Aspartate aminotransferase isozymes in the genusCapsella (Brassicaceae): Subcellular location,gene duplication,and polymorphism

The subcellular location of aspartate aminotransferase isozymes (EC 2.6.1.1) in the genusCapsella(Brassicaceae) was studied. The diploid speciesC. grandiflora andC. rubella have three AAT isozymes, including one located in the plastids. Each locus is duplicated in the tetraploidCapsella bursa-pastoris. Variation at the plastid-coding locus exceeded that at the other loci.C. bursa-pastoris had some unique alleles not detected in the diploid species. Segregation in open-pollinated families revealed thatCapsella grandiflora was outcrossing, whereasC. rubella was highly inbred, with most populations homozygous or uniform at all three loci. Inheritance in the tetraploid colonizerC. bursa-pastoris is disomic. This species was also predominantly selfing with outcrossing rates between 2% and 10%.Financial support by the German Research Foundation DFG is gratefully acknowledged.     

Evolution of the tetraploidCapsella bursa-pastoris (Brassicaceae): Isoelectric focusing analysis of Rubisco

Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco, EC 4.1.1.39) and its subunits (large subunits = LSU, small subunits = SSU) were isolated from threeCapsella spp. by gel electrophoresis and polypeptide composition was analyzed by isoelectric focusing (IEF) in the presence of 8M urea. The described techniques are recommended for large scale systematic studies. Multiple IEF banding patterns of the SSU are probably the outcome of a heterogenous multigene family. The two diploid speciesC. rubella andC. grandiflora show an identical IEF pattern and could be differentiated from the putative allotetraploidC. bursa-pastoris only by the SSU banding pattern. Uniqueness of some SSU bands in the tetraploid and in the two diploid species, respectively, may indicate an ancient alloploid origin of tetraploidC. bursa-pastoris followed by events leading to divergences in the genomes of the allotetraploid and its presumed diploid progenitors after the hybridization event (SSU gene elimination, acquisition of new SSU genes).     

Evolutionary Processes Associated with Biological Invasions in the Brassicaceae

This review highlights evolutionary processes in the Brassicaceae which cause, accompany or are a consequence of biological invasions. Case studies in Capsella and Diplotaxis address the genetics of colonising species. The greatest colonising ability in Capsella bursa-pastoris is associated with polyploidy, predominant selfing, and high genetic diversity. Success of colonial populations seems to be due to the introduction of preadapted genotypes. Colonising species in Diplotaxis reveal contrasting evolutionary patterns. Genetic attributes in D. muralis include annuality, polyploidy, and predominant selfing. Very different from Capsella, D. muralis is nearly devoid of genetic diversity as revealed by molecular markers. In contrast to D. muralis, the colonising D. tenuifolia is perennial, diploid, self-incompatible, and displays high genetic diversity. Hybrid speciation, establishment of the hybrids in man-made habitats, stabilisation of their reproductive system, and reproductive isolation from the parent populations were analysed in Cardamine. The Nasturtium example highlights the importance of hybridisation for the evolution of invasiveness. The last case study concentrates on the evolutionary consequences of hybridisation between native and invading Rorippa species. Introgressive hybridisation between the invasive Rorippa austriaca and native Rorippa species is common and widespread in central Europe, and interspecific gene transfer has led to the formation of a new invasive genotype. Each successful invasion presents new aspects and sound case studies are needed in order to understand the ecology and evolution of the colonisation process and to enable us to assess the evolutionary consequences of biological invasions. This revised version was published online in July 2006 with corrections to the Cover Date.     

Biosystematic studies onCapsella bursa-pastoris (Brassicaceae): Enzyme polymorphism in natural populations

The genetic variability of natural populations ofCapsella bursapastoris in North- and Middle-Europe has been estimated by means of enzyme assays. Zymograms of 81 populations have been developed. 17 loci could be identified, and 8 of them can be heterozygous. Genetic variability is greater between populations than within. No correlation between actual population sizes and genetic heterogeneity could be detected. Some electromorphs shift their frequencies proportionally to increasing adversity of climatic conditions, some appear to be constant over the whole area, and others are characterized by an apparently irregular variation pattern. Marginal populations comprise a significantly higher proportion of heterozygous plants than central ones. Apart from this clinal variation pattern, a mosaic pattern, strongly related to habitat conditions, was observed: genetic heterogeneity is greater in more intensively disturbed sites. The pattern of genetic variation in natural populations ofCapsella bursa-pastoris is rather highly influenced by the breeding system.     

