RFLP analysis of mitochondrial DNA in eggplant and related Solanum species

RFLP analysis of mitochondrial DNA (mtDNA) was performed by the Southern hybridization method using total DNA extracted from eggplant, Solanum melongena L., and six related Solanum species, S. surattense Burm. (i.e. S. virginianum L.), S. torvum Swartz, S. gilo Raddi (i.e. S. aethiopicum), S. integrifolium Poir. (i.e. S. aethiopicum), S. indicum auct. non L. (i.e. S. violaceum Ort.) and S. sanitwongsei Craib. Forty-one fragments were detected by the analysis using 12 combinations of four restriction enzymes and three probes of mtDNA clones from rice. Thirty-four out of the 41 fragments were polymorphic among the species, whereas the other seven were monomorphic. This RFLP analysis of mtDNA is demonstrated to be appropriate for assessing phylogenetic relationships in eggplant and related Solanum species at the interspecific level.     

Evaluating the demographic history of the Seychelles kestrel (Falco araea): Genetic evidence for recovery from a population bottleneck following minimal conservation management

An important requirement for biologists conserving vulnerable species of wildlife and managing genetic problems associated with small population size is to evaluate existing evidence regarding what is known of a species’ recent population history. For endemic island species in particular, current genetic impoverishment could be due to either a recent population crash or a consequence of an evolutionary history of sustained isolation and small effective population size. Interpreting any given case can often be further complicated by incomplete or contradictory evidence from historical field surveys that might suggest a very different demographic history. Here, we use the case of the Seychelles kestrel (Falco araea), an island endemic previously listed as critically-endangered but now relatively common, to illustrate how genetic data from microsatellite genotypes of 100–150-year-old museum specimens reveals a recent and severe population crash since the 1940s to approximately eight individuals, before the population recovered. We re-interpret the historical population trajectory of the Seychelles kestrel in the light of the minimal intervention required for this species to recover. We examine different ecological explanations for the decline and apparently unassisted recovery of the Seychelles kestrel, review the evidence for similarly unaided recoveries elsewhere and discuss the implications of unaided population recoveries for future species conservation programmes. Demographic profiles from historical genetic signatures can provide highly informative evidence when evaluating past and future recovery efforts for endangered species.     

Genetic diversity of core and peripheral Sitka spruce (Picea sitchensis (Bong.) Carr) populations: implications for conservation of widespread species

This study investigated levels of genetic diversity and population differentiation among Sitka spruce (Picea sitchensis) populations classified as core or peripheral based on ecological niche, and continuous or disjunct based on species distribution. Large numbers of trees (N = 200) were sampled from each of eight populations to evaluate the distribution of rare as well as common alleles across the species range. Codominant alleles for eight sequence-tagged site loci were classified based on frequency and geographic distribution in order to develop appropriate sampling strategies to target specific classes of alleles. An important finding of this study is the similarity in genetic diversity as measured by expected heterozygosity between core populations (mean H_E = 0.58) and peripheral populations (mean H_E = 0.56). However, there was significant inbreeding in peripheral (F_IS = 0.17) but not in core (F_IS = 0.03) populations. Large differences in gene flow estimates were observed between core (Nm = 9.0) and peripheral populations (Nm = 3.5). Irrespective of population classification, over 75% of the alleles were common and widespread. Only one allele was classified as rare and localized, and this allele was limited to one core, disjunct and two peripheral, disjunct populations. There was stronger evidence of past bottlenecks in peripheral, disjunct populations than in core, continuous populations. Results are used to suggest sampling strategies for capture of maximum level of genetic diversity and conservation of rare alleles. The conservation of peripheral, particularly disjunct, populations as well as populations in putative glacial refugia may present the best opportunity for conserving rare alleles.     

