Intra- and interspecific genetic differentiation in closely related pines fromPinus subsectionSylvestres (Pinaceae) in the former Soviet Union

Thirty-seven natural populations of four closely related species ofPinus subsect.Sylvestres, P. mugo, P. funebris, P. pallasiana, andP. sylvestris, occurring in the former Soviet Union were investigated by starch-gel electrophoresis. In the populations assayed 127 allelic variants at 25 loci were revealed.Nei's distance coefficient (Dn) was used to estimate the level of genetic differentiation amongP. sylvestris races and among closely related species. A dendrogram constructed using Dn values shows that of the fiveP. sylvestris races analyzed only the geographically isolated var.hamata exhibited sufficient differences at theDia-2 locus (a mean Dn value relative to the other four races is 0.025) to recognize it as a distinct taxon. The remaining races, sylvestris, cretacea, lapponica, and sibirica, have a similar gene pool (Dn values are not greater than 0.010), and they should be regarded as a single taxon,P. sylvestris var.sylvestris. Interspecific comparisons revealed thatP. sylvestris andP. mugo have the closest genetic affinities to each other withNei's genetic distance of 0.108. The dendrogram demonstrates thatP. funebris is closer toP. sylvestris andP. mugo thanP. pallasiana. The available paleontologic data allowed us to conclude thatNei's (1975) time scale estimate for the time of divergence of the taxa was more accurate thanNei's (1971) time scale estimate.     

Allozyme investigations on the genetic differentiation between closely related pines —Pinus sylvestris, P. mugo, P. uncinata, andP. uliginosa (Pinaceae)

In eight natural European populations of four closely related taxa of pines (Pinus sylvestris, P. mugo, P. uncinata, andP. uliginosa) starch-gel electrophoreses revealed altogether 58 alleles at 15 loci from nine enzyme systems. With Nei's genetic distance (D) the largest mean genetic distance (D = 0.171) was found betweenP. sylvestris andP. mugo, a distance corresponding to that between other closely related pine species. Mean genetic distances between the other taxa were less than half that value and characteristic for subspecies or varieties rather than for species. On the basis of our results we suggest that both,P. uliginosa andP. uncinata, could be the result of the ancient hybridization betweenP. sylvestris andP. mugo.     

Genetic relationships between European and Siberian larch,Larix spp. (Pinaceae), studied by allozymes. Is the Polish larch a hybrid between these two species?

Thirteen populations ofLarix decidua subsp.decidua and subsp.polonica, and three populations ofL. sibirica were investigated by starch-gel electrophoresis. In the populations assayed 61 alleles at 17 loci were revealed. The allozyme data support the earlier observations about close relationships between these two larch species. Nei's genetic distances betweenL. decidua andL. sibirica were relatively small (D = 0.057), however, almost five times larger, on average, than those between populations of the same species. Results obtained in this study disagree withBobrov's hypothesis about the hybrid origin of the Polish larch and suggest a direct origin from the European larch.     

Comparison of genetic population structure between two cyprinids, <Emphasis Type="Italic">Hemigrammocypris rasborella</Emphasis> and <Emphasis Type="Italic">Pseudorasbora pumila</Emphasis> subsp., in the Ise Bay basin,central Honshu,Japan

Two small cyprinid fishes, Hemigrammocypris rasborella and Pseudorasbora pumila subsp. (sensu Nakamura 1963), inhabit similar habitats and often occur sympatrically in the Ise Bay basin, central Honshu Island, Japan. Their genetic population structures were revealed, using sequence data from the mitochondrial cytochrome b gene, and then compared. Hemigrammocypris rasborella populations in the Ise Bay area formed a monophyletic group that has been isolated from eastern (Tenryu River) and western (Lake Biwa–Yodo River) populations at least for several hundred thousand years. Pseudorasbora pumila subsp., endemic to the Ise Bay area, was estimated to have become isolated from its sister subspecies, P. p. pumila, about 5 million years ago. Both H. rasborella and P. pumila subsp. had centers of genetic diversity around the Okazaki Plain in the eastern part of the basin and showed trans-bay distribution of haplotypes or haplotype groups. Their common population structure was explained by geological features in the Ise Bay area, in which a large paleo-river system developed in regression periods, suggesting gene flow among populations of each species in the mid to lower reaches of the paleo-river. Based on the estimated expansion or divergence time, however, not all populations experienced gene flow during the Last Glacial. In contrast to the maintenance of high genetic diversity in H. rasborella, almost all populations of P. pumila subsp. have lost mitochondrial DNA genetic diversity. This implies that effective population size of P. pumila subsp. tended to be smaller, probably because of differences in reproductive ecology, even though the two species have been exposed to similar environmental changes. For conservation of the two species, genetic and adaptive differentiation among local populations should be considered, and attention should be paid to inbreeding depression, especially in P. pumila subsp. An erratum to this article can be found at     

