Amplified Fragment Length Polymorphism Variation Research Articles

Phylogeographic structure of Terminalia franchetii (Combretaceae) in southwest China and its implications for drainage geological history

Following the rapid uplift of the Qinghai-Tibetan Plateau, the reorganization of the major river drainages in southwest China was primarily caused by river capture events. However, the impact of these past changes in drainage patterns on the current distribution and genetic structure of the endemic flora of this region remains largely unknown. Here we report a survey of amplified fragment length polymorphism (AFLP) in Terminalia franchetii, an endemic shrub or small tree of the deep and dry-hot river valleys of this region. We surveyed AFLP variation within and among 21 populations (251 individuals) of T. franchetii, distributed disjunctively between northern and southern drainage systems. Using STRUCTURE, principal coordinates analysis, and genetic distance methods, we identified two main population genetic groups (I and II) and four subgroups within the species, as follows: (I) the Upper Jinshajiang Valley (subgroup I((north))) and the Honghe drainage area (subgroup I((south))); (II) the Middle and Lower Jinshajiang and Yalongjiang Valleys (subgroup II((north))) and the Nanpanjiang drainage area (subgroup II((south))). Genetic diversity was lower in group I than in group II. According to the genetic diversity and genetic structure results, we suggest that the modern disjunctive distribution and associated patterns of genetic structure of T. franchetii result from vicariance caused by several historical drainage capture events, involving the separation of the Upper Jinshajiang, Yalongjiang and Daduhe from the Honghe or Nanpanjiang in southwest China.

The extreme Beringian/Atlantic disjunction in Saxifraga rivularis (Saxifragaceae) has formed at least twice

AbstractAim The oceanic Saxifraga rivularis L. presents one of the most extreme disjunctions known in the arctic flora: it has a small amphi‐Beringian range and a larger amphi‐Atlantic one. It was recently suggested to have had a single allopolyploid origin in Beringia at least one glacial cycle ago, followed by gradual expansion in a more humid period and differentiation into two allopatric subspecies (the Atlantic ssp. rivularis and the Beringian ssp. arctolitoralis). Here we explore the history of its extreme disjunction.Location The amphi‐Beringian and northern amphi‐Atlantic regions.Methods We obtained amplified fragment length polymorphisms (AFLPs) and chloroplast DNA sequences from 36 populations (287 individuals) and 13 populations (15 individuals), respectively. The data were analysed using principal coordinates analyses, Bayesian clustering methods, and analyses of molecular variance.Results Two distinctly divergent AFLP groups were observed, corresponding to the two described subspecies, but, surprisingly, four of the West Atlantic populations belonged to the supposedly Beringian endemic ssp. arctolitoralis. This was confirmed by re‐examination of their morphological characteristics. The overall AFLP diversity in the species was low (26.4% polymorphic markers), and there was no variation in the five investigated chloroplast DNA (cpDNA) regions. There was little geographic structuring of the AFLP diversity within each subspecies, even across the extreme disjunction in ssp. arctolitoralis, across the Bering Sea, and across the Atlantic Ocean, except that most plants from the arctic Svalbard archipelago formed a separate genetic group with relatively high diversity.Main conclusions The extreme disjunction in S. rivularis has evidently formed at least twice. The first expansion from Beringia was followed by allopatric differentiation into one Beringian and one Atlantic subspecies, which are distinctly divergent at AFLP loci but still harbour identical cpDNA haplotypes, suggesting that the expansion was quite recent but before the last glaciation. The next expansion from Beringia probably occurred by means of several long‐distance dispersals in the current interglacial, resulting in the colonization of the western Atlantic region by ssp. arctolitoralis. The poor geographic structuring within each subspecies suggests frequent long‐distance dispersals from two main Weichselian refugia, one Beringian and one western‐central European, but it is possible that the genetic group in Svalbard originates from an additional refugium.

Amplified Fragment Length Polymorphism Diversity of Cultivated White Oyster Mushroom Pleurotus ostreatus

Amplified fragment length Polymorphism (AFLP) analysis was performed to study the genetic diversity of fifteen isolates of white oyster mushroom ( Pleurotus ostreatus ) originated from different places in Indonesia (Java, Sumatra, Bali, and Kalimantan) and Thailand. The majority of studied isolates originated from commercial mushroom, and some of them were collected from nature. AFLP analysis revealed 202 loci of positive DNA bands. Heterozygosity value showed polymorphisms of the fifteen studied isolates. Correlation and cluster analyses of the isolates showed that isolate from Thailand (BNK isolate), isolate from Bogor (AMD isolate), and isolate from Purwokerto (USX isolate) were distinctly different from the other isolates indicated by positive bands with DNA size greater than 650 bp. Isolate from Kalimantan (BJM isolate) was distinctly different from Java island isolates. Generally there was no correlation of AFLP variability and geographic distribution of commercial strain isolates.

