Hybridization with invasive Rosa rugosa threatens the genetic integrity of native Rosa mollis

Hybridization between invasive and native species often has severe consequences on fitness and survival of the native relative. We investigated the extent of hybridization between the neophyte Rosa rugosa and native R. mollis, an endangered species in Germany. Rosa mollis is found in only one large population in Germany close to the Baltic coast, which has been heavily invaded by R. rugosa for at least 60 years. We analysed all individuals of R. mollis from this mixed population using microsatellite markers and morphological characters and compared these data with those from allopatric populations of R. mollis and R. rugosa. In the mixed population we identified nine plants (45% of the population) as hybrids between R. mollis (seed parent) and R. rugosa (pollen parent) by the presence of microsatellite alleles private for R. rugosa. These individuals were also morphologically intermediate between the parental species. Gene flow from R. mollis into R. rugosa was negligible. We detected a very low genetic diversity and a low number of seeds per hip in the mixed population of R. mollis, pointing to genetic depletion and low fitness. In the light of these results and the difficulties in removing invasive R. rugosa from European coastlines, we discuss possible conservation strategies for this endangered population. © 2012 The Linnean Society of London, Botanical Journal of the Linnean Society, 2012, 170 , 472–484.     

Low genetic and morphological differentiation in the European species complex of Rosa sherardii,R. mollis and R. villosa (Rosa section Caninae subsection Vestitae)

The polyploid species complex of Rosa villosa sensu lato (Rosa section Caninae subsection Vestitae) consists of three morphologically similar polyploid species: R. sherardii, R. mollis and R. villosa. Whereas R. sherardii is distributed in central Europe, R. mollis and R. villosa represent a vicariant species pair occurring in northern Europe and in mountains of central and eastern Europe, respectively. In this study we analysed multiple data sets (morphology, cytology, microsatellites, AFLP and plastid DNA sequences) to re‐evaluate the systematics of these species and to examine whether cytological differences are reflected in the taxonomy and geographical distributions. Furthermore, these data sets were used to evaluate hypotheses explaining the vicariant distribution of R. mollis and R. villosa. None of these data sets revealed a clear‐cut differentiation between the species. Cytological and molecular data argued for a discrete taxonomic position of the predominantly pentaploid R. sherardii, but these data did not support a separation between the mostly tetraploid R. mollis and R. villosa. Population genetics revealed that samples of the latter species were assembled according to ploidy, but not to species affiliation or geographical distribution. Thus, we assume that the cytologically polymorphic original species had a continuous range prior to the last glaciation period and survived on nunataks or in non‐glaciated coastal regions in northern Europe, but that it failed to recover its former range after the retreat of the ice sheets. © 2013 The Linnean Society of London, Botanical Journal of the Linnean Society, 2014, 174 , 240–256.     

Physical mapping of ribosomal DNA on several species of the subgenus Rosa

The localization of NORs by fluorescent in situ hybridization on metaphase spreads of five diploid (Rosa gigantea Coll., Rosa moschata Herrm., Rosa multiflora Thunb., Rosa rugosa Thunb. and Rosa sempervirens L., 2n=2x=14), one triploid (Rosa chinensis’semperflorens’ Koehne., 2n=3x=21) and one tetraploid (Rosa gallica ’versicolor’ L., 2n=4x=28) ancestral species of modern roses was studied. Two terminal hybridization signals were observed in all diploid species corresponding to a single NOR per genome in these species. Triploid R. chinensis showed three hybridization sites on the short arm of three morphologically similar chromosomes. Six hybridization sites, located at terminal positions of the short arms of three chromosome pairs, were observed in the tetraploid species. These signals corresponded to three pairs of NORs and all of them were located in chromosome pairs of different size. These observations, together with the analysis of meiotic pairing in PMCs, support the view that our specimen of R. chinensis is an autotriploid and that R. gallica’versicolor’ has an alloploidy nature. Received: 27 November 2000 / Accepted: 12 March 2001     

The expression level of <Emphasis Type="Italic">Rosa Terminal Flower 1</Emphasis> (<Emphasis Type="Italic">RTFL1</Emphasis>) is related with recurrent flowering in roses