红松种子园种群表型多样性研究

为揭示红松（Pinus koraiensis Sieb.et Zucc.）不同种群的表型分化程度和变异规律,以吉林省露水河红松种子园6个种群红松为研究对象,采用巢式方差分析、多重比较、变异系数和聚类分析等方法对红松的叶片、球果和种子共15个表型性状进行了系统分析。结果表明：（1）红松15个表型性状在种群间和种群内均存在着极显著的差异,红松种群遗变异比较丰富,在松属植物中属于中等水平,其中纬度最低的露水河种群在其中10个表型性状均值上表现出最高值;（2）红松种群间的表型分化系数（Vst）均值为12.39%,种群内的变异（87.61%）大于种群间的变异（12.39%）,种群内的变异是主要变异来源;（3）各表型性状平均变异系数为13.28%,变幅为6.16%-31.48%,叶片、球果、种子的平均变异系数依次为：球果17.86% > 针叶11.19% > 种子9.87%,种子性状最小,成为最稳定的表型性状,带岭和丰林种群表型性状遗传多样性要高于其他种群;（4）利用欧氏平均距离对红松种群进行UPGMA聚类分析,红松种群的表型性状按照地理距离而聚类,可将红松种子园6个种群分为4类,其表型性状跟地形（东北山脉）有一定的契合。红松种群具有中等水平的表型遗传多样性,种群间和种群内均存在丰富的表型变异,研究结果为顺利开展红松种质资源收集、保存,以及良种选育等工作提供依据。     

Allozyme diversity in Chinese,Seguin and American chestnut (Castanea spp.)

Allozyme genetic variability in three chestnut (Castanea) species was investigated using 19 loci from ten enzyme systems. G-tests of heterogeneity of isozymic allele distribution showed significant differences between the three species at 15 of the 19 loci, and between the 13 C. mollissima populations at 13 of the 19 loci examined. C. mollissima was found to possess a significantly-higher value of mean gene heterozygosity (H=0.3050±0.0419), the percentage of polymorphic loci (P=84.21%) and the average number of alleles per locus (A=2.05), than any other species in the Castanea section Eucastanon. When the genetic variability of populations of C. mollissima from four regions in China was investigated, the population from the Changjiang river region showed a markedly higher mean gene heterozygosity (H=0.3480±0.0436) than populations from the other regions. Genetic relationships among the four regions were assessed by Nei's genetic identity I and standard genetic distance D. An approximately-identical distance between the population from the Changjiang river region and populations from the three other regions was observed, while populations from the latter regions showed almost the same genetic distance from each other. These data, when considered with information existing prior to this study, contribute to an understanding of the possible origin and progenitor of the chestnut species.     

Gene variation and gene flow inOrchis morio (Orchidaceae) from Italy

Data are presented on genetic variation at 27 enzyme loci of the Green-Winged orchid,Orchis morio, in 18 population samples from Italy. The existence in Italy of two subspecies, i.e. subspp.morio andpicta, is not supported by allozyme data. No genetic heterogeneity was found betweenmorio-like andpicta-like samples and specimens. Moreover, morphological transition between the two forms was observed in different Italian populations. The parameters of genetic variability estimated forO. morio populations are consistent with those found among monocotyledon plants, and among those outcrossing, animal-pollinated and with wind-dispersed seeds. Genetic diversity of ItalianO. morio is mostly within populations. Correspondingly, low values of interpopulational genetic distance were found. This appears to be due to high levels of gene flow, which were estimated with different methods. The lack ofO. longicornu from Italian samples, as well as of any hybrid withO. morio (F₁, backcrossed or recombinant individuals) is demonstrated on the basis of genetic data. It is concluded that recurrent reports ofO. longicornu from Italy are due to confusion withO. morio or with otherOrchis species.     