Genetic variation in remnant Festuca hallii populations is weakly differentiated, but geographically associated across the Canadian Prairie

AbstractGenetic diversity is essential for predicting plant evolutionary potential and for formulating conservation strategies. However, little is known about the genetic variation of plains rough fescue [Festuca hallii (Vasey) Piper], a widespread and diagnostic grass in the Fescue Prairie. We used the amplified fragment length polymorphism (AFLP) technique to assess the genetic variation of 30 fragmented populations of F. hallii across the northern Canadian Prairie and its associations with 12 geographical, fragmentation and environmental attributes. Three AFLP primer pairs were used to screen 840 samples, representing approximately 30 samples from each population, and 246 polymorphic bands were scored. The fescue plant was genetically diverse, as revealed by the proportion of polymorphic bands (0.870–0.967), the mean band frequency (0.364–0.457) and the within‐population variation (69.4–85.4). The genetic variation was not highly differentiated, with only 6.5% of the total AFLP variation residing among populations. A Mantel test revealed a significant correlation between genetic and geographical distances (r = 0.39; P < 0.004) and a spatial autocorrelation up to 60 km among populations was detected. The AFLP variation displayed a longitudinal decline and was significantly associated with environmental attributes related to moisture, indicating local adaptation. However, the AFLP variation was not significantly associated with the estimated population size and geographical distance to the nearest neighbor, suggesting that fragmentation has not generated considerable genetic impact on the fescue populations. Implications for fescue conservation, restoration and management are discussed.

Contrasting patterns of genetic diversity at three different genetic markers in a marine mammal metapopulation

Many studies use genetic markers to explore population structure and variability within species. However, only a minority use more than one type of marker and, despite increasing evidence of a link between heterozygosity and individual fitness, few ask whether diversity correlates with population trajectory. To address these issues, we analysed data from the Steller's sea lion, Eumetiopias jubatus, where three stocks are distributed over a vast geographical range and where both genetic samples and detailed demographic data have been collected from many diverse breeding colonies. To previously published mitochondrial DNA (mtDNA) and microsatellite data sets, we have added new data for amplified fragment length polymorphism (AFLP) markers, comprising 238 loci scored in 285 sea lions sampled from 23 natal rookeries. Genotypic diversity was low relative to most vertebrates, with only 37 loci (15.5%) being polymorphic. Moreover, contrasting geographical patterns of genetic diversity were found at the three markers, with Nei's gene diversity tending to be higher for AFLPs and microsatellites in rookeries of the western and Asian stocks, while the highest mtDNA values were found in the eastern stock. Overall, and despite strongly contrasting demographic histories, after applying phylogenetic correction we found little correlation between genetic diversity and either colony size or demography. In contrast, we were able to show a highly significant positive relationship between AFLP diversity and current population size across a range of pinniped species, even though equivalent analyses did not reveal significant trends for either microsatellites or mtDNA.

Comparison of quantitative and molecular genetic variation of native vs. invasive populations of purple loosestrife (Lythrum salicaria L., Lythraceae)

Study of adaptive evolutionary changes in populations of invasive species can be advanced through the joint application of quantitative and population genetic methods. Using purple loosestrife as a model system, we investigated the relative roles of natural selection, genetic drift and gene flow in the invasive process by contrasting phenotypical and neutral genetic differentiation among native European and invasive North American populations (Q(ST) - F(ST) analysis). Our results indicate that invasive and native populations harbour comparable levels of amplified fragment length polymorphism variation, a pattern consistent with multiple independent introductions from a diverse European gene pool. However, it was observed that the genetic variation reduced during subsequent invasion, perhaps by founder effects and genetic drift. Comparison of genetically based quantitative trait differentiation (Q(ST)) with its expectation under neutrality (F(ST)) revealed no evidence of disruptive selection (Q(ST) > F(ST)) or stabilizing selection (Q(ST) < F(ST)). One exception was found for only one trait (the number of stems) showing significant sign of stabilizing selection across all populations. This suggests that there are difficulties in distinguishing the effects of nonadaptive population processes and natural selection. Multiple introductions of purple loosestrife may have created a genetic mixture from diverse source populations and increased population genetic diversity, but its link to the adaptive differentiation of invasive North American populations needs further research.

