Genetic diversity in wild and reared populations of the Japanese bitterling <Emphasis Type="Italic">Tanakia tanago</Emphasis> (Cyprinidae)

 The Japanese bitterling Tanakia tanago is an endangered cyprinid species; thus, captive breeding programs are being conducted in various facilities as ex situ conservation. To examine the genetic diversity in one wild and three reared populations, and its changes during the process of captive breeding, sequences of the mitochondrial cytochrome b gene and control region were determined. The wild population, collected in 1993, was monomorphic. Although the reared population that originated from the wild population was almost monomorphic, a rare haplotype, distinct from all others by a relatively large sequence divergence, was also observed. In the other reared populations, some degree of genetic diversity had been maintained. A reared hybrid population, which originated from a mixture of three distinct populations, showed the greatest genetic diversity. These results suggest considerable genetic diversity within and among populations of T. tanago in the past. Although a loss of genetic diversity was observed in some year-classes of reared populations, there was no tendency for genetic diversity to decrease as a result of captive breeding, probably because offspring were obtained from multi-year-class parents in the captive breeding program. Accordingly, this breeding method should be appropriate for conserving the genetic diversity of T. tanago. Received: June 12, 2002 / Revised: December 3, 2002 / Accepted: December 16, 2002     

黑龙江省强壮水螅遗传多样性的RAPD分析

应用RAPD技术对黑龙江省10个分布区17个种群的强壮水螅(Hydrarobusta)进行了遗传多样性分析。从80个随机引物中筛选出19个引物,共扩增出320条带,分子量在0.3～3.0kb,其中多态性带数299个,占93.44%,表现出高度的遗传多样性。17个种群的遗传相似系数(GI)平均值为0.7158,相应的遗传距离(GD)平均值0.3425;分布区内种群间的差异(GD平均值0.1695)小于各分布区间种群间的差异(GD平均值0.3425),各种群间的遗传距离与它们的地理间距基本一致;但虎头种群与其他种群间的相似度明显偏低(GI平均值0.5413,GD平均值0.6161),视为较特化种群。     

Wild sunflower diversity in Argentina revealed by ISSR and SSR markers: an approach for conservation and breeding programmes

Wild sunflower Helianthus annuus originates from North America and has naturalised in Argentina where it is considered invasive. The present study attempts to assess the genetic diversity using two different molecular marker systems to study the wild genetic patterns and to provide data applicable to conservation and breeding uses. Ten natural populations sampled throughout the wild range and six inbred lines were studied using inter‐simple sequence repeat (ISSR) and simple sequence repeats (SSR) markers. A total of 64 ISSR bands and 29 SSR alleles were produced from 106 wild and cultivated plants. We found 9 ISSR private bands and 21 SSR private alleles in wild accessions, but no private bands/alleles were found in cultivated sunflowers. Molecular variability in wild populations was approximately 60% higher than in inbred lines. Local wild sunflowers kept considerable diversity levels in comparison with populations in the centre of origin (approximately 70%) and therefore they might possess a potential for adaptive evolutionary change. Analysis of molecular variance (AMOVA) indicated population structure with nearly 20% of genetic variability attributable to between‐population differentiation. Principal coordinate analyses (PCO) grouped wild populations from different geographic locations, and a Mantel test showed low congruence between genetic distance (GD) and geographic distances (GGD); hence, molecular data could not rule out multiple wild introduction events. Low correlations were found between ISSR and SSR GD at individual and population levels; thus, divergent evolutionary groups were not evident in local wild sunflowers. Several genetic diversity criteria were utilised to assign conservation value and certain wild populations emerged as interesting sites for more extensive sampling.     

