Analysis of genetic variation in selected stocks of hatchery flounder,Paralichthys olivaceus,using AFLP markers

Amplified fragment length polymorphism (AFLP) analysis was performed in order to evaluate genetic characteristics of one common population and two selective hatchery populations of flounder Paralichthys olivaceus. A group of 60 genotypes belonging to three populations was screened using 10 different AFLP primer combinations. A total of 491 loci were produced in the three studied populations. The loci of 65.78%, 61.47% and 60.92% were polymorphic over all the genotypes tested in common, susceptible and resistant populations, respectively. The number of polymorphic loci detected by single primer combination ranged from 21 to 43. The average heterozygosity of common, susceptible and resistant populations was 0.1656, 0.1609 and 0.1586, respectively, which showed no significant difference. Compared with the common population, the two selective hatchery populations, susceptible and resistant, showed significant genetic differences including a smaller (P < 0.05) number of total loci, a smaller (P < 0.05) number of total polymorphic loci and a smaller (P < 0.05) percentage of low frequency (0–0.2) polymorphic loci. AFLP banding pattern was transformed into binary data and matrices were processed with POPGENE and TFPGA software. Similarity relationships were described graphically by a dendrogram, which clustered the three populations. The AFLP fingerprinting technique was confirmed to be a reproducible and sensitive tool for the study of population genetics of flounder. The present study confirmed that it was important to detect the genetic variability of the selective hatchery populations for the conservation of natural flounder resources.     

Genetic diversity analysis of Perinereis aibuhitensis based on ISSR and SRAP markers of Chinese coast populations

The objective of this study was to obtain an overview of the genetic relationships within Perinereis aibuhitensis using Inter-Simple Sequence Repeat (ISSR) and Sequence-Related Amplified Polymorphism (SRAP) markers that were derived from related populations residing in the Chinese coasts. The percentage of polymorphic bands, Nei's gene diversity and Shannon's information index revealed a high level of genetic diversity at the species level. The analysis of molecular variance revealed that 81.22% (ISSR) and 76.29% (SRAP) of variability were partitioned among individuals within populations, which indicated the coherent trend by Nei's genetic differentiation (Gst) (0.2568/0.2876). The gene flow number (Nm) was 1.4470/1.2385, which indicated that there was limited gene exchange between populations. The phylogenetic tree of the ten P. aibuhitensis populations was separated into four major clusters using the neighbor-joining (NJ) method. These results provide a simple and useful basis for P. aibuhitensis germplasm research and aquaculture breeding.     

Complex genetic structures of Sardinella zunasi in the Northwest Pacific detected by AFLP markers

Seventy-one Sardinella zunasi individuals collected from 5 locations in the Northwest Pacific were used in an AFLP analysis to examine the population genetic structure of S. zunasi. Using 4 selective primer combinations, 414 putative loci were detected, including 340 polymorphic loci (82.13%). Both AMOVA and pairwise F_ST analysis detected significant population genetic structure between Chinese and Japanese populations, supporting separate stocks in this species. The UPGMA trees also detected significant spatial structure within S. zunasi. Pleistocene glaciations are hypothesized to be responsible for the genetic differentiation. The significant genetic differentiation found among S. zunasi populations might also be due to geographic isolation, because a pattern of isolation-by-distance was observed. The weaker but significant structures between the Japanese Aichi and Kagawa populations might be impacted by the semi-enclosed coastal topography there. No significant genetic structure was detected among the Chinese populations, perhaps because of S. zunasi’s migratory behavior and ocean current patterns.     

Genetic diversity and population structure of the red stingray,Dasyatis akajei inferred by AFLP marker

To examine population genetic diversity and variation of the red stingray Dasyatis akajei, samples from 1 freshwater region and 6 coastal localities within its distribution range were analyzed by using amplified fragment length polymorphism (AFLP) technology. A total of 207 loci were identified by 4 primer combinations from 87 individuals, 174 of which were polymorphic (84.1%). A high level of Nei's gene diversity was observed with the overall value of 0.230 ± 0.179. No significant genealogical clusters associated with sampling sites were revealed on the UPGMA tree. Both analysis of molecular variance (AMOVA, Φ_ST = 0.085, P = 0.00) and pairwise F_ST indicated significant genetic differentiation among four marine samples. However, no particular genetic differentiation was detected between marine and the limited sampling freshwater populations (AMOVA, Φ_CT = 0.056, P > 0.05). Except for the TZ vs. WZ (5.193), the gene flow estimates (N_m) demonstrated the effective immigrants were 1.918-2.976, suggesting low level dispersal between pairwise marine populations. Species-specific habits (demersal and sluggish habits) are probably responsible for the population structuring.     

