Interspecific hybridization in Rorippa (Brassicaceae): Patterns and processes

The current knowledge on natural hybridization between European lowland Rorippa species is reviewed. Morphological, cytological and molecular markers provide substantial evidence that four Rorippa species (R. amphibia, R. austriaca, R. palustris and R. sylvestris) are involved in hybridization processes. The main factors promoting initial hybridization events between Rorippa species in Europe are (1) natural and anthropogenic habitat disturbance, (2) the invasion of non‐native R. austriaca into western central Europe and (3) the self‐incompatibility system of R. amphibia, R. austriaca and R. sylvestris coupled with clonal growth. Outcomes of hybridization between European Rorippa species show a broad range from incidental hybrids through the formation of hybrid swarms to the evolution of new hybrid taxa. Rorippa × armoracioides is certainly the most successful Rorippa hybrid. It results from hybridization between R. austriaca and R. sylvestris and is known from many localities in the native range of R. austriaca and in regions invaded by R. austriaca. Within this system hybrid zones of different age provide the rare opportunity to analyse different stages of hybrid formation and hybrid speciation in natural populations.     

Introgressive hybridization in Rorippa (Brassicaceae): gene flow and its consequences in natural and anthropogenic habitats

Introgressive hybridization between three Rorippa species (R. amphibia, R. palustris and R. sylvestris) in northern Germany has been studied using isozymes and noncoding chloroplast DNA (trnL/F spacer). Our results provide substantial evidence for different patterns of gene flow in natural and in anthropogenic environments. Hybridization and bi-directional introgression (chloroplast DNA and allozymes) between R. amphibia and R. sylvestris were detected at the river Elbe, which is one of the last rivers in Central Europe showing a natural dynamic of erosion and sedimentation. The natural dynamic of the Elbe leads to periodic habitat disturbance and the temporal breakdown of ecological isolation barriers between R. amphibia and R. sylvestris. However, the high dynamic does not provide the opportunity for persistence of the morphologically intermediate hybrid R. x anceps (R. amphibia x R. sylvestris). We did not find hybrid zones between R. amphibia and R. sylvestris in the more anthropogenic landscape of northwest Germany. However, contact zones between R. amphibia and R. palustris were detected in drainage ditches in northwest Germany. We found substantial evidence for unidirectional introgression of R. palustris markers (chloroplast DNA and allozymes) into R. amphibia in the man-made habitats. The R. amphibia introgressants in the drainage ditches often showed strongly serrate upper cauline leaves instead of the entire upper cauline leaves typical for R. amphibia. We argue that landscape melioration in northwest Germany, particularly the creation of drainage ditches, favoured both hybrid-zone formation and ecotypic differentiation within R. amphibia.     

中国独行菜族(十字花科)部分属种的分子系统学研究

利用叶绿体DNA的trnL内含子和trnL-F基因间隔区序列测定,对中国独行菜族部分属及相关属植物进行了分子系统学研究,结果表明:独行菜族是多系类群,在分子树中,所研究的中国独行菜族部分属植物被分别聚在了3个不同的并列系中,其中,荠属(Capsella)和亚麻荠属(Camelina)与南芥族的3个属聚在系Ⅰ,认为应将二者分别从独行菜族和大蒜芥族中移出,划入南芥族中;菥蓂属(Thlaspi)和菘蓝属(Isatis)与大蒜芥族的3个属聚成系Ⅱ,支持将二者移出独行菜族,划入大蒜芥族的观点;独行菜属(Lepidium)、臭荠属(Coronopus)和群心菜属(Cardaria)三者关系密切,组成系Ⅲ,其中臭荠嵌入独行菜属中,支持将臭荠属合并到独行菜属的处理意见.     