Determination of genetic relationships among shape Phaseolus vulgaris populations in a conical cross from RAPD marker analyses

Random-amplified polymorphic DNA (RAPD) markers were used to determine genetic relationships among Phaseolus vulgaris breeding populations. Genetic distances were calculated from the distribution of 317 RAPD markers among 8 parents, 10 individuals from 8 cycle-one populations, 10 individuals from 6 cycle-two populations and 10 individuals from 2 cycle-three populations of a conical cross. Genetic distances between populations and parents were consistent with their degree of relationship in the crossing scheme indicating that a RAPD analysis is a sensitive and useful method for categorizing breeding materials according to their genetic similarities. Genetic variation among individuals within populations increased from cycle one to cycle three and variation among populations within the cycles decreased from cycle one to cycle three in the conical cross. The results showed that this crossing scheme can be used to collect the genetic diversity in eight parents into a single plant breeding population. Abbreviations: CBB, common bacterial blight; GD, genetic distance; RAPD, random-amplified polymorphic DNA     

Population genetic structure of the medicinal plant Vitex rotundifolia in China: implications for its use and conservation

Vitexrotundifolia L.is an important plant species used in traditional Chinese medicine.For its efficient use and conservation,genetic diversity and clonal variation of V.rotundifolia populations in China were investigated using inter-simple sequence repeat markers.Fourteen natural populations were included to estimate genetic diversity,and a large population with 135 individuals was used to analyze clonal variation and fine-scale spatial genetic structure.The overall genetic diversity (GD) of V.rotundifolia populations in China was moderate (GD=0.190),with about 40% within-population variation.Across all populations surveyed,the average within-population diversity was moderate (P = 22.6%; GD = 0.086).A relatively high genetic differentiation (Gst=0.587)among populations was detected based on the analysis of molecular variance data.Such characteristics of V.rotundifolia are likely attributed to its sexual/asexual reproduction and limited gene flow.The genotypic diversity (D=0.992) was greater than the average values of a clonal plant,indicating its significant reproduction through seedlings.Spatial autocorrelation analysis showed a clear within-population structure with gene clusters of approximately 20 m.Genetic diversity patterns of V.rotundifolia in China provide a useful guide for its efficient use and conservation by selecting particular populations displaying greater variation that may contain required medicinal compounds,and by sampling individuals in a population at >20 m spatial intervals to avoid collecting individuals with identical or similar genotypes.     

Quantifying and managing the loss of genetic variation in a free-ranging population of takahe through the use of pedigrees

Pedigree analysis has clear benefits for the genetic management of threatened populations through the evaluation of inbreeding, population structure and genetic diversity. The use of pedigrees is usually restricted to captive populations and few examples exist of their exclusive use in managing free-ranging populations. One such example is the management of the takahe (Porphyrio hochstetteri), a highly endangered, flightless New Zealand rail at risk from introduced mammalian predators and habitat loss. During the 1980’s and 90’s, as part of the takahe recovery programme, birds were translocated from the sole remnant population in Fiordland to four offshore islands from which introduced predators had been eradicated. The subsequent “island” population, now numbering 83 and thought to be at carrying capacity, has been closely monitored since founding. Detailed breeding records allow us to analyse the island pedigree, which is up to 7 generations deep. Gene-drop analysis indicated that 7.5% of genetic diversity has been lost over the relatively short timeframe since founding (2.1 generations on average; total genetic founders = 31) due to both a failure to equalise founder representation early on and subsequent disproportionate breeding success (founder equivalents = 12.5; founder genome equivalents = 6.6). A high prevalence of close inbreeding will have also impacted on genetic diversity. Predictions from pedigree modelling suggest that 90% genetic diversity will be maintained for only 12 years, but by introducing a low level of immigration from the Fiordland population and permitting the population to grow, 90% GD could be maintained over the next 100 years. More generally, the results demonstrate the value of maintaining pedigrees for wild populations, especially in the years immediately after a translocation event.     

