Taxonomic significance of pollen morphology for species delimitation in <Emphasis Type="Italic">Psidium</Emphasis> (Myrtaceae)

Previous studies reported Psidium as one of the most difficult genera to delimit within the American Myrtaceae. Even though palynology has improved the taxonomy of Angiosperms, information about the usefulness of pollen morphology for taxonomic purposes in Myrtaceae remains contradictory. Here, we investigate the significance of pollen morphology for Psidium taxonomy with specific focus on its usefulness for determining species groups of taxonomic significance. Pollen traits observed by light and scanning electron microscopy were quantified and examined using cluster and ordination analyses. Average size of pollen grains was visualized by boxplots. Pollen grains of Psidium are isopolar, oblate, peroblate or oblate-spheroidal, 3-syncolporate or 4-syncolporate. The sexine ornamentation is rugulate, granulate or spinulose-granulate and differs between the mesocolpium and apocolpium. Cluster analysis revealed four distinct groups: Psidium cauliflorum (G1) and Psidium oligospermum (G3) as single-species groups; Psidium brownianum, P. oblongatum, P. ovale, P. sartorianum, P. guajava, Psidium sp. 1, Psidium sp. 2 (G2), and Psidium cattleianum, P. longipetiolatum, P. guineense, P. myrtoides (G4). Supported by ordination analysis, three traits better explained these groups: type of exine ornamentation, size of P-EV and pollen shape. The used approach efficiently distinguished related species, as well as explained species groups of taxonomic significance suggesting pollen morphology to be a significant source of information for taxonomic studies in Psidium.     

Pollen morphology of selected crop plants from southern China and testing pollen morphological data in an archaeobotanical study

Identification of pollen grains of cultivated plants is essential in archaeobotanical studies. In this study, we investigated the pollen morphology of 30 species which are representatives of most of the crop plants in southern China, using a light microscope. Our results show that the pollen grains of these species or genera can generally be identified by their size, aperture(s) and exine sculpture. We found that: (1) some cultivated cereals can be distinguished from wild species of Poaceae according to their size frequency combined with their morphological features; (2) the lengths of the equatorial diameter (E), polar axis (P) and the greatest dimension of the lumina (the size of the network sculpturing) of the exine reticulum may be diagnostic features to distinguish some brassicaceous vegetables. There are significant differences between the E and P values among Brassica campestris (B. rapa, oilseed rape, Chinese cabbage), B. alboglabra (B. oleracea var. alboglabra, gai lan, Chinese kale), B. parachinensis (B. rapa var. parachinensis, choy sum, Chinese flowering cabbage) and B. chinensis (B. rapa ssp. chinensis, pak choi), but moderate differences in the longer axis length of the reticulum lumina, which provide potential for identifying species on the basis of pollen grains. We compared the P values and the longer axis length of the lumina of modern specimens of Brassicaceae pollen grains with those of fossil pollen extracted from the Ming-Qing cultural layer in the Fuqikou site at Chongqing, China, and found that the fossil pollen grains of Brassicaceae probably represent vegetable plants related to B. parachinensis. Moreover, we measured the diameters of rice pollen grains from modern paddy fields to assess the pollen size frequency and found that the size range from ~?34 to 38 µm is closely associated with rice pollen in southern China, which can be used to detect pollen signals of human activities in archaeobotanical investigations.     

Morphological variation in pollen grains of <Emphasis Type="Italic">Mucuna</Emphasis> (Leguminosae): new biogeographic and evolutionary patterns