MtDNA genetic diversity and population history of a dwindling raptorial bird, the red kite (Milvus milvus)

The red kite (Milvus milvus) occurs in a relatively small area in the southwestern Palearctic region, with population strongholds in Central Europe. Following strong human persecutions at the beginning of the 20th century, populations have receded, particularly in peripheral areas and islands. In order to describe and compare levels of genetic diversity and phylogeographic patterns throughout its entire distribution in Europe, sequence variation of a 357 bps part of the mitochondrial DNA control region was assessed in eight populations and 105 individuals. Overall, results indicate that population declines have affected red kite mtDNA variation. We found low levels of genetic diversity (values of nucleotide diversity ranging from 0 in Majorca island to 0.0062 in Central Europe), with only 10 distinct haplotypes, separated by low levels of genetic divergence (mean sequence divergence = 0.75%). Highest haplotype and nucleotide diversities match with demographic expectations, and were found in Central European and Central Spanish samples, where present strongholds occur, and lowest values in the declining southern Spanish and insular samples. Φst estimates indicated moderate gene flow between populations. Phylogeographic patterns and mismatch distributions analyses suggest central European regions may have been colonized from southern glacial refugia (in the Italian or Iberian peninsulas). Interspecific phylogenetic comparisons and divergence date estimates indicated the genetic split between the red kite and its closely related species, the black kite (Milvus migrans), might be relatively recent. The low level of genetic variation found in the red kite mitochondrial control region, compared to the black kite, is likely the result of relatively recent divergence (associated with founder events), successive bottlenecks and small population sizes. As there are several ongoing projects aimed at reinforcing populations in countries such as the United Kingdom, Italy or Spain, our results may prove useful for the genetic management of the species.     

Conservation genetics and implications for restoration of Hemigenia exilis (Lamiaceae), a serpentine endemic from Western Australia

Hemigenia exilis (Lamiaceae) is a rare plant endemic to serpentine soils of the Goldfields of Western Australia. The species was presumed extinct until 1995, when it was re-discovered on a nickel ore deposit. To delineate the origin and extent of seed collection for rehabilitation after mining, and to identify the impact of removing one population due to mining, we assessed level and partitioning of genetic variation and differentiation. Twelve populations were sampled for DNA fingerprinting using the random amplified polymorphic DNA (RAPD) technique. Ten primers produced 89 bands, 97% being polymorphic. Genetic diversity within populations ranged from 0.197 to 0.409, averaging 0.38 at the species level, which is high compared with most other endemic species. Heterozygosity within populations ranged from 0.355 to 0.431, averaging 0.27 over the species. AMOVA partitioned over 80% of the total variation within populations. Multidimensional scaling revealed weak but significant differentiation into a northern and southern provenance. Despite selective sampling, the genetic data provided useful information for the management of Hemigenia exilis. For restoration, seed should be collected from a range of habitats of several populations, while keeping the two provenances separate. This strategy is likely to maintain high genetic diversity and locally adapted populations.     

Relationship between population size and genetic diversity in endangered populations of the European bullhead (Cottus gobio): implications for conservation

This study investigated the relationship between the current size of endangered bullhead (Cottus gobio) populations and microsatellite genetic variability. Additionally, the microsatellite data were used to evaluate whether a genetic test for population bottlenecks was able to provide evidence of recent severe population declines. Finally, our results were used to develop conservation priorities and measures. Population size appears to be a crucial parameter in determining the amount of genetic diversity that can be preserved in bullheads, since a significant positive correlation was observed between both variables. Furthermore, in some populations we were able to detect genetic signatures of the documented decline in population size. We suggest that the most immediate goal for bullhead conservation should be to increase the size and the range of the populations, and in doing so minimise or even reverse further genetic erosion. Potential management actions like habitat quality improvement, reduction of river fragmentation and supplementation programmes (translocation, supportive breeding) are discussed.     

