A new pollination system: brood-site pollination by flower bugs in Macaranga (Euphorbiaceae)

Background and Aims Macaranga: (Euphorbiaceae) is a large genus of dioecious trees with approx.260 species. To date, only one pollination study of the genushas reported brood-site pollination by thrips in M. hullettii.In this study, the pollination system of Macaranga tanariusis reported. Methods: The study was conducted on Okinawa and Amami Islands, Japan.Flower visitors on M. tanarius were collected and their pollenload and behaviour on the flowers examined, as well as inflorescencestructure and reward for the pollinators. Key Results: The most abundant flower visitors found on the male and femaleinflorescences were Orius atratus (Anthocoridae, Hemiptera),followed by Decomioides schneirlai (Miridae, Hemiptera). Pollenload on O. atratus from flowering pistillate inflorescenceswas detected as well as from staminate flowers. Orius atratusand D. schneirlai are likely to use the enclosed chambers formedby floral bracts as breeding sites before and during floweranthesis, and feed on nectar on the adaxial surface of flowerbracts. The extrafloral nectary has a ball-shaped structureand the contained nectar is not exposed; the hemipterans piercethe ball to suck out the nectar. Conclusions: The results indicate that the plant is pollinated by flowerbugs breeding on the inflorescences. This study may be the firstreport of pollination systems in which flower bugs are the mainpollinators. Similarity of pollination systems between M. hullettiiand M. tanarius indicates that the two brood-site pollinationsystems have the same origin. The pollinator species belongsto a predacious group, whose major prey includes thrips. Thepollination system might represent a unique example of evolutionfrom predatory flower visitors feeding on the pollinators (thrips)to the main pollinators.     

A chloroplast genealogy of myrmecophytic Macaranga species (Euphorbiaceae) in Southeast Asia reveals hybridization, vicariance and long-distance dispersals

Macaranga (Euphorbiaceae) includes about 280 species with a palaeotropic distribution. The genus not only comprises some of the most prominent pioneer tree species in Southeast Asian lowland dipterocarp forests, it also exhibits a substantial radiation of ant-plants (myrmecophytes). Obligate ant-plant mutualisms are formed by about 30 Macaranga species and 13 ant species of the genera Crematogaster or Camponotus. To improve our understanding of the co-evolution of the ants and their host plants, we aim at reconstructing comparative organellar phylogeographies of both partners across their distributional range. Preliminary evidence indicated that chloroplast DNA introgression among closely related Macaranga species might occur. We therefore constructed a comprehensive chloroplast genealogy based on DNA sequence data from the noncoding ccmp2, ccmp6, and atpB-rbcL regions for 144 individuals from 41 Macaranga species, covering all major evolutionary lineages within the three sections that contain myrmecophytes. A total of 88 chloroplast haplotypes were identified, and grouped into a statistical parsimony network that clearly distinguished sections and well-defined subsectional groups. Within these groups, the arrangement of haplotypes followed geographical rather than taxonomical criteria. Thus, up to six chloroplast haplotypes were found within single species, and up to seven species shared a single haplotype. The spatial distribution of the chloroplast types revealed several dispersals between the Malay Peninsula and Borneo, and a deep split between Sabah and the remainder of Borneo. Our large-scale chloroplast genealogy highlights the complex history of migration, hybridization, and speciation in the myrmecophytes of the genus Macaranga. It will serve as a guideline for adequate sampling and data interpretation in phylogeographic studies of individual Macaranga species and species groups.     

Evolution of myrmecophytism in western Malesian Macaranga (Euphorbiaceae)

Plants inhabited by ants (myrmecophytes) have evolved in a diversity of tropical plant lineages. Macaranga includes approximately 300 paleotropical tree species; in western Malesia there are 26 myrmecophytic species that vary in morphological specializations for ant association. The origin and diversification of myrmecophytism in Macaranga was investigated using phylogenetic analyses of morphological and nuclear ITS DNA characters and studies of character evolution. Despite low ITS variation, the combined analysis resulted in a well-supported hypothesis of relationships. Mapping myrmecophytism on all most parsimonious trees resulting from the combined analysis indicated that the trait evolved independently between two and four times and was lost between one and three times (five changes). This hypothesis was robust when tested against trees constrained to have three or fewer evolutionary transformations, although increased taxon sampling for the ITS analysis is required to confirm this. Mapping morphological traits on the phylogeny indicated that myrmecophytism was not homologous among lineages; each independent origin involved a suite of different specializations for ant-plant association. There was no evidence that myrmecophytic traits underwent sequential change through evolution; self-hollowing domatia evolved independently from ant-excavated domatia, and different food-body production types evolved in different lineages. The multiple origins of myrmecophytism in Macaranga were restricted to one small, exclusively western Malesian lineage of an otherwise large and nonmyrmecophytic genus. Although the evolution of aggregated food-body production and the formation of domatia coincided with the evolution of myrmecophytism in all cases, several morphological, ecological, and biogeographic factors appear to have facilitated and constrained this radiation of ant-plants.     