The influence of sea currents, past disruption of gene flow and species biology on the phylogeographical structure of coastal flowering plants

AbstractAim We investigate the geographical genetic structure of two coastal plant species, Cakile maritima Scop. (Brassicaceae) and Eryngium maritimum L. (Apiaceae), through three sea straits and along one continuous stretch of coast using amplified fragment length polymorphisms (AFLPs). The two species have a similar ecology in that they grow in sandy habitats, but differ in life‐form (annual vs. perennial) and dispersability of seeds by sea water as inferred from floating experiments. The sea straits differ in their geological history and their modern current systems. The primary goal of our study was to test the hypothesis that sea straits have an influence on the geographical patterns of genetic variation at the population level.Location The areas around the Strait of Gibraltar, the Dardanelles, the Bosporus and the Atlantic coast of western France.Methods For both species we investigated AFLP variation in several populations from each area. Bayesian clustering and diversity and differentiation measures were used to analyse the genetic data.Results In most areas the spatial genetic structure was similar between the two species. They share the presence of distinct genetic gaps along the coast through the Strait of Gibraltar and the Bosporus, and these genetic gaps coincide with the straits. Both species show genetic continuity along the coast of western France. A distinct genetic gap was found through the Dardanelles for C. maritima but not for E. maritimum.Main conclusions The study shows that sea straits have an influence on the geographical patterns of genetic variation. Sea currents are inferred to cause the genetic gap through the Strait of Gibraltar. In the Bosporus and, for C. maritima, through the Dardanelles, the genetic gaps found are explained by the past closure of these two straits as well as by present‐day factors. Simulations indicate that the lower differentiation of C. maritima through the Dardanelles than through the Bosporus cannot be explained by the difference in geological history of these two straits. The difference in seed dispersability between the two species is argued to be responsible for the observation that differentiation among genetic clusters is higher in E. maritimum than in C. maritima where a direct comparison is possible.

The phylogeographical history of the Iberian steppe plant Ferula loscosii (Apiaceae): a test of the abundant‐centre hypothesis

The geology and climate of the western Mediterranean area were strongly modified during the Late Tertiary and the Quaternary. These geological and climatic events are thought to have induced changes in the population histories of plants in the Iberian Peninsula. However, fine-scale genetic spatial architecture across western Mediterranean steppe plant refugia has rarely been investigated. A population genetic analysis of amplified fragment length polymorphism variation was conducted on present-day, relict populations of Ferula loscosii (Apiaceae). This species exhibits high individual/population numbers in the middle Ebro river valley and, according to the hypothesis of an abundant-centre distribution, these northern populations might represent a long-standing/ancestral distribution centre. However, our results suggest that the decimated southern and central Iberian populations are more variable and structured than the northeastern ones, representing the likely vestiges of an ancestral distribution centre of the species. Phylogeographical analysis suggests that F. loscosii likely originated in southern Spain and then migrated towards the central and northeastern ranges, further supporting a Late Miocene southern-bound Mediterranean migratory way for its oriental steppe ancestors. In addition, different glacial-induced conditions affected the southern and northern steppe Iberian refugia during the Quaternary. The contrasting genetic homogeneity of the Ebro valley range populations compared to the southern Iberian ones possibly reflects more severe bottlenecks and subsequent genetic drift experienced by populations of the northern Iberia refugium during the Pleistocene, followed by successful postglacial expansion from only a few founder plants.

Assessment and comparison of AFLP and SSR based molecular genetic diversity in Indian isolates of Ascochyta rabiei, a causal agent of Ascochyta blight in chickpea (Cicer arietinum L.)

