The evolution of Spartina anglica G. E. Hubbard (Gramineae): genetic variation and status of the parental species in Britain

The perennial salt marsh grass Spartina anglica is one of the classic examples of allopolyploid speciation. It originated on the south coast of England at the end of the nineteenth century following chromosome doubling in S. × townsmdii , a hybrid between the native British S. marilima and a species introduced from the United States, S. alterniflora. The nature of the origin of S. anglica is beyond doubt; however, it is not known whether it had a single or multiple origin. In order to address this problem we undertook a survey of the genetic variation in the parental species of S. anglica using isozyme electrophoresis. The results show that S. alterniflora has no detectable variation and that S. maritima has extremely low levels of variation. These results, unfortunately, prevent the question of a single or multiple origin from being answered. Possible reasons for the low levels of variation and its influence on the future of the species are discussed. Another problem concerning the parental species is the rapid decline of S. maritima in Britain. It is often assumed that the major factor in this regression is the invasion of its habitats by S. anglica. We have examined the status of S. marilima throughout its range in Britain and have found that S. anglica rarely co-occurs with S. maritima. We propose that the decline of S. maritima is largely due to the physical erosion of its habitats and that this erosion may produce suitable habitats for colonization by S. anglica.     

Molecular investigations in populations of Spartina anglica C.E. Hubbard (Poaceae) invading coastal Brittany (France)

Spartina anglica is a classical example of recent alloploid speciation. It arose during the end of the nineteenth century in England by hybridization between the indigenous Spartina maritima and the introduced East-American Spartina alterniflora. Duplication of the hybrid genome (Spartina x townsendii) gave rise to a vigorous allopolyploid involved in natural and artificial invasions on different continents. Spartina anglica was first recorded in France in 1906, and since then, it has spread all along the western French coast. Earlier studies revealed that native British populations display consistent morphological plasticity and lack of isozyme variation. In this paper, we use different molecular markers (randomly amplified polymorphic DNA, intersimple sequence repeats and restriction patterns from nuclear and chloroplast DNA sequences) to analyse the genetic patterns of the French populations of S. anglica. Our results show that French populations are mainly composed of one "major" multilocus genotype. This genotype is identical to the first-generation hybrid S. x townsendii from England. Losses of few markers from this genotype are observed but are restricted to a few populations from Brittany; it is likely that they appeared independently, subsequent to their introduction. In southern Brittany, no hybrids between S. anglica and S. maritima have been found where the two species co-occur. All French populations of S. anglica display the same chloroplast DNA sequences as S. alterniflora, the maternal genome donor. These findings are consistent with a severe genetic bottleneck at the time of the species formation, as a consequence of a unique origin of the species. Both parental nuclear sequences are present in the allopolyploid populations, revealing that for the markers investigated, no extensive changes have occurred in this young species.     

Origin and genetic diversity of Spartina anglica (Poaceae) using nuclear DNA markers

Spartina alterniflora, introduced into the UK in the 1800s, was the seed parent in an interspecific hybridization with S. maritima. The sterile F1 hybrid S. ×townsendii gave rise to the fertile allopolyploid S. anglica by chromosomal doubling. Previous chromosome, isozyme, and cpDNA surveys did not reveal notable genetic variation within either the parental or the hybrid species. We used nuclear DNA markers (random amplified polymorphic DNA ([RAPD]) and inter-simple sequence repeats (ISSR) to further explore the origin, diversity, and parentage of S. anglica. We found DNA fragments in S. ×townsendii were the aggregate of diagnostic DNA fragments from S. maritima and S. alterniflora, thus confirming its hybrid origin. The S. ×townsendii genotype was identical to most of the S. anglica individuals analyzed, establishing the genetic concordance of these two taxa. We found widespread genetic variation within S. anglica. This could indicate that S. anglica arose several times, from different S. maritima sires. Alternatively, alleles could have been lost through recombination and/or through loss of entire chromosomes in S. anglica. Finally, all but one S. anglica individual had a S. alterniflora component that was indistinguishable from a S. alterniflora plant extant in Marchwood, UK, leaving open the possibility that this plant is the actual seed parent of S. anglica.     