基于种皮形态和叶绿体trnL-F对菘蓝属（十字花科）的系统学位置研究

利用种皮微形态观察与叶绿体DNA的trnL内含子和trnL-F基因间隔区序列测定分析相结合,对十字花科菘蓝属植物及其相关属种的系统学关系进行了探讨.研究结果表明原属独行菜族的菘蓝属植物,其种皮特征与原属鸟头荠族的舟果荠属植物相似,同属一类型.而与独行菜族的模式属独行菜属植物、大蒜芥族的模式属大蒜芥属植物以及鸟头荠族的模式属鸟头荠属植物种皮微形态差异显著;在基于叶绿体DNA的trnL内含子和trnL-F基因间隔区序列所构建的系统发育树中,菘蓝属植物与舟果荠属植物距离最近,而与独行菜属、大蒜芥属、鸟头荠属植物具有一定的间隔,结合形态特征,本研究认为,菘蓝属与舟果荠属植物具有较近的亲缘关系,而不是同族的独行菜属植物.     

拟南芥属的系统位置：种皮及分子证据

拟南芥属（Arabidopsis（L.）Heynhold）拥有现代分子生物学研究的模式植物拟南芥（A.thaliana（L.）Heynhold）,但其属的系统位置及与近缘属关系争议较大。根据对拟南芥属及其相关属种的种皮微形态观察,结合测定分析各属种叶绿体DNA的trnL内含子和trnL-F基因间隔区序列,结果表明,拟南芥属的近缘属种包括荠属（Capsella Medic.）、亚麻荠属（Camelina（L.）Crantz）、须弥芥属（Crucihimalaya Al-Shehbaz et al.）、无苞芥属（Olimarabidopsis Al-Shehbaz et al.）、旗杆芥属（Turritis L.）、南芥属（Arabis L.）和糖芥属（Erysimum Kitagawa）。     

辣根属和豆瓣菜属(十字花科)系统位置的分子证据

对十字花科葶苈族的辣根属、南芥族的豆瓣菜属及相关属种植物的叶绿体DNA的trnL内含子和trnL-F基因间隔区序列进行了测定分析。结果表明,辣根属植物与南芥族的山芥属、蔊菜属、豆瓣菜属、碎米荠属在系统发育树中聚成一支,与葶苈族的模式属葶苈属植物相隔较远,结合形态特征,本研究认为辣根属应从葶苈族移出,其系统位置应靠近山芥属、蔊菜属、豆瓣菜属、碎米荠属植物;此外,系统发育树中,豆瓣菜属植物并入碎米荠属中,表明二者具有更近的亲缘关系,本研究结果不支持《中国植物志》第33卷对辣根属和豆瓣菜属的系统位置的处理。     

Genetic diversity in diploid vs. tetraploid Rorippa amphibia (Brassicaceae)

The frequency of polyploidy increases with latitude in the Northern Hemisphere, especially in deglaciated, recently colonized areas. The cause or causes of this pattern are largely unknown, but a greater genetic diversity of individual polyploid plants due to a doubled genome and/or a hybrid origin is seen as a likely factor underlying selective advantages related to life in extreme climates and/or colonization ability. A history of colonization in itself, as well as a recent origin, and possibly a limited number of polyploidization events would all predict less genetic diversity in polyploids than in diploids. The null hypothesis of higher gene diversity in polyploids has to date hardly been quantified and is here tested in self-incompatible Rorippa amphibia (Brassicaceae). The species occurs in diploid and tetraploid forms and displays clear geographical polyploidy in Europe. On the basis of eight microsatellite loci it can be concluded that the level of gene diversity is higher in tetraploids than in diploids, to an extent that is expected under neutral evolution when taking into account the larger effective population size in the doubled cytotype. There is thus no evidence for reduced genetic diversity in the tetraploids. The evidence presented here may mean that the tetraploids' origin is not recent, has not been affected by bottlenecks and/or that tetraploids were formed multiple times while an effect of introgression may also play a role.     

