云南不同土壤铅背景值下大叶茶种群对铅的吸收积累特征及其遗传分化

在具有不同土壤铅(Pb)背景值的云南大叶茶主产区,分析了11个大叶茶种群（简写为P1, P2, …… P11）所在地的土壤Pb含量,相应各种群的老、嫩叶Pb含量和富集系数,并利用ISSR分子标记研究了这些种群的遗传特征,以期认识不同大叶茶种群在不同Pb背景值下对Pb的吸收积累特征及其遗传分化状况。结果表明：（1）在本研究区域内,大叶茶种群间土壤Pb含量、老叶和嫩叶Pb含量、富集系数差异显著,土壤有效Pb、嫩叶Pb含量分别在0.78—15.20mg/kg和2.03—7.02 mg/kg之间,嫩叶Pb富集系数变化范围为0.001—0.24;种群内差异小,例如P6种群内嫩叶Pb含量在2.82—2.84 mg/kg之间,嫩叶Pb富集系数变幅为0.09—0.10。（2）筛选的10个ISSR引物扩增出81条带,平均多态位点百分率（PPB）为75.25%;Shannon’s指数（I）估算出种群间的变异为34.28%,利用POPGENE软件计算出种群间遗传分化系数GST为0.3116,分子方差分析（AMOVA）也显示种群间变异占35.37%（P<0.001）,表明不同种群的大叶茶出现了遗传分化。（3）UPGMA聚类分析发现,11个种群可分为5个类群,对Pb吸收累积能力高的与能力低的种群在聚类分析中存在明显分异;相关性分析表明,土壤有效Pb含量与PPB、I、Nei’s基因多样性指数（H）的 Pearson相关系数r分别为-0.633,-0.786,-0.581（P<0.05）,土壤有效Pb含量与大叶茶种群遗传多样性水平程度不同呈负相关。讨论分析认为,在土壤Pb高背景值条件下,部分大叶茶种群遗传多样性水平降低,不同种群对Pb的吸收累积能力存在明显差异,种群间出现了显著的遗传分化。对低铅富集的遗传分化现象的深入研究将可能为遴选拒吸收污染物的洁净种质、在污染条件下进行无公害生产提供新途径。     

六种阔叶树种叶片水浸提液对云南松种子萌发生长的化感作用

为了筛选营建云南松抗虫混交林的树种,该研究观察了云南切梢小蠹非寄主植物川滇桤木、缅桂、滇朴、樟树、麻栎和山茶六种阔叶树种叶片不同浓度的水浸提液对云南松种子萌发及幼苗生长的化感作用。结果表明:(1)川滇桤木、滇朴、麻栎和山茶四种阔叶树种叶片的水浸提液对云南松种子萌发和幼苗生长表现出低浓度促进,高浓度抑制效应。(2)在测试浓度范围内,樟树和缅桂叶片水浸提液对云南松种子萌发和幼苗生长存在抑制作用。综上所述,六种阔叶树种对云南松的化感作用敏感性不同,低浓度范围内的川滇桤木、滇朴、麻栎和山茶树叶片水浸提液对云南松生长表现出促进作用,樟树和缅桂叶片水浸提液对云南松生长具有抑制作用。因此,结合营林技术,可选用川滇桤木、滇朴、麻栎等阔叶树种与云南松混交,营建云南松抗虫混交林。     

秃房茶嘌呤生物碱组成特点及生化品质成分的研究

该研究采用分光光度法和高效液相色谱法,分不同叶位对秃房茶(Camellia gymnogyna)的嘌呤生物碱组成特点以及茶多酚、儿茶素组分、游离氨基酸、黄酮、茶氨酸等生化品质成分进行了测定。结果表明:秃房茶的嘌呤生物碱组成及配比显著区别于茶叶植物凤凰单从(C.sinensis),同时具有可可碱、咖啡碱和苦茶碱三种组分,而且可可碱含量最多,为13.46~39.72 mg·g~(-1),咖啡碱含量最低,为0.51~2.02 mg·g~(-1),苦茶碱含量介于两者中间并随芽叶成熟度增加而升高。茶叶植物只存在咖啡碱和可可碱,含量变化分别为22.22~53.13 mg·g~(-1)和0.47~12.82 mg·g~(-1)。在相同叶位中,秃房茶儿茶素组分含量变化规律为EGCGCECGEGCECGCCGGCG,且儿茶素总量、酯型儿茶素含量均低于茶叶植物,而非酯型儿茶素总量接近,保持为40~50 mg·g~(-1)。除黄酮含量在各叶位变化趋势不大外,其他品质成分含量变化基本符合第1叶芽第2叶第3叶第4叶,一芽二叶的含量介于第1叶和第2叶之间的规律,而茶多酚、黄酮、茶氨酸等其它品质成分含量均低于茶叶植物。该研究首次明确了秃房茶主要生化品质成分变化规律,特别是嘌呤生物碱的组成及配比特点,且含有特征性成分—苦茶碱。该研究结果为生物碱代谢机理、特异茶加工、功能成分开发、低咖啡碱资源、选育种等提供了优良材料。     