Intraspecific molecular variation inHieracium sect.Alpina (Asteraceae), an apomictic group

Intraspecific variation of four agamospecies ofHieracium sect.Alpina was studied using RAPD and isozyme techniques. No variation in either multiprimer RAPD or multi-enzyme phenotypes was observed withinH. holosericeum, suggesting that this widespread species consists of only a single genotype. A low level of within-population isozyme variation was seen inH. tenuifrons andH. calenduliflorum, the origin of which appears to be consistent with somatic mutation. Most isozyme and all RAPD variation in these two species was partitioned between populations. A strong correlation with geography suggests that its cause may be due to polytopic (-polyphyletic?) origin or perhaps to mutation and dispersal. The most variable species wasH. alpinum, in which isozyme variation occurred mostly within populations rather than between them, suggesting occasional sexual events or that the parents ofH. alpinum were heterozygous. RAPD variation in this species, in contrast, was partitioned between Scottish and Swiss populations, suggesting the existence of geographical races.     

Isozyme Variation among Biological Species in the Gibberella fujikuroi Species Complex (Fusarium Section Liseola)

Isozyme phenotypes were determined for 101 strains of Gibberella fujikuroi and 2 strains of Gibberella nygamai that represent seven biological species (mating populations) isolated from a variety of plant hosts in dispersed geographic locations. Fourteen enzymes were resolved in one or more of three buffer systems. Two of the enzymes, arylesterase and acid phosphatase, were polymorphic within two or more biological species and are suitable for intraspecific studies of population variation. Six enzymes, alcohol dehydrogenase, aspartate aminotransferase, glucose-6-phosphate dehydrogenase, mannitol dehydrogenase, phosphoglucomutase, and phosphogluconate dehydrogenase, were monomorphic in all of the isolates examined. The remaining six enzymes, fumarase, glucose phosphate isomerase, glutamate dehydrogenase (NADP), isocitrate dehydrogenase (NADP), malate dehydrogenase, and triose-phosphate isomerase, could potentially be used to distinguish the different biological species. Mating populations C and D are the most similar, since the mating population C isolates examined had the same isozyme phenotype as did a subset of the isolates in mating population D. Mating population E is the least similar to the other taxa examined. Unique isozyme phenotypes are present but are composed of banding patterns shared among the biological species. This finding supports the hypothesis that these biological species, with the possible exception of mating populations C and D, are reproductively isolated from one another and that no significant gene flow is occurring between them. Isozyme analysis is a useful method to distinguish these closely related biological species. Examination of isozyme phenotypes is more rapid than the present technique, which is based on sexual crosses; can be applied to strains that are not sexually fertile; and is more sensitive than traditional morphological characters, which cannot distinguish more than three or four morphological groups among the seven biological species. While emphasizing the discreteness of the mating populations as biological entities, our isozyme data also reaffirm the close genetic relationship among these groups.     

Quantitative trait, genetic, environmental, and geographical distances among populations of the C4 grass Trachypogon plumosus in Neotropical savannas

Venezuelan savannas are exposed to land‐use changes and biological invasions which compromise their persistence and function. The native C₄ grass Trachypogon plumosus is the most important component of the savannas under diverse combinations of climate and soils, suggesting substantial interpopulation variation. We examined quantitative traits and isozyme variation of nine populations of this grass and related these estimates to geographical and environmental features of sampled locations. Isozyme diversity estimates were based on 10 polymorphic enzyme systems whereas 21 quantitative traits, from field and controlled growth conditions, were evaluated. Distance matrices for quantitative traits, isozyme, geographical and environmental data were subjected to clustering analysis. Correspondence between quantitative trait distance and genetic distance, and their association to geographical and environmental distances were analysed with Mantel tests. All quantitative traits differed significantly among populations. The average Q_ST calculated for eight quantitative traits measured in the greenhouse was 0.157. Isozyme diversity differed significantly among populations. About 28% of total isozyme variation occurred among populations. Significant positive associations were detected between environmental, quantitative field traits, and geographical distance as well as between the later and genetic distances. Genetic distances did not correspond significantly with quantitative traits nor did environmental distances. Ecologically meaningful associations were detected between field quantitative traits, environmental, and geographical data using cluster analysis. Our results support the hypothesis that processes of the neutral type are mainly responsible for the variation patterns observed in T. plumosus populations in Venezuelan savannas. Variation observed for quantitative traits among populations seems to be due to the effect of environmental conditions on phenotypically plastic traits, and not the result of directional selection favouring different phenotypes in different populations.     