Regeneration growth of the invasive clonal forb Rorippa austriaca (Brassicaceae) in relation to fertilization and interspecific competition

A special type of clonal growth, spread by lateral roots, ishypothesized to be a favourable trait of invasive, opportunistic plant speciesof disturbed habitats. We tested this hypothesis for the invasive forbRorippa austriaca (Brassicaceae). Regenerationfrom root fragments, subsequent vegetative spread and allocation patterns inrelation to varied nutrient supply and intensity and pattern of interspecificcompetition were analyzed in container experiments. Regeneration success fromroot fragments was 100% and clonal spread was rapid but vegetativeperformance was strongly reduced under unfertilized conditions and,particularly, when interspecific competition was present. While the ratio ofabove- to belowground bio-mass did not differ considerably betweentreatments, R. austriaca allocated a high amount ofresources to belowground growth resulting in low aboveground but highbelowground biomass at harvest time relative to the matrix vegetation.Differences in shoot number or biomass between simulated gaps and denselyvegetated quadrants in the containers were (relatively) weak.Reproductive effort was less reduced under low resource levels, and the clonesdid not set seed at all, irrespective of the treatment. Our results show thatclonal growth by lateral roots and plasticity in clonal growth patterns inR. austriaca promote both exploitation of gaps andnutrient-rich microsites and resistance to competitors. Such plasticity,combined with its ability to regenerate from widely-dispersed rootfragments, contribute to the ability of the species to invade and persistwithindisturbed and spatially heterogeneous habitats.     

Hybrid zones between invasive Rorippa austriaca and native R. sylvestris (Brassicaceae) in Germany: ploidy levels and patterns of fitness in the field

Hybrid zones may serve as natural laboratories for evolutionary studies. One common viewpoint is that hybrids may always be less fit than their parents due to genetic discontinuities. An alternative idea is that genotype-environment interactions influence the outcome of natural hybridization. Our comparative study of two different natural hybrid zones between the invasive diploid Rorippa austriaca and the native polyploid R. sylvestris in Germany identified the ploidy level as a major determinant of hybrid fitness. Different ploidy levels and patterns of fitness were detected in different hybrid zones. In one hybrid zone (Mülheim, Ruhr valley) hybrids were pentaploid and showed a relatively high seed set, whereas in the second hybrid zone (Randersacker, Main valley) hybrids were triploid and displayed extremely low fitness values. Analyses of fitness values in different natural hybrid zones between the same two species may lead to very different conclusions about the evolutionary significance of natural hybridization.     

Cultivation shapes genetic novelty in a globally important invader

Acacia saligna is a species complex that has become invasive in a number of countries worldwide where it has caused substantial environmental and economic impacts. Understanding genetic and other factors contributing to its success may allow managers to limit future invasions of closely related species. We used three molecular markers to compare the introduced range (South Africa) to the native range (Western Australia). Nuclear markers showed that invasive populations are divergent from native populations and most closely related to a cultivated population in Western Australia. We also found incongruence between nuclear and chloroplast data that, together with the long history of cultivation of the species, suggest that introgressive hybridization (coupled with chloroplast capture) may have occurred within A. saligna. While we could not definitively prove introgression, the genetic distance between cultivated and native A. saligna populations was comparable to known interspecific divergences among other Acacia species. Therefore, cultivation, multiple large‐scale introductions and possibly introgressive hybridization have rapidly given rise to the divergent genetic entity present in South Africa. This may explain the known global variation in invasiveness and inaccuracy of native bioclimatic models in predicting potential distributions.     

Isolation and characterization of microsatellite loci in the invasive Alliaria petiolata (Brassicaceae)

Eight novel polymorphic microsatellite loci are presented for garlic mustard (Alliaria petiolata, Brassicaceae) a European herb that is a serious invader of North American deciduous forests. The microsatellites will be useful tools to analyse pathways of introduction of garlic mustard, as well as its evolutionary potential in the invasive range.     

