Pollen tube growth and the pollen‐tube pathway of Nymphaea odorata (Nymphaeaceae)

An important aspect of the evolution of carpel closure, or angiospermy, is the relationship between pollen tube growth patterns and internalization of the pollen‐tube pathway. True carpel closure, involving postgenital fusion of inner carpel margins, is inferred to have arisen once within the ancient order Nymphaeales, in the common ancestor of Nymphaeaceae. We studied pollen tube development, from pollination to fertilization, in a natural population of Nymphaea odorata, using hand pollinations and timed flower collections. Pollen germinates in stigmatic secretions within 15 min and pollen tubes enter subdermal transmitting tissue within an hour, following wide intercellular spaces towards the zone of postgenital fusion. At the zone of fusion they turn downwards to grow in narrow spaces between interlocked cells and then wander freely to ovules within ovarian secretions. The pollen‐tube pathway is 2–6 mm long and upper ovules are first penetrated 2.5 h after pollination. Pollen tubes grow at rates of approximately 1 mm/h whether in stigmatic fluid, transmitting tissues or ovarian secretions. Pollen‐tube pathways are structurally diverse across Nymphaeales, yet their pollen tubes have similar morphologies and rapid growth rates. This pattern suggests pollen tube growth innovations preceded and were essential for the evolution of complete carpel closure. © 2010 The Linnean Society of London, Botanical Journal of the Linnean Society, 2010, 162 , 581–593.     

Mode of pollen tube growth in pistils of Ticodendron incognitum (Ticodendraceae,Fagales) and the evolution of chalazogamy

Ticodendron incognitum is the sole species of the Ticodendraceae, which was established as a new family in the Fagales less than 20 years ago. Considering the diverse modes of pollen tube growth observed in other Fagales, we investigated the growth of pollen tubes in the pistil of Ticodendron. At the time of pollination, T. incognitum had four immature ovules in a bilocular ovary, thus exhibiting delayed fertilization, as in other Fagales. During the period when fertilization was delayed, pollen tube growth in the pistil was intermittent, consisting of five steps associated with development of the ovules and embryo sacs. Four cessation sites occurred: in the style, in the tissue of the upper part of the ovary, inside and outside of the funicle and at the chalaza. A single pollen tube eventually reaches a mature embryo sac through the chalaza in one of the four ovules. While both delayed fertilization and intermittent pollen tube growth play a role in male and female gametophyte selection, as in other Fagales, the five‐step process of pollen tube growth through the chalaza (i.e. chalazogamy) is characteristic of lineages of the Casuarinaceae, Ticodendraceae and Betulaceae (the latter with the loss of one step). © 2008 The Linnean Society of London, Botanical Journal of the Linnean Society, 2008, 157 , 621–631.     

Stylar polymorphism on the edge: unusual flower traits in Moroccan Narcissus broussonetii (Amaryllidaceae)

Heterostyly and related polymorphisms (e.g. stigma‐height dimorphism) have been used as model systems for studying the origin and maintenance of plant population variability. Stigma‐height dimorphism frequently occurs in Narcissus and is associated with a particular flower shape. In the present study, we describe a new, peculiar case of stigma‐height dimorphism in Narcissus broussonetii, a species on the margin of the geographical distribution of the genus. We determined the stylar condition of N. broussonetii and its variation across populations, analyzed perianth morphology and its relationship with stylar variation, and compared this species with other stylar dimorphic species of the genus. We also studied the incompatibility system and pollination ecology of the species. Narcissus broussonetii is a style‐dimorphic species, as suggested in early studies that were subsequently neglected, and displays unusual flower morphology, with a long floral tube and a virtual absence of a corona. The species shows a late‐ (ovarian) acting incompatibility system and crosses within and between morphs are fertile. We observed short‐tongued diurnal and long‐tongued nocturnal pollinators. Our findings confirm that the presence of the observed dimorphism across populations is most probably the result of the joint action of a nonheteromorphic incompatibility system, extremely long and narrow floral tubes, and a combined role of short‐ and long‐tongued pollinators. © 2015 The Linnean Society of London, Botanical Journal of the Linnean Society, 2015, 177 , 644–656.     