Phylogeography of the Coccus scale insects inhabiting myrmecophytic Macaranga plants in Southeast Asia

Comparative historical biogeography of multiple symbionts occurring on a common host taxa can shed light on the processes of symbiont diversification. Myrmecophytic Macaranga plants are associated with the obligate mutualistic symbionts: Crematogaster (subgenus Decacrema) ants and Coccus scale insects. We conduct phylogeographic analyses based on mitochondrial cytochrome oxidase I (COI) from 253 scale insects collected from 15 locations in Borneo, Malaya and Sumatra, to investigate the historical biogeography of the scales, and then to draw comparisons with that of the symbiotic, but independently dispersing, Decacrema ants which are not specific to different Coccus lineages. Despite the different mode of ancient diversification, reconstruction of ancestral area and age estimation on the Coccus phylogeny showed that the scales repeatedly migrated between Borneo and Malaya from Pliocene to Pleistocene, which is consistent with the Decacrema ants. Just as with the ants, the highest number of lineages in the scale insects was found in northern northwest Borneo, suggesting that these regions were rainforest refugia during cool dry phases of the Pleistocene. Overall, general congruence between the Plio–Pleistocene diversification histories of the symbiotic scales and ants suggests that they experienced a common history of extinction/migration despite their independent mode of dispersal and host-colonization.     

Slippery ant-plants and skilful climbers: selection and protection of specific ant partners by epicuticular wax blooms in Macaranga (Euphorbiaceae)

 In many ant-plant species of the genus Macaranga in South-East Asia, conspicuous blooms of epicuticular wax crystals cover the stem surface. We found that many ant species were unable to walk on these surfaces. Only the specific ant partners of glaucous Macaranga host plants were capable of moving on the slippery stems without difficulty. Therefore, the epicuticular coatings of Macaranga myrmecophytes appear to have a selective function and protect the associated ants against competitors. The epicuticular aggregates function as a physical barrier; no evidence of chemical repellence was found. The extent to which ”foreign” ant species are excluded from a tree strongly depends on inclination, diameter and length of the glaucous stem sections. The particular growth form of some glaucous Macaranga ant-plants enhances the influence of the wax barriers. The ant associates of glaucous and glossy Macaranga ant-plants (genera Crematogaster and Camponotus) differ strongly in their capacity to adhere to the glaucous stems. For this reason, the wax blooms in Macaranga can act as an ecological isolation mechanism for the sympiotic ants. Within the genus Macaranga, we find a high correspondence between the occurrence of glaucousness and obligatory ant association (50% in ant-plants; 6.7% in non-myrmecophytes). The genus Macaranga thus represents one of the few cases known so far where epicuticular wax crystals are likely to have evolved in relation to insects. Received: 2 January 1997 / Accepted: 9 June 1997     

Pollination systems in pioneer trees of the genus Macaranga (Euphorbiaceae) in Malaysian rainforests

Many species of Macaranga (Euphorbiacae) are fast‐growing pioneer trees with an important role in early succession in south‐east Asian rainforests. Within the genus, diverse types of ant–plant associations exist and it has therefore been a model system for studying mutualistic interactions. Little information existed up to now, however, on its reproductive biology. Our comparative study in the genus Macaranga in Sundaland revealed specific flower characteristics and uncommon brood‐site pollination systems: enclosed inflorescence morphologies with narrow entrances strongly restrict the set of flower visitors in many species. Thysanoptera were the most abundant insects in 20 of the 26 investigated Macaranga species and, in three species, heteropteran adults and larvae were dominant. Both insect groups used the flower chambers as breeding sites and fed on nectar‐producing trichomes inside the bracteoles. Thrips as well as heteropterans are assumed to contribute to pollination. Different Macaranga sections were associated with different flower visitors, suggesting isolation by different pollinators. Thrips pollination and myrmecophyty often occurred in the same sections. The development of enclosed flowers might have facilitated tight ant–plant interactions and prevent ant–pollinator conflicts. However, the complex ecosystems in which the mutualistic systems evolved are rapidly changed with unknown consequences for these specific interactions. © 2011 The Linnean Society of London, Biological Journal of the Linnean Society, 2011, 103 , 935–953.     

