First evidence of late-acting self-incompatibility in the Aristolochiaceae.

Most Aristolochiaceae species studied so far are from temperate regions, bearing self-compatible protogynous trap flowers. Although self-incompatibility has been suggested for tropical species, the causes of self-sterility in this family remain unknown. To fill this gap, we studied the pollination of the tropical Aristolochia esperanzae, including the physical and physiological anti-selfing mechanisms. Floral visitors trapped inside flowers were collected to determine the pollinators. Protogyny was characterized by observing the temporal expression of sexual phases and stigmatic receptivity tests. The breeding system was investigated using hand-pollination treatments. Pollen tube growth was observed using epifluorescence to identify the self-incompatibility mechanism. Flies were the most frequent visitors found inside A. esperanzae trap flowers, with individuals from the family Ulidiidae being potential pollinators since they carried pollen. The characteristic flower odour and presence of larvae indicate that A. esperanzae deceives flies through oviposition-site mimicry. Although this species showed incomplete protogyny, stigmatic receptivity decreased during the male phase, avoiding self-pollination. Fruits developed only after cross- and open pollination, indicating that the population is non-autonomous, non-apomictic, and self-sterile. This occurred through a delay in the growth of geitonogamous pollen tubes to the ovary and lower ovule penetration, indicating a late-acting self-incompatibility mechanism. Our findings expand the number of families in which late-acting self-incompatibility has been reported, demonstrating that it is more widespread than previously thought, especially when considering less-studied tropical species among the basal angiosperms.

Aristolochia mimics stink bugs to repel vertebrate herbivores via TRPA1 activation.

Mimicry is the phenomenon in which one species (the mimic) closely resembles another (the model), enhancing its own fitness by deceiving a third party into interacting with it as if it were the model. In plants, mimicry is used primarily to gain fitness by withholding rewards from mutualists or deterring herbivores cost-effectively. While extensive work has been documented on putative defence mimicry, limited investigation has been conducted in the field of chemical mimicry. In this study, we used field experiments, chemical analyses, behavioural assays, and electrophysiology, to test the hypothesis that the birthwort Aristolochia delavayi employs chemical mimicry by releasing leaf scent that closely resembles stink bug defensive compounds and repels vertebrate herbivores. We show that A. delavayi leaf scent is chemically and functionally similar to the generalized defensive volatiles of stink bugs and that the scent effectively deters vertebrate herbivores, likely through the activation of TRPA1 channels via (E)-2-alkenal compounds. This study provides an unequivocal example of chemical mimicry in plants, revealing intricate dynamics between plants and vertebrate herbivores. Our study underscores the potency of chemical volatiles in countering vertebrate herbivory, urging further research to uncover their potentially underestimated importance.

Pollen ultrastructure in Aristolochia manshuriensis and A. contorta (Aristolochiaceae).

Pollen ultrastructure has been studied in two relict and rare species of the genus Aristolochia, A. contorta Bunge and A. manshuriensis Kom. (Aristolochiaceae). Both species have inaperturate, spheroidal, sometimes distally monocolpate or distally bicolpate pollen grains. The equatorial and polar axes of pollen grain in A. manshuriensis are 48.5 and 44.0 µm, respectively. The percentage of defective pollen grains in A. manshuriensis is 3.4%. The fossulate, perforated exine is up to 2.3 µm in thickness; the sexine and the nexine are almost equal in thickness. In A. contorta, the equatorial axis of pollen grain is 36.6 µm: the defectiveness percentage, 24.5%. The exine is verrucate, up to 0.3 µm in thickness, while the sexine is two to three times thicker than the nexine. The pollen germination experiments have shown that pollen of A. manshuriensis, in contrast to A. contorta, can germinate in 10-20% sucrose at 22°Ð¡. These data and the high percentage of pollen defectiveness in A. contorta indicate that the androecium function in this species is reduced. The reduction of the androecium function is evidenced by a small amount of pollen grains in anthers or empty anthers and a high percentage of defective pollen grains.

Spatio-temporal patterns in pollination of deceptive Aristolochia rotunda L. (Aristolochiaceae).