Morphological characterization of seeds of three Australian wild Citrus species (Rutaceae): Citrus australasica F. Muell., C. inodora F.M. Bailey and C. garrawayi F.M. Bailey

The comparative morphology of the seeds of three Australian Citrus species, C. australasica C. inodora and C. garrawayi, was studied. Their seed characteristics were broadly similar to those of the cultivated species of the genus, when observed under light and scanning electron microscopy. Citrus garrawayi differed in seed shape (rounded to triangular) and seed coat morphology (i.e., thicker with longer epidermal protrusions) from C. australasica and C. inodora (rounded surface with flat underside in shape). The well-developed minute epidermal protrusions on the seed coat of C. garrawayi were more similar to those in the cultivated species, C. × sinensis and C. × aurantium. In contrast, the surface topography of C. australasica and C. inodora seeds was more like that of the cultivated species, C. × aurantifolia and C. × limon. Seed morphology, especially surface topography, was found to be a useful tool for taxonomic identification in Australian wild citrus.     

秦巴山区黄牛群体的微卫星DNA遗传多样性

为分析秦巴山区西镇牛、赤崖牛、陨巴牛和宣汉牛4个黄牛(Bos taurus)品种的遗传多样性,本研究以蜀宣花牛和郏县红牛作对照,利用12对微卫星引物对6个黄牛品种的289头黄牛个体进行了遗传多样性检测,统计了各品种的等位基因及频率、有效等位基因数、遗传杂合度、多态信息含量及各群体间遗传距离,并对其进行聚类分析.结果表明,6个黄牛品种在12个微卫星位点共发现110个等位基因,等位基因频率为0.001 6～0.517 3,总群体各位点平均有效等位基因数为2.787 7～7.132 6;各位点多态信息含量为0.5192～0.895 3;平均杂合度为0.667 2～0.724 1.群体间发现特有等位基因11个,基因频率为0.008 1～0.381 6;优势等位基因(P＞0.4) 19个,基因频率为0.403 2～0.820 0;共有等位基因41个,占全部等位基因的37.27％,仅有13个共有等位基因为优势等位基因(P＞0.4),占37.71％.群体间Nei's遗传一致度为0.596 5～0.840 8,标准遗传距离(Ds)为0.173 5～0.524 7.聚类分析结果显示,6个黄牛品种聚为3类,陨巴牛与宣汉牛首先聚在一起,然后同西镇牛聚在一类;赤崖牛与郏县红牛聚为一类;蜀宣花牛自成一类.研究结果表明,12对微卫星标记可用于秦巴山区黄牛遗传多样性的分析,秦巴山区各黄牛品种遗传多样性丰富,选育程度不同,4个黄牛品种虽然地理分布格局相近,自然环境相似,但亲缘关系并非相近,应为来源不同的品种.因此,西镇牛、赤崖牛、陨巴牛和宣汉牛不宜合并为1个品种.本研究所揭示的秦巴山区黄牛品种的遗传分化特点及亲缘关系,为研究品种的遗传共适应特点,预测杂交优势,制定育种战略等提供了科学依据.     

绿色豌豆蚜不同地理种群的遗传多样性

豌豆蚜是我国苜蓿上危害最为严重的害虫之一。利用微卫星标记研究了我国绿色型豌豆蚜10个地理种群的遗传相似性、基因分化、遗传距离与地理距离和海拔之间的关系及其基因结构。选取15对引物扩增300个个体,共检测到41个多态性条带,多态性条带百分率(PPB)为100%。10个豌豆蚜地理种群观测等位基因数(Na)为1.592 7,有效等位基因数(Ne)为1.356 9,Nei’s基因多样性指数为0.206 6,Shannon-Wiener指数(I)为0.307 6。新疆维吾尔自治区、陕西省、河南省种群的遗传多样性较高,内蒙古自治区、山东省、青海省种群相对较低。然而,10个地理种群豌豆蚜(绿色型)的遗传分化较高(Gst=0.399 6)。种群聚类分析结果显示,全部豌豆蚜种群明显聚为两大类群,山东省、河南省种群为一大类群,其余为另一大类群。Mentel检测表明,遗传分化与地理距离、海拔无显著相关性。我国豌豆蚜种群具有非常丰富的遗传多样性,应加强豌豆蚜的监测和治理。     