Wild genetic diversity preservation in a small-sized first generation breeding population of <Emphasis Type="Italic">Allanblackia floribunda</Emphasis> (Clusiaceae)

Founder group size is of prime importance in tree breeding programs. We determined whether sampling 20-plus trees for breeding in Allanblackia floribunda, a tropical forest tree species that has been recently enrolled in tree improvement program for fruit and seed production, would affect neutral genetic diversity and inbreeding level in both breeding and production populations. Using eight informative microsatellite loci, we: (a) assessed the nuclear genetic diversity of ten natural populations, and of the breeding population in the humid forest zone of Cameroon; (b) investigated temporal effective-size fluctuations in A. floribunda natural populations, with a view to identifying the role of past demographic events in the genetic structure of the studied species; and (c) tested the hypothesis that genetic diversity in a founder group of 20 individuals is not different from that existing in the wild. The eight loci were variable. High levels of genetic diversity (A = 4.96; H _E = 0.59) and moderate differentiation (R _ST = 0.061) were found within and among populations in wild stands. High genetic distances existed between populations ( \textaverage chord distance = 0.\text2769 ±0.00\text554 ) \left( {{\text{average chord distance}} = 0.{\text{2769}} \pm 0.00{\text{554}}} \right) . Eight of the ten surveyed populations showed signs of deviation from mutation-drift equilibrium, suggesting Pleistocene population bottlenecks and fluctuations in effective population size. Mantel tests did not reveal any relationships between genetic and geographic distances. A neighbor-joining dendrogram showed a population structure that could be explained by historical factors. The hypothesis tested has been accepted. However, a slight increase in inbreeding was observed in the breeding population.     

The genetic structure of the European breeding populations of a declining farmland bird,the ortolan bunting (<Emphasis Type="Italic">Emberiza hortulana</Emphasis>), reveals conservation priorities

Anthropogenic activities, such as agricultural intensification, caused large declines in biodiversity, including farmland birds. In addition to demographic consequences, anthropogenic activities can result in loss of genetic diversity, reduction of gene flow and altered genetic structure. We investigated the distribution of the genetic variation of a declining farmland and long-distance migratory bird, the ortolan bunting Emberiza hortulana, across its European breeding range to assess the impact of human-driven population declines on genetic diversity and structure in order to advise conservation priorities. The large population declines observed have not resulted in dramatic loss of genetic diversity, which is moderate to high and constant across all sampled breeding sites. Extensive gene flow occurs across the breeding range, even across a migratory divide, which contributes little to genetic structuring. However, gene flow is asymmetric, with the large eastern populations acting as source populations for the smaller western ones. Furthermore, breeding populations that underwent the largest declines, in Fennoscandia and Baltic countries, appear to be recently isolated, with no gene exchange occurring with the eastern or the western populations. These are signs for concern as declines in the eastern populations could affect the strength of gene flow and in turn affect the western populations. The genetic, and demographic, isolation of the northern populations make them particularly sensitive to loss of genetic diversity and to extinction as no immigration is occurring to counter-act the drastic declines. In such a situation, conservation efforts are needed across the whole breeding range: in particular, protecting the eastern populations due to their key role in maintaining gene flow across the range, and focussing on the northern populations due to their recent isolation and endangered status.     

No evident spatial genetic structuring in the rapidly declining Black-tailed Godwit <Emphasis Type="Italic">Limosa limosa</Emphasis><Emphasis Type="Italic">limosa</Emphasis> in The Netherlands

With 40% of the European Black-tailed Godwit population breeding in The Netherlands, this country harbours internationally significant numbers of this species. However, ongoing agricultural intensification has resulted in the fragmentation of the population and drastic population declines since 1967. By establishing genetic diversity, genetic differentiation and gene flow on the basis of 12 microsatellites, we investigated whether the population genetic structure of the Dutch Black-tailed Godwit bears the marks of these changes. Genetic diversity appeared to be moderate, and Bayesian model-based analysis of individual genotypes revealed no clustering in the Dutch populations. This was supported by pairwise F_ST values and AMOVA, which indicated no differentiation among the nine breeding areas. Gene flow estimates were larger than “one migrant per generation” between sample locations, and no isolation by distance was demonstrated. Our results indicate the maintenance of moderate levels of genetic diversity and genetic connectivity between breeding sites throughout the Dutch Black-tailed Godwit breeding population. We suggest that the Dutch Black-tailed Godwit breeding areas should be managed as a single panmictic unit, much as it is presently done.     

