Dianxiang Zhang

More Miocene Dispersal Between Africa and Asia—the Case of Bridelia (Phyllanthaceae)

Abstract Several hundred angiosperm genera range from Africa to Asia and Australia, among them Bridelia (Phyllanthaceae), with ca. 40 species, including commercially important timber trees. We here use nuclear and chloroplast DNA sequences from herbarium material, plus new collections from China, to test the monophyly of Bridelia and to infer the geologic times when it acquired its disjunct range. For the Southeast Asian mainland, within-species sampling, including material collected close to the type localities, allowed testing current species concepts. Based on a sample of 114 chloroplast matK sequences of Phyllanthaceae, Bridelia is monophyletic and sister to an Asian clade which requires resurrecting an older generic name to make the African Cleistanthus monophyletic. Within Bridelia, gene trees from the combined data (3,177 aligned nucleotides from 25 species of Bridelia plus outgroups) agree with most morphological species boundaries. Exceptions are that B. tomentosa must include B. harmandii and B. curtisii to become monophyletic and that B. fordii is distinct from B. retusa. The topology, together with relaxed clock divergence times, implies that Bridelia dispersed from tropical Asia to Africa once or twice between 10 and 1.85 million years ago (Ma). Australia was reached, probably from New Guinea, at least twice, both times ca. 2 Ma. Together with earlier studies, there are now at least eight cases of Neogene long distance dispersal between Africa and Asia (followed by speciation), with no directional bias apparent so far.

Intrafloral differentiation of stamens in heterantherous flowers.

Flowers that have heteromorphic stamens (heterantherous flowers) have intrigued many researchers ever since the phenomenon was discovered in the 19th century. The morphological differentiation in androecia has been suggested as a reflection of “labor division” in pollination in which one type of stamens attracts pollinators and satisfies their demand for pollen as food and the other satisfies the plants need for safe gamete dispersal. The extent and patterns of stamen differentiation differ notably among taxa with heterantherous flowers. Seven species with heteromorphic stamens in three genera were sampled from Leguminosae and Melastomataceae, and the morphological difference of androecia, pollen content, pollen histochemistry and viability, pollen micro‐morphology, as well as the main pollinators were examined and compared. Pollen number differs significantly between stamen sets of the same flower in most species investigated, and a correlation of pollen number and anther size was substantiated. Higher pollen viabilities were found in the long (pollinating) stamens of Senna alata (L.) Roxb. and S. bicapsularis (L.) Roxb. Dimorphic pollen exine ornamentation is reported here for the first time in Fordiophyton faberi Stapf. The height of stigma and anther tips of the long stamens in natural conditions was proved to be highly correlated, supporting the hypothesis that they contact similar areas of the pollinators body.

Cryptic dioecy in Mussaenda pubescens (Rubiaceae): a species with stigma-height dimorphism

BACKGROUND AND AIMS Evolutionary transitions from heterostyly to dioecy have been proposed in several angiosperm families, particularly in Rubiaceae. These transitions involve the spread of male and female sterility mutations resulting in modifications to the gender of ancestral hermaphrodites. Despite sustained interest in the gender strategies of plants, the structural and developmental bases for transitions in sexual systems are poorly understood. METHODS Here, floral morphology, patterns of fertility, pollen-tube growth and floral development are investigated in two populations of the scandent shrub Mussaenda pubescens (Rubiaceae), native to southern China, by means of experimental and open-pollinations, light microscopy, fluorescence microscopy and scanning electron microscopy combined with paraffin sectioning. KEY RESULTS Mussaenda pubescens has perfect (hermaphroditic) flowers and populations with two style-length morphs but only weak differentiation in anther position (stigma-height dimorphism). Experimental pollinations demonstrated that despite morphological hermaphroditism, the species is functionally dioecious. The long-styled (L) morph possesses sterile pollen and functions as a female, whereas the short-styled (S) morph is female sterile and functions as a male. Self- and intra-morph pollinations of the S-morph were consistent with those expected from dimorphic incompatibility. The two populations investigated were both S-morph (male) biased. Investigations of early stages of floral development indicated patterns typical of hermaphroditic flowers, with no significant differences in organ growth between the floral morphs. Meiosis of microspore mother cells was of the simultaneous type with tetrads isobilateral in shape. The tapetal cells in anther walls of the L-morph became vacuolized during meiosis I, ahead of the uninucleate microspore stage in the S-morph. In the L-morph, the microspore nucleus degenerated at the tetrad stage resulting in male sterility. Microsporogenesis and male gametophyte development was normal in the S-morph. Failure in the formation of megaspore mother cells and/or the development of megagametophytes resulted in female sterility in the S-morph, compared with normal megasporogenesis in the L-morph. CONCLUSIONS In M. pubescens, cryptic dioecy has evolved from stigma-height dimorphism as a result of morph-specific sterility mutations.