Comparative chloroplast DNA phylogeography of two tropical pioneer trees, Macaranga gigantea and Macaranga pearsonii (Euphorbiaceae)

Macaranga (Euphorbiaceae) has received much ecological and evolutionary research attention as a genus that includes some of the most conspicuous pioneer trees of Southeast Asian tropical rainforests and because of its manifold associations with ants, including about 30 species that are obligate ant-plants (myrmecophytes). We used sequence data from three chloroplast DNA loci (ccmp5, ccmp6, atpB-rbcL) to assess phylogeographical patterns in species of Macaranga, section Pruinosae, sampled from various regions of Borneo and the Malay Peninsula. Forty-nine chloroplast DNA haplotypes (HT) were identified among 768 specimens from five species, Macaranga gigantea (N = 329; 23 HT), Macaranga pearsonii (N = 347; 21 HT), Macaranga puberula (N = 24; 4 HT), Macaranga hosei (N = 48; 6 HT), and Macaranga pruinosa (N = 20; 5 HT). Forty-one haplotypes were species-specific, whereas eight haplotypes were shared by two, three, or four species and occupied internal positions in a parsimony network. Population genetic parameters based on haplotype frequencies proved to be in a similar range in the non-myrmecophytic M. gigantea and in the ant-associated M. pearsonii, which have overlapping distributions in northern and eastern Borneo. A comparison of G _ST and N _ST values revealed a strong phylogeographic structure in both species, whereas colonization pathways suggested by the network topology were different. Both species exhibited similar levels of haplotypic diversity and moderate to high levels of population differentiation. There were no obvious indications for an influence of the symbiotic ant partners on the population structure of their host plants.     

Isolation and characterization of 11 new microsatellite markers for Macaranga (Euphorbiaceae)

We provide primer sequences for 11 new polymorphic microsatellite markers developed in the tropical ant-plant genus Macaranga (Euphorbiaceae), after enrichment cloning of Macaranga tanarius and Macaranga hypoleuca. Allele numbers per locus ranged from two to 16 among 20 accessions of M. tanarius, and from three to 10 among 22 accessions of M. hypoleuca. Observed and expected heterozygosities ranged from 0.150 to 0.900 and from 0.375 to 0.894 in M. tanarius, and from 0.545 to 1.000 and from 0.434 to 0.870 in M. hypoleuca, respectively. Six of the 11 primer pairs successfully cross-amplified polymorphic polymerase chain reaction products in Macaranga winkleri.     

The effects of the floral infection by a bacterial pathogen in a dioecious plant,Mallotus japonicus (Euphorbiaceae)

Among all the different organs of a plant, flowers might have one of the most dynamic microbial communities, since many microbes are transmitted during flowering by insects and pollen. However, little is known about how these microbes affect floral characteristics and plant reproduction. Among the microbes transmitted to flowers, pathogens may have highly negative effects on plants' fitness. In this study, we investigated whether a bacterial pathogen, Erwinia mallotivora, occurs on flowers of the host plant Mallotus japonicus, and whether the transmission of the pathogen to flowers can result in systemic infection and/or reduction of fruit production. The pathogen has been reported to infect through leaves, while its ecology on flowers is unknown. We first confirmed the presence of the pathogen on flowers, indicating possible transmission by visitors or pollen. Then, we showed that the bacteria can infect the plant through flowers by inoculating the pathogen to both male and female flowers. Interestingly, the symptoms on leaves appeared earlier on the female plants than on the males. Besides, the inoculation significantly decreased fruit set of the female plants. Our results suggest a higher cost of infection in a female than in a male once the pathogen infected flowers. Although the effects of pathogen infection to flowers have rarely investigated in wild plants, it would be an interesting topic for future study if such sexual differences in the infection cost can cause sexual conflict and intraspecific adaptation load.     

Phenology and fecundity in 11 sympatric pioneer species of Macaranga (Euphorbiaceae)in Borneo

