging bouts, switching randomly between species proportional to their floral preference. In a wider spaced array in which the bouquets reflected natural plant spacing, foraging oligolectic bees often transferred pollen surrogates (fluorescent powders) both between conspecific flowers (geitonogamy and xenogamy) and between the two Balsamorhiza species. The Connoisseur Hypothesis was therefore rejected. Foraging infidelity by these oligolectic Osmia bees will contribute to introgression and hybridization where interfertile species of Balsamorhiza meet and flower together. A literature review reveals that other plant genera whose species hybridize also attract numerous oligolectic bees, providing independent opportunities to test the generality of this conclusion.     

Parasitism <Emphasis Type="Italic">versus</Emphasis> mutualism in the ant-garden parabiosis between <Emphasis Type="Italic">Camponotus femoratus</Emphasis> and <Emphasis Type="Italic">Crematogaster levior</Emphasis>

Ant-gardens represent a special type of association between ants and epiphytes. Frequently, two ant species can share the same nest in a phenomenon known as ‘parabiosis’, but the exact nature (i.e., mutualistic or parasitic) of this interaction is the subject of debate. We thus attempted to clarify the mutual costs and benefits for each partner (ants and plants) in the Crematogaster levior/Camponotus femoratus ant-garden parabiosis. The ants’ response to experimental foliar damage to the epiphytes and to the host tree as well as their behavior and interactions during prey capture were investigated to see if the purported parasitic status of Cr. levior could be demonstrated in either the ant-ant or in the ant-plant interactions. The results show that both species take part in protecting the epiphytes, refuting the role of Cr. levior as a parasite of the ant-garden mutualism. During capture of large prey Ca. femoratus took advantage from the ability of Cr. levior to discover prey; by following Cr. levior trails Ca. femoratus workers discover the prey in turn and usurp them during agonistic interactions. Nevertheless, the trade-off between the costs and benefits of this association seems then to be favorable to both species because it is known that Cr. levior benefits from Ca. femoratus building the common carton nests and furnishing protection from vertebrates. Consequently, parabiosis can then be defined as the only mutualistic association existing between ant species, at least in ant-gardens. Received 31 August 2006 ; revised 8 December 2006 ; accepted 12 December 2006     

Fungal endophytes associated with the mistletoe <Emphasis Type="Italic">Phoradendron perrottettii</Emphasis> and its host tree <Emphasis Type="Italic">Tapirira guianensis</Emphasis>

The endophytic mycobiota of leaves and stems of the mistletoe Phoradendron perrottettii and its host tree Tapirira guianensis, two physiologically connected plant species of the Brazilian savannah in southeastern Brazil, were investigated to evaluate host and organ recurrence among endophytes. Leaves and stems of P. perrottettii and leaves of T. guianensis were sampled in the dry and wet season. Stems of T. guianensis were also sampled in the wet season. Endophytes were isolated by an adapted trituration and particle filtration protocol. A total of 1,615 isolates representing 99 species and 20 sterile morphotypes were recovered; 64 morphospecies occurred as singletons. The number of isolates and species was higher in the wet season. Leaves of P. perrottettii were less densely colonized than other organs studied, but were the most species-rich. Conversely, stems of T. guianensis yielded more isolates but were less species-rich. Both plants were found to harbor similar but distinguishable endophytic assemblages. The Jaccard’s index of similarity between the fungal assemblages of both plants was 0.82, higher than found for other plants in similar habitats. The fungal species composition seemed to be influenced by the collection season and organ type, as demonstrated by multivariate correspondence analysis. Paraconiothyrium brasiliense, P. sporulosum and Verticillium leptobactrum were the dominant species in P. perrottettii. In leaves of T. guianensis, Pseudocercospora sp., Phomopsis sp. 1 and Lecanicillium psalliotae were the most frequent, while Stagonospora sp. 1 and Phomopsis sp. 1 were the dominant endophytes in its stems. The results indicated that some of the dominant endophytic taxa isolated in this study colonize different hosts and plant organs while others seem to exhibit a high degree of host or organ recurrence. This study represents the first evaluation of diversity of fungal endophytes in natural vegetation of the Brazilian savannah and contributes information about the distribution and possible specificity of endophytes in tropical dicotyledoneous plants.     