Genetic variation within and between populations of Potamogeton pusillus agg.

Patterns of isozyme variation were examined in 17 populations of P. pusillus and P. berchtoldii, together with one population of P. trichoides taken for comparison. Both P. pusillus and P. berchtoldii displayed low levels of variation within populations associated with high levels of interpopulation differentiation. This pattern of partitioning of genetic variation within and between populations is attributed to the founder effect, frequent vegetative propagation by turions, dominant self-fertilization and limited seedling recruitment. The mechanism of pollen transfer was investigated in cultivation. Effective pollination takes place in air above the water surface (autogamy, geitonogamy, anemogamy), on the water surface (epihydrogamy) or below water surface (hydroautogamy). The species are self-compatible. The low level of infra-population variation together with rare occurrence of heterozygotes suggest that selfing is the most frequent mode of pollination, although the protogynous flowers may occasionally permit some cross-pollination. Unique enzyme markers were found for P. pusillus and P. berchtoldii, and also for the single population of P. trichoides. All multienzyme phenotypes were species-specific. Isozyme data support the separate position of P. pusillus and P. berchtoldii. UPGMA dendogram based on enzyme data of 133 plant samples revealed three distinct main enzymatic entities perfectly corresponding to the three morphologically defined species.     

三倍体罗汉果及其亲本的同工酶比较研究

为了加快三倍体罗汉果育种的进程和效率,进一步提高其亲本选择性,该文对F1代3x罗汉果与其4x和2x亲本的过氧化物酶(POD)同工酶和酯酶(EST)同工酶进行了比较。结果表明:F1代3x罗汉果与其4x和2x亲本的POD和EST同工酶均有一定差异,3x和4x的POD和EST同工酶不同迁移率的平均酶带数目较2x多,酶带活性也较强;4x的POD同工酶数目较3x的多,但EST同工酶两者差异较小;F1代3x罗汉果的同工酶均出现了与其4x和2x亲本不同的新酶带,预示着其可能具有的杂种优势;聚类分析结果显示,F1代3x罗汉果与其4x母本的遗传距离更近,并且当4x母本间亲缘遗传关系较近时,其F1代3x在亲缘遗传上也相聚较近,说明F1代3x在亲缘遗传上更倾向于其4x母本。由于三倍体罗汉果育种亲本间存在杂种优势,且更倾向于其母本遗传,因此通过该研究可以初步总结出三倍体罗汉果及其父本、母本的遗传规律,并提出罗汉果三倍体良种选育的建议,即在3x罗汉果育种时需要更加关注4x母本的优良性状表现,并以亲本的遗传背景为基础,选择遗传差异较为显著的父本和母本。     

Weedy adaptation in Setaria spp. II. Genetic diversity and population genetic structure in S. glauca,S. geniculata,and S. faberii (Poaceae)