Genetic structures of allopatric and sympatric populations in Pseudorasbora pumila pumila and Pseudorasbora parva

Pseudorasbora pumila pumila was collected to investigate its genetic relationships from 11 localities in Hokkaido and eastern Honshu of Japan with Pseudorasbora parva from 6 localities in Hokkaido, eastern Honshu, and Polovynka River (a tributary of the Amur River in eastern Russia) in 1996–1998. In Hokkaido and Akita prefectures (Honshu), three P. pumila pumila populations were distributed with P. parva sympatrically. Samples of P. pumila pumila and P. parva were analyzed based on 27 allozyme loci. Genetic differentiation between P. pumila pumila and P. parva (D = 0.522–0.622, average 0.601) was almost equal to or greater than that between other congeneric fish species previously reported. In the three sympatric populations of two species, which had arisen from artificial transplantation of the latter into localities of the former, the individual frequency composition of genetic types (P. pumila pumila, P. parva, F₁ hybrid, and later filial generation hybrid) were significantly different from each other. Differing from the previous studies, F₁ hybrids were fertile and coexisting populations of the two species included the F₁ hybrid and the later filial generation hybrid. These results suggest that genetic population dynamics of sympatric populations of the two species would be influenced not only by hybridization but also by ecological and environmental factors. Alternatively, the existence of populations that possessed both the alleles specific to P. pumila pumila and those specific to P. parva may indicate that introgressive hybridization, instead of species displacement, has occurred between the two species under a certain condition.     

Patterns of genetic differentiation in two annual bromegrasses,Bromus lanceolatus andB. hordeaceus (Poaceae)

Infraspecific genetic differentiation was analysed in two tetraploid annual bromegrasses,Bromus lanceolatus (from N Africa) andB. hordeaceus (from N Africa and France). Genetic analysis of populations was based on allozyme polymorphisms at 17 loci. Different fixed heterozygous phenotypes were scored in both species, according to their allopolyploid origin. In N Africa, more variation occurred among populations ofB. lanceolatus than ofB. hordeaceus. The variation was not randomly distributed among populations of both species. InB. lanceolatus, differentiation was correlated with climatic variables rather than with geographic distance between populations. Higher correlation of genetic differentiation with geographic distance occurred inB. hordeaceus, particularly at large geographic scale, between French and N African populations. Within each region, the populations appeared weakly genetically differentiated, even when belonging to different subspecies.     

Phylogenetic relationships of the Japanese minnows,Pseudorasbora (Cyprinidae), as inferred from mitochondrial 16S rRNA gene sequences

The phylogenetic relationships among threePseudorasbora fishes (Cyprinidae, Sarcocheiichthyinae) occurring in Japan (P. parva, P. pumila pumila andP. pumila subsp. sensu Nakamura [1963]) were inferred from nucleotide sequences of the mitochondrial 16S rRNA gene. The sequences. of 1240 bp, were determined and compared for 22 specimens from 2–8 populations for each taxon, with a singlePungtungia herzi specimen as an outgroup. A total of 171 sites (13.8%) were variable among the specimens, but only 0–2 sites within each population. The phylogenetic relationships estimated by neighbor-joining, maximum-parsimony and maximum-likelihood methods confirmed a sister relationship between the twoP. pumila subspecies, with a high level of confidence. However, their genetic distinction from each other (4.1±0.4SD % sequence difference on average) was at a level similar to that between them andP. parva (5.9±0.5%). The geographic distribution of the twoP. pumila subspecies, which are separated by the Fossa Magna region, suggests that the genetic divergence of the two subspecies originated from a vicariant process separating the freshwater ichthyofaunas of eastern and western Honshu.Pseudorasbora parva populations were divided into two genetic groups (1.8±0.2% sequence difference), one group comprising continental and part of the Japanese populations, and the other the remaining Japanese populations. This suggests that at least two genetically divergent lineages had been originally distributed in Japan, but a strong possibility remains that the present situation has resulted from artificial transplantation.     