Ascochyta blight (AB), caused by Ascochyta rabiei (Pass.) Labr. (anamorph), is the most damaging disease of chickpea (Cicer arietinum L.) and is a serious biotic stress constraint for chickpea production. To understand the molecular diversity in A. rabiei populations of India, a total of 64 isolates collected from AB-infected chickpea plants from different agroclimatic regions in the North Western Plain Zone (NWPZ) of India were analyzed with 11 AFLP (amplified fragment length polymorphism) and 20 SSR (simple sequence repeat) markers. A total of 9 polymorphic AFLP primer pairs provided a total of 317 fragments, of which 130 were polymorphic and showed an average PIC value 0.28. Of the SSR markers, 12 showed polymorphism and provided a total of 29 alleles with an average PIC value 0.35. To the best of our knowledge, this is the first report on a comparison of AFLP and SSR diversity estimates in A. rabiei populations. The dendrogram developed based on AFLP and SSR data separately, as well as on the combined marker dataset, grouped the majority of AB isolates as per geographic regions. Model based population structure analysis revealed four distinct populations with varying levels of ancestral admixtures among 64 isolates studied. Interestingly, several AFLP primer combinations and SSR markers showed the locus/allele specific to AB isolates of certain regions, e.g., Hisar, Sriganganagar, Gurdaspur, and Sundarnagar. Genetic variability present in AB isolates of the NWPZ of India suggests the continuous monitoring of changes in A. rabiei population to anticipate the breakdown of AB resistance in chickpea cultivars grown in India.

Strait of Gibraltar: an effective gene‐flow barrier for wind‐pollinated Carex helodes (Cyperaceae) as revealed by DNA sequences, AFLP, and cytogenetic variation

The Strait of Gibraltar is the most important barrier disconnecting the landmasses of Europe and Africa on the western Mediterranean extreme. Carex helodes is a wind-pollinated species endemic to the western Mediterranean. Because molecular and cytogenetic data allow the inference of its evolutionary history, we analyzed variations in chromosome number, including meiotic chromosome behavior, amplified fragment length polymorphism (AFLP) fingerprints, and nucleotide substitutions in plastid and nuclear DNA sequences. Cytogeographic results showed that the African populations have stabilized at a single chromosome number of 2n = 74, whereas the most frequent cytotype in Iberia is 2n = 72. Phylogenetic reconstructions of 17 sequences from nine closely related species revealed that C. helodes is monophyletic and that the Moroccan populations are embedded in the Iberian lineages. The haplotype network is also consistent with a European origin of the northern African haplotype. AFLP analysis also revealed hierarchical levels of genetic variation compatible with a founder effect process responsible for the African populations. All sources of evidence support the hypothesis that the Strait of Gibraltar has been an effective gene-flow barrier, generating two isolated evolutionary lineages after their dispersal. Recent connections between the two lineages appear unlikely, whereas active gene flow occurs among populations within the two lineages.

AFLP variation in 25 Avena species

Current molecular characterization of ex situ plant germplasm has placed more emphasis on cultivated gene pools and less on exotic gene pools representing wild relative species. This study attempted to characterize a selected set of germplasm accessions representing various Avena species with the hope to establish a reference set of exotic oat germplasm for oat breeding and research. The amplified fragment length polymorphism (AFLP) technique was applied to screen 163 accessions of 25 Avena species with diverse geographic origins. For each accession, 413 AFLP polymorphic bands detected by five AFLP primer pairs were scored. The frequencies of polymorphic bands ranged from 0.006 to 0.994 and averaged 0.468. Analysis of molecular variance revealed 59.5% of the total AFLP variation resided among 25 oat species, 45.9% among six assessed sections of the genus, 36.1% among three existing ploidy levels, and 50.8% among eight defined genome types. All the species were clustered together according to their ploidy levels. The C genome diploids appeared to be the most distinct, followed by the Ac genome diploid A. canariensis. The Ac genome seemed to be the oldest in all the A genomes, followed by the As, Al and Ad genomes. The AC genome tetraploids were more related to the ACD genome hexaploids than the AB genome tetraploids. Analysis of AFLP similarity suggested that the AC genome tetraploid A. maroccana was likely derived from the Cp genome diploid A. eriantha and the As genome diploid A. wiestii, and might be the progenitor of the ACD genome hexaploids. These AFLP patterns are significant for our understanding of the evolutionary pathways of Avena species and genomes, for establishing reference sets of exotic oat germplasm, and for exploring new exotic sources of genes for oat improvement.