We examined the relationship between the recurrent flowering character and the expression patterns of TERMINAL FLOWER 1 (TFL1) homologs in roses, using flower buds of Rosa multiflora, R. rugosa, R. chinensis, and six other rose species and nine rose cultivars. RTFL1 (Rosa TFL1) genes were amplified from rose genomic DNA using a combination of degenerate and gene-specific primers by thermal asymmetric interlaced-PCR and normal PCR, respectively. Their copy numbers in different species were determined by Southern blots. We used real-time PCR to analyze the expression patterns of RTFL1 genes at four developmental stages (pre-sprouting, young, mid-aged, and mature flower buds). Our results show that there are at least three RTFL1 homologs in roses; RTFL1a, RTFL1b, and RTFL1c. The sequences of the homologs were more similar among the same homolog in different species than among the different homologs in the same species. For RTFL1a, we detected two copies in R. multiflora, two copies in R. rugosa, and one copy in R. chinensis. For RTFL1c, we detected one copy in R. multiflora, two copies in R. rugosa, and three copies in R. chinensis. We detected only one copy of RTFL1b in R. chinensis. RTFL1c was expressed at high levels at all four flowering stages in R. multiflora and R. rugosa, which are non-recurrent flowering species, whereas it was barely detected in R. chinensis (a recurrent flowering species) at any stage. These results were further verified in six other non-recurrent flowering species and nine recurrent flowering cultivars. These results suggest that the recurrent flowering habit in roses results from lower expression of RTFL1c, which may be related to recurrent flowering character in roses.     

Evidence of apomixis in hemisexual dogroses, Rosa section Caninae

All members of Rosa section Caninae, dogroses are polyploid and characterized by their unbalanced meiosis, which in most cases leads to a pronounced morphological influence from the maternal parent. In a previous investigation on a pair of reciprocal crosses between two species in this section, Rosa dumalis and R. rubiginosa (2n=35), nine offspring plants (approximately 10%) did not receive any of the 21 RAPD markers present in the respective pollen parent. This was interpreted as a possible occurrence of apomixis. These nine plants have now been subjected to a further study with additional markers. Thirteen new RAPD markers showed the same result as in the previous investigation: none of the nine plants inherited any of the pollen donor markers. The reproducibility of the RAPD markers was checked by mixing DNA samples to obtain a series of artificial hybrids between the two parent plants. Twelve RAPD markers gave the expected result, whereas one marker appeared only 50% of the time. In addition, pollen viability, mean number of seeds per hip, mean seed weight, and mean weight of fruit flesh per hip have been studied on the four progeny groups: R. dumalis×R. rubiginosa plants which received pollen donor markers (PM plants), R. dumalis×R. rubiginosa plants which did not receive any pollen donor markers (NPM plants), R. rubiginosa×R. dumalis PM plants and R. rubiginosa×R. dumalis NPM plants. A canonical discriminant analysis based on these four reproductive characters separated the four progeny groups. There were significant differences between the two PM groups in all investigated characters, and also between the PM and the NPM groups in pollen viability. The result from the RAPD markers together with the differences in pollen viability between the PM and NPM progeny groups is taken as an indication that apomixis occurs within the Caninae section. Received: 13 October 1999 / Revision accepted: 28 January 2000     

Genetic structure in chloroplast and nuclear microsatellites in Rosa rugosa around sea straits in northern Japan

Sea‐strait geological history and seawater dispersal patterns can affect the genetic structure of coastal plants. We investigated microsatellite (simple sequence repeat, SSR) variations in chloroplast (cp) and nuclear (n) DNA of the coastal shrub R osa rugosa, native to northeastern Asia, and three species closely related to it. In northern Japan, we sampled R. rugosa along coasts around the Soya and Tsugaru straits with different histories of the Quaternary land formation as a result of different water depths. Both cpSSR and nSSR variations suggest rare hybridization between R . rugosa and the closely related species. Variations in one of two cpSSRs showed genetic differentiation between eastern and western Hokkaido. This genetic structure may result from introgression from R. davurica in eastern Hokkaido and/or isolation between the eastern and western coastlines of Hokkaido as a result of the Quaternary land formation in the Soya Strait. On the other hand, variations in 10 nSSRs were geographically homogeneous with weak isolation by distance along coastlines. These results suggest that extensive gene flow has been homogenizing the genetic structure of R . rugosa in northern Japan.     