Pollination success of highly specialised flowers is susceptible to fluctuations of the pollinator fauna. Mediterranean Aristolochia rotunda has deceptive trap flowers exhibiting a highly specialised pollination system. The sole pollinators are kleptoparasitic flies in search of food. This study investigates these pollinators on a spatio-temporal scale and the impact of weather conditions on their availability. Two potential strategies of the plants to cope with pollinator limitation, i.e. autonomous selfing and an increased floral life span, were tested. A total of 6156 flowers were investigated for entrapped pollinators in 10 Croatian populations. Availability of the main pollinator was correlated to meteorological data. Artificial pollination experiments were conducted and the floral life span was recorded in two populations according to pollinator availability. Trachysiphonella ruficeps (Chloropidae) was identified as dominant pollinator, along with less abundant species of Chloropidae, Ceratopogonidae and Milichiidae. Pollinator compositions varied among populations. Weather conditions 15-30 days before pollination had a significant effect on availability of the main pollinator. Flowers were not autonomously selfing, and the floral life span exhibited considerable plasticity depending on pollinator availability. A. rotunda flowers rely on insect pollen vectors. Plants are specialised on a guild of kleptoparasitic flies, rather than on a single species. Pollinator variability may result in differing selection pressures among populations. The availability/abundance of pollinators depends on weather conditions during their larval development. Flowers show a prolonged trapping flower stage that likely increases outcrossing success during periods of pollinator limitation.

Case study of building of conservation coalitions to conserve ecological interactions.

We engaged experts in various fields of study (pollination ecology, chemical ecology, and ethnobotany), invited community participation, and provided environmental education in an effort to conserve an endangered birthwort (Aristolochia delavayi) and a vulnerable pipevine swallowtail (Byasa daemonius). Scientists studied the uptake and sequestration of the secondary metabolites aristolochic acids from A. delavayi leaves by different stages of pipevine swallowtail as a defense mechanism; low fruit set of the myophilous A. delavayi due to pollinator limitation; and the emission of chemical signals that attract parasitic wasps by the prepupae of B. daemonius. The results of these studies were part of an education program delivered by personnel of non-governmental organizations. The program was devised to deliver information to the public about the health risks of consuming A. delavayi individuals (aristolochic-acid-associated cancers) and to establish a bridge between the public and scientific research. Following delivery of the program, the behavior of residents changed considerably. Community residents were involved in management activities, including participation in a program to promote understanding of ecological interactions between A. delavayi and B. daemonius; designing an in situ conservation site; monitoring A. delavayi and B. daemonius individuals; and promoting the natural fruit set of A. delavayi by scattering animal excrement to attract fly pollinators. The integration of scientific information and community participation appears to have resulted in an increase in abundance of threatened A. delavayi and B. daemonius populations. We believe the involvement of local people in conservation is necessary for successful species conservation.

Time-scale dependency of host plant biomass- and trait-mediated indirect effects of deer herbivory on a swallowtail butterfly.

Despite recent attempts to quantify the relative strength of density- and trait-mediated indirect effects, rarely has the issue been properly addressed at the population level. Most research is based on short-term small-scale experiments in which behavioural and/or physiological responses prevail. Here, we estimated the time-scales during which density- and trait-mediated effects manifest, as well as the strength of these effects, using an interaction chain with three organisms (deer-plant-butterfly). A hierarchical Bayesian model was performed by using a long-term data set of deer density in the Boso Peninsula, central Japan (where local densities differ spatially and temporally) as well as densities of the swallowtail butterfly Byasa alcinous and its host plant Aristolochia kaempferi. The time-scale effect of deer on plant quantity and quality was estimated according to the degree of carry-over effects. The negative influence on leaf density showed a temporal saturation pattern over the long term, while the positive influence on leaf quality due to resprouting of leaves after deer browsing showed no clear temporal trend. The net indirect effect changed from positive to negative with time, with the negative density-mediated effect becoming prominent in the long term. Our novel approach is widely applicable in assessing the dynamic impacts of wildlife if the spatio-temporal variability of expansion and/or invasion history is known.

Aristolochia clematitis, the herb responsible for aristolochic acid nephropathy, in an uncultivated piece of land of an Italian nephrologist.

BACKGROUND: Aristolochia clematitis (AC), a herbaceous plant that belongs to the family of Aristolochiaceae, is today considered as being responsible for Balkan endemic nephropathy (BEN). Very scarce information is available in the medical literature about the presence of AC outside Balkan area. This article reports on the finding of AC in Northwest Italy and the results of a questionnaire delivered to locals on their knowledge about AC. METHODS: AC was found in an uncultivated piece of land of a hilly area of Northwest Italy. It was identified by matching it with images available in the literature and Internet. The questionnaire, which was delivered with a set of 12 photographs and a bunch of true AC, contained 15 questions aimed at collecting information on the knowledge of the respondents about the existence, name, distribution and possible uses of AC. RESULTS: A total of 23 locals, mostly farmers, were interviewed. Among them, 22 (95.6%) had already seen AC, mostly in uncultivated areas; 4 (18%) had a name for it; 21 (95.4%) considered it as a weed and denied any personal use of it; 18 (81.8%) stated that breeding animals disliked AC and no one was aware that AC might damage kidneys. CONCLUSIONS: This study demonstrates that AC can be found outside the Balkan region and that people know it but today do not make any use of it. Other studies carried out by nephrologists in other geographic areas could expand our knowledge about AC outside the basin of BEN.