Setaria glauca (yellow foxtail), S. geniculata (knotroot foxtail), and S. faberii (giant foxtail) are important cosmopolitan weeds of temperate and tropical regions. Isozyme markers were used to investigate genetic diversity and population genetic structure in 94 accessions of yellow foxtail, 24 accessions of knotroot foxtail, and 51 accessions of giant foxtail, collected mainly from North America and Eurasia. Giant foxtail populations were nearly identical genetically, with only one population exhibiting isozyme polymorphism. Yellow and knotroot foxtail populations had low genetic diversity but marked population differentiation. Although the latter species are similar morphologically, they are readily distinguished electrophoretically, with Nei's genetic identity being 0.83. In both species, genetic divergence between accessions from Eurasia and North America was minimal. Populations from the native ranges had slightly greater genetic diversity than those from the respective introduced ranges. Yellow foxtail populations genetically clustered into Asian, European, and North American groups. Within North America, yellow foxtail populations from Iowa were genetically diverse whereas populations collected from other North American locations were nearly monomorphic for the same multilocus genotype. Knotroot foxtail populations in North America were genetically differentiated into northern and southern groups on either side of a line at ≈37° N latitude. No genetic patterning was evident in knotroot foxtail populations from Eurasia. In both yellow and knotroot foxtail, patterns of population genetic structure have been influenced by several factors, including genetic bottlenecks associated with founder events, genetic drift, and natural selection.     

Isozyme uniformity in a wild extinct insular plant, Lysimachia minoricensis J.J. Rodr. (Primulaceae)

Isozyme analyses were conducted to evaluate the genetic diversity of seed accessions of Lysimachia minoricensis (a Balearic endemic plant that became extinct in the wild) provided by 10 European botanical gardens. No isozyme variation was detected after examination of the electrophoretic patterns of 22 putative loci in more than 150 plants. The lack of genetic variation in L. minoricensis is probably due to the shortage of sample propagules originally recovered in the field before its extinction. Extant plants of L. minoricensis are believed to have originated from a single source, therefore limiting their use as seed stocks for restoration projects.     

Conservation genetics of the endangered endemic Hawaiian genus Brighamia (Campanulaceae)

The endemic Hawaiian genus Brighamia (Campanulaceae) comprises two federally endangered, morphologically similar species, B. insignis from Kaua`i and Ni`ihau and B. rockii from Moloka`i. To assist the design of conservation management programs for these taxa, isozyme analyses were performed to assess the levels of genetic diversity at the population and species levels, including comparisons within and among seven natural populations and one ex situ collection each of B. insignis and B. rockii. Our sampling (N = 80) represents ~41% of all known individuals in the wild. Isozyme analyses revealed levels of genetic variation comparable to those reported for other Hawaiian flowering plant taxa but low levels of genetic variation at the population and species levels when compared to flowering plants in general. Ex situ individuals (N = 61) were genetically representative of natural populations and hence may appropriately serve as stock for population augmentations. The two morphologically similar Brighamia species were highly distinct genetically. The combination of morphological and ecological similarity with allozymic dissimilarity observed in Brighamia is unique among the Hawaiian taxa studied to date.     

Isozyme variation under different modes of reproduction in two clonal winter annuals, Sedum rosulato-bulbosum and Sedum bulbiferum (Crassulaceae)

We studied isozyme variation in two annual species that produce bulbils, Sedum rosulato-bulbosum , which includes both sexually reproducing plants and obligate clonal plants that result from triploidy (fertile and sterile S. rosulato-bulbosum , respectively), and an obligate clonal plant, Sedum bulbiferum , to examine the relationship between reproductive mode and isozyme variation. The sterile S. rosulato-bulbosum population exhibited no genotypic variation, but showed high genetic variation (gene diversity, H _e  = 0.60) because five of the six loci that we analyzed were heterozygous. Almost all ramets of S. bulbiferum across 20 populations shared an identical isozyme phenotype, although we could not identify the genetic basis of the phenotype. In contrast, fertile S. rosulato-bulbosum exhibited genotypic variation across the species, but comprised genotypically uniform and polymorphic populations whose genotypic variations correlated positively with the genetic variations within the populations ( H _e at the genet level per population ranged from 0.08 to 0.37). Genetic drift and habitat conditions inhibiting seedling recruitment may have caused this among-population variation. The results for sterile and fertile S. rosulato-bulbosum suggest that exclusive clonal reproduction causes low genotypic variation, but maintains genetic variation within individuals. Factors that affect the maintenance of genetic variation in these plants are discussed on the basis of these findings.     