Maintenance of sympatric floral tube‐length variation in a Cape irid

Transition zones between morphologically different individuals of the same species provide insights into the evolution and maintenance of gene‐flow barriers. Here we investigate Tritoniopsis revoluta, which has geographically variable tube lengths, thought to be adaptations to insects with different proboscis lengths. We found a narrow transition zone between plants differing by three‐fold in perianth tube length. We determined whether strong gene‐flow barriers result from assortative mating arising from different pollinators, a high prevalence of selfing, or post‐pollination incompatibilities between plants with different tube lengths. We found that there was little evidence to support assortative mating through different pollinators. Both short‐ and long‐tubed plants were mainly visited by bees with short proboscides. Selfing is unlikely to contribute significantly to seed set, plants with different tube lengths were interfertile and hybrid plants were fertile. We conclude that the contact zone is unstable because these ecotypes have not accrued enough allopatric differences to translate into strong gene‐flow barriers, or, alternatively, bimodality is not a consequence of secondary contact but the result of a novel mutation for short tubes spreading through a long‐tubed population. © 2011 The Linnean Society of London, Biological Journal of the Linnean Society, 2011, 104 , 129–137.     

New Streptocarpus species (Gesneriaceae) from South Africa

Two new rosulate species of Streptocarpus are described from the eastern seaboard of South Africa. The first is endemic to Mpumalanga Province. This species has almost actinomorphic corollas with small cylindrical tubes and was previously included within Streptocarpus parviflorus. However, molecular and morphological data and habitat preference do not support this classification. The second species is from the Msikaba River Gorge in the Eastern Cape Province. It adds to the already impressive list of endemic plant species from this region and is allied to other rosulate species of the Eastern Cape. It approaches Streptocarpus rexii in flower size but differs in its much shorter corolla tubes, which lack purple nectar guides. In addition, the corolla floors are marked with yellow bars reminiscent of Streptocarpus cyaneus and also seen in sympatric populations of the small‐flowered Streptocarpus modestus. © 2008 The Linnean Society of London, Botanical Journal of the Linnean Society, 2008, 158 , 743–748.     

Novel organelle associations in photoreceptors of a serpulid polychaete worm

Cerebral and branchial eyes of serpulid polychaetes have been studied by electron microscopy for the first time. In one species both eye types possess a novel ultrastructure. The receptor cells of the simple cerebral ocelli are rhabdomeric and display a close structural and functional relationship between ciliary rootlets and mitochondria. The receptors of the compound branchial eyes contain both a rhabdom and a stack of photosensitive ciliary membranes.     

Comparative morphology,biology and phylogeny of terminal‐instar larvae of the European species of Toryminae (Hym., Chalcidoidea,Torymidae) parasitoids of gall wasps (Hym. Cynipidae)

We present a phylogenetic and taxonomic study of the morphology and biology of the terminal‐instar larval stage of 19 species representing all the genera of Torymidae parasitoids of gall wasps in Europe, with the single exception of Megastigmus. The genera studied include Adontomerus Nikol'skaya, Idiomacromerus Crawford, Chalcimerus Steffan & Andriescu, Glyphomerus Förster, Pseudotorymus Masi and Torymus Dalman. We primarily used chaetotaxy and some head structures. The terminal‐instar larvae of all studied species are thoroughly described for the first time and illustrated with SEM images. We provide diagnostic characters for the family and the genera studied, and keys to genera and species for the identification of torymid larvae associated with cynipid galls. The majority of the torymid larvae studied are solitary monophagous parasitoids. Finally, to assess the potential use of larval characters in systematic studies of the family, a phylogenetic analysis of the studied taxa based on 42 larval morphological characters is proposed and compared with the current taxonomy of Torymidae. Our results suggest that body chaetotaxy, and characters of the head and mouthparts could be used for genera and species discrimination. © 2008 The Linnean Society of London, Zoological Journal of the Linnean Society, 2008, 154 , 676–721.     