The betrayed thief - the extraordinary strategy of Aristolochia rotunda to deceive its pollinators.

Pollination of several angiosperms is based on deceit. In such systems, the flowers advertise a reward that ultimately is not provided. We report on a previously unknown pollination/mimicry system discovered in deceptive Aristolochia rotunda (Aristolochiaceae). Pollinators were collected in the natural habitat and identified. Flower scent and the volatiles of insects (models) potentially mimicked were analyzed by chemical analytical techniques. Electrophysiological and behavioral tests on the pollinators identified the components that mediate the plant-pollinator interaction and revealed the model of the mimicry system. The main pollinators of A. rotunda were female Chloropidae. They are food thieves that feed on secretions of true bugs (Miridae) while these are eaten by arthropod predators. Freshly killed mirids and Aristolochia flowers released the same scent components that chloropids use to find their food sources. Aristolochia exploits these components to deceive their chloropid pollinators. Aristolochia and other trap flowers were believed to lure saprophilous flies and mimic brood sites of pollinators. We demonstrate for A. rotunda, and hypothesize for other deceptive angiosperms, the evolution of a different, kleptomyiophilous pollination strategy. It involves scent mimicry and the exploitation of kleptoparasitic flies as pollinators. Our findings suggest a reconsideration of plants assumed to show sapromyiophilous pollination.

Diurnal pattern of stomatal conductance in the large-leaved temperate liana Aristolochia macrophylla depends on spatial position within the leaf lamina.

BACKGROUND AND AIMS: The large distance between peripheral leaf regions and the petiole in large leaves is expected to cause stronger negative water potentials at the leaf apex and marginal zones compared with more central or basal leaf regions. Leaf zone-specific differences in water supply and/or gas exchange may therefore be anticipated. In this study, an investigation was made to see whether zonal differences in gas exchange regulation can be detected in large leaves. METHODS: The diurnal course of stomatal conductance, gs, was monitored at defined lamina zones during two consecutive vegetation periods in the liana Aristolochia macrophylla that has large leaves. Local climate and stem water potential were also monitored to include parameters involved in stomatal response. Additionally, leaf zonal vein densities were measured to assess possible trends in local hydraulic supply. KEY RESULTS: It was found that the diurnal pattern of gs depends on the position within a leaf in A. macrophylla. The highest values during the early morning were shown by the apical region, with subsequent decline later in the morning and a further gradual decline towards the evening. The diurnal pattern of gs at the marginal regions was similar to that of the leaf tip but showed a time lag of about 1 h. At the leaf base, the diurnal pattern of gs was similar to that of the margins but with lower maximum gs. At the the leaf centre regions, gs tended to show quite constant moderate values during most of the day. Densities of minor veins were lower at the margin and tip compared with the centre and base. CONCLUSIONS: Gas exchange regulation appears to be zone specific in A. macrophylla leaves. It is suggested that the spatial-diurnal pattern of gs expressed by A. macrophylla leaves represents a strategy to prevent leaf zonal water stress and subsequent vein embolism.

Characterization of the basal angiosperm Aristolochia fimbriata: a potential experimental system for genetic studies.

BACKGROUND: Previous studies in basal angiosperms have provided insight into the diversity within the angiosperm lineage and helped to polarize analyses of flowering plant evolution. However, there is still not an experimental system for genetic studies among basal angiosperms to facilitate comparative studies and functional investigation. It would be desirable to identify a basal angiosperm experimental system that possesses many of the features found in existing plant model systems (e.g., Arabidopsis and Oryza). RESULTS: We have considered all basal angiosperm families for general characteristics important for experimental systems, including availability to the scientific community, growth habit, and membership in a large basal angiosperm group that displays a wide spectrum of phenotypic diversity. Most basal angiosperms are woody or aquatic, thus are not well-suited for large scale cultivation, and were excluded. We further investigated members of Aristolochiaceae for ease of culture, life cycle, genome size, and chromosome number. We demonstrated self-compatibility for Aristolochia elegans and A. fimbriata, and transformation with a GFP reporter construct for Saruma henryi and A. fimbriata. Furthermore, A. fimbriata was easily cultivated with a life cycle of just three months, could be regenerated in a tissue culture system, and had one of the smallest genomes among basal angiosperms. An extensive multi-tissue EST dataset was produced for A. fimbriata that includes over 3.8 million 454 sequence reads. CONCLUSIONS: Aristolochia fimbriata has numerous features that facilitate genetic studies and is suggested as a potential model system for use with a wide variety of technologies. Emerging genetic and genomic tools for A. fimbriata and closely related species can aid the investigation of floral biology, developmental genetics, biochemical pathways important in plant-insect interactions as well as human health, and various other features present in early angiosperms.

