Effects of alien invasion by <Emphasis Type="Italic">Bombus terrestris</Emphasis> L. (Apidae) on the visitation patterns of native bumblebees in coastal plants in northern Japan

When alien pollinator species enter a native community of pollinators in which resource partitioning has been established, the pollination network between plants and pollinators may be modified through the interactions between the pollinators over the use of floral resources. We observed the floral-use patterns of native (Bombus hypocrita and B. deuteronymus) and alien (B. terrestris) bumblebee species in a coastal grassland in northern Japan. We analyzed the factors determining resource partitioning patterns. B. hypocrita tended to visit flowers with shallow or wide open corollas, such as Rosa rugosa, whereas B. deuteronymus visited flowers with complex or deeper corollas, such as Lathyrus japonicus. Given the wider floral preference of B. terrestris, floral use by the alien bumblebees consistently overlapped with that of native bumblebees. The visitation of B. terrestris to R. rugosa flowers was positively correlated with that of B. hypocrita. These bumblebee species frequently used similar floral resources, in part because of the large overlap in the seasonality of their foraging activity. The visitation frequency of B. deuteronymus to L. japonicus flowers was independent of the visitation frequency of other bumblebee species. The major visitation periods of the bumblebees to L. japonicus flowers reciprocally differed between B. deuteronymus and B. terrestris, suggesting phenological resource partitioning between these species. Our study suggests that phenological niche partitioning is more common in specialized flowers (L. japonicus) than in generalized flowers (R. rugosa).     

<Emphasis Type="Italic">Tamarix</Emphasis> (Tamaricaceae) hybrids: the dominant invasive genotype in southern Africa

Hybridization is regarded as a rapid mechanism for increasing genetic variation that can potentially enhance invasiveness. Tamarix hybrids appear to be the dominant genotypes in their invasions. Exotic Tamarix are declared invasive in South Africa and the exotic T. chinensis and T. ramosissima are known to hybridize between themselves, and with the indigenous T. usneoides. However, until now, it was not known which species or hybrid is the most prevalent in the invasion. With a biocontrol programme being considered as a way of suppressing the alien Tamarix populations, it is important to document the population genetic dynamics of all species in the region. This investigation sought to identify Tamarix species in southern Africa and their hybrids, describe their population structure, and reveal the geographic origin of the invasive species. To achieve this, nuclear Internal Transcribed Spacer (ITS) sequence data and the multilocus Amplified Fragment Length Polymorphisms (AFLPs) markers were used. Phylogenetic analysis and population genetic structure confirmed the presence of three species in South Africa (T. chinensis, T. ramosissima and T. usneoides) with their hybrids. The indigenous T. usneoides is clearly genetically distant from the alien species T. chinensis and T. ramosissima. Interestingly, the Tamarix infestation in South Africa is dominated (64.7 %) by hybrids between T. chinensis and T. ramosissima. The exotic species match their counterparts from their places of origin in Eurasia, as well as those forming part of the invasion in the US.     

<Emphasis Type="Italic">Bruguiera hainesii</Emphasis>, a critically endangered mangrove species,is a hybrid between <Emphasis Type="Italic">B. cylindrica</Emphasis> and <Emphasis Type="Italic">B. gymnorhiza</Emphasis> (Rhizophoraceae)

Bruguiera hainesii (Rhizophoraceae) is one of the two Critically Endangered mangrove species listed in the IUCN Red List of Threatened Species. Although the species is vulnerable to extinction, its genetic diversity and the evolutionary relationships with other Bruguiera species are not well understood. Also, intermediate morphological characters imply that the species might be of hybrid origin. To clarify the genetic relationship between B. hainesii and other Bruguiera species, we conducted molecular analyses including all six Bruguiera species using DNA sequences of two nuclear genes (CesA and UNK) and three chloroplast regions (intergenic spacer regions of trnL-trnF, trnS-trnG and atpB-rbcL). For nuclear DNA markers, all nine B. hainesii samples from five populations were heterozygous at both loci, with one allele was shared with B. cylindrica, and the other with B. gymnorhiza. For chloroplast DNA markers, the two haplotypes found in B. hainesii were shared only by B. cylindrica. These results suggested that B. hainesii is a hybrid between B. cylindrica as the maternal parent and B. gymnorhiza as the paternal one. Furthermore, chloroplast DNA haplotypes found in B. hainesii suggest that hybridization has occurred independently in regions where the distribution ranges of the parental species meet. As the IUCN Red List of Threatened Species currently excludes hybrids (except for apomictic plant hybrids), the conservation status of B. hainesii should be reconsidered.     