Genetic differentiation in the genus Lithops L. (Ruschioideae, Aizoaceae) reveals a high level of convergent evolution and reflects geographic distribution

Southern Africa is one of the hot spots for plant biodiversity, with ca. 80% of species endemic to this area. Rapid and recent radiations in Southern African plant genera were triggered by fine-scale differences in climate, topography and geology. The genus Lithops (Ruschioideae, Aizoaceae) contains 37 species and is widely distributed in Southern Africa. Species delimitation within the genus is challenging because the limited number of morphological characters in these reduced succulents varies intensely between populations, presumably as adaptations to local geological environments. We analysed phylogenetic relationships within Lithops using non-coding chloroplast DNA (trnS-trnG intergenic spacer), nuclear ribosomal internal transcribed spacer (nrITS) sequences and AFLP data. Genetic variability of the sequence data was very low, but AFLP data detected nine clades within Lithops that do not fit current morphology-based taxonomy. Two of these clades are separated by their distribution on the northern and eastern border of the distribution area, and four clades are found in the Gariep Centre in the estuary of the Orange River. Morphological similarities, especially colour of leaves, evolved repeatedly within the clades, thus we hypothesise that closely related species became adapted to different soil types in a mosaic-like geological environment. One-third of the species are found in the Gariep Centre, characterised by extremely diverse edaphic habitats.     

Lack of trnL intron in the thermoacidophilic red alga Galdieria sulphuraria (Galdieri) Merola

Abstract

The plastid trnC‐trnL(UAA)‐ilvH region from Galdieria sulphuraria was cloned and sequenced with the aim of verifying the absence of the trnL intron. The sequence alignment shows both the absence of a trnL intron and the colinearity of the whole region of the plastidial DNA of G. sulphuraria with that of the other thermoacidophilic red algae.     

New perspectives in diet analysis based on DNA barcoding and parallel pyrosequencing: the trnL approach

The development of DNA barcoding (species identification using a standardized DNA sequence), and the availability of recent DNA sequencing techniques offer new possibilities in diet analysis. DNA fragments shorter than 100-150 bp remain in a much higher proportion in degraded DNA samples and can be recovered from faeces. As a consequence, by using universal primers that amplify a very short but informative DNA fragment, it is possible to reliably identify the plant taxon that has been eaten. According to our experience and using this identification system, about 50% of the taxa can be identified to species using the trnL approach, that is, using the P6 loop of the chloroplast trnL (UAA) intron. We demonstrated that this new method is fast, simple to implement, and very robust. It can be applied for diet analyses of a wide range of phytophagous species at large scales. We also demonstrated that our approach is efficient for mammals, birds, insects and molluscs. This method opens new perspectives in ecology, not only by allowing large-scale studies on diet, but also by enhancing studies on resource partitioning among competing species, and describing food webs in ecosystems.     

Phylogenetic analysis of Nymphaeales using fast-evolving and noncoding chloroplast markers

The Nymphaeales (water-lilies and relatives) represent one of the earliest branching lineages of angiosperms and comprise about 70 aquatic species. Here, we present a comprehensive study of phylogenetic relationships within the Nymphaeales from a dataset containing 24 representatives of the order, including all currently recognized genera and all subgenera of the genus Nymphaea , plus 5 outgroup taxa. Nine different regions of the chloroplast genome − comprising spacers, group II introns, a group I intron, and a protein coding gene − were analysed. This resulted in a character matrix of 6597 positions and an additional 369 characters obtained from coded length mutations. Maximum parsimony and Bayesian analyses of the complete dataset yielded congruent, fully resolved and well-supported trees. Our data confirm the monophyly of the Cabombaceae but do not provide convincing support for the monophyly of Nymphaeaceae with respect to Nuphar . Moreover, the genus Nymphaea is inferred to be paraphyletic with respect to Ondinea , Victoria and Euryale . In fact, the Australian endemic Ondinea forms a highly supported clade with members of the Australian Nymphaea subgenus Anecphya . In addition, Victoria and Euryale are inferred to be closely related to a clade comprising all night-blooming water-lilies ( Nymphaea subgenera Hydrocallis and Lotos ). An experimental approach showed taxon sampling to be of influence on the nodes reconstructed in core Nymphaeaceae. The results underscore that more diverse genera, if not clearly known to be monophyletic, should be represented by all major lineages.  © 2007 The Linnean Society of London, Botanical Journal of the Linnean Society , 2007, 154 , 141–163.     