Development of a genomic in situ hybridization method using Technovit 7100 sections of early wheat embryo

It is difficult to observe the behavior of chromosomes in early wheat embryos because they are wrapped in several cell layers of the ovary. Here we conducted genomic in situ hybridization on sections of ovary embedded in Technovit 7100, a resinous compound suitable for in situ hybridization of mRNA in sectioned tissues. With this resin it is possible to make thin sections with high resolution, no autofluorescence, and good water permeability. These features enable histochemical study using fluorescence microscopy. We established the most suitable conditions for the denaturation of target DNA embedded in Technovit resin, and performed GISH on them. Using this method, we identified Leymus mollis chromosomes in the young ovary of F₁ hybrids between wheat and L. mollis. Furthermore, we observed the behavior of maize chromosomes in early wheat × maize hybrid embryos.     

Karyotype and genomic in situ hybridization pattern in ×Brassicoraphanus, an intergeneric hybrid between Brassica campestris ssp. pekinensis and Raphanus sativus

The karyotype and genomic in situ hybridization (GISH) of an intergeneric hybrid Baemoochae, ×Brassicoraphanus, which originated from hybridization between Chinese cabbage, Brassica campestris (synonym, rapa) ssp. pekinensis, and radish, Raphanus sativus, were analyzed to determine its chromosome complement. In the karyotype analysis, B. campestris was verified to have 2n = 20 chromosomes, including a particular pair of the subtelomeric chromosomes with the nucleolar organizer; R. sativus to have 2n = 18 chromosomes, including a particular pair of the submetacentric chromosomes with the secondary constriction of nucleolar organizer; and ×Brassicoraphanus to have 2n = 38 chromosomes, including both the subtelomeric chromosomes of Brassica and the secondary constriction chromosome pair of Raphanus. These findings indicate that ×Brassicoraphanus is a polyploid between Brassica and Raphanus. In the GISH analysis using chromosomes of B. campestris and R. sativus as the probe and blocking DNA, respectively, only 20 chromosomes of Brassica had hybridization signals. This result reveals that ×Brassicoraphanus is an intergeneric hybrid consisting of the complete genomes of both Brassica and Raphanus. However, the nucleolar organizers of Brassica and Raphanus were not identified because the hybridization signals appeared to be centering mainly around the centromere, becoming weak at the edges.     

Habitat invasion risk assessment based on Landsat 5 data,exemplified by the shrub Rosa rubiginosa in southern Argentina

Prediction of invasive species spread helps to plan management actions. We performed a risk assessment by quantifying habitat invasibility, predicted the potential distribution of an invasive species using the Maxent modelling program and confirmed patterns using detailed field studies. Our study was conducted in southern Argentina, Patagonia, where large areas are already invaded by the European shrub Rosa rubiginosa. A total of 163 R. rubiginosa locations served as ground truth data, and predictors were obtained from the spaceborne sensor Landsat 5. Based on the Maxent Method (area under the receiver operating characteristic curve 0.8), the habitat invasibility map covered about 5000 km². Our model revealed that R. rubiginosa has the potential to invade 36% of the area along a steep precipitation gradient (target region 600–1400 mm per year). The Tasseled Cap brightness index and the normalized vegetation index explained most of the variance in our model, followed by the Tasseled Cap greenness and wetness indices, which can be interpreted as indicators of disturbance. Highest levels of invasibility were predicted for urban areas, along roads and rivers, on pastures, in Austrocedrus chilensis forests and inside Nothofagus dombeyi forest gaps. Detailed field assessments of rose cover performed in seven habitat types supported these results: rose cover significantly decreased with increasing tree cover (P < 0.01). Our data revealed that the occurrence of R. rubiginosa is not connected to a certain habitat type, but that it thrives in open patches following habitat disturbance. Our approach is a widely applicable, cost‐free remote sensing method that can serve as a risk assessment tool for alien plant species invasion of habitats.     