Phylogeography of the heathers Erica arborea and E. trimera in the afro-alpine ‘sky islands’ inferred from AFLPs and plastid DNA sequences

The ericaceous vegetation zone of the unique and highly fragmented afro-alpine environment in the eastern African high mountains is typically dominated by the heather Erica arborea, often in combination with its close relative E. trimera. Both species are shrubs or small trees with tiny seeds, potentially capable of dispersal by wind over long distances. While E. arborea is widely distributed in Africa, the Middle East and Europe, E. trimera is endemic to the afro-alpine region where it is restricted to higher altitudes than E. arborea. We used Amplified Fragment Length Polymorphisms (AFLPs) and variation in non-coding plastid DNA sequences to test whether these two morphologically and ecologically very similar species display similar phylogeographic patterns in the afro-alpine region. We predict that the more high-altitudinal E. trimera shows more distinct genetic structuring than E. arborea, because dispersal of the latter may have been facilitated by formation of interglacial forest bridges between mountains. Based on extensive field sampling in most of the high mountains of Ethiopia and East Africa, we show that the two species are clearly distinct at AFLP and plastid DNA loci. Both showed low levels of overall AFLP diversity, suggesting bottlenecking in small refugial populations during unfavourable climatic periods. However, their genetic structuring and inferred phylogeographic histories were conspicuously different. The more high-altitudinal E. trimera consisted of three to four distinct AFLP groups, which also had different plastid DNA haplotypes and different geographic distributions, suggesting long-term restriction to several refugia (at least one in Ethiopia and two in East Africa). In contrast, E. arborea showed little geographic structuring at AFLP loci and only a single, widespread plastid DNA haplotype, which may suggest recent colonization of the entire study area from a single source population, likely via a combination of gradual expansion via forest bridges and long-distance dispersals. The source population of E. arborea may be situated in (or north of) Ethiopia, which harbours most genetic diversity.     

Genetic variation of coreoleuciscus splendidus populations (Teleostei; Cypriniformes) from four major river drainage systems in South Korea as assessed by AFLP markers

Amplified fragment length polymorphism (AFLP) technique was used to assess genetic variation and population structuring of Coreoleuciscus splendidus (Teleostei; Cypriniformes) from four major river drainage systems in South Korea (i.e. the Han and Geum Rivers belonging to the West Korea Subdistrict and the Seomjin and Nakdong Rivers belonging to the South Korea Subdistrict). Four combinations of selective primers generated 152–186 DNA fragments, of which 45–59 were polymorphic. The Geum River population had the lowest genetic variation in terms of average heterozygosity. The pairwise genetic differentiation (F _ST) estimates showed that the four populations of C. splendidus were genetically differentiated (P < 0.05). The F _ST values were higher between population groups of the West and South Subdistricts than between populations from the two river systems within each subdistrict. A dendrogram based on the distance matrix revealed that all C. splendidus individuals bifurcated into two subdistrict population groups and were further divided into two river populations within each subdistrict. Individuals from each population consistently clustered together with high statistical support. The analysis of molecular variance after partitioning of C. splendidus populations into two groups (i.e. the West and South Korea Subdistricts) revealed very strong geographic structuring between them with 74.01% of total variance (P < 0.001). Taken together, the AFLP markers clearly distinguished not only between two population groups of the West and South Subdistricts but also between populations from two river systems within each subdistrict. Morphological differences between the two subdistrict population groups in body color and the presence of black array(s) on dorsal, anal, and caudal fin rays agreed with their high degree of genetic variation. Our findings strongly indicate that restricted dispersal and gene flow are apparent among four geographically separate populations of C. splendidus, and each population should be treated as an evolutionarily significant unit; this should be an a priori consideration for developing their conservation strategies.     

