Genetic signals of ancient decline in Aleppo pine populations at the species' southwestern margins in the Mediterranean Basin

Microsatellite markers were used to characterize the structure of genetic diversity in natural Moroccan Aleppo pine (Pinus halepensis Mill.) populations, the most southwesterly marginal populations of the species in the Mediterranean Basin. Twenty-two natural populations and one artificial population, located in four regions covering most of the natural range of P. halepensis in the country, were sampled. Across this range, towards the south and west (and towards high altitudes) the populations become increasingly discrete and discontinuous. The nuclear microsatellite marker analysis suggests that a large proportion of the Aleppo pines in Morocco have derived from a single genetic lineage, represented by a central group of 11 of the examined populations located in the High and Middle Atlas Mountains. In addition, two smaller groups, represented by the marginal southwestern High Atlas populations, and three still smaller north / northeastern groups of populations located in the Rif and northeast Middle Atlas Mountains, could be genetically distinguished. Further, coalescence analysis of historical demographic population patterns suggests that ancient bottlenecks occurred in all of the natural populations. However, the population differentiation and genetic diversity levels we found were good (F(st) =15.47), presumably because of the species' good potential for long-distance dispersal of seeds and high invasive capacity, which appear to have maintained a state of stable near-equilibrium, meta-population dynamics since ancient times.     

Microsatellite markers reveal high genetic diversity in date palm (<Emphasis Type="Italic">Phoenix dactylifera</Emphasis> L.) germplasm from Sudan

Genetic diversity in date palm germplasm from Sudan representing 37 female and 23 male accessions was investigated using 16 loci of microsatellite (SSR) primers. Eight female accessions from Morocco were included as reference material. The tested SSR markers showed a high level of polymorphism. A total of 343 alleles were detected at the 16 loci. The number of alleles per marker ranged from 14 to 44 with an average of 21.4 per locus. A high level of expected heterozygosity was observed among Sudan cultivars (0.841), Morocco cultivars (0.820) and male accessions (0.799). The results indicate that the genetic groups of the Sudan cultivars and/or males do not follow a clear geographic pattern. However, the morocco group showed significant differentiation in relation to the Sudan groups, as measured by F (ST) values and genetic distances. The effect of the methods of pollination and cultivar selection on the genetic structure was clearly detected by the weak clustering association that was observed for the majority of accessions originating from Sudan and Morocco as well. This suggests the need for further investigation on the genetic diversity of Sudanese date palm germplasm. A deeper insight will be revealed by a detailed analysis of populations originating from different geographic locations.     

Rangewide Genetic Diversity in Natural Populations of Chinese Pine (Pinus tabulaeformis)

Thirteen natural populations from throughout the range of the Chinese pine (Pinus tabulaeformis Carr.) were examined using inter-simple sequence repeat markers to characterize the genetic structure at the species level and to compare the extent and distribution of genetic variation among central, intermediate, and marginal populations. Although the total genetic variation in the Chinese pine was mainly maintained within populations, the genetic differentiation among populations was significant (P < 0.001). The genetic divergence was significantly correlated with geographic distance (P < 0.05). Genetic diversity tended to decrease from the central to intermediate and marginal populations. The marginal populations had significantly lower intrapopulation genetic diversity than central populations (P < 0.05). Cluster analysis based on Nei’s unbiased genetic distances confirmed the difference among four central populations and the rest. Both historical and contemporary factors may have played key roles in shaping the spatial genetic structure of this species.     

Phylogeography of North African Atlas cedar (Cedrus atlantica, Pinaceae): Combined molecular and fossil data reveal a complex Quaternary history

Northwest Africa is a major hotspot of plant biodiversity, but very little is known about the Quaternary range dynamics of plant species in this region. Here we investigate the range-wide population structure and phylogeography of Atlas cedar (Cedrus atlantica), an emblematic forest tree endemic to Morocco and Algeria. We genotyped 261 individuals from 11 populations using AFLP markers. Data were analyzed using both conventional F(ST)-based techniques and Bayesian clustering. Overall population differentiation was high (F(ST) = 0.25). Two major groups of populations were identified, one distributed through the Rif and Middle Atlas mountains in Morocco and the other through the Algerian Tell Atlas and Aurès mountains as well as the Middle Atlas. Combined molecular and fossil data indicate that C. atlantica survived the Last Glacial Maximum in at least three disjunct refugia along the coast of the Mediterranean Sea, whereas the Middle Atlas, today the core of the species range, has been colonized relatively recently (<10000 yr BP). The colonization history of individual populations has left clear imprints in their present-day diversity, which may vary greatly even between nearby stands. Our study illustrates how integrating different data sources and analytical approaches can help elucidate complex range dynamics that would otherwise remain undeciphered.     