Nectarless flowers with deep corolla tubes in Pedicularis: does long pistil length provide an arena for male competition?

Pollinator‐mediated selection does not seem to have a direct influence on the evolution of a long corolla tube in a nectarless flower. We hypothesized that the long pistil length of the nectarless flower with a deep corolla tube provided an opportunity for male competition. Pedicularis siphonantha, a nectarless and partially self‐incompatible lousewort with substantial variation in corolla tube length, was used to test the hypothesis. We tested whether and how corolla tube length affected seed production per capsule and seed germination rate with different pollination treatments. Flowers were hand‐pollinated with pollen from one self donor and one outcross donor and mixed pollen grains consisting of equal amounts from the two donor types, respectively. Additionally, seeds from open‐pollinated flowers with different corolla tube lengths were collected separately for measurement of germination rate. Pollination treatment and corolla tube length significantly affected number of seeds per capsule. Moreover, a significant positive relationship between seeds per capsule and corolla tube length was found when mixed hand pollination was conducted. Seeds of self hand‐pollinated flowers had a lower germination rate than those from outcross‐pollinated flowers. Under open pollination, seeds from flowers with longer corolla tubes tended to have higher germination rate. In P. siphonantha, outcross pollen may have a higher probability of contributing to the next generation when transferred to flowers with longer corolla tubes. The pistil length, therefore, should be seen as a female choice mechanism, which provides an arena for male‐to‐male competition. © 2015 The Linnean Society of London, Botanical Journal of the Linnean Society, 2015, 179 , 526–532.     

Interactions between hawkmoths and flowering plants in East Africa: polyphagy and evolutionary specialization in an ecological context

Hawkmoths (Lepidoptera, Sphingidae) are considered important pollinators in tropical regions, but the frequency and degree of reciprocal specialization of interactions between hawkmoths and flowers remain poorly understood. Detailed observations at two sites in Kenya over a two‐year period indicate that adult hawkmoths are routinely polyphagous and opportunistic, regardless of their proboscis length. About 700 individuals of 13 hawkmoth species were observed visiting a wide range of plant species at the study sites, including 25 taxa that appear to be specifically adapted for pollination by hawkmoths. We estimate that 277 plant species in Kenya (c. 4.61% of the total angiosperm flora) are adapted for pollination by hawkmoths. Floral tube lengths of these plants have a bimodal distribution, reflecting the existence of two hawkmoth guilds differing in tongue length. Hawkmoths exhibited strongly crepuscular foraging patterns with activity confined to a 20‐min period at dusk and, in some cases, a similar period just before dawn. Corolla tube length appears to act as a mechanical filter as the longest‐tubed plants were visited by the fewest hawkmoth species and these were exclusively from the long‐tongued guild. Tube length showed a strong positive relationship with nectar volume, even after phylogenetic correction, which implies that plants with long corolla tubes are under selection to offer relatively large amounts of nectar to entice visits by polyphagous long‐tongued hawkmoths. Our study shows that diffusely co‐evolved pollination systems involving long‐tongued hawkmoths are clearly asymmetrical, with plants exhibiting a high degree of floral specialization, while hawkmoths exhibit polyphagous behaviour. © 2013 The Linnean Society of London, Biological Journal of the Linnean Society, 2013, 110 , 199–213.     

Commensal symbiosis between agglutinated polychaetes and sulfate‐reducing bacteria

Pendant bioconstructions occur within submerged caves in the Plemmirio Marine Protected Area in SE Sicily, Italy. These rigid structures, here termed biostalactites, were synsedimentarily lithified by clotted‐peloidal microbial carbonate that has a high bacterial lipid biomarker content with abundant compounds derived from sulfate‐reducing bacteria. The main framework builders are polychaete serpulid worms, mainly Protula with subordinate Semivermilia and Josephella. These polychaetes have lamellar and/or fibrillar wall structure. In contrast, small agglutinated terebellid tubes, which are a minor component of the biostalactites, are discontinuous and irregular with a peloidal micritic microfabric. The peloids, formed by bacterial sulfate reduction, appear to have been utilized by terebellids to construct tubes in an environment where other particulate sediment is scarce. We suggest that the bacteria obtained food from the worms in the form of fecal material and/or from the decaying tissue of surrounding organisms and that the worms obtained peloidal micrite with which to construct their tubes, either as grains and/or as tube encompassing biofilm. Peloidal worm tubes have rarely been reported in the recent but closely resemble examples in the geological record that extend back at least to the early Carboniferous. This suggests a long‐lived commensal relationship between some polychaete worms and heterotrophic, especially sulfate‐reducing, bacteria.     