Evaluation of genetic diversity of Panicum turgidum Forssk from Saudi Arabia

The genetic diversity of 177 accessions of Panicum turgidum Forssk, representing ten populations collected from four geographical regions in Saudi Arabia, was analyzed using amplified fragment length polymorphism (AFLP) markers. A set of four primer-pairs with two/three selective nucleotides scored 836 AFLP amplified fragments (putative loci/genome landmarks), all of which were polymorphic. Populations collected from the southern region of the country showed the highest genetic diversity parameters, whereas those collected from the central regions showed the lowest values. Analysis of molecular variance (AMOVA) revealed that 78% of the genetic variability was attributable to differences within populations. Pairwise values for population differentiation and genetic structure were statistically significant for all variances. The UPGMA dendrogram, validated by principal coordinate analysis-grouped accessions, corresponded to the geographical origin of the accessions. Mantel’s test showed that there was a significant correlation between the genetic and geographical distances (r = 0.35, P < 0.04). In summary, the AFLP assay demonstrated the existence of substantial genetic variation in P. turgidum. The relationship between the genetic diversity and geographical source of P. turgidum populations of Saudi Arabia, as revealed through this comprehensive study, will enable effective resource management and restoration of new areas without compromising adaptation and genetic diversity.     

Genetic diversity of the African poplar (<Emphasis Type="Italic">Populus ilicifolia</Emphasis>) populations in Kenya

We evaluated the genetic diversity of the African poplar (Populus ilicifolia) populations found in Kenya compared with reference samples of five poplar species from North America and one species introduced in Kenya from India (KEFRI-Kenya). Amplified fragment length polymorphism (AFLP) was used with the objective of providing important information for breeding and in situ/ex situ conservation of this species. Samples collected from three locations along the species’ natural range (Athi, Ewaso Nyiro, and Tana rivers) were compared with four samples of locally planted Populus deltoides stand introduced from India and ten reference samples from North America. Six AFLP primer combinations produced 521 clear bands for analysis. The percentage polymorphic loci were lowest in Tana (20.4 %) and highest in Athi (40.6 %). The average heterozygosity across the studied populations was between 0.07 and 0.3. AMOVA revealed more genetic variation partitioning within population (87 %; P?<?0.01) than among populations (13 %; P?<?0.01) suggesting significant genetic variation between populations. Further, UPGMA delineation showed two clusters of the Tana, Athi, and Ewaso Nyiro populations clustered together compared to the North America and India/KEFRI reference samples. Moreover, the study showed that the Athi population is more diverse than those of Tana and Ewaso Nyiro and may be important for conservation, domestication, and improvement studies. The genetic differentiation (F _ST ?=?0.134) among Kenyan P. ilicifolia populations suggests limited possibility of gene flow between these populations.     

Genetic diversity and structure of the noxious alien grass Praxelis clematidea in southern China

Praxelis clematidea (Asteraceae), a plant species native to South America, is a noxious weed in southern China. We examined the genetic variation and population structure of 12 populations (76 individuals) of P. clematidea from Fujian, Guangdong, and Hainan Provinces in China using inter-simple sequence repeat (ISSR) analysis. From an initial set of 69 ISSR primers, 10 were selected which yielded 80 reproducible bands. Polymorphic bands (P) were 100%, Shannon's information index (I) was 0.4226, and Nei's gene diversity (H) was 0.2791. We infer that the high levels of genetic diversity exhibited by P. clematidea may have contributed to its invasiveness. Gene flow among populations was 2.4930, which has led to homogenization. The coefficient of population differentiation (Gst = 0.1671) indicated low levels of genetic variation among populations and high levels of genetic polymorphism within populations. There was a negative correlation between population elevation and genetic diversity, while there was a significant positive correlation between genetic distance and geographic distance based on a Mantel test (r = 0.5820, P < 0.01). Some populations from different provinces clustered together in principal coordinate and UPGMA analyses indicating that human-mediated events may have contributed to the dispersal of the species.     

台湾海峡鲐鱼种群遗传结构

以往研究表明,台湾海峡的鲐鱼分属2个地理种群,即东海种群和闽南——粤东地方种群.为研究这2个种群的遗传结构,对鲐鱼闽东(30尾)和闽南(30尾)种群进行了AFLP分析,8对选择性引物在2个种群60个个体中,共扩增出497个位点,其中多态位点343个.闽东和闽南种群的多态位点比例、Nei遗传多样性指数和Shannon遗传多样性指数分别为57.75％、64.59％,0.1779、0.2123,0.2725和0.3228,2个种群的遗传多样性处于同一水平.与其他鱼类对比显示,台湾海峡鲐鱼种群的遗传多样性水平高.生境广及生命周期短被认为是台湾海峡鲐鱼具有较高遗传变异水平的原因;基因分化系数Gst、Shannon遗传多样性指数和AMOVA分析均显示鲐鱼的遗传变异主要来源于种群内,而种群间无明显的遗传分化.Nm显示2个种群间基因交流频繁.种群的显性基因型频率分布显示2个种群有基本相同的种群遗传结构.结果表明,鲐鱼闽东和闽南种群间无明显的遗传差异.幼体较强的扩散能力、海洋环流及洄游特性可能是造成台湾海峡鲐鱼种群间遗传同质性较高的原因.     