Two new spontaneous hybrids of American hard pines from Pinus sect. Trifoliae (Pinaceae) found in the unique Russian Sochi Arboretum

In the 1990s, an extraordinarily abundant live collection of the genus Pinus, including 49 species, 4 subspecies, and 7 interspecific hybrids, found in the Sochi Arboretum in Russia on the northeast coast of the Black Sea, was taxonomically revised. Among these taxa, two previously unknown interspecific hybrids of American hard pine species from P. sect. Trifoliae had arisen spontaneously in culture. These are described here as the new hybrid species Pinus × transamericana (P. muricata × P. rigida) and Pinus × critchfieldii (P. patula × P. taeda). All known natural or artificial interspecific hybrids within P. sect. Trifoliae are listed, and hybridization between American hard pine species is discussed. (© 2012 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Functional diversification in the Nudix hydrolase gene family drives sesquiterpene biosynthesis in Rosa × wichurana

Roses use a non‐canonical pathway involving a Nudix hydrolase, RhNUDX1, to synthesize their monoterpenes, especially geraniol. Here we report the characterization of another expressed NUDX1 gene from the rose cultivar Rosa x wichurana, RwNUDX1‐2. In order to study the function of the RwNUDX1‐2 protein, we analyzed the volatile profiles of an F₁ progeny generated by crossing R. chinensis cv. ‘Old Blush’ with R. x wichurana. A correlation test of the volatilomes with gene expression data revealed that RwNUDX1‐2 is involved in the biosynthesis of a group of sesquiterpenoids, especially E,E‐farnesol, in addition to other sesquiterpenes. In vitro enzyme assays and heterologous in planta functional characterization of the RwNUDX1‐2 gene corroborated this result. A quantitative trait locus (QTL) analysis was performed using the data of E,E‐farnesol contents in the progeny and a genetic map was constructed based on gene markers. The RwNUDX1‐2 gene co‐localized with the QTL for E,E‐farnesol content, thereby confirming its function in sesquiterpenoid biosynthesis in R. x wichurana. Finally, in order to understand the structural bases for the substrate specificity of rose NUDX proteins, the RhNUDX1 protein was crystallized, and its structure was refined to 1.7 Å. By molecular modeling of different rose NUDX1 protein complexes with their respective substrates, a structural basis for substrate discrimination by rose NUDX1 proteins is proposed.     

和田沙漠玫瑰品种系统发育关系初步研究

该研究基于DNA分子数据分析和田沙漠玫瑰品种与丰花玫瑰、紫枝玫瑰、大马士革玫瑰品种的关系,构建和田沙漠玫瑰的系统发育关系,以探讨其杂交起源.结果表明:(1)和田沙漠玫瑰具有独特的遗传组成,品种内遗传距离(0.017±0.003)小于品种间的遗传距离(分别为0.029±0.005、0.024±0.004、0.022±0....     

Identification of fungal diseases of Rosa damascena in Kerman province of Iran

The oil rose, Rosa damascena Mill. (Rosaceae) is an agricultural crop cultivated in various countries of the northern hemisphere, such as Turkey, Bulgaria, Morocco, Iran, Egypt, France, China and India. Iran, presently, is the largest producer of rose water in world. The major production areas in Iran are Kashan, Fars, Kerman and Azerbaijan. Kerman province with 2297 hectares ha of rose gardens and 6198 tons of flower production is one of the important rose production regions. The productions of this region are organic and do not use anychemical compounds such as pesticides and fertilisers. The major fungal pathogens were studied during 2008–2010 in oil rose production areas in Kerman province, Iran. Verticillium dahlias, Rosellinia necatrix, Alternaria alternata, Seimatosporium fusisporum and Podosphaera pannosa have been detected in the oil rose from different regions in Kerman province. A. alternata has the most isolates and infected plants per cent in the oil rose. This is the first report fromVerticillium dahliae, R. necatrix, A. alternata, S. fusisporum on oil roses (R.damascena) in the world.     