A new species of Chirita (Gesneriaceae) from Yunnan,China

A new species of Gesneriaceae, Chirita maguanensis Z.Y.Li, H.Jiang & H.Xu, is described from Southern Yunnan, China. It is similar to C. eburnea Hance in its large, fleshy, white and parallel‐veined bracts, but differs by having broad and oblique leaves with a repand–crenate or crenate margin, suborbicular bracts, membranous, white calyx, purple corolla‐tube, 2‐parted lower lip of stigma and lower disc. © 2008 The Linnean Society of London, Botanical Journal of the Linnean Society, 2008, 158 , 269–273.     

Variation in nectar–sugar profile of Anchusa and allied genera (Boraginaceae)

The nectar–sugar profile (fructose, glucose and sucrose) of 14 species of Anchusa and five members of the allied genera Anchusella, Cynoglottis, Hormuzakia and Lycopsis (Boraginaceae: tribe Boragineae) was determined. Most of the species examined (c. 74%) produce sucrose‐dominant nectar, whereas the remaining taxa produce sucrose‐rich nectars. Little variation in nectar–sugar composition was found in some species, even when sampling was repeated in different years and/or localities. Average sucrose concentration was 57.75% (coefficient of variation 19.1%). The only floral morphological character that was correlated with the nectar–sugar profile is the length of the corolla tube, as taxa with relatively long floral tubes produce nectar with lower glucose concentrations. The flowering period is also related to sugar composition, as nectar of late‐flowering species contains lower sucrose concentrations. However, small differences in sugar profiles do not reflect phylogenetic relationships based on molecular studies. It would appear that dry habitats and time of flowering are the main determinants of nectar–sugar composition in the genus Anchusa sensu lato. © 2010 The Linnean Society of London, Botanical Journal of the Linnean Society, 2010, 162 , 616–627.     

Ovarian self‐incompatibility in Narcissus papyraceus (Amaryllidaceae) is the result of a pre‐zygotic response

Ovarian self‐incompatibility, including pre‐ and post‐zygotic reactions, is a complex mechanism for which we still lack many details relating to its function and significance. The joint presence of ovarian self‐incompatibility with style polymorphism is a rare combination that is found in the genus Narcissus. Usually, style polymorphic species have heteromorphic (diallelic and linked to style length locus) incompatibility, which prevents fertilization between individuals of the same morph, thereby helping to maintain equal proportions of floral morphs in populations. However, when present, self‐incompatibility in Narcissus is not linked to style polymorphism and cross‐fertilization within each morph is possible. Hence, self‐incompatibility in Narcissus is of particular interest when attempting to unravel the nature of the rejection reaction and aiming to assess possible cryptic differences in the fertilization process in intra‐ and inter‐morph crosses, which might ultimately explain the wide variation of morph‐ratio in the field. We examined the breeding system of Narcissus papyraceus, a style‐dimorphic species that has biased morph ratios in most of its populations. We studied pollen‐tube growth in the pistil and ovule fate after experimentally controlled hand pollinations. The growth of pollen tubes in self‐ and intra‐ and inter‐morph crosses was similar up to the point of micropyle penetration in both morphs but, subsequently, a pre‐zygotic failure appeared to affect male and female gametophytes in selfed pistils. A high proportion of ovules from self‐pollinated flowers showed signs of collapse and self‐pollen tubes were blocked or behaved abnormally before entering the embryo sac. Self‐incompatibility was stronger in the long‐styled morph than in the short‐styled morph. We did not find any conclusive sign of differential functioning between intra‐ and inter‐morph cross‐pollinations in any morph. These results enable us to rule out the possible effects of pollen–pistil interactions in N. papyraceus as a cause of morph‐ratio biases and confirm the exceptional nature of the self‐incompatibility mechanism in this polymorphic species. © 2015 The Linnean Society of London, Botanical Journal of the Linnean Society, 2015, 177 , 629–643.     