Stable,fertile somatic hybrids between <Emphasis Type="Italic">Sinapis alba</Emphasis> and <Emphasis Type="Italic">Brassica juncea</Emphasis> show resistance to <Emphasis Type="Italic">Alternaria brassicae</Emphasis> and heat stress

Wild relatives of Brassica are a rich reservoir of genes that are invaluable for the improvement of cultivated species. Sinapis alba is a close relative of crop Brassicas that possesses several desirable traits such as tolerance to Alternaria black spot disease, heat stress, insect pests and nematodes. This study is aimed at developing and characterizing hybrids between Brassica juncea and S. alba with the ultimate goal of transferring genes for tolerance to Alternaria brassicae and heat stress, the traits that are lacking in cultivated Brassica. We generated three hybrids between B. juncea and S. alba through protoplast fusion. The hybridity was confirmed through cytology and molecular markers. While two of the hybrids were symmetric, the third one was asymmetric and had greater resemblance to B. juncea. Hybrids showed some characteristic features of the parents and were fully male and female fertile and also set seeds upon back crossing with the parent species. In vitro leaf assay and field inoculation studies revealed that the hybrids are highly resistant to A. brassicae. Besides, hybrids set seeds at temperature of >?38 °C when parents failed to produce seeds indicating that hybrids possess heat tolerance. These stable hybrids provide a reliable genetic resource for transfer of genes from S. alba into cultivated Brassica species.     

<Emphasis Type="Italic">Calcarisporium xylariicola</Emphasis> sp. nov. and introduction of <Emphasis Type="Italic">Calcarisporiaceae</Emphasis> fam. nov. in Hypocreales

During a survey of fungicolous fungi, a novel taxon from the surface of stroma of an unidentified Xylaria species was collected. Phylogenetic analyses showed that this taxon clustered with Calcarisporium sp. and C. arbuscula isolates, but was resolved as a distinct species. A detailed morphological examination coupled with phylogenetic analysis indicated that the taxon represented a new species. Calcarisporium xylariicola sp. nov. is thus introduced. The new taxon is characterized by short conidiophores with swollen bases and less length/width ratio of conidia that distinguish it from other Calcarisporium species. Calcarisporium is presently placed in Hypocreales genera, incertae sedis genus. Species in the genus are largely fungicolous, or occasionally caulicolous or foliicolous, and have hyaline, erect, verticillate conidiophores and sympodial, polyblastic conidiation. A phylogenetic analysis of combined SSU, ITS, LSU, TEF and RPB2 sequence data from Calcarisporium species and other taxa in Hypocreales indicate that Calcarisporium is a distinct lineage from other families. Therefore, a new family, Calcarisporiaceae, in Hypocreales is introduced.     

Molecular and cytological analyses of A and C genomes at meiosis in synthetic allotriploid <Emphasis Type="Italic">Brassica</Emphasis> hybrids (ACC) between <Emphasis Type="Italic">B.napus</Emphasis> (AACC) and <Emphasis Type="Italic">B.oleracea</Emphasis> (CC)