Lack of evidence on hybrid swarm in the sympatric population of Pinus mugo and P. sylvestris

Natural hybridisation between the taxa from Pinus mugo complex and P. sylvestris was postulated in several sympatric populations of the species in Europe. However, due to the absence of precise methods for identification of hybrid seeds and hybrid trees, the frequency of hybridisation and its influence on the genetic structure of relict P. mugo populations has not been clarified so far. In the present study, the species-specific chloroplast DNA (cpDNA) marker and isozymes were applied to test the hybridisation hypothesis in a postulated hybrid swarm population of the species from “Bór na Czerwonem” reserve at the northern foothills of Tatra Mts., Poland. The cpDNA marker was used to identify hybrids within two groups of polycormic (multi-stemmed) and monocormic (single-stemmed) pines from this population. Allelic frequencies at isozyme loci from both groups were compared to frequencies found in allopatric populations of the species. Additionally, cpDNA haplotypes of seedlings derived from open pollinated seeds were studied to detect the possibility of successful cross-pollination. The mixture of seedlings with P. sylvestris and P. mugo cpDNA haplotypes was derived from one parental tree that indicates hybridisation. However, all the mature polycormic pines had cpDNA haplotypes species specific to P. mugo and the isozyme frequencies were similar to frequencies found in three allopatric populations of P. mugo from Tatra Mts. (mean genetic distance, Dn=.027). The differences were much larger in comparisons with monocormic pines from this area (Dn=.085) and two P. sylvestris samples from distant allopatric populations (Dn=.077). Nearly all monocormic pines had cpDNA species specific to P. sylvestris and isozyme frequencies similar to other populations of this species (mean Dn=.004). Only one P. sylvestris-like monocormic tree had cpDNA of P. mugo and can be considered as a hybrid. The results do not provide evidence supporting the hybrid swarm hypothesis. Rather, the results suggest that mature hybrids have a low frequency within this population and (rare) hybridisation is not reciprocal but unidirectional with P. mugo as pollen donor.     

Genetic Diversity and Genetic Structure of an Endangered Species, Trillium tschonoskii

The genetic diversity and genetic structure of Trillium tschonoskii (Maxim) were investigated using amplified fragment length polymorphism markers. Eight primer combinations were carried out on 105 different individuals sampled from seven populations. Of the 619 discernible DNA fragments generated, 169 (27.3%) were polymorphic. The percentage of polymorphic bands within populations ranged from 4.52 to 10.50. Genetic diversity (H_E) within populations ranged from 0.0130 to 0.0379, averaging 0.0536 at the species level. Genetic differentiation among populations was detected based on Nei's genetic diversity analysis (53.03%) and analysis of molecular variance (AMOVA) (52.43%). AMOVA indicated significant genetic differentiation among populations (52.43% of the variance) and within populations (47.57% of the variance) (p < 0.0002). Gene flow was low (0.4429) among populations. Species breeding system and limited gene flow among populations are plausible reasons for the high genetic differentiation observed for this species. We propose an appropriate strategy for conserving the genetic resources of T. tschonoskii in China.     

A comparison of the genetic diversity in Dipteronia sinensis Oliv. and Dipteronia dyeriana Henry

Dipteronia is an endemic genus to China and includes only two species, Dipteronia sinensis and D. dyeriana. Based on random amplified polymorphic DNA (RAPD) markers, a comparative study of the genetic diversity and genetic structure of Dipteronia was performed. In total, 128 and 103 loci were detected in 17 D. sinensis populations and 4 D. dyeriana populations, respectively, using 18 random primers. These results showed that the proportions of polymorphic loci for the two species were 92.97% and 81.55%, respectively, indicating that the genetic diversity of D. sinensis was higher than that of D. dyeriana. Analysis, based on similarity coefficients, Shannon diversity index and Nei gene diversity index, also confirmed this result. AMOVA analysis demonstrated that the genetic variation of D. sinensis within and among populations accounted for 56.89% and 43.11% of the total variation, respectively, and that of D. dyeriana was 57.86% and 42.14%, respectively. The Shannon diversity index and Nei gene diversity index showed similar results. The abovementioned characteristics indicated that the genetic diversity levels of these two species were extremely similar and that the interpopulational genetic differentiation within both species was relatively high. Analysis of the genetic distance among populations also supported this conclusion. Low levels of interpopulational gene flow within both species were believed to be among the leading causes for the above-mentioned phenomenon. The correlation analysis between genetic and geographical distances showed the existence of a remarkably significant correlation between the genetic distance and the longitudinal difference among populations of D. sinensis (p < 0.01), while no significant correlation was found between genetic and geographical distances among populations of D. dyeriana. This indicated that genetic distance was correlated with geographical distances on a large scale rather than on a small scale. This result may be related to differences in the selection pressure on species by their habitats with different distribution ranges. We suggest that in situ conservation efforts should focus on establishing more sites to protect the natural populations and their habitats. Ex situ conservation efforts should focus on enhancing the exchange of seeds and seedlings among populations to facilitate gene exchange and recombination, and to help conserve genetic diversity. __________ Translated from Acta Phytoecologica Sinica, 2005, 29(5): 785–792 [译自: 植物生态学报, 2005, 29(5): 785–792]     