The origin of cornbelt maize: The isozyme evidence

Historical records show t hat the Midwestern dent corns of the United States originated from hybridization of two landraces, Northern Flint and Southern Dent. We examined the origin of Southern and Midwestern Dents by means of isozyme electrophoresis. Isozyme genotypes were determined for 23 loci in 12 plants each of 32 accessions of Southern Dent. Previously published isozyme data for maize landraces of Mexico and North America and for U.S. Midwestern Dents were included for comparative purposes. The data show that Northern Flint and Southern Dent are among the isozymically most divergent maize landraces. Nei’s genetic identities between populations of these two landraces are very low for conspecific populations (ca. 0.80). Southern Dent of the southeastern U.S. appears closely related to similar dent corns of southern Mexico, supporting a previously published hypothesis that U.S. Southern Dent is largely derived from the dent corns of southern Mexico. The Midwestern Dents, which resulted from crosses of Southern Dent and Northern Flint, are much more like Southern Dent than Northern Flint in their isozyme profile. Similarly, public inbreds show greater affinity to Southern Dent with the exception of sweet corn lines, which resemble Northern Flint in their isozyme allele frequencies. North American public inbreds do not contain appreciable isozymic variation beyond that found in Northern Flint and Southern Dent.     

Isozyme Variability in Plants Regenerated from Calli of Cereus peruvianus (Cactaceae)

Morphological and isozyme variation was observed among plants regenerated from callus cultures of Cereus peruvianus. Different morphological types of shoots (68%) were observed in 4-year-old regenerated plants, while no distinct morphological variants were observed in plants grown from germinated seeds. Isozyme patterns of 633 plants regenerated from calli and of 261 plants grown from germinated seeds showed no variation in isocitrate dehydrogenase isozyme, and the differential sorbitol dehydrogenase, alcohol dehydrogenase, malate dehydrogenase, acid phosphatase, and peroxidase isozyme patterns observed in regenerated plants were attributed to nonallelic variation. Allelic variation was detected at three isoesterase loci. The proportion of polymorphic loci for both populations was 13.6% and the deviation from Hardy–Weinberg equilibrium for the Est-1 and Est-7 loci observed in somaclones was attributed to the manner in which the regenerant population was established. The high values for genetic identity among regenerant and seed-grown plant populations are in accordance with the low levels of interpopulation genetic divergence. In somaclones of C. peruvianus, morphological divergence was achieved within a short time but was not associated with any isozyme changes and also was not accompanied by biochemical genetic divergence.     

Isozyme studies in provenance research of forest trees

Summary The nature and origin of the isozymes and the techniques for their detection in forest trees are briefly reviewed. The theories used to interpret the isozyme variation are summarized. Recent isozyme variation studies in provenance research are discussed in relation to known variation pattern detected by classical nursery and field tests. The populations of a tree species can be sometimes, but not always, distinguished by their isozyme patterns. For a number of species, relationships between environment of origin of the provenances and some isozyme frequencies have been statistically established. In one case (Picea sitchensis) where direct comparison between the variation pattern detected by isozymes and the genetic variation of silviculturally important traits was possible, no meaningful relationships between both patterns could be detected. Nei's genetic distances and indices of gene diversity do not appear to be useful in provenance research. The concept of genetic distance based on gene frequencies is probably not very useful in provenance research either.     

Molecular tests of the proposed diploid hybrid originof Gilia achilleifolia (Polemoniaceae)