Success of interspecific hybridization relies mostly on the adequate similarity between the implicated genomes to ensure synapsis, pairing and recombination between appropriate chromosomes during meiosis in allopolyploid species. Allotetraploid Brassica napus (AACC) is a model of natural hybridization between Brassica rapa (AA) and Brassica oleracea (CC), which are originally derived from a common ancestor, but genomic constitution of the same chromosomes probably varied among these species through time after establishment, giving rise to cytogenetic difference in the synthetic hybrids. Herein we investigated meiotic behaviors of A and C chromosomes of synthetic allotriploid Brassica hybrids (ACC) at molecular and cytological levels, which result from the interspecific cross between natural B. napus (AACC) and B.oleracea (CC), and the results showed that meiosis course was significantly aberrant in allotriploid Brassica hybrids, and chromosomes aligned chaotically at metaphase I, chromosome bridges and lags were frequently observed from later metaphase I to anaphase II during meiosis. Simultaneously, we also noticed that meiosis-related genes were abruptly down-regulated in allotriploid Brassica hybrids, which likely accounted for irregular scenario of meiosis observed in these synthetic hybrids. Therefore, these results indicated that inter-genomic exchanges of A and C chromosomes could occur frequently in synthetic Brassica hybrids, and provided an efficient approach for genetic changes of homeologous chromosomes during meiosis in polyploid B.napus breeding program.     

Chromosomal distribution of H3K4me2, H3K9me2 and 5-methylcytosine: variations associated with polyploidy and hybridization in <Emphasis Type="Italic">Brachiaria</Emphasis> (Poaceae)

Key message

Assessment of chromosomal distribution of modified histones and 5-methylcytosine shown that there are diversification of chromosomal types among species of Brachiaria and its interspecific hybrids.

Abstract

Histone post-translational modifications and DNA methylation are epigenetic processes that are involved in structural and functional organization of the genome. This study compared the chromosomal distribution of modified histones and 5-methylcytosine (5-mCyt) in species and interspecific hybrids of Brachiaria with different ploidy levels and reproduction modes. The relation between H3K9me2 and 5-mCyt was observed in the nucleolus organizer region, centromeric central domain and pericentromeric region. H3K4me2 was detected in euchromatic domains, mainly in the terminal chromosomal regions. Comparison of chromosomal distribution among species and hybrids showed greater variation of chromosomal types for the H3K9me2 in B. decumbens (tetraploid and apomictic species) and the 963 hybrid, while, for the H3K4me2, the variation was higher in B. brizantha and B. decumbens (tetraploid and apomictic species) and 963 hybrid. The chromosome distribution of 5-mCyt was similar between B. brizantha and B. decumbens, which differ from the distribution observed in B. ruziziensis (diploid and sexual species). Significant alterations in DNA methylation were observed in the artificially tetraploidized B. ruziziensis and in the interspecific hybrids, possibly as result of hybridization and polyploidization processes. The monitoring of histone modifications and DNA methylation allowed categorizing nuclear and chromosomal distribution of these epigenetic marks, thus contributing to the knowledge of composition and structure of the genome/epigenome of Brachiaria species and hybrids. These data can be useful for speciation and genome evolution studies in genus Brachiaria, and represent important markers to explore relationships between genomes.

     

Fossil wood of <Emphasis Type="Italic">Betula erkovetskiensis</Emphasis> sp. nov. (Betulaceae) from the Miocene of the Erkovetskii Brown Coal Field,Amur Region,Russia

A new species, Betula erkovetskiensis Blokhina et O.V. Bondarenko (Betulaceae), from the deposits of the Sazanka Formation (upper Middle?Upper Miocene) of the Erkovetskii Brown Coal Field (Amur Region, Russia) is described based on anatomical features of fossil wood. The new species shows some wood anatomical characters of the extant birch subgenus Betula, B. davurica, B. nigra (section Dahuricae), and B. papyrifera (section Betula). Fossil wood of Betula is found in the Amur Region for the first time.     