Population structure and genetic diversity distribution in wild and cultivated populations of the traditional Chinese medicinal plant Magnolia officinalis subsp. biloba (Magnoliaceae)

Magnolia officinalis subsp. biloba, a traditional Chinese medicinal plant, experienced severe declines in the number of populations and the number of individuals in the late 20th century due to the widespread harvest of the subspecies. A large-scale cultivation program was initiated and cultivated populations rapidly recovered the loss in individual plant numbers, but wild populations remained small as a consequence of cutting. In this study, the levels of genetic variation and genetic structure of seven wild populations and five domestic populations of M. officinalis subsp. biloba were estimated employing an AFLP methodology. The plant exhibited a relatively high level of intra-population genetic diversity (h = 0.208 and H _j = 0.268). The cultivated populations maintained approximately 95% of the variation exhibited in wild populations, indicating a slight genetic bottleneck in the cultivated populations. The analysis of genetic differentiation revealed that most of the AFLP diversity resided within populations both for the wild group (78.22%) and the cultivated group (85.92%). Genetic differentiation among populations in the wild group was significant (F _ST = 0.1092, P < 0.005), suggesting wild population level genetic structure. Principal coordinates analysis (PCO) did not discern among wild and cultivated populations, indicating that alleles from the wild population were maintained in the cultivated gene pool. Results from the present study provide important baseline data for effectively conserving the genetic resources of this medicinal subspecies.     

Conservation genetics of endangeredBrasenia schreberi based on RAPD and AFLP markers

Brasenia schreberi J.F. Gmelin is a declared endangered species found in the lakes and ponds of South Korea. For planning its conservation strategy, we examined the genetic diversity within and among six populations, using randomly amplified polymorphic DNA (RAPD) and amplified fragment length polymorphism (AFLP). Polymorphisms were more frequently detected per loci with AFLP (69.3%) than RAPD (36.8%). High genetic diversity was recognized within populations: polymorphic loci (PPL) values ranged from 36.3% in the CJM population to 74.5% in the GGT population, with a mean value of 47.8% based on AFLP markers. Great genetic differentiation (θ^B) was detected among the six populations (0.670 on RAPD and 0.196 on AFLP), and we calculated a low rate of gene flow (N_em), i.e., 0.116 on RAPD and 0.977 on AFLP. Furthermore, a Mantel test revealed that no correlation existed between genetic distances and geographical distances among the six local populations, based on RAPD or AFLP markers. These results are attributed to a number of factors, including an insufficient length of time for genetic diversity to be reduced following a natural decline in population size and isolation, adaptation of the genetic system to small population conditions, and a restricted gene flow rate. Based on both its genetic diversity and population structure, we suggest that a strategy for conserving and restoringB. schreberi must focus on maintaining historical processes, such as high levels of outbreeding, while monitoring increased gene flow among populations. This is because a reduction in genetic diversity as a result of genetic drift is undesirable.     

Applying AFLPs in Aizoaceae: The Delosperma herbeum complex as a case study

Amplified fragment length polymorphic (AFLP) markers were used to discriminate between populations of Delosperma herbeum as well as other species of Delosperma occurring in Gauteng and adjacent provinces in South Africa. A total of 99 individuals representing 30 populations were examined for 158 polymorphic AFLP markers using the EcoRI and MseI restriction enzymes. Cluster analysis of AFLP marker scores, visualized through Neighbor-Joining and Neighbor-Net trees as well as Principal Coordinate Analysis, provides an effective means of discriminating between different species in the complex. The lack of bootstrap support for Delosperma davyi in the Neighbor-Joining tree and its position in the Principal Coordinate Analysis probably signifies it to be a superfluous name for D. herbeum. The clustering of the other species also suggests that flower colour is not a reliable feature for grouping species in Delosperma. A number of clusters of D. herbeum were correlated to a degree with distribution.     