Mucuna comprises 105 species with an overall pantropical distribution and is divided into three subgenera: M. subg. Mucuna, M. subg. Stizolobium and M. subg. Macrocarpa. Although phylogenetic studies have supported the occurrence of three main clades, evolutionary relationships among them are not fully resolved. The objective of this study was to examine pollen grain morphology from representatives of all three subgenera and map these onto the phylogenetic trees generated by analysis of other characters. Pollen grain surface, form, size, and aperture number were compared. A Bayesian inference tree using matK sequences was constructed. The results indicate that the representatives of M. subg. Macrocarpa have the smallest pollen grains in the genus (a synapomorphic character here identified for this subgenus) and that species of subgenus Mucuna (those with umbelliform inflorescences) have the largest pollen grains. Additional morphological diversity of the pollen grain surface was noted: reticulate and/or micro-reticulate (in all three subgenera), perforate, gemmate or verrucose (only in M. subg. Mucuna). For all studied taxa, the pollen grains are triaperturate, except for two species of M. subg. Mucuna, which have tetraperturate pollen. The phylogenetic tree obtained using the matK marker resolved M. subg. Stizolobium as the earliest diverging lineage in Mucuna. Based on this phylogeny, a reticulate ornamentation pattern of the pollen surface may represent the ancestral state for the genus, while the larger pollen size and the foraminate, gemmate, and verrucose ornamentations are derived characteristics within the genus. These putative derived ornamentations have been observed only in neotropical species.     

Pollen ultrastructure in <Emphasis Type="Italic">Aristolochia manshuriensis</Emphasis> and <Emphasis Type="Italic">A. contorta</Emphasis> (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°С. 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.     

Morphology and variability of pollen of the genus <Emphasis Type="Italic">Spiraea</Emphasis> L. (Rosaceae) in Siberia and the Far East

Pollen of 12 species of the genus Spirea L. growing in different regions of Siberia and the Far East was investigated. The ultrasculpture of pollen grains is described. On the basis of aperture morphology and exine sculpture, pollen grains are typified, with key indices given for their identification. The species of the Spiraria section are distinguished by large grains. Pollen grains of S. media and S. alpina considerably vary in size. The level of natural polymorphism of palynological indices has been established.     

Identification of birch pollen species using FTIR spectroscopy

In this study, the morphology and chemical composition of pollen grains of six birch species (Betula utilis Doorenbos, B. dahurica, B. maximowicziana, B. pendula, B. pubescens and B. humilis) were examined to verify which of these features allow distinguishing them in a more unambiguous way. For this purpose, scanning electron microscopy and light microscopy, as well as Fourier transform infrared (FTIR) spectroscopy and curve-fitting analysis of amide I profile, were performed. The microscopy images show that the pollen grains of B. pubescens, B. pendula and B. humilis are similar in diameter and significantly smaller than those of others species, with the largest diameter observed for B. utilis Doorenbos. However, the results obtained from FTIR spectroscopy indicate that the chemical compositions of B. pubescens and B. pendula are similar, but B. humilis is outlaying. Summarizing, it is not possible to unambiguously state, which feature or which technique is the best for differentiating between the six chosen birch species. However, the study showed that both techniques have potential for identification of birch pollen species.     

Pollen morphology of Microstachys (Euphorbiaceae) with emphasis on neotropical species

Abstract

In Euphorbiaceae tribe Hippomaneae is the pantropical genus Microstachys, comprised by 24 species, mostly from Brazil. Palynological studies with several representatives of the genus are scarce. The purpose of this study was to describe the pollen morphology of Microstachys and find possible differentiating characteristics between its species. Ten species of the genus were analysed, represented by 21 herbarium collections. Their pollen grains were acetolysed, measured and photographed under light microscopy (LM) and scanning electron microscopy (SEM). The analysed species possess pollen in monads, of small to medium size, isopolars, amb distincticly trilobed, oblate-spheroidal, prolate-spheroidal or subprolate, tricolporate with narrow and very long colpi, almost fused at the poles sometimes forming an invagination with tapering ends and margo of the colporus wide to very wide and psilate; endoapertures lalongate to very lalongate with tapering ends, presence of a narrow costae distinct in the endoaperture; exine microreticulate homobrochate, walls with simple columella; at the mesocolpus the sexine separates from the nexine forming cavea; and sexine always thicker than the nexine. Microstachys is a stenopalynous genus, despite its species varying in pollen size, shape and width of the cavea, margo of the colporus, and length of the endoapertures, demonstrating taxonomic value, and indicating a contribution towards future systematic analyses.     