Cryptic self-incompatibility and distyly in Hedyotis acutangula Champ. (Rubiaceae)

Distyly, floral polymorphism frequently associated with reciprocal herkogamy, self- and intramorph incompatibility and secondary dimorphism, constitutes an important sexual system in the Rubiaceae. Here we report an unusual kind of distyly associated with self- and/or intramorph compatibility in a perennial herb, Hedyotis acutangula. Floral morphology, ancillary dimorphisms and compatibility of the two morphs were studied. H. acutangula did not exhibit precise reciprocal herkogamy, but this did not affect the equality of floral morphs in the population, as usually found in distylous plants. Both pin and thrum pollen retained relatively high viability for 8 h. The pollen to ovule ratio was 72.5 in pin flowers and 54.4 in thrum flowers. Pistils of pin flowers remained receptive for longer than those of thrum flowers. No apparent difference in the germination rate of pin and thrum pollen grains was observed when cultured in vitro, although growth of thrum pollen tubes was much faster than that of pin pollen tubes. Artificial pollination revealed that pollen tube growth in legitimate intermorph crosses was faster than in either intramorph crosses or self-pollination, suggesting the occurrence of cryptic self-incompatibility in this species. Cryptic self-incompatibility functioned differently in the two morphs, with pollen tube growth rates after legitimate and illegitimate pollination much more highly differentiated in pin flowers than in thrum flowers. No fruit was produced in emasculated netted flowers, suggesting the absence of apomixis. Our results indicate that H. acutangula is distylous, with a cryptic self-incompatibility breeding system.

Duodichogamy and androdioecy in the Chinese Phyllanthaceae Bridelia tomentosa.

Flowering plants commonly separate male and female function in time, but rarely are the two stages synchronized within and among individuals. One such temporal mating system is duodichogamy in which each plant produces two batches of male flowers that are temporally separated by a batch of female flowers, with within-individual synchrony and among-individual asynchrony to ensure mating partners. Duodichogamy is known only from a few species in four genera in unrelated families. We report on duodichogamy in the Chinese tree species Bridelia tomentosa (Phyllanthaceae), a common colonizer of disturbed habitats. In three populations monitored over 2 yr, most trees flowered in the order male → female → male, and resting periods between flowering bouts precluded selfing almost completely. Individuals flowered for several weeks, with the onset of flowering slightly asynchronous among trees. Pollination was by flies, and experimental pollen supplementation of a subset of a trees flowers did not increase fruit set, suggesting high levels of insect visitation and possible resource limitation. Nineteen percent of the 166 trees monitored skipped the first male phase, and another 13% skipped the female phase, remaining male in both years (and also a third year). The regular presence of pure males, if genetically fixed, would make B. tomentosa androdioecious in addition to duodichogamous. Comparison of duodichogamous taxa known so far shows that all have few ovules, fitting with the hypothesis that duodichogamy may result from male competition for access to a small supply of ovules.

A large-scale chloroplast phylogeny of the Lamiaceae sheds new light on its subfamilial classification

Lamiaceae, the sixth largest angiosperm family, contains more than 7000 species distributed all over the world. However, although considerable progress has been made in the last two decades, its phylogenetic backbone has never been well resolved. In the present study, a large-scale phylogenetic reconstruction of Lamiaceae using chloroplast sequences was carried out with the most comprehensive sampling of the family to date (288 species in 191 genera, representing approximately 78% of the genera of Lamiaceae). Twelve strongly supported primary clades were inferred, which form the phylogenetic backbone of Lamiaceae. Six of the primary clades correspond to the current recognized subfamilies Ajugoideae, Lamioideae, Nepetoideae, Prostantheroideae, Scutellarioideae, and Symphorematoideae, and one corresponds to a portion of Viticoideae. The other five clades comprise: 1) Acrymia and Cymaria; 2) Hymenopyramis, Petraeovitex, Peronema, and Garrettia; 3) Premna, Gmelina, and Cornutia; 4) Callicarpa; and 5) Tectona. Based on these results, three new subfamilies—Cymarioideae, Peronematoideae, and Premnoideae—are described, and the compositions of other subfamilies are updated based on new findings from the last decade. Furthermore, our analyses revealed five strongly supported, more inclusive clades that contain subfamilies, and we give them phylogenetically defined, unranked names: Cymalamiina, Scutelamiina, Perolamiina, Viticisymphorina, and Calliprostantherina.