Potential allelopathic effects of the tropical legume <Emphasis Type="Italic">Sesbania virgata</Emphasis> on the alien <Emphasis Type="Italic">Leucaena leucocephala</Emphasis> related to seed carbohydrate metabolism

Allelopathy has been considered a key mechanism to explain the invasiveness of some species. It is well known that invasive plants can affect native plants by producing novel allelochemicals but some exotic plant species may be also sensitive to allelochemicals released by native species, providing a tool to reduce growth and impacts of invasive exotic species. Here, using growth chamber experiments we tested the mutual potential allelopathic effects of Sesbania virgata (a native dominant species) and the alien Leucaena leucocephala seeds. S. virgata was unaffected by seed leachates of L. leucocephala, indicating that, under lab conditions, this legume presents resistance to the phytotoxic compounds produced by seeds of this alien species. In contrast, germination and seedling growth of L. leucocephala were strongly affected by the phytochemicals produced by seeds of S. virgata. A delay in endospermic mobilization of storage carbohydrates (raffinose-family oligosaccharides and galactomannan) was observed in the alien species. These potential allelopathic effects could not be attributed sole to the presence neither of the phytoxic catechin nor of ABA in seed leachates of S. virgata. Our findings indicate that the in vitro behavior of S. virgata is consistent with its aggressiveness in natural environment and suggest sesbanimide as a potential candidate as implicated in the noxious effects of S. virgata on the alien species.     

New and little known species of the dance-fly subgenus <Emphasis Type="Italic">Xanthempis</Emphasis> Bezzi,genus <Emphasis Type="Italic">Empis</Emphasis> L. (Diptera,Empididae), from the Caucasus

Five new species of the subgenus Xanthempis Bezzi are described from the Caucasus: Empis (Xanthempis) annae sp. n. (Russia: Krasnodar Territory), E. (X.) grichanovi sp. n. (Russia: Krasnodar Territory; Georgia), E. (X.) pseudoconcolor sp. n. (Russia: Krasnodar and Stavropol territories; Georgia: Abkhazia), E. (X.) teberdaensis sp. n. (Russia: Karachay-Cherkessia), and E. (X.) zamotajlovi sp. n. (Russia: Krasnodar Territory and Adygea). The females of E. (X.) alanica Shamshev and E. (X.) kovalevi Shamshev are described for the first time. New data on the distribution of some previously described species are reported. The geographical distribution of Xanthempis is discussed. A key to Xanthempis species from the Caucasus is compiled.     

On the ecology of some species of genus <Emphasis Type="Italic">Stuckenia</Emphasis> (Potamogetonaceae) in lakes of Zabaykalsky krai and the Republic of Buryatia

This paper clarifies the distribution and ecology of representatives of the genus Stuckenia (S. chakassiensis (Kaschina) Klinkova, S. pectinata (L.) Börner, S. macrocarpa (Dobroch.) Tzvelev, and S. vaginata (Turcz.) Holub) in brackish and saline lakes in eastern Russia. It is shown that S. chakassiensis grows in most Transbaikal lakes with a high salinity instead of S. pectinata, as was described earlier. Therefore, the range of this interesting and still insufficiently known taxon significantly extends to the east of Russia. Being very common in the mineralized waters of West Siberia, S. macrocarpa was not observed in Transbaikalia. In soda waters of Transbaikalia, S. chakassiensis forms thickets with a rather high productivity.     

An overlooked hybrid between the two diploid <Emphasis Type="Italic">Chenopodium</Emphasis> species in Central Europe determined by microsatellite and morphological analysis

The presence and extent of hybridization within the Chenopodium album aggregate (Amaranthaceae) is still unclear. Although many hybrid combinations have been described, their existence in the field has never been systematically studied and verified. The main aim of this study was to ascertain the extent of interspecific hybridization between the diploid species C. ficifolium and C. suecicum using highly variable nuclear microsatellite markers. Due to the absence of such kind of molecular markers for the whole C. album group, we divided the analysis into two steps: (1) Eleven microsatellite loci designed for the closely related species C. quinoa were cross-amplified in five Eurasian species of the C. album diploid–polyploid complex, i.e. C. album s.s. (6x), C. striatiforme (4x), C. strictum (4x), C. ficifolium (2x) and C. suecicum (2x); (2) For the detection of interspecific hybridization between C. ficifolium and C. suecicum, we sampled 480 individuals from five localities in Central Europe. We also investigated morphological differences between the parental taxa and their hybrid and devised a key for their determination. Analysis of variation in microsatellite loci using Bayesian methods, PCoA and Neighbour-joining tree identified 32 F1 hybrids. These F1 hybrids, described here as C. paradoxum Mandák, formed a cluster between well-differentiated parental species, combining the morphological characters of both their parents. Moreover, genetic analyses also recognized several F2 or backcross hybrids, whose delimitation, mainly from C. suecicum and F1 hybrids, based on morphological characters, is problematic.     