Conservation genetics of Heritiera littoralis (Sterculiaceae), a threatened mangrove in China,based on AFLP and ISSR markers

AFLP and ISSR markers were used to determine the genetic variations in eight mangrove and non-mangrove populations of Heritiera littoralis (Sterculiaceae), a threatened species in China. Our results showed a moderate to high level of genetic variation in this species (P = 63.69%, H_T = 0.20 for AFLP; P = 76.07%, H_T = 0.22 for ISSR), and a relatively high level of genetic differentiation among populations (G_ST = 0.24 for AFLP and 0.27 for ISSR). Life history traits, long-distance dispersal of floating seeds, and local environments may play important roles in shaping the genetic diversity and genetic structure of this species. Investigation of the plant’s reproductive capacity showed that the natural seed production of H. littoralis was low, as was the germination rate and the transformation rate from juvenile to adult. H. littoralis is seriously threatened and is in urgent need of conservation in China.     

Genetic diversity in natural populations of Jacaranda decurrens Cham. determined using RAPD and AFLP markers

Jacaranda decurrens (Bignoniaceae) is an endemic species of the Cerrado with validated antitumoral activity. The genetic diversity of six populations of J. decurrens located in the State of São Paulo was determined in this study by using molecular markers for randomly amplified polymorphic DNA (RAPD) and amplified fragment length polymorphism (AFLP). Following optimization of the amplification reaction, 10 selected primers generated 78 reproducible RAPD fragments that were mostly (69.2%) polymorphic. Two hundred and five reproducible AFLP fragments were generated by using four selected primer combinations; 46.3% of these fragments were polymorphic, indicating a considerable level of genetic diversity. Analysis of molecular variance (AMOVA) using these two groups of markers indicated that variability was strongly structured amongst populations. The unweighted pair group method with arithmatic mean (UPGMA) and Pearson''s correlation coefficient (RAPD -0.16, p = 0.2082; AFLP 0.37, p = 0.1006) between genetic matrices and geographic distances suggested that the population structure followed an island model in which a single population of infinite size gave rise to the current populations of J. decurrens, independently of their spatial position. The results of this study indicate that RAPD and AFLP markers were similarly efficient in measuring the genetic variability amongst natural populations of J. decurrens. These data may be useful for developing strategies for the preservation of this medicinal species in the Cerrado.     

Genetic diversity of the endangered Chinese endemic plant Monimopetalum chinense revealed by amplified fragment length polymorphism (AFLP)

Monimopetalum chinense Rehd. is an endangered woody vine endemic to eastern China. Using amplified fragment length polymorphism (AFLP) markers, we examined levels of genetic variation within and among eleven populations located across the species’ distribution. Although modest levels of heterozygosity were detected, other measures of genetic diversity registered relatively high levels of variability, both at the species level (P = 91.0%, H_E = 0.232, I_S = 0.365) and at the population level (P = 53.0%, H_E = 0.155, I_S = 0.239). Populations also exhibited high levels of genetic differentiation (Nei’s genetic diversity analysis, G_ST = 0.330), corresponding to isolation-by-distance and hierarchical population structure. These results indicate that, despite low levels of gene flow, populations of M. chinense still harbor substantial amounts of genetic diversity. Management plans for the species should include measures that ensure genetic diversity remains high within and among extant populations.     

Assessing genetic diversity of populations of topmouth culter (Culter alburnus) in China using AFLP markers

Genetic diversity of five wild populations and a cultured population of topmouth culter (Culter alburnus) was investigated using amplified fragment length polymorphism (AFLP). A total of 373 reproducible bands amplified with seven AFLP primer combinations were obtained from 163 fish. The percentage of polymorphic loci ranged widely from 37.0% to 69.2% within distinct populations. The cultured population appeared to have a lower level of polymorphism (37.0%), gene diversity (0.121 ± 0.188) and Shannon's Information index (0.183 ± 0.263) than the wild populations. Analysis of molecular variance (AMOVA) revealed that average F_ST value overall loci was 0.2671, and the percentage of variation within population (73.29%) was larger than among populations (26.71%) (P < 0.01). The six populations were clustered into two major clades with UPGMA. The results from analysis of population pairwise gene flow indicated moderate gene flow among populations. Our study indicated that the genetic diversity of the cultured population was reduced compared with the wild populations. Geographic isolation, habitat, and artificial selection all may have played important roles in population differentiation. The information may be beneficial to future broodstock selection and defining conservation management for the different populations of topmouth culter.     