Mitochondrial and chloroplast DNA-based phylogeny of Pelargonium (Geraniaceae)

Overall phylogenetic relationships within the genus Pelargonium (Geraniaceae) were inferred based on DNA sequences from mitochondrial(mt)-encoded nad1 b/c exons and from chloroplast(cp)-encoded trnL (UAA) 5' exon-trnF (GAA) exon regions using two species of Geranium and Sarcocaulon vanderetiae as outgroups. The group II intron between nad1 exons b and c was found to be absent from the Pelargonium, Geranium, and Sarcocaulon sequences presented here as well as from Erodium, which is the first recorded loss of this intron in angiosperms. Separate phylogenetic analyses of the mtDNA and cpDNA data sets produced largely congruent topologies, indicating linkage between mitochondrial and chloroplast genome inheritance. Simultaneous analysis of the combined data sets yielded a well-resolved topology with high clade support exhibiting a basic split into small and large chromosome species, the first group containing two lineages and the latter three. One large chromosome lineage (x = 11) comprises species from sections Myrrhidium and Chorisma and is sister to a lineage comprising P. mutans (x = 11) and species from section Jenkinsonia (x = 9). Sister to these two lineages is a lineage comprising species from sections Ciconium (x = 9) and Subsucculentia (x = 10). Cladistic evaluation of this pattern suggests that x = 11 is the ancestral basic chromosome number for the genus.     

Hybridization among sympatric species of Rhododendron (Ericaceae) in Turkey: morphological and molecular evidence

Rhododendron (Ericaceae) is a large genus in which barriers to hybridization are especially weak, but many species are maintained in sympatry. Hybridization among four species of Rhododendron subsect. Pontica, which occur in sympatry in Turkey, was investigated. Material of R. ponticum, R. smirnovii, R. ungernii, and R. caucasicum and their putative hybrids was collected from the wild. Based on morphology, chloroplast DNA and nuclear ribosomal DNA restriction fragment length polymorphism (RFLP) profiles, each accession was identified as a species or hybrid combination. Five of the six possible hybrid combinations among the four species were detected. Rhododendron ponticum × R. smirnovii was represented by a single individual and R. caucasicum × R. smirnovii by one small group of hybrid plants. The combinations R. ponticum × R. ungernii and R. ungernii × R. smirnovii showed evidence of frequent backcrossing, while R. ponticum × R. caucasicum appeared unusual in that an intermediate hybrid type was abundant, whereas hybrids with phenotypes approaching either parent were rare. Possible explanations of this latter situation are discussed. The results suggest that natural hybridization among Rhododendron species is common and that ecological factors are important in maintaining integrity when species occur in sympatry.     

A combination of molecular markers identifies evolutionarily significant units in Cedrela odorata L. (Meliaceae) in Costa Rica

The necessity for conservation of the geneticcomponent of biodiversity is now widelyrecognised. A broad genetic base is required tomaintain evolutionary potential and thepopulation erosion occurring in much of theworld's forests threatens the genetic integrityof many tree species. Spanish Cedar (Cedrela odorata L.) has been under severepressure for generations and is now the focusof a study aimed at assessing the levels anddistribution of genetic diversity in remainingpopulations. Ten Costa Rican populations wereanalysed using chloroplast and AFLP markers.The overall level of diversity was as expectedfor an outcrossing, long-lived, woody species(H _T = 0.27). However, this concealeda deep divergence within the species, forchloroplast and AFLP (_CT = 0.83)markers. Populations were differentiated in twogroups that exhibited contrasting habitatpreferences and two ecotypes, wet and dry, wereidentified. Within the ecotypes, all but onepopulation were fixed for a single chloroplasthaplotype and within populations, total genomicdiversity levels were low (H _S= 0.03–0.13). Populations possessing the dryecotype maintained significantly more diversitythan those from wet regions. Within the wetecotype group, pairwise genetic distancebetween populations fitted an isolation bydistance model. The group was stronglysubdivided and showed isolation by distancearound the southern edge of the centralmountain ranges. The genetic divergence of thetwo ecotypes, observed at both organellar andnuclear loci, identifies evolutionarilysignificant units that, taken together withprevious studies of the species, provide arational basis on which to build a conservationpolicy for the species.