Nuclear genetic variation of <Emphasis Type="Italic">Rosa odorata</Emphasis> var. <Emphasis Type="Italic">gigantea</Emphasis> (Rosaceae): population structure and conservation implications

Rosa odorata var. gigantea is one of the most important ancestors of modern roses, which owns many merit traits including large flower, early flowering, and tea scent. Due to habitat loss and fragmentation, it has been listed as a rare and endangered species in China. In this study, a total of 424 accessions from 27 locations across its major distribution range were sampled. Its genetic diversity and population structure were assessed using a combination of seven nuclear microsatellite markers and one single-copy nuclear gene. Moderate to high within-population genetic diversity and moderate differentiation among populations were revealed despite its narrow distribution. The sampled 27 populations were resolved into two genetic clusters with limited contemporary and historical gene flows. The Red River Fault Zone was inferred to be a physical or geographical barrier to gene flow between these two genetic clusters. Genetic distances were significantly associated with geographic distances, indicating the isolation-by-distance model. Our ecological niche modeling indicated that R. odorata var. gigantea had high current potential areas in the central Yunnan province and substantial losses of high potential distribution areas in the western Yunnan in the future. Two detected clusters showed significant genetic differentiation and represented two separate evolutionary lineages, which should be recognized as two evolutionary significant units (ESUs) for conservation concerns. These results could be applied for the decision-making and conservation planning for this important species.     

Seasonal cold hardiness in maritime pine assessed by different methods

Three screening methods—visual scoring (V), relative conductivity (C) and fluorometry (F)—were used to study the genetic variation in cold hardiness among six populations of maritime pine (Pinus pinaster Ait.) comprising both Atlantic and Mediterranean origins. Freezing damage assessments were carried out in three organs—needles, stems and buds—in two seasons, spring and autumn. We found high levels of genetic differentiation among populations for cold hardiness in autumn, but not in spring. Within populations, differences were always significant (p?<?0.05) no matter which organ or screening method was used. Measuring F was the fastest and most easily replicated method to estimate cold hardiness and was as reliable as V and C for predicting the species performance. In autumn, there was a positive correlation between the damage measured in all three types of organs assessed, whereas in spring, correlation among organs was weak. We conclude that sampling date in spring has a crucial impact to detect genetic differences in maritime pine populations, whereas autumn sampling allows more stable comparisons. We also conclude that the fluorometry method provides a more efficient and stable comparison of cold hardiness in maritime pine.     

Allozyme Variation in Turkmenian Populations of Wild Barley, Hordeum spontaneum Koch.

The extent and structure of genetic variation in 720 individualsrepresenting 36 populations of wild barley, Hordeum spontaneum,from Central Asia (Turkmenistan) were determined using starchgel electrophoresis of eight water soluble leaf proteins encodedby 13 loci. The populations were grouped into seven ecogeographicregions. The study found: (a) a similar amount of within populationgenetic diversity (H_e = 0.106), but lower total genetic diversity(H_T = 0.166) to that reported for Middle East populations ofH. spontaneum; (b) of the total genetic diversity, 61% was attributableto variation within populations, 27% between populations ofa region, and 12% among regions; (c) of the 42 alleles found,11 were ubiquitous, 22 were widespread and common, three localand common and seven local and rare; (d) there was a poor correlationbetween population genetic and geographic distances; and (e)the frequencies of only a few alleles correlated significantlywith climatic or geographic parameters. The extent and structureof genetic variation of Turkmenian populations, which representthe Central Asian part of the species' range, were significantlydifferent in some important aspects from Middle Eastern andeastern Mediterranean populations. The mosaic pattern of geneticvariation found in wild barley in the Middle East is less pronouncedin populations from Central Asia where there is less geneticdifferentiation among populations and regions, and more ubiquitousor common and fewer localized alleles. Copyright 2001 Annalsof Botany Company Allozyme, Central Asia, genetic diversity, Hordeum spontaneum, wild barley     

Analyses of genetic population structure of two ecologically important mangrove tree species, Bruguiera gymnorrhiza and Kandelia obovata from different river basins of Iriomote Island of the Ryukyu Archipelago, Japan