Glacial history of Saxifraga paniculata (Saxifragaceae): molecular biogeography of a disjunct arctic-alpine species from Europe and North America

The molecular biogeography of the disjunctly distributed and morphologically highly variable species Saxifraga paniculata Mill. was analysed using amplified fragment length polymorphism (AFLP) and chloroplast microsatellites. The study comprised 77 samples from mountain regions in Europe and North America throughout the complete range of distribution. AFLP data revealed clear genetic differentiation between samples from the Arctic, the Caucasus, and the eastern European mountains. Samples from the Alps were divided into two groups. One group clustered with the samples from central Europe and the Pyrenees, whereas another group with individuals from southern Norway. AFLP diversity was lowest in the Arctic and highest in the Alps. Chloroplast microsatellite analysis revealed eight haplotypes but no unequivocal phylogeographical pattern. However, haplotype diversity was highest in the Alps and central Europe whereas, in the Arctic, only few widespread haplotypes could be found. The results indicate in situ survival of S. paniculata in the Caucasus, the eastern European mountains, and the Alps. The Arctic has presumably been colonized postglacially from North American refugia south of the ice shield. Southern Norway and the Pyrenees have most likely been colonized from two phylogeographically different groups in the Alps. The origin of the central European samples remains ambiguous. In situ survival seems to be as possible as several postglacial recolonization events from the Alps. The obtained molecular data clearly support the subdivision of S. paniculata into three subspecies: ssp. cartilaginea from the Caucasus, ssp. laestadii from northern Norway, Iceland, and North America, and ssp. paniculata from the other geographical regions.

Genetic Diversity in Populations of Phytophthora citricola Associated with Horticultural Crops in California.

California populations of the plant pathogen Phytophthora citricola were examined for amplified fragment length polymorphism (AFLP), pathogenicity on almond, and sensitivity to mefenoxam. The characterizations of AFLP variation and mefenoxam sensitivity were based on 86 isolates (44 from almond, 11 from avocado, 3 from strawberry, 18 from walnut, and 10 from six other hosts). Cluster analysis of the AFLP data using the unweighted pair group method indicated a high level of genetic diversity among the isolates, and four main clusters were identified-one dominated by isolates from almond, another including all isolates from avocado, and two including isolates from several hosts other than avocado. Analysis of molecular variance revealed that 38.4 and 24.9% of the AFLP variation were associated with host and geographical factors, respectively. Of 24 isolates, including those from almond, avocado, strawberry, and walnut, 22 were aggressive on almond shoots; there was no evidence of host specificity. All but 1 of the 86 isolates grew at different rates on V8 juice medium amended with mefenoxam at 1 ppm, indicating partial tolerance to the fungicide. Isolates of P. citricola from California populations are genetically diverse, and much of the variation is associated with host and geography. These populations are all potentially pathogenic on almond and tolerant to mefenoxam.

Crossability and amplified fragment length polymorphism variation among four types of Panicum coloratum L. and meiotic chromosome paring of their F1 hybrids

AbstractThe objective of this study was to determine the genetic relationships among four types (A, B, C and D) of Panicum coloratum L. on the basis of the crossability and the amplified fragment length polymorphism (AFLP) variations among the types, and the meiotic chromosome pairing and fertility in F1 hybrids obtained from crosses between different types. F1 hybrids were obtained from crosses between P. coloratum L. var. makarikariense cv. Burnett (A) and P. coloratum L. PI 404632 (D), and between P. coloratum L. CPI 16794 (B) and P. coloratum L. cv. Solai (C). Other crosses failed to produce F1 hybrid seeds. F1 hybrids between Burnett (A) and PI 404632 (D) had 2n = 36 chromosomes and regular meiotic chromosome pairing with 18 bivalents per cell. They produced approximately 80% stainable pollen and normally set seeds. F1 hybrids between Solai (C) and CPI 16794 (B) had 2n = 45 chromosomes and a mean chromosome pairing of 9.50 univalents, 17.67 bivalents and 0.05 trivalents. The most frequent paring behavior was nine univalents and 18 bivalents, suggesting that Solai has two genomes homologous to those of CPI 16794 and one different genome. The F1 hybrids showed approximately 80% stainable pollen and approximately 20% seed fertility. Cluster analysis of polymorphic AFLP in five plants of each of the four types yielded two major clusters. Burnett (A) and PI 404632 (D) clustered together, while Solai (C) clustered with CPI 16794 (B). The results of hybridization studies and AFLP analysis of the four types of P. coloratum, and meiotic examination of the derived F1 hybrids demonstrate that types A and D are genetically differentiated from types B and C, and are reproductively isolated from these two types.