Mitochondrial DNA sequence, morphology and ecology yield contrasting conservation implications for two threatened buckmoths (Hemileuca: Saturniidae)

Taxa of conservation interest are frequently identified using morphological or ecological characters. These characters are assumed to represent evolutionary importance, population structure and/or phylogenetic relationships in such organisms. We tested this assumption using two species complexes of the moth genus Hemileuca (Saturniidae). Both have populations threatened by habitat loss and need conservation protection. Legislation protects one taxon with apparent ecological differences. We sequenced 624 base pairs of mtDNA from the COI gene for geographically distant populations of the Hemileuca maia species complex and the H. electra species complex. Resultant phylogenies contradict prior assumptions about relationships in both species complexes. The legislatively protected Bog Buckmoth is paraphyletic with widespread H. maia, and its use of a novel hostplant seems to be a local adaptation. Divergent morphology and hostplant use among H. electra subspecies are associated with modest genetic divergence (0.48%). However, a group of unrecognized populations that are morphologically similar and geographically close to H. electra electra have mtDNA that is divergent by an average of 4.1%. There is disagreement regarding prioritization of ecological divergence over neutral genetic distance in conservation. We place ecological variation in a phylogenetic context and recommend that exploration of genetic relationships be undertaken when populations are threatened. Adaptive ecological variation should be evaluated in a phylogenetic context to understand its conservation importance. This study illustrates the importance both of phylogenetic context and the use of independent characters in assessing biodiversity for conservation prioritization.     

Cross-amplification and characterization of microsatellite loci for three species of <Emphasis Type="Italic">Theobroma</Emphasis> (Sterculiaceae) from the Brazilian Amazon

This study reports on the cross-species amplification of 23 microsatellite markers previously developed for Theobroma cacao L. (Sterculiaceae), source of chocolate in three economically important Amazonian species of Theobroma (T. grandiflorum, T. subincanum, T. sylvestre). Thirteen of the 23 microsatellite loci tested were polymorphic across the three species at 2–13 alleles per locus. The observed heterozygosity per locus varied from 0.18 to 0.84 and expected heterozygosity ranged from 0.28 to 0.87. The high level of transferability and genetic information content of these microsatellite loci indicate their usefulness for population genetic, mating system and breeding studies of these economically important Amazonian fruit trees.     

A wide range of genetic diversity in pear (<Emphasis Type="Italic">Pyrus ussuriensis</Emphasis> var. <Emphasis Type="Italic">aromatica</Emphasis>) genetic resources from Iwate,Japan revealed by SSR and chloroplast DNA markers

Iwateyamanashi (Pyrus ussuriensis var. aromatica) is one of the Pyrus species which grows wild in Japan. The number of Iwateyamanashi trees has been decreasing, so conservation and evaluation is urgently needed. Over 500 accessions of Pyrus species collected from Iwate in northern Tohoku region are maintained at Kobe University as an Iwateyamanashi germplasm collection. In order to investigate the genetic diversity, five SSR (simple sequence repeat) markers, developed from Japanese and European pear were examined for 86 Pyrus individuals including 58 accessions from Iwate. These SSR loci could discriminate between all the Iwate accessions except for 10 that bear seedless fruit, as well as determine the genetic diversity in Iwateyamanashi germplasms. High levels of variation were detected in 41 alleles and the mean observed heterozygosity across 5 loci was 0.50 for the Iwate accessions. Seedless accessions sharing identical SSR genotype with the local pear variety “Iwatetanenashi” were supposed to have been propagated vegetatively via grafting. In an UPGMA phenogram, Japanese pear varieties (P. pyrifolia) were clustered into two groups with some Iwate accessions including seedless ones. Another 38 Iwate accessions were not clustered clearly, and there was no clear relationship between these accessions and geographical distribution or morphological characters. Allele frequency revealed that the Iwate accessions were genetically more divergent than the Japanese pear varieties. Most Japanese pears possessed a 219 bp deletion at a spacer region between the accD and psaI genes in the chloroplast DNA (cpDNA), but other Pyrus species and two Iwateyamanashi trees did not. In the Iwate accessions, 79.3% had a deletion type cpDNA and others had a standard type cpDNA without deletion. These results are indicative of the wide range of genetic diversity in the Iwate accessions which include Japanese pear varieties. A combination of SSR and cpDNA analyses revealed high heterogeneity in Iwateyamanashi and coexistence of Iwateyamanashi and hybrid progeny with P. pyrifolia. These could be reasons for the wide range of continuous morphological variation described previously.     