The origin and genetic diversity of Pinus radiata in Australia

Summary Despite the fact that forest trees are in early stages of domestication there has been little direct evaluation of either the origin of, or genetic diversity within the breeding material in tree improvement programs. Allozyme variation was used to compare the total genetic diversity in the breeding programs of P. radiata within Australia and the five wild populations in North America. The current breeding populations were very similar genetically and were essentially homogenous with only 1.8% of the variation among programs. The total genetic diversity in the species was 0.12, which is a low estimate compared to most conifers. Overall in the Australian material the genetic diversity was somewhat less. The comparison of allelic frequencies in the five native populations with the Australian material indicates that the Monterey and Año Nuevo populations were probably the major source of the original introductions and that a substantial portion of the genetic diversity in the two populations has been captured in current breeding programs. The three southern populations do not appear to be currently represented in the breeding programs. The implications for future breeding strategies are discussed.     

Population structure and genetic diversity of wild <Emphasis Type="Italic">Helianthus</Emphasis> species from Mozambique

The production of sunflower suffered a major decline in Mozambique after its independence in 1975. Civil war, human activities and environmental damage subjected the species to an ecological stress contributing to reduce the number and size of wild populations. As this reduction is often related to a loss of genetic variation we estimated the genetic diversity within and among populations of wild Helianthus from five districts of Mozambique using RAPD markers. The 44 accessions studied grouped into four major clusters exhibiting structured variability with regard to geographic origin. A high level of genetic diversity (He = 0.350 and I = 0.527) was retained at the population level. The genetic variation among populations was high (59.7%), which is consistent with low gene flow (Nm = 0.338). The proportion of total genetic diversity residing among these populations should be kept in mind to devise different conservation strategies in order to preserve these populations. Currently wild Helianthus genetic resources present in Maputo and Sofala are on the edge of extinction mainly due to excessive urbanization. Therefore, conservation of what remains of this plant genetic diversity is essential for sustainable utilization and can be useful for breeding programs.     

Turnover and post‐bottleneck genetic structure in a recovering population of Peregrine Falcons Falco peregrinus

Dispersal is a process that increases genetic diversity and genetic connectivity of populations. We studied the turnover rate of breeding adults and genetic population structure to estimate dispersal in Peregrine Falcons in Finland. We used relatedness estimates among Finnish Peregrine Falcons over a 5‐year period, genotyping over 500 nestlings with 10 microsatellite loci to reveal the rate of turnover. Our results reveal a high turnover rate (21.7%) that does not seem to be correlated with the breeding success of the previous year. The extent of population genetic structure and diversity, and possible signs of the population crash during the 1970s, was assessed with a reduced dataset, excluding relatives. We found genetic diversity to be similar to previously studied falcon populations (expected heterozygosity of 0.581) and no population genetic structuring among our sampled populations. We did not find a genetic imprint of the past population bottleneck that the Finnish Peregrine population experienced. We conclude that high dispersal rates are likely to have contributed to maintaining genetic diversity across the landscape, by mixing individuals within the species’ distribution in Finland and thus preventing genetic structuring and negative effects associated with the population decline in the 1970s.     