Population decline despite high genetic diversity in the new allopolyploid speciesSenecio cambrensis(Asteraceae)

Senecio cambrensis (Welsh groundsel) is a new allohexaploid species, which originated in Wales, UK, in the early part of the 20th century following hybridization between the native tetraploid groundsel (Senecio vulgaris) and the introduced diploid Oxford ragwort (Senecio squalidus). A survey of the number of populations and flowering individuals per population of S. cambrensis in Wales was conducted at peak flowering time in June 2002, 2003 and 2004. The results show a dramatic decrease in both population number and population size of the species since the 1980s when the last population census was conducted. A survey of amplified fragment length polymorphism (AFLP) variation showed that this decline has occurred despite the fact that S. cambrensis contains a high level of genetic diversity with each individual screened possessing a unique multilocus phenotype. The level of variance within the species was similar to that found in one parent (S. vulgaris) and slightly greater than that among samples of the other parent (S. squalidus). Only a small proportion (5%) of AFLP diversity was partitioned among populations indicating a lack of population structure and possibly high levels of gene flow via seed dispersal in what is predominantly a selfing species. Senecio cambrensis showed closer similarity in AFLP phenotype to S. vulgaris than to S. squalidus. Possible causes of this and also the high level of AFLP diversity found in S. cambrensis are discussed. It is suggested that intergenomic recombination following occasional multivalent formation during meiosis in S. cambrensis is likely to be an important cause of both phenomena, although other causes are not ruled out.

Genetic diversity and biogeography of the traditional Chinese medicine, Gardenia jasminoides, based on AFLP markers

Gardenia jasminoides Ellis is used in traditional Chinese medicine (TCM) in China. Levels of genetic variation and patterns of population structure within and among eight wild or cultivated populations of G. jasminoides Ellis in China were investigated using amplified fragment length polymorphism (AFLP) markers. Of the 11 primers screened, four produced highly reproducible AFLP bands. Using these primers, 244 discernible DNA fragments were generated with 165 bands (67.6%), were polymorphic, indicating considerable genetic variation at the species level. In contrast, there were relatively low levels of polymorphism at the population level with the percentage of polymorphic bands (PPB) ranging from 36.89% to 59.43%. Genetic diversity within populations ranged from 0.2086 to 0.3108, averaging 0.2392 at the species level. A high level of genetic differentiation among populations was detected based on Nei's genetic diversity analysis (76.59%), Shannon's index analysis (64.8%) and AMOVA analysis (72.75%). No significant statistical differences (analysis of molecular variance [AMOVA], p = 0.0639) in AFLP variation were found between regions. However, the variance among populations and within populations differed significantly ( p < 0.001). An indirect estimate of historical levels of gene flow ( N m = 1.7448) was consistent with the high mean genetic identity (mean I = 0.9263) found among populations. There is an association between geographic and genetic distances between populations. Presently gene change exists between populations.

Genetic diversity ofSinapis albagermplasm as revealed by AFLP markers

Sinapis albaL. is a major specialty crop grown as a condiment in western Canada, but little is known about its genetic diversity. The objective of this study was to assess the level and pattern of genetic diversity in a collection of 127S. albaaccessions held at Plant Gene Resources of Canada using amplified fragment length polymorphism (AFLP) markers. Five AFLP primer pairs were applied, and 134 polymorphic bands were scored for each accession. These scored bands had frequencies of occurrence ranging from 0.02 to 0.99 with an average of 0.69. More AFLP variation was found within single (79.1%) than between (20.9%)S. albaaccessions. A small degree of AFLP difference (1.7%) was observed among the accessions of various regions, while relatively large variation (9.2%) existed among the accessions of various countries. A large AFLP difference (15.6%) also existed between the yellow- and brown-seeded accessions, but only 6.2% difference was observed between the cultivar and landrace accessions. Two distinct groups ofS. albagermplasm were identified on the basis of the seed colour (yellow or brown), although a few mixtures also existed. No apparent ‘duplicated’ accessions were observed. The most diverse accessions were from Italy, Spain, France and Greece. Among the most genetically distinct accessions were SA97 from Portugal, SA89 and SA88 from France, SA83 from Russia and SA57 from Italy. These findings are significant not only for managingS. albagermplasm, but also for identifying diverse germplasm that can be used by plant breeders to improveS. albaseed yield and quality parameters.