Evidence for autonomous selfing in grassland Protea species (Proteaceae)

It has been assumed that species of the large African genus Protea have strong self‐incompatibility systems. However, this assumption was based largely on studies conducted on a clade of bird‐pollinated species that occur in the shrubby fynbos vegetation of the Cape region of southern Africa. To test whether self‐incompatibility occurs in a grassland/savanna Protea clade, which is largely insect‐pollinated, we performed controlled pollination experiments on four species, P. caffra, P. dracomontana, P. simplex and P. welwitschii. Although pollen–ovule ratios of all four species fall within the range for outcrossers, all four species are self‐compatible and capable of autonomous seed production. Using fluorescence microscopy, we found that self‐pollen tubes had the same probability of reaching ovules as cross‐pollen tubes. In the small tree P. caffra, selfed progeny had rates of germination and survivorship that were identical to those of crossed progeny. The grassland Protea spp. studied are likely to have mixed mating systems on account of being both visited by insects and capable of autonomous selfing. If one assumes previous reports of self‐incompatibility in Protea to be reliable, there have been at least five losses of self‐incompatibility and two gains of autonomous selfing in this genus. However, earlier studies in the genus were often methodologically flawed and a thorough re‐analysis of breeding systems in Protea is required. © 2012 The Linnean Society of London, Botanical Journal of the Linnean Society, 2012, 169 , 433–446.     

Benthic Invertebrates Associated with a Serpulid Polychaete Assemblage in a Temperate Estuary

Hydroides dianthus (Verrill), a serpulid polychaete which secretes calcareous tubes, provides microhabitats for a variety of benthic invertebrates in Delaware Bay. These microhabitats function as biologically generated refuges and physical heterogeneities for at least 54 species. Dominance was measured by the biological index value which yielded 12 taxa that comprised 90% by number of the serpulid assemblage. These taxa included seven polychaetes, two amphipods, Nematoda, Oligochaeta, and Harpacticoida. In addition to the refuges provided by the serpulids, it was concluded that this temperate serpulid assemblage was comparable in function but not scale to calcareous biogenic assemblages in lower latitudes.     

Breeding systems in Clivia (Amaryllidaceae): late‐acting self‐incompatibility and its functional consequences

Late‐acting (ovarian) self‐incompatibility, characterized by minimal or zero seed production following self‐pollen tube growth to the ovules, is expected to show phylogenetic clustering, but can otherwise be difficult to distinguish from early‐acting inbreeding depression. In Amaryllidaceae, late‐acting self‐incompatibility has been proposed for Narcissus (Narcisseae) and Cyrtanthus (Cyrtantheae). Here, we investigate whether it occurs in the horticulturally important genus Clivia (Haemantheae) and test whether species in this genus experience ovule discounting in wild populations. Seed‐set results following controlled hand pollinations revealed that Clivia miniata and C. gardenii are largely self‐sterile. Self‐ and cross‐pollinated flowers of both species had similar proportions of pollen tubes entering the ovary, and those of C. gardenii also did not differ in the proportions of pollen tubes that penetrated ovules, thus ruling out classical gametophytic self‐incompatibility acting in the style, but not early inbreeding depression. Flowers that received equal mixtures of self‐ and cross‐pollen set fewer seeds than those that received cross‐pollen only, but it was unclear whether this effect was a result of ovule discounting or interactions on the stigma. The prevention of self‐pollination by the emasculation of either single flowers or whole inflorescences in wild populations did not affect seed set, suggesting that ovule discounting is not a major natural limitation on seed production. Flowers typically produce one to three large fleshy seeds from approximately 16 available ovules, even when supplementally hand pollinated, suggesting that fecundity is mostly resource limited. The results of this study suggest that Clivia spp. are largely self‐sterile as a result of either a late‐acting self‐incompatibility system or severe early inbreeding depression, but ovule discounting caused by self‐pollination is not a major constraint on fecundity. © 2014 The Linnean Society of London, Botanical Journal of the Linnean Society, 2014, 175 , 155–168.     