Heat shock effects on seed germination of five Brazilian savanna species.

Fire is considered an important factor in influencing the physiognomy, dynamics and composition of Neotropical savannas. Species of diverse physiognomies exhibit different responses to fire, such as population persistence and seed mortality, according to the fire frequency to which they are submitted. The aim of this study is to investigate the effects of heat shocks on seed germination of Anadenanthera macrocarpa (Benth.) Brenan, Dalbergia miscolobium Benth., Aristolochia galeata Mart. & Zucc., Kielmeyera coriacea (Spreng.) Mart. and Guazuma ulmifolia Lam., which are native species of the Brazilian savanna. The temperatures and exposure times to which the seeds were submitted were established according to data obtained in the field during a prescribed fire: 60 °C (10, 20 and 40 min), 80 °C (5, 10 and 20 min) and 100 °C (2, 5 and 10 min). Untreated seeds were used as controls. Seeds of A. galeata and K. coriacea showed high tolerance to most heat treatments, and seeds of A. macrocarpa showed a significant reduction in germination percentage after treatments of 80 °C and 100 °C. Treatments of 100 °C for 10 min reduced germination percentage for all species except G. ulmifolia, which has dormant seeds. For this species, germination was accelerated by heat treatments. The high temperatures applied did not interfere with the time to 50% germination (T(50) ) of the tolerant seeds. Seeds of the savanna species K. coriacea and A. galeata were more tolerant to heat shocks than seeds of the forest species A. macrocarpa. Guazuma ulmifolia, the forest species with seeds that germinate after heat shock, also occurs in savanna physiognomies. Overall, the high temperatures applied did not affect the germination rate of the tolerant seeds.

Escaping the lianoid habit: evolution of shrub-like growth forms in Aristolochia subgenus Isotrema (Aristolochiaceae).

PREMISE OF THE STUDY: A large range of growth forms is a notable aspect of angiosperm diversity and arguably a key element of their success. However, few studies within a phylogenetic context have explored how anatomical, developmental, and biomechanical traits are linked with growth form evolution. Aristolochia (∼500 species) consists predominantly of climbers, but a handful of shrub-like species are known from Aristolochia subgenus Isotrema (hereafter, shortened to Isotrema). We test hypotheses proposing that the establishment of functional traits linked to lianescence might limit the ability to evolve structurally diverse growth forms, particularly self-supporting forms. • METHODS: We focus on the origin of the shrub habit in Isotrema, from which we sampled representatives from climbing to self-supporting forms. Morphological, anatomical, and biomechanical characters are optimized on a chloroplast- and nuclear-derived phylogeny. • KEY RESULTS: Character-state reconstructions revealed that the climbing habit is plesiomorphic in Isotrema and shrub-like forms are derived from climbers. However, shrubs do not constitute a monophyletic group. Both shrubs and climbers show large multiseriate rays, but differ in terms of vessel size and proportion of fibers and soft tissues. • CONCLUSION: We suggest that while shrub-like species might have partly escaped from the constraints of life as lianas; their height size and stability are not typical of self-supporting shrubs and trees. Shrubs retained lianoid stem characters that are known to promote flexibility such as ray parenchyma. The transitions to a shrub-like form likely involved relatively simple, developmental changes that may be attributed to heterochronic processes.

Family matters: effect of host plant variation in chemical and mechanical defenses on a sequestering specialist herbivore.