Molecular assessment of Ulva (Ulvales,Chlorophyta) diversity in Vietnam including the new species U. vietnamensis

Species diversity of Ulva in Vietnam was investigated using three commonly used genetic markers, the nuclear encoded rDNA ITS region and the plastid encoded rbcL and tufA genes. Single locus species delimitation methods, complemented with morphological and ecological information resulted in the delimitation of 19 species. This diversity is largely incongruent with the traditional understanding of Ulva diversity in Vietnam. Only four species identified in this study, U. lactuca, U. reticulata, U. spinulosa, and U. flexuosa, have been previously reported, and seven species, U. ohnoi, U. tepida, U. chaugulii, U. kraftiorum, U. meridionalis, U. limnetica, and U. aragoënsis, are recorded for the first time from Vietnam. Seven genetic clusters could not be associated with species names with certainty. A new species, U. vietnamensis, is described from marine to brackish coastal areas from southern Vietnam based on its morphological and molecular distinctiveness from the currently known Ulva species. A comparison with recent molecular-based studies of Ulva diversity showed that species composition in Vietnam is similar to that of adjacent countries, including Japan, China, as well as Australia. Our study emphasizes the importance of molecular data in the assessment of Ulva diversity, and indicates that a lot of diversity may still remain to be discovered, especially in tropical regions.     

Genetic diversity and geographic differentiation analysis of duckweed using inter-simple sequence repeat markers

Duckweed, with rapid growth rate and high starch content, is a new alternate feedstock for bioethanol production. The genetic diversity among 27 duckweed populations of seven species in genus Lemna and Spirodela from China and Vietnam was analyzed by ISSR-PCR. Eight ISSR primers generating a reproducible amplification banding pattern had been screened. 89 polymorphic bands were scored out of the 92 banding patterns of 16 Lemna populations, accounting for 96.74% of the polymorphism. 98 polymorphic bands of 11 Spirodela populations were scored out of 99 banding patterns, and the polymorphism was 98.43%. The genetic distance of Lemna varied from 0.127 to 0.784, and from 0.138 to 0.902 for Spirodela, which indicated a high level of genetic variation among the populations studied. The unweighted pair group method with arithmetic average (UPGMA) cluster analysis corresponded well with the genetic distance. Populations from Sichuan China grouped together and so did the populations from Vietnam, which illuminated populations collected from the same region clustered into one group. Especially, the only one population from Tibet was included in subgroup A2 alone. Clustering analysis indicated that the geographic differentiation of collected sites correlated closely with the genetic differentiation of duckweeds. The results suggested that geographic differentiation had great influence on genetic diversity of duckweed in China and Vietnam at the regional scale. This study provided primary guidelines for collection, conservation, characterization of duckweed resources for bioethanol production etc.     

The Genus Rhyacophila Pictet (Trichoptera: Rhyacophilidae) in Vietnam

Currently, 20 species of the genus Rhyacophila are known from Vietnam. Of these, 17 species are distributed among eight species groups, with three species unassigned. Examination of material collected by personnel of the Royal Ontario Museum in Toronto, Canada has revealed six new species and five new records of Rhyacophila from Vietnam, distributed among six species groups, four of which are new to Vietnam. Newly described species include: Rhyacophila smithi, Rh. curriei, Rh. laocai, Rh. meyi, Rh. olahi, and Rh. fernandi. Rhyacophila bidens Kimmins, Rh. inaequalis Denning and Schmid, Rh. lhabu Schmid, Rh. malayana Banks, and Rh. rhombica Martynov are recorded from Vietnam for the first time. The females of four species, Rh. bidens, Rh. immaculata Mey, Rh. malayana Banks, and Rh. tamdaona Oláh, are described for the first time. All 31 species of Rhyacophila now known from Vietnam are included in this work, organized by branch and species group.     