Close-Range Host Searching Behavior of the Stemborer Parasitoids <Emphasis Type="Italic">Cotesia sesamiae</Emphasis> and <Emphasis Type="Italic">Dentichasmias busseolae</Emphasis>: Influence of a Non-Host Plant <Emphasis Type="Italic">Melinis minutiflora</Emphasis>

Studies were conducted on the host searching behavior of the larval parasitoid Cotesia sesamiae (Cameron) (Hymenoptera: Braconidae) and the pupal parasitoid Dentichasmias busseolae Heinrich (Hymenoptera: Ichneumonidae), both of which attack lepidopteran (Crambidae, Noctuidae) cereal stemborers. The behavior of D. busseolae was observed in a diversified habitat that consisted of stemborer host plants (maize, Zea mays L. and sorghum, Sorghum bicolor (L). Moench (Poaceae)) and a non-host plant (molasses grass, Melinis minutiflora Beauv. (Poaceae)), while C. sesamiae was observed separately on host plants and molasses grass. In previous olfactometer studies, C. sesamiae was attracted to molasses grass volatiles while hboxD. busseolae was repelled. The aim of the present study was to investigate the influence of molasses grass on close-range foraging behavior of the parasitoids in an arena that included infested and uninfested host plants. Dentichasmias busseolae strongly discriminated between host and non-host plants, with female wasps spending most of the time on infested host plants and least time on molasses grass. Likewise, C. sesamiae spent more time on uninfested and infested host plants than it did on molasses grass in single choice bioassays. While on infested plants, the wasps spent more time foraging on the stem, the site of damage, than on other areas of the plant. Overall, the results indicate that presence of the non-host plant does not hinder close range foraging activities of either parasitoid.     

Woodrat (<Emphasis Type="Italic">Neotoma</Emphasis>) herbivores maintain nitrogen balance on a low-nitrogen,high-phenolic forage, <Emphasis Type="Italic">Juniperus monosperma</Emphasis>

The acquisition of adequate quantities of nitrogen is a challenge for herbivorous vertebrates because many plants are in low nitrogen and contain secondary metabolites that reduce nitrogen digestibility. To investigate whether herbivores maintain nitrogen balance on plant diets low in nitrogen and high in secondary compounds, we studied the effect of juniper (Juniperus monosperma) ingestion on the nitrogen balance of two species of herbivorous woodrats (Neotoma stephensi and N. albigula). These woodrat species feed on the foliage of juniper: N. stephensi is a juniper specialist, whereas N. albigula is a generalist that incorporates some juniper in its diet. Based on the nitrogen contents of the natural diets of these woodrats, we predicted that the generalist would be in negative nitrogen balance on a juniper diet whereas the specialist would not be affected. We found that both species of woodrat had low-nitrogen requirements (334.2 mg N/kg^0.75/day) and that a diet of 50% juniper did not result in negative nitrogen balance for either species. However, excretion patterns of nitrogen were altered; on the 50% juniper diet, fecal nitrogen losses increased ~38% and urinary nitrogen losses were half that of the control diet. The results suggest that absorption and detoxification of juniper secondary compounds may be more important for restricting juniper intake by the generalist than nitrogen imbalance.     

Absence of Learning and Local Specialization on Host Plant Selection by <Emphasis Type="BoldItalic">Heliconius erato</Emphasis>

There is considerable interspecific variation in larval host plants (Passifloraceae) used among Heliconius erato (Lepidoptera: Nymphalidae) populations. This study evaluates the role of learning and the influence of interspecific variation in host plant attributes on such local specialization in H. erato host plant choices. Experiments were conducted under laboratory, insectary, and field conditions, with the two most widely used host plants in southern Brazil (Passiflora suberosa Linnaeus and Passiflora misera Humbold, Bonpland et Kunth). Larval feeding preference and induction were evaluated through choice tests for all instars. Oviposition was evaluated in relation to host plant preference, Hopkins host-selection principle, and conditioning time (for 3, 7, 11, or 15 days). Also, oviposition choice was tested regarding density, intemode length, and presence of intact terminal bud on P. suberosa and P. misera shoots. Both larvae and adults of H. erato phyllis showed preference for P. misera compared to P. suberosa, under all conditions. Larval feeding preference could not be induced for most instars. The Hopkins effect was not detected and oviposition choice could not be conditioned. Females alternated use of host plant species as a function of variation in either density or presence of terminal buds on shoots. Thus, our data indicate host plant preference in H. erato phyllis is not learned but innate. Therefore, we concluded that variation in local use of host plant by this butterfly in southern Brazil results from qualitative and quantitative variation of the passion vine species.     

RETRACTED ARTICLE: Foraging of Host-Habitat and Superparasitism in <Emphasis Type="Italic">Cotesia glomerata</Emphasis>: A Gregarious Parasitoid of <Emphasis Type="Italic">Pieris brassicae</Emphasis>

The study investigates differences in the oviposition pattern of a braconid parasitoid, Cotesia glomerata (Linn.) in Pieris brassicae (Linn.) in relation to their use of different cruciferous food plants. The response of P. brassicae to superparasitism and consequences for the parasitoid were examined in order to elucidate the ecological significance of this behaviour. Female parasitoid located various crucifers and searched for host more frequently almost on all the host plants tested i.e. cabbage, cauliflower, Chinese cabbage, broccoli and radish. According to the estimated relative number of female locating hosts, cabbage was the most attractive plant for C. glomerata and total number of eggs laid in host larvae feeding on it was higher than in larvae feeding on other plants. Laboratory experiments demonstrated that superparasitism reduced survivorship of P. brassicae larvae. Superparasitism lengthened parasitoid development and prolonged the feeding period of host larvae. Sex ratio and the body weight of emergent wasps correlated negatively with brood size. Despite a trade-off between maximising brood size and optimising the fitness of individual offspring, two or three ovipositions on P. brassicae larvae resulted in a greater female dry mass than did a single oviposition on the host. Thus, superparasitism might be of adaptive significance under certain circumstances, especially when host density is low and unparasitized hosts are rare in a habitat.     