Comparison of the pollen content on the body and in the gut of hoverflies (Diptera,Syrphidae)

We analyzed the pollen content in the gut and on the body of hoverflies (Diptera: Syrphidae), mostly from three genera (Eristalis, Syrphus s. l., and Sphaerophoria) that visited flowers of two jointly flowering species of Apiaceae with indistinguishable pollen (Anthriscus sylvestris and Aegopodium podagraria) and of Bunias orientalis (Brassicaceae). Pollen compositions depended on the plant from which the flies were collected rather than on the insect genus. At the same time, individual preferences of the studied flies to food plant species were confirmed. The pollen compositions in the gut and on the body were weakly correlated. Two possible reasons for such a weak correlation are discussed: individual differences in the fly feeding habits and different time of pollen preservation in the gut and on the body.     

Pollen of Castaneoideae (Fagaceae) from the Holocene sediments of the Sea of Japan

Fossil pollen grains of Castaneoideae from Holocene sediments from the southern Sea of Japan (J-3) at a depth of 200–202, 165–167, and 120–122 cm were studied with a scanning electron microscope. Pollen of some extant species of Castaneoideae, native to the region (Castanopsis cuspidata, C. sclerophylla, and Lithocarpus glaber) were also studied. As a result, four types of fossil pollen grains were revealed; type I is identified to species as Castanopsis cuspidata, type II is referred to the genus Castanopsis, type III is assigned to the genus Lithocarpus, and type IV is only identifined to subfamily (Castaneoideae type).     

Pollen morphology of selected species of the genera <Emphasis Type="Italic">Chrysodracon</Emphasis> and <Emphasis Type="Italic">Dracaena</Emphasis> (Asparagaceae,subfamily Nolinoideae) and its systematic implications

Pollen grains of 3 species from the genus Chrysodracon and 20 species from the genus Dracaena were examined by light, scanning electron and fluorescence microscopy. The basic shape of the pollen grains is subprolate and prolate, but prolate-spheroidal pollen grains may also be found. Pollen of Chrysodracon and Dracaeana is dispersed as monads, rarely as dyads. In terms of size, the pollen grains studied were classified as medium-sized and large. There are two different types of apertures: monosulcate and monoulcerate in Dracaena and monoulcerate in Chrysodracon. The surface of the non-apertural areas is psilate-perforate, irregularly folded, microreticulate and fossulate. Ornamentation of the apertural region is microreticulate, microreticulate-baculate, baculate, psilate-perforate, psilate-perforate-verrucate, granulate and irregularly folded. Irregular perforations are present, and the wall structure is tectate-columellate. Our results suggest that some species of Chrysodracon and Dracaena may be separated based on their pollen grain micromorphology on the distal region.     

Differentiation of the pollen size in five representatives of <Emphasis Type="Italic">Taraxacum</Emphasis> sect. <Emphasis Type="Italic">Palustria</Emphasis>

Measurements of the pollen size in 5 species of Taraxacum sect. Palustria at three levels of ploidy: 2n = 3x = 24 (T. paucilobum), 2n = 4x = 32 (T. vindobonense, T. trilobifolium), 2n = 5x = 40 (T. mendax) and one taxon of unknown number of chromosomes 2n = ? (T. portentosum) are presented in this paper. Obtained results indicate a lack of distinct positive correlation between the pollen size and ploidy in the studied group of plants. Distinct relationship was, however, found between ploidy and the range of pollen size and shape variability. Most variable were the pollen grains of triploid T. paucilobum and the least — those in pentaploid T. mendax. Ranges of pollen variability in tetraploid T. trilobifolium and T. vindobonense and in T. portentosum of unknown number of chromosomes showed intermediate values.     