Insect herbivores contend with various plant traits that are presumed to function as feeding deterrents. Paradoxically, some specialist insect herbivores might benefit from some of these plant traits, for example by sequestering plant chemical defenses that herbivores then use as their own defense against natural enemies. Larvae of the butterfly species Battus philenor (L.) (Papilionidae) sequester toxic alkaloids (aristolochic acids) from their Aristolochia host plants, rendering larvae and adults unpalatable to a broad range of predators. We studied the importance of two putative defensive traits in Aristolochia erecta: leaf toughness and aristolochic acid content, and we examined the effect of intra- and interplant chemical variation on the chemical phenotype of B. philenor larvae. It has been proposed that genetic variation for sequestration ability is "invisible to natural selection" because intra- and interindividual variation in host-plant chemistry will largely eliminate a role for herbivore genetic variation in determining an herbivore's chemical phenotype. We found substantial intra- and interplant variation in leaf toughness and in the aristolochic acid chemistry in A. erecta. Based on field observations and laboratory experiments, we showed that first-instar larvae preferentially fed on less tough, younger leaves and avoided tougher, older leaves, and we found no evidence that aristolochic acid content influenced first-instar larval foraging. We found that the majority of variation in the amount of aristolochic acid sequestered by larvae was explained by larval family, not by host-plant aristolochic acid content. Heritable variation for sequestration is the predominant determinant of larval, and likely adult, chemical phenotype. This study shows that for these highly specialized herbivores that sequester chemical defenses, traits that offer mechanical resistance, such as leaf toughness, might be more important determinants of early-instar larval foraging behavior and development compared to plant chemical defenses.

Underdeveloped embryos and germination in Aristolochia galeata seeds.

Aristolochiaceae have been described as having seeds with underdeveloped embryos and morphological or morphophysiological dormancy. Aristolochia galeata is a native climber found in the Cerrado biome, associated with road and gallery forest edges. The aims of this study were to investigate: embryo growth rate, morphology and seed germination parameters under different treatments. Embryos were excised to obtain embryo length at four stages: initial, seeds after coat rupture, radicle tip protrusion and cotyledon emergence from the seed coat. Germination tests were conducted at 30 °C under three nitrate concentrations (1, 10 and 20 mM), fluctuating temperature (27/20 °C) and light and dark conditions. We found that seeds have underdeveloped embryos, which take about 301 ± 178 h (±SD) to achieve seed coat rupture, another 205 ± 126 h to reach radicle protrusion and 176 ± 76 h more to the final stage of cotyledon emergence. Germinability was above 52% in all treatments, except in the dark (15%). For all treatments, average germination time was above 290 ± 123 h. Potassium nitrate increased germinability to >87%. No particular treatment was required for embryo development, but seeds in the population that continued to germinate after 1 month were probably in various states of non-deep, simple morphophysiological dormancy. Increased germinability in nitrate treatments and light requirement for germination could prevent germination under unsuitable environmental conditions and be a strategy to increase seedling establishment in the cerrado.

Persistently low fruiting success in the Mediterranean pipevine Aristolochia baetica (Aristolochiaceae): a multi-year study.

Low fruit set is common in many plant species and may be caused by a variety of factors, such as predation, resource limitation or deficient pollination, or it may be an evolutionary strategy. In this paper, we investigate factors that affect fruit set in Aristolochia baetica (Aristolochiaceae), a Mediterranean pipevine found in southwest Spain. Fruit production was monitored in two populations over 4 years (2002-2005), and the causes of flower or fruit loss were determined. Experimental hand-pollinations were performed, and germinated pollen grains on the stigmas of open-pollinated flowers were quantified. Fruit set was always very low (4-14%). Floral abscission initially reduced reproductive output by more than 50%; then herbivory (6-12%) and fruit abortion (8-26%) caused further reductions. Given that the number of efficiently pollinated flowers was always higher than that of ripe fruits, and that xenogamous hand-pollination did not increase fruit set in relation to open-pollination, the final fruit production of A. baetica seems not to be pollen-limited. Fruit abortion of effectively pollinated flowers supports the idea that resource availability limits fruit set. In A. baetica, fruit abortion could lead to mate selection of the best quality fruits. Moreover, the initiated fruits that finally abort could also satiate predators, contributing to increase progeny fitness.

Structure and biomechanics of trapping flower trichomes and their role in the pollination biology of Aristolochia plants (Aristolochiaceae).