Pollination ofNuphar (Nymphaeaceae) in Europe: Flies and bees rather thanDonacia beetles

Nuphar (Nymphaeaceae) comprises a small holarctic group of aquatic perennials whose flowers are pollinated by flies, bees, and beetles. We studied pollination in different populations of the two European speciesN. lutea andN. pumila in Norway and in Germany. Flowers are self-compatible and protogynous, preventing automatic selfing, and insect pollination is required for seed production. Sympatric populations were studied in Vest-Agder county in Norway to determine whetherN. lutea andN. pumila have the same or different pollinators. Allopatric populations ofN. lutea in Germany and Norway were then compared to determine whether their pollinator spectra differ as would be expected in an open flower with seemingly easily accessible pollen and nectar. Results of the present as well as previous studies of the pollination ofN. lutea andN. pumila show that both species are mainly pollinated by flies, including apparentNuphar specialists, such as the scatophagidHydromyza livens and the ephydridsHydrellia andNotiphila, the last also a long-known pollinator ofN. advena in Florida. Pollinator overlap between sympatric heterospecific populations was small, while allopatric conspecific populations had similar visitor and pollinator spectra. We found no evidence of pollination byDonacia beetles as reported from some North American populations ofNuphar. Dedicated to Univ.-Prof. DrF. Ehrendorfer on the occasion of his 70th birthday     

An AFLP analysis of genetic diversity and structure of Caragana microphylla populations in Inner Mongolia steppe, China

Genetic diversity and structure of five natural populations of Caragana microphylla from the Inner Mongolia steppe were estimated using AFLP markers. Five pairs of primers generated a total of 312 bands among 90 individuals, with percentage of polymorphic bands (PPB) being 63% at the population level and 76% at the species level, respectively. The genetic diversity within populations was correlated significantly with the soil N:P ratio. AMOVA analysis revealed high genetic variations within populations (95.5%). The estimated number of migrants per generation (Nm) was 10.72, indicating a high level of gene flow among populations. There was no significant correlation (r = 0.36) between genetic distance and geographical distance. These results were discussed in terms of eco-geographical variations among populations, together with the life history traits and breeding system of the species. The knowledge obtained may have important implications for better conservation and wise use of the vegetation dominate by C. microphylla.     

Genetic variation within the endangered mangrove species Sonneratia paracaseolaris (Sonneratiaceae) in China detected by inter-simple sequence repeats analysis

Sonneratia paracaseolaris, is a critically endangered mangrove species in China. Using inter-simple sequence repeats (ISSR) markers, we compared the genetic variation of introduced populations with that of natural populations to check whether the genetic diversity has been conserved. At the species level, genetic diversity was relatively high (P = 81.37%, He = 0.2241, and SI = 0.3501). Genetic variation in introduced populations (P = 75.78%, He = 0.2291, and SI = 0.3500) was more than that in natural populations (P = 70.81%, He = 0.1903, and SI = 0.2980). Based on Nei's G_ST value, more genetic differentiation among natural populations was detected (G_ST = 0.3591). Our data show that the genetic diversity of S. paracaseolaris was conserved in introduced populations to some extent, however, owing to the small natural populations and the threats they encountered, more plants should be planted to enlarge and restore the populations.     