Comparative exine development from the post-tetrad stage in the early-divergent lineages of Ranunculales: the genera <Emphasis Type="Italic">Euptelea</Emphasis> and <Emphasis Type="Italic">Pteridophyllum</Emphasis>

Studies of pollen wall development produce a great deal of morphological data that supplies useful information regarding taxonomy and systematics. We present the exine development of Euptelea and Pteridophyllum, two taxa whose pollen wall development has never previously been studied using transmission electron microscopy. Both genera are representatives of the two earliest-diverging families of the order Ranunculales and their pollen data are important for the diagnosis of the ancestral pollen features in eudicots. Our observations show these genera are defined by having microechinate microreticulate exine ornamentation, perforate tectum, columellate morphology of the infratectum and the existence of a foot layer and endexine. The presence of lamellations is detected during the early stages of development in the nexine of both genera, especially in the apertures. Euptelea presents remains of the primexine layer during the whole maturation process, a very thin foot layer, and a laminate exinous oncus in the apertural region formed by ectexine and endexine elements. Pteridophyllum has a thicker tectum than Euptelea, a continuous foot layer and a thicker endexine. In the apertures, the exinous oncus is formed by islets and granules of endexine, in contrast to the Euptelea apertures. The secretory tapetum produces orbicules in both genera, but they have different morphology and electron-density. Comparisons with pollen data from related orders and families confirm the ancestral states for the pollen of eudicots proposed in previous studies: reticulate and echinate surfaces, columellate infractectum and a thin foot layer relative to the thickness of the ectexine. According to our observations, we propose considering the possibility of a polymorphic state for the aperture number in the ancestor of Ranunculales, and suggest the development of orbicules as the ancestral state in this order.     

Pollen heteromorphism is pervasive in <Emphasis Type="Italic">Thalictrum</Emphasis> (Ranunculaceae)

Among the angiosperms, features of pollen morphology such as grain size, aperture number and surface ornamentation display striking variation. It is less well appreciated that pollen morphology may vary within and among populations of the same species as well as within individual plants. In some species, individual plants produce multiple types of fertile pollen grains (called pollen heteromorphism). Aspects of pollen morphology, such as aperture number, are likely to affect fertilization success with different morphologies favored in different local competitive and ecological environments. This study surveys variation in pollen grain morphology among species throughout the genus Thalictrum. Pollen from individuals of 36 species was rehydrated from herbarium specimens, and light microscopy was used to quantify pollen grain aperture number and size. I find that pollen aperture-number heteromorphism is present within all Thalictrum species studied, and distributions of aperture-number morphs vary both within and among species. This study provides an example of significant pollen heteromorphism within a genus that also varies widely for pollination mode and sexual system.     

Proline synthesis in developing microspores is required for pollen development and fertility

Background

In many plants, the amino acid proline is strongly accumulated in pollen and disruption of proline synthesis caused abortion of microspore development in Arabidopsis. So far, it was unclear whether local biosynthesis or transport of proline determines the success of fertile pollen development.

Results

We analyzed the expression pattern of the proline biosynthetic genes PYRROLINE-5-CARBOXYLATE SYNTHETASE 1 & 2 (P5CS1 & 2) in Arabidopsis anthers and both isoforms were strongly expressed in developing microspores and pollen grains but only inconsistently in surrounding sporophytic tissues. We introduced in a p5cs1/p5cs1 p5cs2/P5CS2 mutant background an additional copy of P5CS2 under the control of the Cauliflower Mosaic Virus (CaMV) 35S promoter, the tapetum-specific LIPID TRANSFER PROTEIN 12 (Ltp12) promoter or the pollen-specific At5g17340 promoter to determine in which site proline biosynthesis can restore the fertility of proline-deficient microspores. The specificity of these promoters was confirmed by β-glucuronidase (GUS) analysis, and by direct proline measurement in pollen grains and stage-9/10 anthers. Expression of P5CS2 under control of the At5g17340 promoter fully rescued proline content and normal morphology and fertility of mutant pollen. In contrast, expression of P5CS2 driven by either the Ltp12 or CaMV35S promoter caused only partial restoration of pollen development with little effect on pollen fertility.

Conclusions

Overall, our results indicate that proline transport is not able to fulfill the demand of the cells of the male germ line. Pollen development and fertility depend on local proline biosynthesis during late stages of microspore development and in mature pollen grains.