Fish and seafood traceability based on AFLP markers: Elaboration of a species database

Several sociological, health and conservation arguments request a correct labelling of seafood products. Nowadays, molecular genetics is a useful tool for food chain traceability, particularly in regards to species identification. Among the variety of PCR-based molecular markers, AFLPs (Amplified Fragment Length Polymorphisms) have recently been used to investigate genomes of different complexities. This paper assesses the potential use of the AFLP technology to determine fish and seafood species in processed commercial products and domestic stocks. In particular a species database of fish, molluscs and crustaceans has been created with the aim to identify species of origin of seafood products by previously defined AFLP patterns. Different EcoRI and TaqI primer combinations were selected from 20 screened combinations in relation to the total number of detected fragments and polymorphic ones. Most informative combinations were E32/T32, E32/T33, E33/T33, E33/T37, E33/T38, E40/T33, E40/T37, E42/T32, E42/T37. The comparison of informative markers between unknown frozen or fresh products and reference samples has enabled the accurate identification of 32 different species. The taxonomic characterization has been performed either at the species or at the population level depending on the number of available individuals. AFLP variation at the population level is particularly helpful for the stock traceability of domestic strains. Size homoplasy was also investigated in one species to assess the rate of non-homologous comigrating fragments and to detect additional polymorphic markers to be used in stock identification. Results of Band Sharing Index (BSI) and percentage of polymorphic fragments are presented and are discussed in relation to the wide applicability of AFLPs both for fish and seafood safety and authenticity testing in such fields as food traceability and restocking management. The database, available upon request at nonnis@biol.unipr.it, will be continuously updated.

Isolation and genetic diversity of endangered grey nurse shark (Carcharias taurus) populations

Anthropogenic impacts are believed to be the primary threats to the eastern Australian population of grey nurse sharks (Carcharias taurus), which is listed as critically endangered, and the most threatened population globally. Analyses of 235 polymorphic amplified fragment length polymorphisms (AFLP) loci and 700 base pairs of mitochondrial DNA control region provide the first account of genetic variation and geographical partitioning (east and west coasts of Australia, South Africa) in C. taurus. Assignment tests, analysis of relatedness and Fst values all indicate that the Australian populations are isolated from South Africa, with negligible migration between the east and west Australian coasts. There are significant differences in levels of genetic variation among regions. Australian C. taurus, particularly the eastern population, has significantly less AFLP variation than the other sampling localities. Further, the eastern Australian sharks possess only a single mitochondrial haplotype, also suggesting a small number of founding individuals. Therefore, historical, rather than anthropogenic processes most likely account for their depauperate genetic variation. These findings have implications for the viability of the eastern Australian population of grey nurse sharks.

Genetic and Pathogenic Variation in Phytophthora cactorum Affecting Fruit and Nut Crops in California

Isolates of Phytophthora cactorum and 15 other species of Phytophthora were characterized according to their genomic DNA, pathogenicity, and sensitivity to mefenoxam. Amplified fragment length polymorphism (AFLP) analysis was completed for 132 isolates of P. cactorum (30 from almond, 86 from strawberry, 5 from walnut, and 11 from other hosts) and 22 isolates of 15 other Phytophthora spp. from various hosts. All 16 Phytophthora spp. were distinguishable by unique AFLP banding patterns. Cluster analysis of the AFLP data revealed high coefficients of genetic similarity (>0.9) among all California isolates of P. cactorum. Analysis of molecular variance indicated that, among all 132 isolates of P. cactorum, 30.8 and 24.5% of the AFLP variation was associated with hosts and geographical sources of isolates, respectively, whereas 15.0% of the variation was associated with isolate niche (i.e., an aerial plant part, portion of the root system, or soil). Among the 86 isolates of P. cactorum from strawberry, characterization by source in the production system (i.e., fruiting field or plant nursery) did not account for a significant proportion of the variation (0.6%, P = 0.204). In pathogenicity tests on strawberry plants (cv. Diamante) in a greenhouse, isolates of P. cactorum from hosts other than strawberry and an isolate from a strawberry fruit caused only negligible amounts of disease, but isolates from strawberry root systems were highly aggressive. On excised shoot segments of almond (cv. Drake), all isolates of P. cactorum originally from almond were pathogenic, and 8 of 17 isolates of the pathogen from other hosts caused significantly less disease than the almond isolates. All 132 isolates of P. cactorum were sensitive to mefenoxam at 1 ppm. Populations of P. cactorum in California apparently are mefenoxam sensitive and exhibit host specificity with relatively minor variation in genomic DNA. The genetic variation observed in P. cactorum included significant geographical and host origin components, which has implications for disease management approaches.