We analyzed nuclear and chloroplast microsatellite makers to assess genetic diversity and examine genetic structure of two mangrove tree species, Bruguiera gymnorrhiza and Kandelia obovata recovered from nine major river basins of Iriomote Island of the Ryukyu Archipelago, Japan. The average number of alleles per nuclear locus per population was 2.6 in B. gymnorrhiza and 1.7 in K. obovata. Bayesian clustering analysis using InStruct identified two genetic clusters in B. gymnorrhiza and three clusters in K. obovata. Chloroplast microsatellites revealed two dominant haplotypes from B. gymnorrhiza and three haplotypes from K. obovata. The overall result suggests low genetic diversity for both species. AMOVA for nuclear microsatellites showed 9.3?% of population variation in B. gymnorrhiza. Although genetic differentiation between several populations was significant in this species, F _ST suggested low to moderate level of differentiations between most of the populations. Distribution of genetic clusters and chloroplast haplotypes also suggested weak differentiations among B. gymnorrhiza populations. In K. obovata, population variation was, however, relatively high (27.8?%). The high differentiation between K. obovata populations was also suggested from the F _ST and genetic clusters from nuclear microsatellites, and chloroplast haplotypes. A significant correlation between chloroplast genetic distances and coastline distances as well as haplotype distributions for both species suggest that propagules from each species mostly disperse to the neighboring river basins. While significant F _IS and higher percentage of admixed clusters from nuclear microsatellites suggested inbreeding, continual gene exchange by propagule dispersal among populations especially among neighboring populations was suggested for both species from maternally inherited chloroplast microsatellites analyses.     

Variation in chloroplast single-sequence repeats in Portuguese maritime pine (Pinus pinaster Ait.)

Genetic variation in 12 Pinus pinaster (maritime pine) populations spanning most of the distribution range of the species in Portugal was evaluated using six polymorphic chloroplast microsatellite (cpSSR) loci. Thirty-two haplotypes were found. There were indications of very weak differentiation among populations (Weir’s θcoefficient, 0.023), and the R _ST value, derived from the stepwise mutation model (SMM), was not significantly different from zero. The pattern, in which similarities in allele size, in base pairs, do not contribute to the genetic structure, may be due to the recent mixing of genetic material from different stands through plantations. Overall, a high level of haplotypic variation within populations was detected. Using the SMM estimator (mean genetic distance of individuals within populations, D ² _sh –––) we divided the populations into two groups, with above and below average values. The first group contained 5 populations, mainly from the central part of the country, which possess, in general, high levels of haplotypic diversity. Among them, 2 populations were divergent from the others based on the pair-wise Nei’s distance. The results indicate that there is no discernible geographic genetic pattern for the Portuguese populations of P. pinaster investigated. The history of expansion of the species range in Portugal during the twentieth century (mainly due to human activity) and extensive gene flow among populations associated with the expansion could explain this finding. Received: 15 February 2000 / Accepted: 14 April 2000     

Variability of nuclear microsatellite loci in the populations of Siberian dwarf pine (<Emphasis Type="Italic">Pinus pumila</Emphasis> (Pallas) Regel) from the Russian part of the range

Variability of nuclear microsatellite loci was examined in Siberian dwarf pine. Six microsatellite loci (RPS2, RPS6, RPS12, RPS124, RPS127, Pc18) demonstrated different polymorphism levels in ten populations of Siberian dwarf pine. The average number of alleles per locus was 4.88, the average observed heterozygosity was 0.465, and the average expected heterozygosity was 0.510. About 13% of total genetic variability was explained by the genetic differences between the populations (F _ST = 0.129). Genetic distances between the examined populations of Pinus pumila inferred from the data on the SSR marker frequencies statistically significantly correlated with the geographical distances between the population samples. The level of genetic variability of the populations from Kamchatka Peninsula was lower than that demonstrated by continental and island populations. The genetic differentiation of the Kamchatka–Magadan and other populations of Siberian dwarf pine observed in our study can be explained in terms of their formation from different Pleistocene refugial centers.     