Gilia achilleifolia is a putative diploid hybrid species. Hybrid origin was hypothesized based on traditional biosystematicevidence (i.e., morphological, cytological, and crossability data),which may be insufficient to establish genealogical history. Here,phylogenetic analysis of sequence data from the internal transcribedspacer (ITS) regions is used to examine the relationship between theputative hybrid species and its proposed parents. Isozyme variation isassayed to test for genetic additivity in the putative hybrid taxon andmorphological data are analyzed cladistically to evaluate the charactersthat led to the original hypothesis of hybrid origin. The ITS-basedgene tree placed G. achilleifolia in two divergent clades, eachsister to one of the putative parental lineages. Little isozymeadditivity was observed and G. achilleifolia possessed sixunique alleles among 42 alleles observed. However, ITS and isozymetrees differed in their placement of the two lineages of G.achilleifolia; both lineages are closer to a third putative parentin the isozyme tree. Also, G. achilleifolia is intermediate orpolymorphic for all nine morphological characteristics differentiatingthe parental species. Sorting of ancestral polymorphisms cannot easilyaccount for expression patterns of seven of these characters. In ourview, these results fail to distinguish between alternative hypothesesof ancient hybrid origin and divergent evolution, belying the difficultyof detecting ancient hybrids.     

松属植物遗传多样性研究进展

研究松属遗传多样性的方法涉及表型、同工酶、染色体、DNA等多层面。 松树表型性状变异广泛,其不同树种不同性状的遗传力（或遗传率）均存在差异。到目前为止,同工酶仍是检测松树遗传多样性的最常用方法,一般而言,松属树种群体内等位酶多样性程度高,群体间分化较低,但各树种的情形也不尽相同。松属树种染色体水平的变异很低,其核型高度一致。核DNA组较一般阔叶树大,遗传多样性丰富,但叶绿体等质体DNA则多样性较低。影响遗传多样性的因素很多,其中自身的交配系统和外部的生长环境是影响它的两个主要因素。最后,回顾了松树的起源及其遗传多样性保护策略等方面的研究。 Abstract:The ways of probing genetic diversity of pines involve many aspects,such as morphology,chromosome,isozyme,DNA,etc.The phenotypic characteristics in pines vary widely and the differences of inheritability(h2) are obvious among characteristics and among species.Up to now,isozyme is still the most common means to measure genetic diversity of pines.Generally,there are high allozyme diversity within populations and low differentiation coefficient among populations,but differences exist between species in Pinus.The variations of chromosome among pines are very low and the karyotypes of pines are consentaneous,but the genomes of pines in cell nucleus are much larger than that of broadleaves.Diversity of pines are abundant at nucleus DNA level but are poor at plastid DNA level,such as ctDNA.There are many factors that will affect genetic diversity of pines,in which mating system and environment are two main factors.Finally,we reviewed the research on origin of Pinus and conservation strategy of genetic diversity,etc.     

Biochemical genetic variation in populations of golden trout, Salmo aguabonita: evidence of the threatened Little Kern River golden trout, S.a. whitei.

Eight wild populations of the High Sierra golden trout, Salmo aguabonita, and one domestic stock of rainbow trout, Salmo gairdneri, were examined for biochemical-genetic variation in eight protein systems. Variation within the eight systems was determined by at least 10 loci in both golden and rainbow trout and all the alleles identified in rainbow trout were observed as electro-phoretically identical phenotypes in golden trout. Variation was observed at an average of 51 percent of the loci in the golden trout samples and for five of the 10 loci in the rainbow trout. Average heterozygosity ranged from 12.6 to 13.9 percent for seven of the golden trout populations with one showing a low value of 5.4 percent. A comparable estimate of 12.1 percent was found for the rainbow stock. On the basis of genetic variation and allele frequencies at three loci, the eight golden trout populations were divided into two distinct groups. Three populations sampled from the Little Kern River basin tended to be genetically distinct from two additional Little Kern River basin populations and from three geographically distinct populations sampled from the eastern Kern River area. The former three populations were hypothesized to be of a recent rainbow-golden hybrid origin. Trout in the other two Little Kern River basin populations, sampled in head-waters of a stream tributary to the Little Kern River, were considered to be the threatened Little Kern golden trout, S. a. whitei Evermann, due to their high degree of genetic similarity to the geographically distinct subspecies S. a. aguabonita sampled from the eastern Kern River area. The finding of substantial genetic variation in the wild golden trout populations indicates that this threatened species is not at present genetically impoverished and thus does not appear to be in immediate danger of extinction through lack of adaptive capability.