Genetic polymorphism,haplotype distribution,and phylogeny of <Emphasis Type="Italic">Daphnia</Emphasis> (Cladocera: Anomopoda) species from the water bodies of russia as inferred from the 16S mtDNA gene sequencing

The data on the genetic polymorphism of the most widespread Daphnia species occupying different water bodies of Russia are presented. The phylogenetic relationships between the examined species were established, and the haplotype networks were constructed. A fragment of the 16S mitochondrial DNA gene was used as a genetic marker. The results of molecular phylogenetic analysis generally coincided with modern concepts in the systematics of the genus Daphnia. The representatives of the divergent mitochondrial lineages within the D. longispina, D. pulex, and D. magna complex remain poorly investigated morphologically. For D. dentifera, a new habitat on the territory of Russia, namely, the water bodies of the Lake Baikal basin, was identified. A conclusion was made that the 16S mtDNA gene could be successfully used in phylogeographic analysis of the genus Daphnia.     

Genomic clues to the parental origin of the wild flowering cherry <Emphasis Type="Italic">Prunus yedoensis</Emphasis> var. <Emphasis Type="Italic">nudiflora</Emphasis> (Rosaceae)

Prunus yedoensis Matsumura is one of the popular ornamental flowering cherry trees native to northeastern Asia, and its wild populations have only been found on Jeju Island, Korea. Previous studies suggested that wild P. yedoensis (P. yedoensis var. nudiflora) is a hybrid species; however, there is no solid evidence on its exact parental origin and genomic organization. In this study, we developed a total of 38 nuclear gene-based DNA markers that can be universally amplifiable in the Prunus species using 586 Prunus Conserved Orthologous Gene Set (Prunus COS). Using the Prunus COS markers, we investigated the genetic structure of wild P. yedoensis populations and evaluated the putative parental species of wild P. yedoensis. Population structure and phylogenetic analysis of 73 wild P. yedoensis accessions and 54 accessions of other Prunus species revealed that the wild P. yedoensis on Jeju Island is a natural homoploid hybrid. Sequence-level comparison of Prunus COS markers between species suggested that wild P. yedoensis might originate from a cross between maternal P. pendula f. ascendens and paternal P. jamasakura. Moreover, approximately 81% of the wild P. yedoensis accessions examined were likely F1 hybrids, whereas the remaining 19% were backcross hybrids resulting from additional asymmetric introgression of parental genotypes. These findings suggest that complex hybridization of the Prunus species on Jeju Island can produce a range of variable hybrid offspring. Overall, this study makes a significant contribution to address issues of the origin, nomenclature, and genetic relationship of ornamental P. yedoensis.     

Genetic analysis of Japanese and American specimens of <Emphasis Type="Italic">Scirpus hattorianus</Emphasis> suggests its introduction from North America

Scirpus hattorianus is a possible alien species in Japan, and a clarification of its unclear taxonomy is required to reveal its origin. It is not known whether the plants initially described from Japan represent the same species distributed in North America. To clarify the origin of the species, we attempted to sequence old specimens collected about 80 years ago using newly designed primer pairs specific for short sequences, including the variable sites. Chloroplast sequences of ndhF were compared among Japanese and North American S. hattorianus, and the closely related species, S. atrovirens, S. flaccidifolius, and S. georgianus. We succeeded in sequencing all samples, and two haplotypes were detected in S. hattorianus: one was unique to the species and the other, detected from specimens potentially collected from the same population as the types, was shared by both North American S. hattorianus and two closely related species, S. atrovirens and S. flaccidifolius. Our results suggest that Japanese S. hattorianus is an alien species that was introduced from North America at least twice.     