Genetic diversity and population structure in natural populations of Moroccan Atlas cedar (Cedrus atlantica; Pinaceae) determined with cpSSR markers

Atlas cedar (Cedrus atlantica) is an ecologically and economically important forest tree species of northern Africa and is considered one of the endangered conifer species in the region. Chloroplast microsatellites (cpSSR) were used to study genetic variation within and among populations and geographical structure in natural populations of C. atlantica throughout its entire distribution range in Morocco. A total of 25 chloroplast haplotypes and 66 cpSSR alleles were found among 162 individuals. The cpSSRs indicate that C. atlantica appears to maintain a high level of genetic diversity (mean H(e) = 0.95), as observed in most coniferous species. Values of mean pairwise distance within a population (D(2)(SH)) were related to the size and location of the populations. AMOVA analysis showed that most of the variation in C. atlantica occurs within populations and confirmed the general tendency of gymnosperms to display lower values of population differentiation than angiosperms. The distance-based clustering method (PCoA and neighbor-joining analysis) and the geographical structure revealed a poor structure among the six populations of Cedrus atlantica. Also, a Mantel test indicated a weak correlation between geographic and genetic distances (P = 0.106, r = 0.363). These results are also interpreted in the context of postglacial history of the region plus human impacts.     

Evolutionary History of the Morocco lizard-Fingered Geckos of the <Emphasis Type="Italic">Saurodactylus brosseti</Emphasis> Complex

Studies of biodiversity in the Maghreb have revealed high genetic diversity and divergent genetic lineages among many taxa including squamates. Geographic barriers such as the Atlas Mountains are one of the key factors promoting genetic differentiation and the high levels of endemism. The lizard-fingered gecko Saurodactylus brosseti is endemic to Morocco. Its range includes both sides of the Atlas Mountains, and although high diversity was previously identified within the species, much of the range was unsampled. To understand the evolutionary and biogeographical history of this species, we used mitochondrial and nuclear DNA sequence data from 64 populations sampled across the entire species range. We employed phylogenetic methods based on gene trees and species trees as well as a time calibrated Bayesian genealogy and coalescent species delimitation approaches. We uncovered four highly divergent and allopatric mitochondrial lineages that did not share haplotypes at variable nuclear loci, suggesting the four groups have been evolving independently since the Miocene, according to our molecular dating estimates. These results coupled with the geographic pattern of genetic diversity suggest a possible role of the Atlas Mountains for the divergence observed between the four lineages of S. brosseti, while each lineage probably later underwent several episodes of fragmentation followed by (re-) expansion during Pleistocene climatic oscillations. Bayesian species delimitation analysis indicates that the four lineages may well be distinct species but we suggest that detailed morphological analyses are needed prior to taxonomic changes. The four lineages represent ancient independent evolutionary units, and deserve conservation management as distinct entities.     

Low genetic diversity and limited gene flow in a dominant mangrove tree species (Rhizophora stylosa) at its northern biogeographical limit across the chain of three Sakishima islands of the Japanese archipelago as revealed by chloroplast and nuclear SSR analysis

We examined the genetic diversity, population structure and gene flow in a dominant mangrove tree (Rhizophora stylosa) at its northern biogeographical limit in Sakishima islands of the Japanese archipelago. Simple sequence repeat (SSR) markers from chloroplast (cpSSR) and nuclear DNA were used to analyze 16 populations recovered from various river basins across the chain of three Sakishima islands—Iriomote, Ishigaki and Miyako. The average number of alleles (1.7–2.7) and observed heterozygosities (0.031–0.216) at nuclear SSR and haploid diversity (0.000–0.489) at cpSSR across the populations suggested low genetic diversity in R. stylosa in Sakishima islands. cpSSR analysis identified two haplotypes, and Bayesian clustering analysis (nuclear SSR) revealed two genetic clusters. Analysis of molecular variance (nuclear SSR) showed significant population differentiations. Pairwise tests consistently revealed significant differentiation between most of the population pairs; however, the degrees of differentiations are generally correspondent to the relative geographical distances as suggested from pairwise F _ST and cpSSR genetic distances. Moreover, Mantel tests showed some signals of correlations between genetic distances (nuclear and chloroplast) and geographical distances. These results suggest that combined contribution of gene flow via pollen and propagule dispersal in R. stylosa mostly occurred between neighboring river basins. The appearances of two cpSSR haplotypes (maternal lineages) as well as two nuclear genetic clusters (putative ancestral lineages) at various river basins support the hypothesis that present-day R. stylosa populations across the Sakishima islands were established from few identical founders; however, significant differentiations among various river basins most likely resulted from the limited gene flow and high inbreeding.     