Stipa×brozhiana (Poaceae) nothosp. nov. from the western Pamir Alai Mts (middle Asia) and taxonomical notes on Stipa×tzvelevii

Stipa ×brozhiana M. Nobis nothosp. nov. (Poaceae) is described and illustrated. The taxon belongs to sect. Smirnovia Tzvel. and originated from the hybridization of species belonging to sect. Smirnovia and Barbatae Junge. Stipa ×brozhiana is morphologically close to Stipa lipskyi Roshev., but is easily distinguished by its much shorter hairs on the seta, thinner, uni‐ or indistinctly bigeniculate awns, differently shaped callus and longer ligules of vegetative shoots. Characters distinguishing S. ×brozhiana from the parental species and other similar hybrid taxa belonging to sect. Smirnovia and occurring in the Pamir Alai Mts are discussed. In addition, the taxonomical status of S. ×tzvelevii Ikonn. pro sp., another taxon that has originated from hybridization between species belonging to sect. Smirnovia and Barbatae, is discussed. The taxon was originally described as a distinct species, but is now considered to be the hybrid S. caucasica×S. orientalis. The main characters distinguishing S. ×tzvelevii from parental species are given.     

The fate of ribosomal RNA genes in spontaneous polyploid dogrose hybrids [Rosa L. sect. Caninae (DC.) Ser.] exhibiting non‐symmetrical meiosis

Dogroses represent an exceptional system for studying the effects of genome doubling and hybridization: their asymmetrical meiosis enables recombination in bi‐parentally inherited chromosomes but prevents it in maternally inherited ones. We employed fluorescent in situ hybridization, genome skimming, amplicon sequencing of genomic and cDNA as well as conventional cloning of nuclear ribosomal DNA in two phylogenetically distinct pentaploid (2n = 5x = 35) species, Rosa canina and Rosa inodora, and their naturally occurring reciprocal hybrids, Rosa dumalis (5x) and Rosa agrestis (5x, 6x). Both progenitor species differed in composition, meiotic behaviour and expression of rDNA loci: R. canina (five 18S and 5–8 5S loci) was dominated by the Canina ribotypes, but R. inodora (four 18S loci and 7–8 5S loci) by the Rubiginosa ribotype. The co‐localized 5S/18S loci occurred on either bivalent‐forming (R. canina) or univalent‐forming (R. inodora) chromosomes. Ribosomal DNA loci were additively inherited; however, the Canina ribotypes were dominantly expressed, even in genotypes with relatively low copy number of these genes. Moreover, we observed rDNA homogenization towards the paternally transmitted Canina ribotype in 6x R. agrestis. The here‐observed variation in arrangement and composition of rDNA types between R. canina and R. inodora suggests the involvement of different genomes in bivalent formation. This results supports the hypothesis that the asymmetrical meiosis arose at least twice by independent ancient hybridization events.     

Long term dynamics of Rosa multiflora in a successional system

Long term studies of invasion dynamics are critical in developing a more complete understanding of the factors that influence species spread. To address this issue, the dynamics of the non-native invasive plant, Rosa multiflora, were examined using a 40-year record of successional change. The roles of biotic and abiotic factors in regulating R. multiflora invasion were also assessed. The invasion showed an initial 9-year time lag, followed by a 20-year period of population expansion and an ultimate decline as succession progressed. During all phases of R. multiflora’s invasion, there was continuous turnover within plots. Rainfall during the previous season was found to increase R. multiflora colonization during population expansion while tree species inhibited the invader’s growth. During expansion and decline of R. multiflora, common associated species were often positively or negatively correlated with changes in R. multiflora cover. Though early population dynamics were regulated by propagule pressure, the major influence on R. multiflora late in succession was canopy closure. Although the invasion of this species was largely self-limiting in this system, the species is likely to persist within late successional systems and may require management intervention.     