Genetic differentiation among populations ofArabis serrata (Brassicaceae) along its geographic distribution

Levels and distribution of genetic variation were investigated in ten populations of the perennialArabis serrata distributed along a latitudinal gradient throughout Japan. Populations of this endemic species occupy predominantly three types of habitats: limestone and derived soils, volcanic and disturbed sites. Previous studies showed that plants ofA. serrata are differentiated in morphological and ecological traits under both natural conditions and common garden experiments suggesting genetic differentiation among populations. To assess the degree of genetic differentiation under different habitats ofA. serrata populations, we analyzed the isozyme genetic structure. Ten populations, located in mountains of central and northern Honshu and Hokkaido, were analyzed by starch gel enzyme electrophoresis. Fourteen loci of eight enzymes were resolved and six loci were monomorphic for all the populations. Populations sampled maintain low levels of genetic variation (P=0.16;H=0.05;A=1.16) compared to that maintained by other outcrossing seed plants. In some cases, few or no heterozygous individuals were detected, and consequently, mean observed heterozygosity was zero or near zero. Six (29%) of the fixation indices,F, estimated deviated significantly from Hardy Weinberg genotypic expectations indicating a deficiency of heterozygotes in most of the cases. The mean genetic identity (Nei'sI) between population pairs was 0.852 and indicates a moderate level of genetic differentiation among populations.Arabis serrata has most of its genetic variation distributed among rather than within populations. The among-population component of the total genetic diversity (G _ST mean value) was 0.416, indicating genetic differentiation between populations. There groups of populations were recognized in an unrooted tree generated by the Neighbor-Joining method. These results suggest groups of populations differentiated regionally. Estimates of interpopulational gene flow (Nm) were very variable (range 0.049–3.718) with a meanNm=1.203 for all populations.     

Systematic implications of chloroplast DNA variation inLepidium sectionsCardamon,Lepiocardamon andLepia (Brassicaceae)

Chloroplast DNA restriction site variation was examined amongLepidium taxa usually classified in sects.Lepia, Lepiocardamon andCardamon. By using 15 restriction endonucleases, filter hybridization experiments, and comparative mapping procedures, a total of 119 variable restriction sites was detected. Of these, 56 were phylogenetically informative and were used in a cladistic analysis. The resulting phylogenetic tree agrees with results derived from morphology, isozyme electrophoresis and the analysis of glucosinolates. The data suggest: 1) The recognition of sect.Lepia, and 2) that sect.Lepiocardamon has close relationships to sect.Cardamon and that both should be amalgamated. Low rates of interspecific chloroplast DNA sequence divergence in sect.Lepia correlated well with morphological data and isoelectric focusing analysis of Rubisco, suggesting that taxa of sect.Lepia are closely related and most likely have diversified recently.     

Erysimum cephalonicum (Brassicaceae), ein neuer Endemit der Insel Kefallinia (Griechenland)

Erysimum cephalonicum Polatschek is a new taxon, endemic to the isle of Kefallinia. Its closest allies areErysimum pusillum Bory etChaub. andE. trichophyllum Heldr. ex sched.

     

Electrophoretic relationships and phylogeny of Nordic polyploids inDraba (Brassicaceae)

104 populations of 15 Nordic species (2x–16x) of the taxonomically complex genusDraba were investigated using enzyme electrophoresis. The polyploids were genetic alloploids showing high levels of fixed heterozygosity and electrophoretic variation; the diploids were homozygous and genetically depauperate. Thus, the data suggest that alloploidy in arctic-alpineDraba serves as an escape from genetic depauperation caused by inbreeding at the diploid level. Although some populations probably have local alloploid origins, electrophoretic data indicate that several polyploids have migrated repeatedly into the Nordic area.Draba crassifolia (2n = 40) is probably octoploid based on x = 5. A hypothesis on the evolutionary history of the polyploids based on x = 8 is presented. Diploids contributing to numerous polyploid genomes and multiple origins of polyploids have seriously blurred taxonomic relationships. Relationships inferred from genetic data do not always correspond to those based on morphology; two morphologically very similar polyploids,D. alpina andD. oxycarpa, were, for example, genetically distant and probably represent independent lineages.     

Evolutionary patterns and processes in the genus Rosa (Rosaceae) and their implications for host-parasite co-evolution

The genus Rosa (Rosaceae) is a remarkable genus with respect to systematic biology. Multiple reproductive strategies ranging from apomixis to outcrossing including hybridisation, as well as different modes of character inheritance like matroclinal, paternal or sex unrelated ones. This complexity makes the genus a model in which simple concepts of radiation, speciation and taxonomy come to their limits. Analyses of the evolutionary patterns and processes in Rosa give clear evidence for the stochastic character of this specific evolutionary scenario.     