Catabolic and genetic microbial indices,and levels of nitrate,ammonium and organic carbon in soil from the black locust (<Emphasis Type="Italic"> Robinia pseudo-acacia</Emphasis>) and tulip poplar (<Emphasis Type="Italic"> Liriodendron tulipifera</Emphasis>) trees in a Pennsylvania forest

Soil samples were collected from the upper soil horizon within 4 m of black locust ( Robinia pseudo-acacia) and tulip poplar trees ( Liriodendron tulipifera) from the same mixed forest in south-central Pennsylvania. The soil samples were analyzed for organic C levels, pH, NO₃ ^-, NH₄ ⁺, catabolic diversity (Shannon diversity index; catabolic H), catabolic evenness (Simpson-Yule index; catabolic E), genetic H, and genetic E. The catabolic H and genetic H of microbes in these soils were found to correlate well with the levels of mineralized N, organic C, and pH. Significant variations in these parameters were found between the soils from near black locust and tulip poplar trees. Conditions in the soil near the black locust trees were more favorable to nitrification as indicated by the elevated pH, organic C, NO₃ ^-, and total mineral N levels, along with lower NH₄ ⁺ levels. The microbial genetic H and E were greater and the catabolic H and E were lower in the black locust soils than in the tulip poplar soils. This suggests that a more specialized environment exists in the soil near the black locust trees which selects for enhanced nitrification and the use of fewer, but preferred catabolic pathways by a more genetically diverse group of microbes that grow to a greater biomass. Conversely, the soils from near the tulip poplar trees are such that they do not select for some dominant catabolic pathways, rather they allow for the use of a greater variety of catabolic pathways by a less diverse microbial population, which appear to grow to a lower biomass. We believe that the combined application of the microbial genetic and catabolic diversity analyses, microbial biomass estimates, and traditional physico-chemical characteristics in soil studies provides information not easily available that can be useful during assessment of soil processes in different terrestrial habitats.     

Sex differences in population genetics, home range size and habitat use of the parti-colored bat (Vespertilio murinus, Linnaeus 1758) in Switzerland and their consequences for conservation

When habitats are declining, niche segregation by demographic groups, such as the two sexes, can have a profound impact on the extinction risk of a species as a whole. Thus, differences in the requirements of demographic groups are of importance in conservation. We combined behavioural and genetic data to investigate whether the sexually segregated parti-colored bat (Vespertilio murinus) exhibits sex-specific niche partitioning. We use our data to evaluate implications for conservation of this potentially vulnerable species in Switzerland, the western boundary of its range. Using radio-telemetry, we found sex-specific differences in habitat use. Foraging females strongly relied on lakes while foraging males displayed more flexibility in their habitat use. Moreover, males covered significantly larger foraging areas than females. Sequencing 341 base pairs of the mitochondrial D-loop of 247 individuals revealed sex-specific differences in the genetic structure of colonies, but no such difference was observed for three nuclear micro-satellite markers. We found high mtDNA diversity in two Swiss male colonies and one German female colony, but low mtDNA diversity in two Swiss female colonies. Our genetic data suggest that considerable gene flow occurs via male dispersal and mating. At the same time immigration of females into the existing female colonies in Switzerland is rare compared to the immigration of new males into male colonies. Since we found the sexes in Vespertilio murinus to differ markedly in their ecology, population genetics, and behaviour, we conclude that sex-specific conservation plans are required to protect this species efficiently.     