Evaluation and comparison of the genetic structure of <Emphasis Type="Italic">Bunias orientalis</Emphasis> populations in their native range and two non-native ranges

We studied the invasive warty cabbage Bunias orientalis (Brassicaceae) in three geographically distinct areas. Using inter-simple sequence repeat fingerprinting, we analyzed warty cabbages, including non-native populations, from the eastern Baltic and western Siberian regions and native populations from southwestern Russia. The eastern Baltic region and western Siberia represent the two opposite directions of B. orientalis spread in climatically different zones. The genetic structures of the native and non-native B. orientalis populations were assessed through analysis of molecular variance (AMOVA) and the Bayesian clustering method and by determining the main measures of genetic diversity. AMOVA revealed considerable population differentiation in both the native and invasive ranges. Our results did not indicate a decrease in genetic diversity in the non-native populations of B. orientalis. Similar measures of genetic diversity and genetic structure were determined in the invasive populations in two geographically and ecologically distinct, non-native regions located in Europe and Asia. In both of these regions, higher genetic diversity was detected in the non-native populations than in the native region populations, which may be due to multiple introductions. However, Bayesian clustering analysis revealed slightly different sources of invasive populations in the two non-native regions. Genetic diversity patterns revealed the lack of isolation by distance between populations and confirmed the influence of anthropogenic factors on the spread of B. orientalis. The significance of native populations as germplasm resources for breeding is discussed.     

Evaluation of the genetic management of the endangered black‐footed ferret (Mustela nigripes)

Empirical support for the genetic management strategies employed by captive breeding and reintroduction programs is scarce. We evaluated the genetic management plan for the highly endangered black‐footed ferret (Mustela nigripes) developed by the American Zoo and Aquarium Associations (AZA) as a part of the species survival plan (SSP). We contrasted data collected from five microsatellite loci to predictions from a pedigree‐based kinship matrix analysis of the captive black‐footed ferret population. We compared genetic diversity among captive populations managed for continued captive breeding or reintroduction, and among wild‐born individuals from two reintroduced populations. Microsatellite data gave an accurate but only moderately precise estimate of heterozygosity. Genetic diversity was similar in captive populations maintained for breeding and release, and it appears that the recovery program will achieve its goal of maintaining 80% of the genetic diversity of the founder population over 25 years. Wild‐born individuals from reintroduced populations maintained genetic diversity and avoided close inbreeding. We detected small but measurable genetic differentiation between the reintroduced populations. The model of random mating predicted only slightly lower levels of heterozygosity retention compared to the SSP strategy. The random mating strategy may be a viable alternative for managing large, stable, captive populations such as that of the black‐footed ferret. Zoo Biol 22:287–298, 2003. © 2003 Wiley‐Liss, Inc.     

Newly developed SSR markers reveal genetic diversity and geographical clustering in spinach (<Emphasis Type="Italic">Spinacia oleracea</Emphasis>)

Spinach is a popular leafy green vegetable due to its nutritional composition. It contains high concentrations of vitamins A, E, C, and K, and folic acid. Development of genetic markers for spinach is important for diversity and breeding studies. In this work, Next Generation Sequencing (NGS) technology was used to develop genomic simple sequence repeat (SSR) markers. After cleaning and contig assembly, the sequence encompassed 2.5% of the 980 Mb spinach genome. The contigs were mined for SSRs. A total of 3852 SSRs were detected. Of these, 100 primer pairs were tested and 85% were found to yield clear, reproducible amplicons. These 85 markers were then applied to 48 spinach accessions from worldwide origins, resulting in 389 alleles with 89% polymorphism. The average gene diversity (GD) value of the markers (based on a GD calculation that ranges from 0 to 0.5) was 0.25. Our results demonstrated that the newly developed SSR markers are suitable for assessing genetic diversity and population structure of spinach germplasm. The markers also revealed clustering of the accessions based on geographical origin with clear separation of Far Eastern accessions which had the overall highest genetic diversity when compared with accessions from Persia, Turkey, Europe, and the USA. Thus, the SSR markers have good potential to provide valuable information for spinach breeding and germplasm management. Also they will be helpful for genome mapping and core collection establishment.     