Reproductive strategies of diploid and polyploid populations of arcticDraba (Brassicaceae)

132 cultivated populations (2x–16x) of 15 arctic-alpine species ofDraba were investigated to clarify a possible relationship between reproductive strategies and polyploid evolution in the genus. The populations were exclusively sexual and produced viable seed after spontaneous self-pollination, but showed large variation both in traits promoting cross-pollination and in autogamous fruit and seed set. Traits promoting cross-pollination, e.g., floral display, protogyny, and delayed selfing, were positively correlated, and these traits were negatively correlated with autogamous fruit and seed set. All diploid and many polyploid populations had high autogamous seed set and small, unscented, non-protogynous, and rapidly selfing flowers. In contrast, all populations with low autogamous seed set and large, scented, and strongly protogynous flowers with distinctly delayed selfing were polyploid. These results are consistent with those previously obtained from enzyme electrophoresis, suggesting that the genetically depauperate diploids are extreme inbreeders and that the highly fixed-heterozygous polyploids vary from extreme inbreeders to mixed maters. The reproductive data lend additional support to the hypothesis that allopolyploidy in arcticDraba serves as an escape from genetic depauperation caused by uniparental inbreeding at the diploid level.     

In vitro multiplication ofVella lucentina M. B. crespo (Brassicaceae), a threatened Spanish endemic species

Summary Vella lucentina M. B. Crespo is a threatened Spanish species that is endemic to a small area in eastern Alicante Province (SE Spain). Micropropagation techniques were applied forex situ conservation of this plant. Aseptic epicotyls bearing the apical bud were grown in Murashige and Skoog medium supplemented with 6-furfurylaminopurine (Kin), N⁶-benzyladenine (BA) or 6-(γ,γ,-dimethilalylamino) purine (2iP). High multiplication rates were obtained with 0.5, 1, or 2 mg·liter⁻¹ BA, or 1 or 2 mg·liter⁻¹ 2iP. Indole-3-acetic acid and indole-3-butyric acid were utilized for rooting in half-strength Murashige and Skoog medium. Regenerated plants were transferred to a potting mix and gradually acclimated to field conditions. No morphological differences were observed amongin vitro andin vivo plants.     

蚓果芥乙烯受体基因的克隆与进化分析

采用已报道的十字花科植物乙烯受体基因(Ethylene Receptor 1,ETR1)通用引物,以蚓果芥基因组DNA为模板,经PCR分析BhETR1的多态性,进而从分子水平上阐明蚓果芥的进化特征.结果表明:(1)已克隆得到的14条ETR1基因序列,长度为1 644～1 661 bp;经Bioedit软件比对,14条核苷酸序列之间相似性为93％～98％,蛋白氨基酸序列相似度为78％;采用DnaSP v5程序分别统计BhETR1序列多态性,发现14条BhETR1序列形成了14种单倍型.(2)从GenBank数据库下载蚓果芥近缘物种ETR1序列进行进化分析,系统发育分析表明蚓果芥14条ETR1序列形成3大分支;根据分支进化关系,每支中分别选取克隆2(c2)、克隆29(c29)、克隆35(c35)与拟南芥ETR1(AtETR1)和琴叶鼠耳芥ETR1(AlETR1)的序列进行比对,其一致性均为78％,说明BhETR1是一个乙烯受体类似(ETR1-like)基因,而蚓果芥自身14条序列的高度异质性表明蚓果芥的遗传背景相对复杂.     

Molecular systematics and biogeography of theCardamine pratensis complex (Brassicaceae)

Representatives of theC. pratensis complex were analysed for allozymes, ITS, non-coding cpDNA, and RAPDs to elucidate phylogenetic relationships and the historical biogeography of this species group. Our concepts differ in some important aspects from current ideas. Two diploid species from southeastern Europe form the Basal Group of the complex. A diploid from the Iberian Peninsula represents another old lineage. The phylogenetically younger Derived Group comprises diploid taxa and all known polyploid taxa. The two old lineages represent pleistocene relicts which were not involved in the formation of the Derived Group. All polyploids evolved in postglacial time from diploids of the Derived Group which may have survived the glaciations in refugia centered around and within the Alps. The arctic-circumpolarC. nymanii is of young age and migrated to Scandinavia in postglacial times from south to north.     