Determination of Genetic Diversity of Vitis vinifera cv. Kabarcik Populations from the Coruh Valley Using SSR Markers

Northeastern Turkey is recognized as one of the most important germplasm centers for the grape in the world. In the present study, simple sequence repeat markers were used to investigate the genetic diversity between four Vitis vinifera cv. Kabarcik populations sampled from the Coruh Valley in Turkey, at altitudes of 800-1,150 m. The mean observed number of alleles per locus varied from 2 (loci VVMD7 and VVMD24) to 6 (VVS2) among populations. The population from the highest altitude showed the greatest average number of alleles, 4.5. With regard to the six loci examined in all populations, the mean observed heterozygosity was higher than the expected heterozygosity. Among the loci, VVS2 (probability of identity = 0.137) was found to be the most informative among populations. Genetic distances between populations ranged from 0.072 to 0.216. Genetic differentiation among populations was strongly related to geographic distances in all populations.     

Genetic diversity and population structure of the endangered medicinal plant Phellodendron amurense in China revealed by SSR markers

Phellodendron amurense is an endangered tree with important medicinal and economic value in China. In this study, eight nuclear SSR primer pairs were employed to assess the genetic diversity and structure of 22 natural populations, including 516 individuals. A total of 66 alleles were detected with an average of 8.3 alleles per locus ranging from 3 to 17. The expected heterozygosity (He) of each SSR locus varied from 0.347 to 0.877 (average 0.627). Analysis of molecular variance (AMOVA) revealed that the main variation component existed within populations (95.11%) rather than among populations (4.89%). The Wilcoxon's sign-rank tests did not show any recent bottleneck effect in any population. A Mantel test displayed a significant correlation between the geographic distances and genetic distances for all populations (r = 0.566, P = 0.0001), indicating conformity to the isolation by distance model. Bayesian clustering and UPGMA supported grouping the populations into two groups. The present genetic structure of P. amurense may be explained by geographical isolation. The lack of genetic structure and genetic diversity decreased with increasing latitude within the Northeast China group may be due to postglacial northward expansion from a single refugium. Proper conservation measures are proposed for this species.     

Phylogeographic patterns in Hypochaeris section Hypochaeris (Asteraceae, Lactuceae) of the western Mediterranean

Aim  To analyse phylogeographic patterns in the four species of Hypochaeris sect. Hypochaeris , evaluating possible areas of origin and the microevolutionary processes that have shaped their morphology, genetics and distribution.
Location  Western Mediterranean area.
Methods  We applied amplified fragment length polymorphism (AFLP) markers to a total of 494 individuals belonging to 82 populations of Hypochaeris arachnoidea , H. glabra , H. radicata and H. salzmanniana to determine population structure.
Results  Populations with the largest proportion of private and rare AFLP fragments were found in Morocco. This region was consequently inferred to be the ancestral area for H. arachnoidea , H. glabra , H. radicata and H. salzmanniana . The Guadalquivir River (southern Spain) was inferred to be an effective dispersal barrier for H. glabra and H. radicata. The Strait of Gibraltar was inferred to be a somewhat weaker barrier than the Guadalquivir River for H. radicata and a much weaker barrier for H. glabra . The main barrier for H. salzmanniana coincides with the extension of the Rif Mountains to the Atlantic coast in Morocco, and the Strait of Gibraltar is a much weaker barrier for this species. Hypochaeris arachnoidea appears to have originated in the Atlas Mountains.
Main conclusions  The highest levels of genetic variation in La Mamora forest ( H. glabra and H. salzmanniana ) or the adjacent central Middle Atlas ( H. arachnoidea and H. radicata ) in Morocco suggest that these areas were a centre of origin of Hypochaeris sect. Hypochaeris . All three potential barriers – the Guadalquivir River, the Strait of Gibraltar, and the Rif Mountains – have been important in shaping genetic diversity in species of section Hypochaeris .     

Genetic variation and differentiation in Juniperus excelsa M. Bieb. populations in Turkey

Genetic variation within and among six populations of Juniperus excelsa M. Bieb., in a common garden in Lakes District of Turkey, was analyzed using four nuclear microsatellite primer pairs originally developed for J. communis. A total of five loci were observed as Jc037 amplified two distinct size ranges. The number of alleles observed for the species varied from 2 to 13, with an average of 4.5 alleles per locus. The mean expected heterozygosity (H _e) of populations was 0.584, after correction for null alleles. The mean F _IS value (?0.014) was close to zero showing no significant deviation from Hardy–Weinberg equilibrium. A low level of genetic differentiation was observed among populations (F _ST = 0.028; p < 0.001) and Nei’s genetic distance ranged from 0.014 to 0.120 between population pairs. Furthermore, there was no significant correlation between genetic distances on the one hand and geographic distances and trait differentiation on the other hand. However, the eastern populations Bey?ehir and Sorgun showed very similar genotypic structures and were differentiated from all other populations. A continuous monitoring of phenotypic traits and the association between nucleotide variation in functional genes and adaptive traits such as drought and frost tolerance of J. excelsa populations in common gardens will be useful to design effective conservation strategies in the future.     