*Catching insects to ensure pollination is one of the most elaborate and specialized mechanisms of insect-plant interactions. Phylogenetically, Aristolochiaceae represent the first angiosperm lineage that developed trap flowers. Here we report the structure and function of specific trichomes contributing to the highly specialized trapping devices. *Investigations were carried out on six Mediterranean Aristolochia species. The morphology and arrangement of the trapping trichomes were investigated by scanning electron microscopy (SEM) and cryo-SEM. To demonstrate frictional anisotropy of the trapping trichome array, a microtribological approach was used. *The results of our experiments support a hypothesis long proposed, but never tested, regarding the trapping mechanism in proterogynous Aristolochia flowers: that an array of highly specialized trichomes arranged eccentrically to the underlying surface is responsible for the easy entrance of insects into flowers but impedes their escape. As they enter the male stage of anthesis, flowers significantly modify their inner surface characteristics, allowing insects to leave. *We have demonstrated the substantial contribution of trapping trichomes to the capture, retention and release of pollinators, an important prerequisite for making cross-pollination possible in most Aristolochia species. Finally, we compare trapping trichomes of Aristolochia with similar structures found in other trapping flowers as well as in pitchers of carnivorous plants not optimized for insect release.

Pollinators, flowering phenology and floral longevity in two Mediterranean Aristolochia species, with a review of flower visitor records for the genus.

The pollination of Aristolochia involves the temporary confinement of visitors inside the flower. A literature review has shown that some species are visited by one or a few dipteran families, while others are visited by a wider variety of dipterans, but only some of these are effective pollinators. We observed flowering phenology and temporal patterns of pollinator attendance in diverse populations of Aristolochia baetica and A. paucinervis, two species that grow in SW Spain, frequently in mixed populations. The two species had overlapping floral phenologies, extended flowering periods and long-lived flowers. A. baetica attracted a higher number of visitors than A. paucinervis. Drosophilids and, to a lesser extent, phorids, were the main pollinators of A. baetica, whereas in A. paucinervis, phorids were the only pollinators. Attendance to A. paucinervis flowers by phorids in mixed populations was markedly lower than in pure populations. This effect was more evident in years with lower pollinator density. Our results suggest that A. baetica and A. paucinervis may compete for pollinators in mixed populations.

[Specific features of pollination in the Manchurian birthwort Aristolochia manshuriensis].

An analysis of pollination system in Aristolochia manshuriensis has shown that flower structure in this species is strictly adapted to cross-pollination, but the possibility of an autogamous or geitonogamous type of self-pollination with the involvement of insects is not excluded. The flowers of A. manshuriensis are most frequently visited by flies of the family Anthomyiidae, which markedly contribute to their pollination. Males account for 65% of pollinator insects collected from the flowers.

A novel trade-off of insect diapause affecting a sequestered chemical defense.

Diapause allows insects to temporally avoid conditions that are unfavorable for development and reproduction. However, diapause may incur a cost in the form of reduced metabolic energy reserves, reduced potential fecundity, and missed reproductive opportunities. This study investigated a hitherto ignored consequence of diapause: trade-offs involving sequestered chemical defense. We examined the aristolochic acid defenses of diapausing and non-diapausing pipevine swallowtail butterflies, Battus philenor. Pipevine swallowtail larvae acquire these chemical defenses from their host plants. Butterflies that emerge following pupal diapause have significantly less fat, a female fitness correlate, compared to those that do not diapause. However, butterflies emerging from diapaused pupae are more chemically defended compared to those that have not undergone diapause. Furthermore, non-diapausing butterflies are confronted with older, lower quality host plants on which to oviposit. Thus, a trade-off exists where butterflies may have greater energy reserves at the cost of less chemical defense and sub-optimal food resources for their larvae, or have substantially less energetic reserves with the benefit of greater chemical defense and plentiful larval food resources.

APETALA3 and PISTILLATA homologs exhibit novel expression patterns in the unique perianth of Aristolochia (Aristolochiaceae).

Several lines of evidence suggest that sterile floral organs, collectively known as the perianth, have evolved multiple times during the evolution of the angiosperms. In the family Aristolochiaceae, the perianth is formed by two whorls of organs in the genus Saruma but by only one whorl in the remaining genera, including Aristolochia. Although the morphology of Saruma is similar in appearance to the core eudicot perianth, with leaf-like sepals and showy colored petals, the unipartite perianth of Aristolochia combines morphological aspects of both calyx and corolla. To investigate the organ identity program functioning in the novel perianth of Aristolochia, we identified homologs of the B-class genes APETALA3 (AP3) and PISTILLATA (PI) in both Saruma and Aristolochia. The expression patterns of these genes in Saruma indicate they are functioning in the development of the second whorl petaloid organs and third whorl stamens. In Aristolochia, however, the expression of AP3 and PI homologs in the perianth does not suggest a role in organ identity but, rather, in promoting late aspects of cell differentiation. The implications of these findings for the evolution of both petaloidy and B gene function are discussed.