Epigenetic Variability in the Genetically Uniform Forest Tree Species Pinus pinea L

There is an increasing interest in understanding the role of epigenetic variability in forest species and how it may contribute to their rapid adaptation to changing environments. In this study we have conducted a genome-wide analysis of cytosine methylation pattern in Pinus pinea, a species characterized by very low levels of genetic variation and a remarkable degree of phenotypic plasticity. DNA methylation profiles of different vegetatively propagated trees from representative natural Spanish populations of P. pinea were analyzed with the Methylation Sensitive Amplified Polymorphism (MSAP) technique. A high degree of cytosine methylation was detected (64.36% of all scored DNA fragments). Furthermore, high levels of epigenetic variation were observed among the studied individuals. This high epigenetic variation found in P. pinea contrasted with the lack of genetic variation based on Amplified Fragment Length Polymorphism (AFLP) data. In this manner, variable epigenetic markers clearly discriminate individuals and differentiates two well represented populations while the lack of genetic variation revealed with the AFLP markers fail to differentiate at both, individual or population levels. In addition, the use of different replicated trees allowed identifying common polymorphic methylation sensitive MSAP markers among replicates of a given propagated tree. This set of MSAPs allowed discrimination of the 70% of the analyzed trees.     

Genetic diversity and relationships among species of the genus Thymus L. (section Serpyllum)

Amplified fragment length polymorphism (AFLP) markers were used to assess genetic relations of 32 Thymus populations belonging to seven species of genus Thymus, section Serpyllum, from Serbia. The collected species belong to three subsections: (1) Alternantes Klokov (Th. pulegioides L.), (2) Isolepides (Borbás) Halácsy (Th. glabrescens Willd., Th. marschallianus Willd., Th. pannonicus All.), and (3) Pseudomarginati (Braun ex Borbás) Jalas (Th. balcanus Borbás, Th. moesiacus Velen., Th. praecox Opiz). Analysis of molecular variance (AMOVA) revealed that most of the genetic diversity was attributable to differences among individuals within populations, while the remaining variability was almost equally distributed among species and among populations within species. The pairwise AMOVA analyses further showed that species differentiation was significant in all cases, except between Th. marschallianus and Th. pannonicus. All the individuals belonging to Th. pulegioides (subsection Alternantes) formed a well-supported clade, the most divergent from all other species in this study. This genetic differentiation of Th. pulegioides is supported by its morphological features because it is the only analyzed taxon having hairs only on the stem edges, and can easily be recognized by this character. According to our results, the subsection Isolepides is polyphyletic. Th. glabrescens is clearly separated from the other two species of this subsection (Th. marschallianus, Th. pannonicus). Although morphologically distinguishable by leaf indumentum, individuals belonging to Th. marschallianus and Th. pannonicus could not be differentiated at the molecular level. The species belonging to subsection Pseudomarginati formed a monophyletic clade, although not supported by high bootstrap value. The clade is further divided into three well-supported clades representing distinct species (Th. balcanus, Th. praecox and Th. moesiacus). In our study, the AFLP markers demonstrated that they could be suitable for studying complex genetic relationships, including frequent interspecies hybridization events, although strict reticulate evolutionary history could not be determined. All the population genetic parameters in the section Serpyllum suggest that the reproductive incompatibility between the species is very weak, which indicates a strong reticulating system, especially in the case when two or more species occur sympatrically.     

Genetic variation in Banksia saxicola (Proteaceae), a rare Australian plant with a markedly disjunct distribution

 Banksia saxicola A.S. George (Proteaceae) is a rare Australian endemic, found in only two locations in Victoria that are separated by approximately 500 kms: the Grampians and Wilson's Promontory National Parks. The organisation of genetic variation between and within populations at the two locations was assessed using the Amplified Fragment Length Polymorphism (AFLP) technique. Two populations were sampled in the Grampians National Park and one population was sampled at Wilson's Promontory. The three populations were genetically divergent from each other, in particular the Grampians and Wilson's Promontory populations, and this relates to the ancient geographic isolation of these two regions. The Wilson's Promontory population had lower genetic diversity than either Grampians populations, consistent with its smaller population size. The findings are important for strategies to conserve B. saxicola. Received October 3, 2000 Accepted January 3, 2001