Identification and polymorphism of pectinase genes <Emphasis Type="Italic">PGU</Emphasis> in the <Emphasis Type="Italic">Saccharomyces bayanus</Emphasis> complex

Pectinase (endo-polygalacturonase) is the key enzyme splitting plant pectin. The corresponding single gene PGU1 is documented for the yeast S. cerevisiae. On the basis of phylogenetic analysis of the PGU nucleotide sequence available in the GenBank, a family of divergent PGU genes is found in the species complex S. bayanus: S. bayanus var. uvarum, S. eubayanus, and hybrid taxon S. pastorianus. The PGU genes have different chromosome localization.     

Molecular evidence for hybridization between invasive <Emphasis Type="Italic">Solidago canadensis</Emphasis> and native <Emphasis Type="Italic">S</Emphasis>. <Emphasis Type="Italic">virgaurea</Emphasis>

Hybridization between alien and native species is biologically very important and could lead to genetic erosion of native taxa. Solidago × niederederi was discovered over a century ago in Austria and described by Khek as a natural hybrid between the alien (nowadays regarded also as invasive) S. canadensis and native S. virgaurea. Although interspecific hybridization in the genus Solidago is considered to be relatively common, hybrid nature of S. × niederederi has not been independently proven using molecular tools, to date. Because proper identification of the parentage for the hybrid Solidago individuals solely based on morphological features can be misleading, in this paper we report an additive polymorphism pattern expressed in the ITS sequences obtained from individuals representing S. × niederederi, and confirm the previous hypothesis that the parental species of this hybrid are S. canadensis and S. virgaurea. Additionally, based on variability at the cpDNA rpl32-trnL locus, we showed that in natural populations hybridization occurs in both directions.     

Genetic Diversity in the Worldwide <Emphasis Type="Italic">Botrychium lunaria</Emphasis> (Ophioglossaceae) Complex,with New Species and New Combinations

The Botrychium lunaria (Ophioglossaceae) complex worldwide includes the named species B. lunaria, B. crenulatum, B. tunux, and B. yaaxudakeit. These species have been distinguished from each other morphologically and genetically. This study further investigates the genetic diversity and geographic distribution of this complex, examining a large number of plants worldwide. Enzyme electrophoresis was used to examine allelic variation of 22 loci for 1574 plants of putative B. lunaria, B. crenulatum and B. tunux from North America, Eurasia, and New Zealand, and B. dusenii from the Falkland Islands. Variation in allelic composition assessed by genetic identity and cluster analysis using the programs PopGene and STRUCTURE as well as morphology and geography indicated that the complex is composed of six distinct entities; two of which warrant recognition as new species, B. neolunaria, endemic to North America, and B. nordicum, sister to the B. lunaria complex, from Iceland and Norway; and a new combination, B. lunaria var. melzeri , endemic to Greenland, Iceland, and Norway. The new taxa are described in this paper. Three entities within B. tunux are discussed but not proposed for recognition at this time. Botrychium lanceolatum, included in this study, is composed of three morphologically and genetically distinct entities warranting taxonomic recognition.     

Biosystematic study of Berlandiera (Compositae)

Berlandiera is revised to includeB. subacaulis, B. pumila, B. texana, andB. lyrata with its varietieslyrata andmacrophylla. All species may be induced to interbreed readily, producing F₁ progeny which are vigorous but with reduced fertility. Sympatric species produce natural hybrids which are proposed here asB. Xhumilis (=B. pumila XB. subacaulis) andB. Xbetonicifolia (=B. pumila XB. texana). All taxa investigated hadn = 15 chromosomes, including new counts forB. subacaulis and all six F₁ hybrids.