     

Hunting for healthy microbiomes: determining the core microbiomes of <Emphasis Type="Italic">Ceratina,Megalopta</Emphasis>, and <Emphasis Type="Italic">Apis</Emphasis> bees and how they associate with microbes in bee collected pollen

Social corbiculate bees such as honey bees and bumble bees maintain a specific beneficial core microbiome which is absent in wild bees. It has been suggested that maintaining this microbiome can prevent disease and keep bees healthy. The main aim of our study was to identify if there are any core bacterial groups in the non-corbiculate bees Ceratina and Megalopta that have been previously overlooked. We additionally test for associations between the core bee microbes and pollen provisions to look for potential transmission between the two. We identify three enterotypes in Ceratina samples, with thirteen core bacterial phylotypes in Ceratina females: Rosenbergiella, Pseudomonas, Gilliamella, Lactobacillus, Caulobacter, Snodgrassella, Acinetobacter, Corynebacterium, Sphingomonas, Commensalibacter, Methylobacterium, Massilia, and Stenotrophomonas, plus 19 in pollen (6 of which are shared by bees). Unlike Apis bees, whose gut microbial community differs compared to their pollen, Ceratina adults and pollen largely share a similar microbial composition and enterotype difference was largely explained by pollen age. Megalopta displays a highly diverse composition of microbes throughout all adults, yet Lactobacillus and Saccharibacter were prevalent in 90% of adults as core bacteria. Only Lactobacillus was both a core bee and pollen provision microbe in all three species. The consequences of such diversity in core microbiota between bee genera and their associations with pollen are discussed in relation to identifying potentially beneficial microbial taxa in wild bees to aid the conservation of wild, understudied, non-model bee species.     

The genus ixocactus (Loranthaceae,s.s.): Description of its first species

The first species ofIxocactus Rizzini,I. hutchisonii Kuijt, is described and illustrated. The genus is unique in the family in that it is squamate and lacks inflorescences. Other remarkable features are the dense covering of spines, tetracolpate pollen, the small number of pollen grains per anther, the small size of the flower, and the occurrence of this parasite on stems and roots of other mistletoes.Ixocactus is believed to be related toOryctanthus.     

An ultrastructural study of spore wall development and septal pores in species of the <Emphasis Type="Italic">Pachyphlodes</Emphasis> (Pezizaceae,Pezizales) lineage,with a description of the new species <Emphasis Type="Italic">Pachyphlodes annagardnerae</Emphasis>

Pachyphlodes (Pezizaceae) is a genus of truffle-like fungi that is distributed across the Northern Hemisphere. These fungi form ectomycorrhizae primarily with trees in the Fagaceae family, and occasionally with other host plants. The genus Plicariella (= Scabropezia) is phylogenetically inferred as an ally of, or within, the Pachyphlodes lineage. Despite molecular phylogenetic analyses that show the close relationships of species in these two genera, morphological differences in ascomata shape and color, spore ornamentation, and ascus shape are profound. Here, we studied spore wall development to better understand affinities within the Pachyphlodes–Plicariella lineages. Electron microscopy studies indicate that the initial spore wall development is similar across six Pachyphlodes species and a Plicariella species, despite striking differences in mature spore ornamentation among species. Ultrastructural analyses reveal that differences in spore ornamentation among Pachyphlodes species are due to unique developmental events at the final stages of spore wall deposition. Septal pore ultrastructure in Pachyphlodes species is similar to other Pezizaceae that have been studied. Molecular analyses of the five species studied indicate that four of them have not been previously described. The new species Pachyphlodes annagardnerae is here described, and the ultrastructural features of species of Pachyphlodes, Plicariella, and other Pezizales are compared and discussed.     