五个红罗非鱼群体的遗传多样性分析

实验采用微卫星标记技术,选用22对微卫星引物对5个红罗非鱼群体进行遗传多样性分析。经PCR扩增,16个微卫星位点扩增产物在关岛红罗非鱼（GD）、珍珠白红罗非鱼（ZZ）、佛罗里达红罗非鱼（FL）、明月红罗非鱼（MY）、马来西亚红罗非鱼（ML）中均获得了清晰稳定的条带。分析结果显示:16个微卫星标记共检测到146个等位基因。5个群体的平均等位基因数（N_a）在6.5625-8.5625,平均有效等位基因数（N_e）在4.1495-6.1330,平均杂合度（H_e）在0.7491-0.8247,平均多态信息含量（PIC）在0.6939-0.7840,说明它们的遗传多态性丰富。卡方检验表明5个红罗非鱼群体的大部分位点偏离Hardy-Weinberg平衡。在5个群体中,关岛红罗非鱼（GD）与珍珠白红罗非鱼（ZZ）遗传相似系数（0.6171）最小,遗传距离（0.4827）最大,说明两者亲缘关系最远;佛罗里达红罗非鱼（FL）与马来西亚红罗非鱼（ML）遗传相似系数（0.9069）最大,遗传距离（0.0977）最小,可推断两者亲缘关系最近。采用UPGMA进行聚类分析,结果表明:佛罗里达与马来西亚先聚成一支,二者再与珍珠白聚在一起,接着三者与明月聚在一起,最后,四者与关岛聚到一起。聚类结果说明关岛群体与其他4个群体亲缘关系最远;佛罗里达与马来西亚亲缘关系最近,珍珠白群体次之,明月群体再次之。以上结果可推断5个红罗非鱼群体遗传多态性丰富,具有较大的选育潜力。     

陕西省林麝mtDNA D-loop区序列结构和种群遗传多样性

林麝(Moschus berezovskii)曾广泛分布于中国,由于盗猎和栖息地缩小,秦岭地区野生种群数量迅速下降,圈养繁殖种群已成立了几十年,但大多数圈养种群的遗传背景不清,种群规模增长非常缓慢。为了给这一物种的保护和管理提供有用的信息,调查了陕西省林麝1个圈养种群3个野生种群线粒体DNA(mt DNA)D-Loop 632 bp片段的遗传多样性和种群结构。在69个个体中其碱基组成为A+T的平均含量63.2%高于G+C含量36.8%,共检测到变异位点171个(约占总位点数的27.05%)。核苷酸多样性(Pi)为0.04424,平均核苷酸差异数(K)为19.908。69个个体分属32个单倍型,单倍型间的平均遗传距离(P)为0.070。32个单倍型构建的NJ系统树聚为3个分支,4个林麝群体中的单倍型是随机分布的。4个群体的平均遗传距离为0.043(标准误SE为0.005),凤县养殖场群体与留坝和陇县群体的亲缘关系较远。单倍型间的平均遗传距离为0.043,可见其遗传分化尚未达到种群分化的水平。结果表明,陕西省林麝群体mt DNA D-loop区序列存在着较丰富的变异和遗传多样性,凤县野生群体和凤县养殖场群体的核苷酸多样性和单倍型多样较高,养殖场种群没有出现近亲繁殖及遗传多样性下降的情况。凤县野生群体和凤县养殖场群体两者遗传分化较小,存在着较高的基因流水平。     

Conservation genetics of an endemic and threatened amphibian (<Emphasis Type="Italic">Capensibufo rosei</Emphasis>): a leap towards establishing a genetic monitoring framework