Flavonol glycosides acylated with 3-hydroxy-3-methylglutaric acid as systematic characters in Rosa

LC–UV–MS/MS analysis of leaf extracts from 146 accessions of 71 species of Rosa revealed that some taxa accumulated flavonol O-glycosides acylated with 3-hydroxy-3-methylglutaric acid, which are relatively uncommon in plants. The structures of two previously unrecorded examples isolated from Rosa spinosissima L. (syn. Rosa pimpinellifolia L.) were elucidated using spectroscopic and chemical methods as the 3-O-α-l-rhamnopyranosyl-(1 → 2)-[6-O-(3-hydroxy-3-methylglutaryl)-β-d-galactopyranosides] of kaempferol (3,5,7,4′-tetrahydroxyflavone) and quercetin (3,5,7,3′,4′-pentahydroxyflavone). The corresponding 3-O-[6-O-(3-hydroxy-3-methylglutaryl)-β-d-galactopyranoside] of quercetin was also present in R. spinosissima, but at lower levels, together with 17 other flavonol O-glycosides for which structures were assigned using LC–UV–MS/MS. The distribution of flavonol 3-hydroxy-3-methylglutarylgalactosides in Rosa was limited to some species of subgenus Rosa section Pimpinellifoliae and Rosa roxburghii Sw. of the monotypic subgenus Platyrhodon, indicating that this character could be of value in phylogenetic analyses of the genus.     

Does genetic distance between parental species influence outcomes of hybridization among coral reef butterflyfishes?

Christmas Island is located at the overlap of the Indian and Pacific Ocean marine provinces and is a hot spot for marine hybridization. Here, we evaluate the ecological framework and genetic consequences of hybridization between butterflyfishes Chaetodon guttatissimus and Chaetodon punctatofasciatus. Further, we compare our current findings to those from a previous study of hybridization between Chaetodon trifasciatus and Chaetodon lunulatus. For both species groups, habitat and dietary overlap between parental species facilitate frequent heterospecific encounters. Low abundance of potential mates promotes heterospecific pair formation and the breakdown of assortative mating. Despite similarities in ecological frameworks, the population genetic signatures of hybridization differ between the species groups. Mitochondrial and nuclear data from C. guttatissimus × C. punctatofasciatus (1% divergence at cyt b) show bidirectional maternal contributions and relatively high levels of introgression, both inside and outside the Christmas Island hybrid zone. In contrast, C. trifasciatus × C. lunulatus (5% cyt b divergence) exhibit unidirectional mitochondrial inheritance and almost no introgression. Back‐crossing of hybrid C. guttatissimus × C. punctatofasciatus and parental genotypes may eventually confound species‐specific signals within the hybrid zone. In contrast, hybrids of C. trifasciatus and C. lunulatus may coexist with and remain genetically distinct from the parents. Our results, and comparisons with hybridization studies in other reef fish families, indicate that genetic distance between hybridizing species may be a factor influencing outcomes of hybridization in reef fish, which is consistent with predictions from terrestrially derived hybridization theory.     

Genetic evidence for hybridization between the native Spartina maritima and the introduced Spartina alterniflora (Poaceae) in South-West France: Spartina × neyrautii re-examined

 Spartina alterniflora, a perennial grass native to the North American Atlantic coast, was introduced during the 19th century in western Europe (Southern England and western France) where it hybridized with the native Spartina maritima. In England, the sterile hybrid S. × townsendii gave rise by chromosome doubling to the highly fertile allopolyploid Spartina anglica, which has now invaded many salt marshes and estuaries in western Europe, and has been introduced in several continents. In South-West France, another sterile hybrid was discovered in 1892 in the Bidassoa Estuary, and named Spartina × neyrautii. According to their morphology, some authors suggested that S. × neyrautii and S. × townsendii result from reciprocal crosses. During the 20th century, the hybridization site was severely disturbed, and surviving of S. × neyrautii was questioned. In this paper, various Spartina populations are investigated in the Basque region (France and Spain), and compared to the hybrid taxa formed in England (S. × townsendii and S. anglica). The samples were analyzed using molecular fingerprinting (RAPD and ISSR) and Chloroplast DNA sequence (trnL-trnT spacer, trnL intron and trnL-trnF spacer). In the Bidassoa estuary, a hybrid isolated clone has been found, that displays additive species-specific nuclear markers of S. maritima and S. alterniflora, and that is subsequently considered as a surviving clone of S. × neyrautii. The molecular analyses indicate that S. × neyrautii and S. × townsendii share the same maternal (S. alterniflora), and paternal (S. maritima) parental species, but also that the two independent hybridization events have involved different parental (nuclear) genotypes in England and in South-West France. Received July 12, 2002; accepted October 4, 2002 Published online: March 20, 2003