Geographic host use variability and host range evolutionary dynamics in the phytophagous insect <Emphasis Type="Italic">Apagomerella versicolor</Emphasis> (Cerambycidae)

The high diversity of phytophagous insects has been explained by the tendency of the group towards specialization; however, generalism may be advantageous in some environments. The cerambycid Apagomerella versicolor exhibits intraspecific geographical variation in host use. In northern Argentina it is highly specialized on the herb Pluchea sagittalis (Asteraceae), while in central and southern areas it uses seven Asteraceae species. To study host species geographical variation from ecological and evolutionary perspectives, we investigated field host availability and use across a wide latitudinal range, and performed laboratory studies on insect oviposition preference and larval performance and mitochondrial DNA (mtDNA) variation in a phylogeographical framework. Geographic variation in host use was unrelated to host availability but was highly associated with laboratory oviposition preference, larval performance, and mtDNA variation. Genetic studies revealed three geographic races of A. versicolor with gene flow restriction and recent geographic expansion. Trophic generalism and oligophagy within A. versicolor seem to have evolved as adaptations to seasonal and spatial unavailability of the preferred host P. sagittalis in cooler areas of the species’ geographic range. No single genotype is successful in all environments; specialization may be advantageous in environments with uniform temporal and spatial host availability, while being a trophic generalist may provide an adaptive advantage in host-constrained environments.     

Harvest-inducibility of the promoter of alfalfa<Emphasis Type="Italic"> S</Emphasis>-adenosyl-<Emphasis Type="SmallCaps">l</Emphasis>-methionine:<Emphasis Type="Italic"> trans</Emphasis>-caffeoyl-CoA3-<Emphasis Type="Italic">O</Emphasis>-methyltransferase gene

A major limitation on the expression of some foreign proteins in transgenic plants is the toxic effect of such proteins on the host plant resulting in inhibition of normal growth and development. A solution to this problem is to control the expression of genes for such proteins by means of inducible promoters, as is frequently done in microbial systems. A cDNA clone was obtained from subtractive hybridization of non-harvested and harvested alfalfa leaf tissue, named hi12. The hi12 cDNA was identified as part of the S-adenosyl-l-methionine: trans-caffeoyl-CoA3-O-methyltransferase gene of alfalfa, a gene encoding an essential key enzyme in lignin synthesis. The hi12 gene was strongly induced by harvesting and wounding but not by heat shock. The promoter of the hi12 gene, isolated by genomic walking, contained several stress response cis-elements. Transgenic plants of tobacco and Medicago truncatula containing the GUS gene driven by the promoter showed GUS expression following harvesting, demonstrating the activity of these regulatory regions in other plant species.     

Tissue culture of <Emphasis Type="Italic">Sinningia speciosa</Emphasis> and analysis of the <Emphasis Type="Italic">in vitro</Emphasis>-generated <Emphasis Type="Italic">tricussate whorled phyllotaxis</Emphasis> (<Emphasis Type="Italic">twp</Emphasis>) variant

Two protocols were developed for the efficient regeneration of Sinningia speciosa from leaf explants via two developmental pathways. The first method involved formation of callus and buds, followed by subsequent root growth, in Murashige and Skoog medium (MS) containing 2.0 mg l⁻¹ 6-benzylaminopurine (BA) and 0.2 mg l⁻¹ α-naphthalene acetic acid (NAA), with a regeneration efficiency of 99.0%. The second method involved producing callus and roots, followed by subsequent formation of buds, in MS medium supplemented with 1.0–5.0 mg l⁻¹ NAA, and resulted in a regeneration efficiency of 90.4%. Our experiments indicate that the root-first pathway resulted in a lower plant regeneration efficiency. Through five continual generations using the buds-first method, a total of 215 regenerated plants were obtained in the last generation, and eight exhibited a phenotype we named tricussate whorled phyllotaxis (twp). Six of the regenerated twp variant plants maintained their tricussate whorled phyllotaxis phenotype, showing no other abnormalities, while one reverted to a wild type before flowering and another formed two rounds of sepals. Physiological analysis revealed that the twp plants responded differently than wild type to exogenous NAA and 2,3,5-triiodobenzoic acid (TIBA), while high-performance liquid chromatography (HPLC) analysis showed that the levels of endogenous indole-3-acetic acid (IAA) and gibberellin (GA) were lower in twp than wild-type plants. These results suggest that the formation of the twp mutant may be related to phytohormones and that the twp variant could be an important material for investigating the molecular mechanism of plant phyllotaxis patterning.