Tripleurospermum ziganaense (Asteraceae,Anthemideae), a new species from north‐east Anatolia,Turkey

A new species of Tripleurospermum Sch.Bip., Tripleurospermum ziganaense Inceer & Hay?rl?oglu‐Ayaz (Asteraceae, Anthemideae), is described and illustrated. The species grows in open places, on rocky slopes and on roadsides in north‐east Anatolia, Turkey. The diagnostic morphological characters that distinguish it from closely related taxa are discussed, and its conservation status is indicated. © 2008 The Linnean Society of London, Botanical Journal of the Linnean Society, 2008, 158 , 696–700.     

Fruit structure and systematics of Monimiaceae s.s. (Laurales)

Fruit structure (anatomy) was studied in 27 species of 15 genera of Monimiaceae s.s. Almost all have apocarpous gynoecia, with the carpels more or less surrounded by a floral cup. The fruitlets are presented on the opened floral cup, which, depending on its pre‐ and post‐floral development, differentially contributes to the attractive part of the mature fruit. Morphologically similar fruits may differ conspicuously in anatomical structure. Based on anatomical characters two different fruit forms were found: drupe(let)s (with compact sclerenchymatic endocarp forming a stone: putamen) and berry(let)s (with parenchymatic endocarp, and mesocarp parenchyma containing isolated sclereid nests). Four types of drupelets differing by the endocarp structure were tentatively distinguished: (1) the Monimia‐type has a many‐cell‐layered putamen of large isodiametric sclereids, interrupted on the ventral side by few radial rows of small sclereids; (2) the Hortonia‐type has a few‐cell‐layered putamen of isodiametric, especially thick‐walled sclereids – it may be composed of two lateral halves, i.e. with the sclerenchyma partially interrupted on the ventral and dorsal sides (but without rows of small sclereids); (3) the Mollinedia‐type has a few‐cell‐layered putamen, with more or less radially elongate sclereids with wavy cell walls; and (4) the Hedycarya‐type has a one‐cell‐layered putamen of pronouncedly radially elongate sclereids with wavy cell walls. Drupelets of some taxa with a single‐cell‐layered endocarp with only weakly thickened cell walls may represent a transition from drupelets to berrylets. The fruit structure supports three major clades recognized earlier by morphological studies and by molecular phylogenetic analyses: (1) Monimioideae (Monimia‐type drupelets), (2) Hortonieae of Mollinedioideae (Hortonia‐type drupelets), and (3) the remainder of Mollinedioideae (Hedycarya‐ and Mollinedia‐types) and berrylets. Fruit structure also supports the close relationship of Monimiaceae and Lauraceae. © 2007 The Linnean Society of London, Botanical Journal of the Linnean Society, 2007, 153 , 265–285.     

Microscopic evidence of serpulid affinities of the problematic fossil tube ‘Serpula’ etalensis from the Lower Jurassic of Germany

Tube structure, ultrastructure and mineralogy support serpulid affinities of the problematic worm fossil ‘Serpula’etalensis from the Lower Jurassic of Germany. The original tube mineralogy of ‘Serpula’etalensis is purely aragonitic and is preserved in Upper Pliensbachian specimens from eastern Germany. ‘Serpula’etalensis represent the earliest record of aragonitic mineralogy for serpulids. The tube is formed of irregularly oriented prismatic crystals that are 3–6 µm in length and 0.5–1.0 µm in diameter. Calcitic specimens of ‘Serpula’etalensis from Upper Sinemurian of southwestern Germany were recrystallized during the diagenesis and lack the original tube ultrastructure.