Using phylogenetic, genetic and demographic evidence for setting conservation priorities for Mexican rare pines

Phylogenetic, genetic and demographic information are key issues for establishing priorities for conservation. In this study we explored the Vane-Wrigth measure of phylogenetic diversity for establishing criteria for conservation, when intra as well as interpopulation level demographic (population size) and genetic (heterozygosity, inbreeding and genetic distances among populations) parameters are taken into consideration. The data were standardized and analyzed independently and integratively enabling the calculation of indices or criteria for conservation priorities. We illustrate the application of this approach in populations of four Mexican pine species that have been considered rare and endangered, Pinus rzedowzkii, P. pinceana, P. lagunae and P. muricata. The application of several independent diversity indices did not allow us to resolve prioritization. However, the integration of all indices generated different values of importance to conservation, and suggested that the populations with the highest priorities for conservation are: two for Pinus rzedowskii, P. pinceana, and P. lagunae, and one for P. muricata. These populations have intermediate sizes, are the most divergent in the phylogeny and contain genetic variation that is representative of the gene pool of each species. Finally, we demonstrated the functionality of some genetic and demographic parameters; the genetic structure, recruitment rate, geographic distribution and demographic stochasticity, as complementary indicators for evaluating the conservation priority among populations and species.     

Population genetic structure of Ceriops tagal (Rhizophoraceae) in Thailand and China

Knowledge of the amount and patterns of genetic variation within and among populations of mangrove trees is essential for devising optimum genetic management strategies for their conservation and sustainable utilization. Ceriops tagal is a widespread viviparous mangrove. Genetic diversity in the species was examined with inter-simple sequence repeat (ISSR). Nine natural populations were collected from Thailand and China. The estimates of genetic variation were extremely low (H_T = 0.0179 ± 0.005, H_S = 0.0084 ± 0.001), and only 47% of the total gene diversity was maintained within populations (G_ST = 0.529). The eastern coastal populations of Thailand were more similar to populations from China than to populations from the western coastline of Thailand. A high level of Nei's genetic identity exists between populations of C. tagal (I = 0.989), suggesting their common ancestry. The low levels of genetic diversity in the species may result from a series of genetic bottlenecks during several glacial epochs.     

Correlations between the ages ofAlnus host species and the genetic diversity of associated endosymbioticFrankia strains from nodules

Nodule samples were collected from four alder species:Alnus nepalensis, A. sibirica, A. tinctoria andA. mandshurica growing in different environments on Gaoligong Mountains, Yunnan Province of Southwest China and on Changbai Mountains, Jilin Province of Northeast China. PCR-RFLP analysis of the IGS betweennifD andnifK genes was directly applied to unculturedFrankia strains in the nodules. A total of 21 restriction patterns were obtained. TheFrankia population in the nodules ofA. nepalensis had the highest genetic diversity among all fourFrankia populations; by contrast, the population in the nodules ofA. mandshurica had the lowest degree of divergence; the ones in the nodules ofA. sibirica andA. tinctoria were intermediate. A dendrogram, which was constructed based on the genetic distance between the restriction patterns, indicated thatFrankia strains fromA. sibirica andA. tinctoria had a close genetic relationship.Frankia strains fromA. nepalensis might be the ancestor ofFrankia strains infecting otherAlnus species. From these results and the inference of the ages ofAlnus host species, it is deduced that there was a co-evolution betweenAlnus and its microsymbiontFrankia in China.

     

Mating system and genetic differentiation inDasypyrum villosum (Poaceae) in Italy

Genetic differentiation at the morphological, isozyme, and DNA levels among sevenDasypyrum villosum (Poaceae, Triticeae, 2n = 14, VV genomes) populations from Italy was studied. A measure of the mating system was also obtained. Genetic diversity was mainly distributed within populations (90%) rather than among populations (10%), typical for most allogamous species. Interpopulation diversity, however, was greatest between the most geographically distant populations (about 750 km apart, 911 m altitude difference). The mating system was estimated to vary from 55 to 100% outcrossing. One population (I-16, Bomarzo) deviated from the others in its uniform early flowering habit and presence of isozyme alleles not found in other populations. It had genetic diversity similar to other populations for isozyme (Got-V2, Got-V3, andEst-VF) and ribosomal RNA (Nor-Vl) loci. This population is believed to be the product of a few migrant founder seeds and its unique characters point out that generalized population biology parameters of genetic diversity are not sufficient for describing species variation nor for developing conservation strategies.