Gelechiidae Moths Are Capable of Chemically Dissolving the Pollen of Their Host Plants: First Documented Sporopollenin Breakdown by an Animal

Background Many insects feed on pollen surface lipids and contents accessible through the germination pores. Pollen walls, however, are not broken down because they consist of sporopollenin and are highly resistant to physical and enzymatic damage. Here we report that certain Microlepidoptera chemically dissolve pollen grains with exudates from their mouthparts. Methodology/Principal Findings Field observations and experiments in tropical China revealed that two species of Deltophora (Gelechioidea) are the exclusive pollinators of two species of Phyllanthus (Phyllanthaceae) on which their larvae develop and from which the adults take pollen and nectar. DNA sequences placed the moths and plants phylogenetically and confirmed that larvae were those of the pollinating moths; molecular clock dating suggests that the moth clade is younger than the plant clade. Captive moths with pollen on their mouthparts after 2-3 days of starvation no longer carried intact grains, and SEM photographs showed exine fragments on their proboscises. GC-MS revealed cis-β-ocimene as the dominant volatile in leaves and flowers, but GC-MS analyses of proboscis extracts failed to reveal an obvious sporopollenin-dissolving compound. A candidate is ethanolamine, which occurs in insect hemolymphs and is used to dissolve sporopollenin by palynologists. Conclusions/Significance This is the first report of any insect and indeed any animal chemically dissolving pollen.

Nuclear ITS Sequences Help Disentangle Phyllanthus reticulatus (Phyllanthaceae), an Asian Species not Occurring in Africa, but Introduced to Jamaica

Abstract Phyllanthus comprises approximately 1,270 species, making it one of the worlds largest plant genera. Molecular data so far are of limited value for understanding Phyllanthus because of the sheer size of the genus. They can, however, help sort-out cryptic species and provide information on the origin of suspected introduced species. One of the seemingly most widespread species is P. reticulatus, which has been recorded from Asia, Australia, Africa, and Jamaica. The name is based on a mixed collection from tropical Asia now in the Lamarck herbarium, and we lectotypify it here. We use nuclear ITS sequences to test the broad treatment of P. reticulatus in recent floras, identify records of “P. reticulatus” from Africa, and investigate the origin of P. reticulatus on Jamaica. A maximum likelihood tree for accessions of P. reticulatus from throughout its supposed range (plus relevant outgroups) shows that the Jamaican plants represent the Asian species, that the African plants called P. reticulatus belong to a separate clade for which P. polyspermus is the oldest available name, and that the sensu lato treatment of P. reticulatus in recent floras is unjustified. Treating the Asian entities P. reticulatus and P. microcarpus as separate species appears justified, and identical ITS sequences in Asian and Jamaican P. reticulatus indicate a recent introduction. The first “island botanists,” J. Macfadyen and N. Wilson, introduced many plants from India in the 1800s, and theirs are the oldest Jamaican collections of P. reticulatus. Since the species was introduced to Jamaica without its obligate Epicephala moth pollinators, it does not set fruit, persisting instead by vegetative growth.

Low genetic diversity and high genetic divergence caused by inbreeding and geographical isolation in the populations of endangered species Loropetalum subcordatum (Hamamelidaceae) endemic to China.

Loropetalum subcordatum (Hamamelidaceae) is one of the most endangered angiosperm species in China. It is narrowly distributed in a few localities in the evergreen broadleaved forest of southern China. Up to now only a few dozen remnant individuals have been found in the four extant populations. In this project, we studied its genetic diversity and population genetic structure using the high resolution molecular marker of amplified fragment length polymorphism. In total, 47 individuals from all the four populations (including all individuals in three populations) were analyzed. Comparably low genetic diversity within populations was revealed and significantly high genetic differentiation among the populations was detected. Four independent groups were identified which corresponded with their geographical ranges. Autogamy is considered to be the major factor contributing to the low genetic variation and high genetic divergence within this species. In addition, small population size, restricted distribution range, geographical isolation and limited seed dispersal may also contribute to the low genetic diversity and high population genetic differentiation. Clonal reproduction was inferred to occur in the two island populations. Suggestions for conservation strategies are provided to preserve the genetic resources of this species.

Pre- and post-zygotic reproductive isolation between co-occurring Mussaenda pubescens var. alba and M. shikokiana (Rubiaceae).

Reproductive isolation is a fundamental requirement for speciation and includes several sequential stages. Few studies have determined the relative contributions of pre- and post-zygotic reproductive isolation in plants, especially between relative species with clear differentiation in flower form. To investigate the mechanisms responsible for reproductive isolation in sympatric Mussaenda pubescens var. alba and Mussaenda shikokiana (Rubiaceae) in Guangxi Province, China, we made observations of flowering phenology, patterns of insect visitation, and conducted pollination experiments, including artificial hybridization. The two species had overlapping flowering times and were pollinated by overlapping pollinators; however, their relative importance differed significantly with M. pubescens visited more commonly by bees and M. shikokiana more frequently by butterflies. Using vegetative and floral characters and molecular evidence based on nuclear ribosomal internal and external transcribed spacer regions we detected seven naturally occurring hybrids among a sample of approximately 125 individuals. Hybrids were characterized by morphologies that most closely resembled their maternal parents based on chloroplast evidence. Studies of artificially synthesized and natural hybrids demonstrated that hybrid seed had very low germination rates and naturally occurring hybrids exhibited pollen sterility. Post-zygotic reproductive isolating mechanisms play a primary role in limiting gene exchange between co-occurring species and maintaining species integrity in areas of sympatry.