Reproductive traits of tropical tree species vary predictably in relation to successional stage, but this variation may be due to the species' phylogenetic histories rather than selective pressures imposed by regeneration requirements. Reproductive phenology, tree size at the onset of reproduction, and fecundity of 11 sympatric, closely related Macaranga species were studied to investigate within-species variation in reproductive traits in relation to resource availability, and among-species variation in relation to other life-history traits (shade tolerance, seed size and maximum tree size, H(max)) and consequently the requirements for forest-gap colonization. Nine species reproduced in synchronous episodes, and two species reproduced continuously over 32 mo. Episodic reproduction was most intense in 1992 following a severe drought. For several species, reproductive trees had greater light availability, lower fecundity in lower light levels, and lower growth rates than nonreproductive trees, reflecting resource-limited reproduction. Among species, H(max) was negatively correlated with shade tolerance and seed size. Tree size at the onset of reproduction and fecundity was strongly linked to this axis of life-history variation, but phenological pattern was not. Absolute tree size at the onset of reproduction was positively correlated with H(max) and negatively correlated with shade tolerance. Relative size at reproductive onset was not correlated with shade tolerance or H(max). Fecundity ranged four orders of magnitude among species and was correlated positively with H(max) and negatively with seed size and shade tolerance. The interrelationships among these reproductive and other life-history traits are strongly correlated with the species' requirements for gap colonization.     

Evolution of obligate pollination mutualism in New Caledonian Phyllanthus (Euphorbiaceae)

About half a dozen obligate pollination mutualisms between plants and their seed-consuming pollinators are currently recognized, including fig-fig wasp, yucca-yucca moth, and the recently discovered Glochidion tree-Epicephala moth mutualisms. A common principle among these interactions is that the pollinators consume only a limited amount of the seed crop within a developing fruit (or fig in the case of fig-fig wasp mutualism), thereby ensuring a net benefit to plant reproduction. A novel obligate, seed-parasitic pollination mutualism between two species of New Caledonian Phyllanthus (Euphorbiaceae), a close relative of Glochidion, and Epicephala moths (Gracillariidae) is an exception to this principle. The highly specialized flowers of Phyllanthus are actively and exclusively pollinated by species-specific Epicephala moths, whose larvae consume all six ovules of the developing fruit. Some flowers pollinated by the moths remain untouched, and thus a fraction of the fruits is left intact. Additional evidence for a similar association of Epicephala moths in other Phyllanthus species suggests that this interaction is a coevolved, species-specific pollination mutualism. Implications for the evolutionary stability of the system, as well as differences in mode of interaction with respect to the Glochidion-Epicephala mutualism, are discussed.     

The significance of ant and plant traits for ant pollination in Leporella fimbriata

Summary Ant metapleural glands secrete surface antibiotics that affect pollen as well as bacteria and fungi. This may be one reason why ant pollination is rare. It is predicted that pollination by ants is possible only in the presence of certain ant and/or plant traits. Two traits are investigated; first, absence of the metapleural glands, and second, the presence of stigmatic secretions that insulate pollen from the ant integument. The pollinator of the orchid Leporella fimbriata is the ant Myrmecia urens. Only one caste is involved, the winged males, and they differ significantly from the queen and worker castes in that they do not possess metapleural glands. This paper reports experiments which test for differential effects on pollen between the males and other castes and evaluates the importance of stigmatic secretions. The results show that the absence of metapleural glands makes no difference as all three castes have strong disruptive effect on pollen artificially applied to the integument. However, during pollination the orchid secures the pollen mass to the ant surface by stigmatic secretions and normal pollen function, fruit production and seed set occur. It appears that both ant and plant traits are pre-adaptive having evolved for functions other than ant pollination.This is contribution number 98 to the Research Unit for Biodiversity and Bioresources, Macquarie University     

Development and characterization of chloroplast microsatellite markers in Macaranga (Euphorbiaceae).

As part of our study on the phylogeography of the ant-plant genus Macaranga, we have screened for polymorphic regions in the chloroplast genome. Initially, ten universal PCR primer pairs targeted at chloroplast microsatellite loci were applied to a small set of specimens, covering various taxonomic levels from intrafamilial to intraspecific. Eight primer pairs produced PCR fragments that behaved as single and discrete bands on agarose gels. The five most promising candidate pairs were further analysed with an extended set of DNA templates, and PCR products were separated on sequencing gels. The number of size variants per locus varied from two to eight, combining into 17 haplotypes among 29 Macaranga accessions from 10 species. Comparative sequencing demonstrated that microsatellites were responsible for the observed size variation at three of five loci, whereas variation at the other loci was caused by larger insertions and (or) deletions (indels). In addition to poly(A) and poly(T) repeats, which are typically found in chloroplast DNA, we also identified a variable (CT)n repeat, with n = 4 to n = 8. Sequencing revealed three examples of size homoplasy, one of which was caused by a single base substitution that raised the actual number of haplotypes to 18. Relationships between haplotypes were assessed by phenetic analyses of size variants and by constructing a parsimony network based on sequence variation. For both types of analysis, the distribution of haplotypes correlated with geographically circumscribed regions rather than with taxonomic boundaries.     