Given the ever-increasing anthropogenic changes to natural ecosystems, it is imperative that temporal changes in genetic diversity be monitored to help safeguard the future viability of species. Capensibufo rosei is a small, range-restricted bufonid from South Africa, believed to have experienced an enigmatic decline likely due to the suppression of natural fires and the loss of grazing animals from some areas. Without these disturbances, their habitat becomes overgrown, which might affect the characteristics of their breeding pools. Since the 1980s, four breeding sites have been lost, presumably due to loss of breeding habitat through encroachment of vegetation. Currently, there are only two known populations [Cape of Good Hope (CGH) and Silvermine nature reserves] both within Table Mountain National Park. Consequently, this species may be vulnerable to stochastic events and genetic erosion through the loss of metapopulation connectivity. To assess the genetic status of this species, genetic diversity within both populations was quantified for two time periods using 11 microsatellite markers. Despite evidence of severe population bottlenecks, both populations possess levels of diversity similar to other anurans, and Silvermine has greater diversity than CGH. A close examination of the data revealed both populations to be genetically dynamic through time, with the loss and gain of rare alleles. Both populations also experienced a slight increase in overall diversity between sampling periods. While the latter was not statistically significant, the monitoring period was perhaps too short to understand changes in diversity over time. These results will form the baseline for future monitoring to better understand this threatened and declining species and to track genetic erosion or recovery.     

Genetic diversity of African maize inbred lines revealed by SSR markers

Knowledge of genetic diversity (GD) and relationships among maize inbred lines is indispensable in a breeding program. Our objectives were to (1) investigate the level of genetic diversity among maize inbred lines and (2) assess their genetic structures by applying simple sequence repeat (SSR) markers. Fifty-six highland and mid-altitude maize inbred lines obtained from CIMMYT programs in Ethiopia and Zimbabwe were genotyped using 27 SSR loci. All of the genotypes studied could unequivocally be distinguished with the combination of the SSRs used. In total, 104 SSR alleles were identified, with a mean of 3.85 alleles per locus. The average polymorphism information content (PIC) was 0.58. GD expressed as Euclidean distance, varied from 0.28 to 0.73 with an average of 0.59. Cluster analysis using unweighted pair group method with arithmetic average (UPGMA) suggested five groups among the inbred lines. Most of the inbred lines adapted to the highlands and the mid-altitudes were positioned in different clusters with a few discrepancies. The pattern of groupings of the inbred lines was mostly consistent with available pedigree information. The variability detected using SSR markers could potentially contribute towards effective utilization of the inbred lines for the exploitation of heterosis and formation of genetically diverse source populations in Ethiopian maize improvement programs.     

Genetic diversity of <Emphasis Type="Italic">Rhynchosporium secalis</Emphasis> in Tunisia as revealed by pathotype,AFLP, and microsatellite analyses

Genetic variability among 122 Rhynchosporium secalis isolates collected from barley in three regions of Tunisia was investigated using host differentials, amplified fragment length polymorphism (AFLP), and microsatellite markers. The isolates were collected from a widely grown scald-susceptible barley cultivar Rihane and a range of local landrace cultivars in geographically distinct regions with different agroclimatic conditions. Pathotypic diversity (the proportion of unique pathotypes) was high in R. secalis populations from the high (100% diversity), moderate (95%), and low (100%) rainfall areas of Tunisia, and from both Rihane (which is the sole variety grown in the high rainfall region) and local landraces (which predominate in the low rainfall area). This may reflect a general adaptability for aggressiveness and suggests that the widely grown cultivar Rihane has exerted little or no selection pressure on the pathogen population since its release in 1983. Genotypic diversity (GD), defined as the probability that two individuals taken at random had different genotypes, was high for populations from Rihane, local landraces, and different agro-ecological zones (GD = 0.96–0.99). There was low genetic differentiation among pathogen populations from different host populations (G _ST ≤ 0.08, θ ≤ 0.12) and agro-ecological zones (G _ST ≤ 0.05, θ ≤ 0.04), which may be partly explained by gene flow due to the movement of infected stubble around the country. There was no correlation (r = 0.06, P = 0.39) between virulence phenotype and AFLP haplotype. A phenetic tree revealed groups with low bootstrap values that did not reflect the grouping of isolates based on host, pathotype, or agro-ecological region. The implications of these findings for R. secalis evolutionary potential and scald-resistance breeding in Tunisia are discussed.