Molecular analysis of the DNA polymorphism of wild barley (Hordeum spontaneum) germplasm using the polymerase chain reaction

Summary Hordeum spontaneum is the progenitor of cultivated barley (H. vulgare) and is an important source of genetic variation for barley breeding programs. The genetic diversity ofH. spontaneum in the Australian germplasm collection was investigated using the polymerase chain reaction with random and semi-random primers. This approach was found to be robust in respect of reaction conditions. Genetic dissimilarity values between genotypes were used to produce a phenogram of the relationships among the accessions using the unweighted pair group method with arithmetic mean. The largest divergence was observed among Israeli accessions, whereas the Turkish and Iranian samples clustered as distinct subsets, each apparently related to portion of the Israeli material. The results indicate that the genetic diversity of the wild barleys is broadly correlated with geographic distribution.     

Influence of habitat fragmentation on the genetic structure of Polyommatus coridon (Lepidoptera: Lycaenidae): implications for conservation

Conservation biologists pay increasing attention to the interdependence between habitat structure and genetic patterns of species and to the importance of conserving genetic diversity. A suitable model species for such a study is Polyommatus coridon, a butterfly of dry calcareous grassland. We studied the population genetic structure of 22 western German populations of this species (874 individuals). We observed no congruence between the observed genetic structure and the geographic arrangement of the studied populations. Genetic diversity was slightly higher in large populations than in small populations. Differentiation among populations was weak or even missing. Populations of an area with high habitat density showed no significant differentiation, whereas areas with low habitat densities showed significant genetic differentiation among populations. At present, hierarchical variance analysis revealed no differentiation between four distinguished limestone areas. The combination of all analytical results allow the definition of management units for P. coridon. Conservation measures are proposed from which a large number of rarer species occurring together with P. coridon will benefit.     

Genetic diversity of <Emphasis Type="Italic">Guizotia abyssinica</Emphasis> (L. f.) Cass. (Asteraceae) from Ethiopia as revealed by random amplified polymorphic DNA (RAPD)

Genetic diversity of 70 populations of niger (Guizotia abyssinica) representing all its growing regions in Ethiopia was investigated using random amplified polymorphic DNA (RAPD) to reveal the extent of its populations genetic diversity. Ninety-seven percent of the loci studied was revealed to be polymorphic for the whole data set. The within population diversity estimated by Shannon diversity index and Nei gene diversity estimates was revealed to be 0.395 and 0.158, respectively. The extent of genetic variation of populations from major niger producing regions was significantly lower than that of populations from other regions; however, it is distributed regardless of altitude of growth. Genetic differentiation between populations was estimated with Shannon index as G^′ _ST (0.432), Nei’s G _ST (0.242) and AMOVA based F _ST (0.350) and appears to be equivalent to the average values calculated from various RAPD based studies on outcrossing species. Higher proportion of the variation detected by AMOVA resided within populations (64.58%) relative to the amount of variation among populations (35.42%). UPGMA cluster analysis showed that most of the populations were clustered according to their region of origin. However, some populations were genetically distant from the majority and seem to have unique genetic properties. It is concluded that the crop has a wide genetic basis that may be used for the improvement of the species through conventional breeding and/or marker assisted selection. Collection of germplasm from areas not yet covered and/or underrepresented is the opportunity to broaden the genetic basis of genebank collection.     

Oleaster (var. sylvestris) and subsp. cuspidata are suitable genetic resources for improvement of the olive (Olea europaea subsp. europaea var. europaea)