Consequences of artificial and natural chromosome doubling on DNA,RNA and protein contents inCochlearia (Brassicaceae)

The effects of chromosome doubling on macromolecular composition, i.e. DNA, RNA and protein, were examined in diploidCochlearia pyrenaica DC., its established natural autotetraploidC. officinalis L., and their newly colchicine-induced autotetraploid and autooctoploid derivatives. DNA, RNA and protein contents increase from lower to higher ploidy levels (2x > 4x > 8x). The established natural autotetraploids and newly induced autotetraploids also differ, as DNA, RNA and protein have been reduced during the course of establishment. Net synthesis of RNA and protein per unit DNA does not change significantly neither from lower to higher ploidy levels nor between the autotetraploids of newly induced and established natural origins.     

Systematics of AustralianLepidium species (Brassicaceae) and implications for their origin: Evidence from IEF analysis of Rubisco

Variation in subunit polypeptide composition of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) from native AustralianLepidium taxa was analyzed by isoelectric focusing (IEF). Species of grexMonoplocoidea on the one hand, and species of gregesPseudoruderalia andPapillosa on the other, were well separated by Rubisco IEF patterns. The protein data suggest grexMonoplocoidea to be placed in sect.Monoploca whereas gregesPseudoruderalia andPapillosa should be integrated into sect.Dileptium. We suggest that the Rubisco type of American and/or AsianLepidium species was contained in those plants migrating to Australia and evolving into the recent native Australian species. Fossil pollen records and vegetation history indicate that the origin ofLepidium in Australia was restricted to the late Tertiary of Quaternary. Data presented in this study fit best with an immigration by long-distance dispersal of seeds by birds from Asia and/or from South America.     

Hybridization and evolution inCardamine (Brassicaceae) at Urnerboden, Central Switzerland: Biosystematic and molecular evidence

Hybridization between two diploid (2n = 2x = 16) species ofBrassicaceae, Cardamine rivularis andC. amara, at Urnerboden, Central Switzerland, resulted in the rather unusual triploid hybridC. insueta (2n = 3x = 24), and later on in the amphiploidC. schulzii (2n = 6x = 48). The hybrid and the neopolyploid species colonized successfully some man-made biotopes. Plants ofC. insueta are mostly functional females with non-dehiscent anthers, but true hermaphrodite individuals with partly sterile pollen grains also occur within the population. Analyses of cpDNA and nuclear DNA permitted to establish the parentage of the hybrid: the maternal parent which contributed unreduced egg cells proved to beC. rivularis whereas the normally reduced pollen originated fromC. amara. The pronounced genetic variability inC. insueta revealed by isozyme and RAPD analyses, at variance with the polarized segregation, heterogamy and strong vegetative reproduction of the hybrid, is possibly influenced by recurrent formation ofC. insueta which party results from backcrosses betweenC. insueta andC. rivularis but may also proceed by other pathways. The amphiploidCardamine schulzii has normally developed anthers but its pollen is sometimes highly sterile. The surprisingly uniform genetic make-up of the new amphiploid species might be related to its possible monotopic origin and/or young phylogenetic age but should be further assessed. Site management seems to be very important to a further development of hybridogenous populations and their parent species. In conclusion, the evolution at Urnerboden is discussed in the context of the traditional concept of multiple plant origins.     

Diversification and the evolution of dispersal ability in the tribe Brassiceae (Brassicaceae)

MethodsThe phylogenetic analysis included 72 taxa sampled from across the Brassiceae and included both nuclear and chloroplast markers. Dispersal-related fruit characters were scored and climate information for each taxon was retrieved from a database. Correlations between fruit traits, seed characters, habitat, range and climate were determined, together with trait-dependent diversification rates.ConclusionsThis study provides evidence that the evolution of increased dispersal ability and larger seed size, which may increase establishment ability, can also influence macro-evolutionary processes, possibly by increasing the propensity for long-distance dispersal. In particular, it may increase speciation and consequent diversification rates by increasing the likelihood of geographic and thereby reproductive isolation.     

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.     

Species relationships within Diplotaxis (Brassicaceae) and the phylogenetic origin of D. muralis

Nineteen from the ca. 30 Diplotaxis species including all known haploid chromosome numbers have been analysed for isoelectric focusing patterns of Rubisco, allozymes and RAPDs. D. erucoides (n=7) was clearly separated from all other species as were D. harra and D. crassifolia (n=13 each). Taxa with n=8 had different IEF patterns, but allozyme data grouped D. siettiana, D. ibicensis and D. brevisiliqua together. Species with n=9 were characterised by different IEF patterns, and their position was neither resolved in the allozyme nor in the RAPD tree. Only the D. catholica accessions were strongly clustered together. D. viminea and D. siifolia (both n=10) were kept separate, whereas the n=11 taxa D. tenuifolia, D. cretacea and D. simplex grouped together. Data confirm D. viminea as maternal parent of the allotetraploid D. muralis, and D. tenuifolia as the likely parent.