<Emphasis Type="Italic">Cannabis</Emphasis> is indigenous to Europe and cultivation began during the Copper or Bronze age: a probabilistic synthesis of fossil pollen studies

Conventional wisdom states Cannabis sativa originated in Asia and its dispersal to Europe depended upon human transport. Various Neolithic or Bronze age groups have been named as pioneer cultivators. These theses were tested by examining fossil pollen studies (FPSs), obtained from the European Pollen Database. Many FPSs report Cannabis or Humulus (C/H) with collective names (e.g. Cannabis/Humulus or Cannabaceae). To dissect these aggregate data, we used ecological proxies to differentiate C/H pollen, as follows: unknown C/H pollen that appeared in a pollen assemblage suggestive of steppe (Poaceae, Artemisia, Chenopodiaceae) we interpreted as wild-type Cannabis. C/H pollen in a mesophytic forest assemblage (Alnus, Salix, Populus) we interpreted as Humulus. C/H pollen curves that upsurged and appeared de novo alongside crop pollen grains we interpreted as cultivated hemp. FPSs were mapped and compared to the territories of archaeological cultures. We analysed 479 FPSs from the Holocene/Late Glacial, plus 36 FPSs from older strata. The results showed C/H pollen consistent with wild-type C. sativa in steppe and dry tundra landscapes throughout Europe during the early Holocene, Late Glacial, and previous glaciations. During the warm and wet Holocene Climactic Optimum, forests replaced steppe, and Humulus dominated. Cannabis retreated to steppe refugia. C/H pollen consistent with cultivated hemp first appeared in the Pontic-Caspian steppe refugium. GIS mapping linked cultivation with the Copper age Varna/Gumelni?a culture, and the Bronze age Yamnaya and Terramara cultures. An Iron age steppe culture, the Scythians, likely introduced hemp cultivation to Celtic and Proto-Slavic cultures.     

Revision of the shield-bug genera <Emphasis Type="Italic">Holcostethus</Emphasis> Fieber and <Emphasis Type="Italic">Peribalus</Emphasis> Mulsant et Rey (Heteroptera,Pentatomidae) of the Palaearctic Region

A complex of the heteropteran genera centering around Peribalus Mulsant et Rey and Holcostethus Fieber is considered. The genus Dryadocoris Kirkaldy reveals no relationship with the above genera and is believed to represent a separate clade of the family Pentatomidae. The genera Peribalus and Holcostethus are revised. The former includes three subgenera: Peribalus s. str. with two species, Asioperibalus subgen. n. (type species Cimex inclusus Dohrn) with six species, and Tianocoris subgen. n. (type species Holcostethus manifestus Kiritshenko) with two species. Holcostethus embraces two subgenera: Holcostethus s. str. and the monotypic Enigmocoris subgen. n. (type species H. fissiceps Horváth). Two new species are described: Peribalus tianshanicus sp. n. from the Tien Shan Mts. and P. przewalskii sp. n. from the northern part of China (Huan He River). P. capitatus Jakovlev and P. vernalis (Wolff) are downgraded to subspecies of P. strictus (F.). P. ovatus Jakovlev is synonymized with P. inclusus (Dohrn). Two new monotypic genera related to the revised complex of genera are established, Paraholcostethus gen. n. (type species Peribalus breviceps Horváth) and Himalayastethus gen. n. (type species H. pilosus sp. n. from Kashmir). A key to, and morphometric characters for all the taxa considered are provided. The key characters, including both male and female genitalia, are illustrated, and distributional maps are given.     

Pollen morphology in delimiting subgenera and species of the genus <Emphasis Type="Italic">Cybianthus</Emphasis> s.l. (Myrsinoideae–Primulaceae)

Pollen grains of species of the subgenera of Cybianthus s.l. were analyzed in order to determine if pollen morphology is consistent with the currently accepted infrageneric classification, and if it can provide characters useful for distinguishing species or groups of species. Acetolyzed pollen grains were analyzed, measured, described and photomicrographed, while non-acetolyzed pollen grains were analyzed using scanning electron microscopy. The pollen grains of all species investigated were isopolar monads, small- to medium-sized, subprolate or prolate, 3-colporate, and rugulate or rugulate-perforate. The different analytical methods (statistical analyses, exploratory analysis of the data and cluster analysis) used demonstrated that pollen morphology is informative in delimiting this genus. However, the pollen characters are generally quite uniform among the subgenres of Cybianthus s.l. Thus, the use of the pollen attributes in the separation of the subordinate species to different subgenera is not effective. Pollen morphology did not give support to current subgeneric systematics, but it did support the genus Cybianthus s.l.