The olive belongs to Olea, a complex genus and to a species with six subspecies. Subsp. europaea includes both the cultivated olive and the oleaster, the wild ancestor of the olive. Little is known on the phenotype of fruits from subsp. cuspidata. We aimed to compare europaea and cuspidata trees for oil content and composition and to verify whether natural hybrids may exist between the two subspecies. Specimens were from Kenya and putative natural hybrids between cuspidata and europaea were from Stellenbosch (South Africa). Cultivar and oleaster trees were sampled in France (continental and Corsica), Italy (Continental and Sardinia) and Tunisia. We have examined the fruit (drupe) structure between subsp. cuspidata and europaea and extracted oil from the pulp, seed and total drupe. Comparison for oil content was made between the cultivated olive and the oleaster and some cuspidata trees from Kenya. A few of cuspidata individuals from Botanical gardens do not enable phenotyping for oil content and composition. Oil composition analyses were performed for the main fatty acids to compare the three taxa. We used microsatellite markers at 11 loci to compute genetic distances between cuspidata, oleaster and cultivar trees and to reveal eventual hybrids. The SSR polymorphisms were huge between the two subspecies and they revealed that putative hybrids were true hybrids sampled around olive orchards. The whole comparison of oil content shows that olive cultivars display higher oil content than the subsp. cuspidata and that the oleaster trees are intermediate, whereas for oil composition of the drupe, cuspidata from Kenya shows less oleic acid than europaea. However, the cuspidata trees show seed oil composition similarities with the other two taxa. The discussion deals with possible reasons to explain the differences and of the possible uses of these taxa for breeding both the olive and brown olive. H. Hannachi and H. Sommerlatte have equal contribution in the article.     

Analysis of the genetic structure of red-legged partridge (Alectoris rufa, Galliformes) populations by means of mitochondrial DNA and RAPD markers: a study from central Italy

The strong hunting pressure on the red-legged partridge, Alectoris rufa, warranted its inclusion into the list of species of European conservation concern. During the last decades, restocking plans with farmed specimens have counterbalanced the hunting drawings from wild populations. Our concern was the study of A. rufa in the easternmost part of its range, the central Italy, to gain insights into the effects of this compensation practice on the genetic structure of its populations. Partridges from both a geographically isolated, long-time protected, wild population (Pianosa island, Tuscan Archipelago National Park) and two Tuscan farms (Bieri and Scarlino) were investigated. All the specimens were very similar in outward appearance, looking much like to A. rufa. Ninety-six sequences of both Cytochrome b and D-loop Control Region of mitochondrial DNA (mtDNA) were analysed to get evidence of ancestry at the population level, whereas, the Random Amplified Polymorphic DNA (RAPD) technique was employed to get fingerprinting at the individual level. Pianosa and Bieri populations showed both the A. rufa and Alectoris chukar mtDNA lineages, whereas the Scarlino one only the A. rufa-mtDNA line. However, a spread overall pattern of A. rufa × A. chukar hybridisation among specimens, whatever their mtDNA lineage could result to be, was disclosed by means of RAPD species-specific markers. This is the first genetically documented record of the A. rufa × A. chukar hybrids. The occurrence of the pure, native A. rufa genome in the easternmost part of the species’ geographical range may be guessed to be virtual.     

Phylogeographic study based on intraspecific sequence variation of chloroplast DNA for the conservation of genetic diversity in the Japanese endangered species Primula sieboldii

To gather information on intraspecific phylogeography for use in conservation programs for the endangered species Primula sieboldii in Japan, we analyzed sequence variation in five noncoding regions of chloroplast DNA. Twenty-two distinct haplotypes were recognized in total. The distribution of most haplotypes was geographically confined, but one haplotype was widely distributed throughout northern Japan, and several haplotypes were found in geographically distant regions. Three major clades were revealed by phylogenetic analysis of the haplotypes. Clade I was distributed in Kyushu and central Honshu, clade II in western Honshu and Hokkaido, and clade III from central Honshu to Hokkaido. According to analysis of molecular variance, 59.9% of the total cpDNA variation existed among regions, 32.5% among populations within regions, and 7.6% within populations. Therefore, if genetic conservation of the species is valued, transplanting of P. sieboldii among regions should be avoided. Multiple lineages often existed even in geographically narrow areas (e.g., within a 20-km range), so transplantation between adjacent populations in restoration activities should be carefully designed so as not to change the gene pool of local populations significantly. Also, the geographical distribution of cpDNA haplotypes may allow us to confirm the origin of plants collected for commercial purposes.