山地五月茶的蝇类传粉研究

观测了山地五月茶Antidesma montanum的开花物候、开花动态、访花者种类和访花行为,并对其繁育系统、花粉组织化学、花粉胚珠比（P／O）、花粉活力进行了检测。结果表明：山地五月茶是雌雄异株植物;其总状花序花期可长达7天;雄花／雌花花序数目为140．33±27．79／208．33±33．65（n=6）,雌雄花的颜色为很淡的黄绿色小型花;雄花单花花期为2天,花药开裂当天有活力,花粉为非淀粉型,花粉胚珠比为3333．33±607．18;单花花蜜量可达0．34±0．03μL,花蜜含糖量3．69％±0．30％。花的结构和开花式样适合蝇类传粉。芳香的气味是吸引蝇类的直接物质：花蜜是传粉者的报酬。主要的传粉昆虫为双翅目Diptera丽蝇科Calliphoridae的Chrysomya megacephala、Chrysomyasp．、寄蝇科Tachinidae的Drino sp．和蝇科Muscidae的Spilogona sp．、Mitroplatia sp．。套网不授粉的处理不结实,表明山地五月茶不存在无融合生殖,人工辅助授粉的坐果率（39．1％）略高于自然坐果率（36．7％）,二者无显著差异,表明其坐果率主要受自身资源分配限制。还讨论了蝇类传粉与雌雄异株性系统以及蝇类传粉与热带林中小型黄绿色花植物的相关性。     

Food bodies and their significance for obligate ant-association in the tree genus Macaranga (Euphorbiaceae)

FIALA, B. & MASCHWITZ, U., 1992. Food bodies and their significance for obligate ant-association in the tree genus Macaranga (Euphorbiaceae). The production of extrafloral nectar and food bodies plays an important role in many tropical ant-plant mutualisms. In Malaysia, a close association exists between ants and some species of the pioneer tree genus Macaranga (Euphorbiaceae). Macaranga is a very diverse genus which exhibits all stages of interaction with ants, from facultative to obligatory associations. The ants nest inside the hollow internodes and feed mainly on food bodies provided by the plants. Food body production had previously been reported only in myrmecophytic Macaranga species, where it is usually concentrated on protected parts of the plants such as recurved stipules. We found that non-myrmecophytic Macaranga species also produce food bodies on leaves and stems, where they are collected by a variety of ants. Levels of food body production differ between facultatively and obligatorily ant-associated species but also among the various non-myrmecophytes. This may be related to the degree of interaction with ants. Food body production starts at a younger age in the myrmecophytic species than in the transitional or non-myrmeccophytic Macaranga. Although food bodies of the non-inhabited Macaranga species are collected by a variety of ants, there is no evidence of association with specific ant species. Our observations suggest that food bodies enhance the evolution of ant-plant interactions. Production of food bodies alone, however, does not appear to be the most important factor for the development of obligate myrmecophytism in Macaranga.     

A revision of Ptychopyxis (Euphorbiaceae) in southeast Asia

Ptychopyxis is a genus comprised of 11 species. Nine are found in southeast Asia mainland and west Malesia and two species are endemic to New Guinea. One of the endemic New Guinean species is newly described here, viz. P. esdras Stoops sp. nov.     

东南亚兰科植物的物种多样性、生活习性及其传粉系统（附表）

兰科(Orchidaceae)植物广布于除两极和极端沙漠地区外的各种陆地生态系统,包括5个亚科800多属28 000多种。东南亚地区兰科植物种数约占世界的1/3,是兰科植物生物多样性热点区域之一。通过查阅文献及书籍等资料,该文系统整理了东南亚兰科植物物种种类及其扩散演化历史,并对其生活习性和传粉系统进行了归类。结果表明:(1)东南亚兰科植物8 855种,分属5亚科17族26亚族240属;(2)主要生活型为附生的有127属6 000种以上,地生97属2 000种以上,腐生13属约100种,藤本4属40余种;(3)根据整理出的东南亚79个属的兰科植物传粉系统发现,有44个属含有自动自交的物种,具报酬物的传粉系统有花粉(仅见于拟兰亚科)、芳香类物质(仅见于香荚兰亚科)和花蜜(5个亚科均有)等报酬物类型。欺骗性传粉系统广泛存在于各个亚科,包括食源性欺骗、性拟态、繁殖地拟态和信息素拟态等类型。东南亚兰科植物在物种、生活习性及传粉系统都展现出极高的多样性,对这些生物学特点的总结将为兰科植物的保育提供一定的理论基础和本底资料。