Assessing the allozyme variation in cultivars and Swedish landraces of rye (Secale cereale L.)

Genetic interpretation and diversity of 9 isozyme loci have been estimated in 7 improved varieties and 19 landraces from Sweden by means of starch gel electrophoresis. The isozyme systems were ACO, DIA, GPI, MDH, PGD and PGM. For the statistic analysis we used the following measures: average number of alleles per locus, percentage of polymorphic loci, average heterozygosity direct count and average heterozygosity Hardy-Weinberg expected unbiased estimate. The measures were made on species and population levels. The distribution of the total genetic diversity among populations was also calculated. To illustrate the genetic relationships among populations, genetic distances were measured and principal component analysis performed. As expected in a cross-pollinated crop we found high genetic diversity and a larger variation within than among the populations. Somewhat unexpectedly, however, we found that the currently used varieties have the same high level of heterozygosity as the landraces but in the dendrogram the two groups are separated. The dendrogram showed three main clusters. The large cluster included 21 populations and the two small clusters were clearly distinguishable from the rest. The landrace spring-type could not be separated from the landraces winter-type, but we did detect a difference between different spring types. A few populations had unique alleles for certain loci.     

Invasion genetics of <Emphasis Type="Italic">Senecio vulgaris</Emphasis>: loss of genetic diversity characterizes the invasion of a selfing annual,despite multiple introductions

Genetic variation in invasive populations is affected by a variety of processes including stochastic forces, multiple introductions, population dynamics and mating system. Here, we compare genetic diversity between native and invasive populations of the selfing, annual plant Senecio vulgaris to infer the relative importance of genetic bottlenecks, multiple introductions, post-introduction genetic drift and gene flow to genetic diversity in invasive populations. We scored multilocus genotypes at eight microsatellite loci from nine native European and 19 Chinese introduced populations and compared heterozygosity and number of alleles between continents. We inferred possible source populations for introduced populations by performing assignment analyses and evaluated the relative contributions of gene flow and genetic drift to genetic diversity based on correlations of pairwise genetic and geographic distance. Genetic diversity within Chinese populations was significantly reduced compared to European populations indicating genetic bottlenecks accompanying invasion. Assignment tests provided support for multiple introductions with populations from Central China and southwestern China descended from genotypes matching those from Switzerland and the UK, respectively. Genetic differentiation among populations in China and Europe was not correlated with geographic distance. However, European populations exhibited less variation in the relation between G _ST and geographical distance than populations in China. These results suggest that gene flow probably plays a more significant role in structuring genetic diversity in native populations, whereas genetic drift appears to predominate in introduced populations. High rates of selfing in Chinese populations may restrict opportunities for pollen-mediated gene flow. Repeated colonization-extinction cycles associated with ongoing invasion is likely to maintain low genetic diversity in Chinese populations.     

四种松毛虫不同地理种群遗传多样性的等位酶分析

【目的】采用等位酶电泳技术对中国松毛虫属Dendrolimus 4种共9个地理种群进行遗传多样性和遗传分化研究。【方法】对6种等位酶系统乳酸脱氢酶（LDH）、苹果酸脱氢酶（MDH）、苹果酸酶（ME）、乙醇脱氢酶（ADH）、甲酸脱氢酶（FDH）、谷氨酸脱氢酶（GDH）进行聚丙烯酰胺凝胶电泳分析。【结果】在4种松毛虫9个地理居群中共检测到10个基因位点,其中4个位点为多态位点,检测到17个等位基因; 种群总体水平多态位点比率P=40%,平均有效基因数A=1.700,平均期望杂合度He=0.151,种群平均遗传距离为0.001～0.285; 其中马尾松毛虫指名亚种Dendrolimu punctatus Walker 6个居群的遗传分化度Fst=0.265,基因流Nm=0.692。4种松毛虫之间遗传关系最近的是落叶松毛虫D. superans Butler和马尾松毛虫的地理亚种赤松毛虫D. punctatus spectabilis Butler,遗传关系最远的是落叶松毛虫D. superans Butler和云南松毛虫D. houi Lajonquiere。【结论】 马尾松毛虫居群间遗传分化程度较大,基因交流较少,遗传漂变已经成为导致该物种种群分化的主要原因之一;遗传距离与地理距离存在一定相关性。