A comparison of genetic variation among wild and cultivated Morus Species (Moraceae: Morus) as revealed by ISSR and SSR markers

In this study, inter-simple sequence repeats (ISSR) ans simple sequence repeat (SSR) markers were used to investigate genetic diversity of 27 mulberry accessions including 19 cultivated accessions (six M. multicaulis, three M. alba, two M. atropurpurea, two M. bombycis, one M. australis, two M. rotundiloba, one M. alba var. pendula, one M. alba var. macrophylla, and one M. alba var. venose) and 8 wild accessions (two M. cathayana, two M. laevigata, two M. wittiorum, one M. nigra and one M. mongolica). ISSRs and SSRs were compared in terms of their informativeness and efficiency in a study of genetic diversity and relationships among 27 mulberry genotypes. SSRs presented a higher level of polymorphism and greater information content. All index values of genetic diversity both markers analyzed using Popgene 32 software indicated that within wild species had higher genetic diversity than within cultivated species. Cultivation may caused the lose of genetic diversity of mulberry compared with wild species revealed by ISSR and SSR markers. The mean genetic similarity coefficients among all mulberry genotypes ascribed by ISSR and SSR matrices were 0.7677 and 0.6131, respectively. For all markers a high similarity in dendrogram topologies was obtained although some differences were observed. Cluster analysis of ISSR and SSR using UPGMA method revealed that the wild species are genetically distant from the domesticated species studied here. The correlation coefficients of similarity were statistically significant for both marker systems used. Principal coordinates analysis (PCA) for ISSR and SSR data also supports their UPGMA clustering. These results have an important implication for mulberry germplasm characterization, improvement, molecular systematics and conservation.     

Inheritance and quantitative trait loci analyses of the anthocyanins and catechins of Camellia sinensis cultivar ‘Ziyan’ with dark-purple leaves

Owing to the potential health benefits, anthocyanin-rich teas (Camellia sinensis) have attracted interest over the past decade. Previously, we developed the cultivar ‘Ziyan,’ which has dark-purple leaves because of the accumulation of a high amount of anthocyanins. In this study, we performed a genetic analysis of this anthocyanin-rich tea cultivar and 176 of its naturally pollinated offspring. For two consecutive years, we quantified the anthocyanins and catechins of ‘Ziyan’ and the offspring population. While >60% of the offspring accumulated less than half of the amount of anthocyanins of ‘Ziyan,’ 17 (2018) and 15 (2019) individuals exceeded ‘Ziyan’ in anthocyanin content. A negative correlation between anthocyanin and total catechin content (r = −0.59, P < 0.001) was observed. The population was genotyped with 131 SSR markers spanning all linkage groups of the C. sinensis genome. Kruskal-Wallis tests identified 10 markers significantly associated with anthocyanins, catechins and their ratios in both years. Quantitative trait locus (QTL) analyses using the interval mapping method detected 13 QTLs, suggesting the dark-purple trait of ‘Ziyan’ is because of the pyramiding of anthocyanin-promoting alleles on at least five linkage groups. Two genetic loci reversely related to anthocyanin and total catechin contents were identified. This study provides valuable information for genetic improvement of purple tea cultivars and for fine-mapping related genes.     

DNA sequences of the mitochondrial COI gene have low levels of divergence among deep-sea octocorals (Cnidaria: Anthozoa)

We are analyzing genetic diversity in deep-seamount octocorals with the ultimate goal of studying the effect of retention and dispersal of larvae on genetic population structure. Here we report on the sequence diversity of the mitochondrial cytochrome oxidase I (COI) gene among 11 species. Uncorrected pairwise sequence divergences ranged from 0.4–10.3% for comparisons among species spanning the intrageneric to interordinal levels. Relative to other invertebrates, these divergences are very low, suggesting that COI may not be useful as a genetic marker for studying dispersal among deep-sea octocoral populations. Possible explanations for the reduced rates of divergence observed include a lower rate of evolution for octocoral mitochondrial genomes and the presence of a gene, mtMSH, which may code for a mitochondrial DNA mismatch-repair system. We report the finding of mtMSH in three deep-sea octocorals (Acanthogorgia sp., Corallium ducale, and Paramuricea sp.), which brings the total published observations of this gene to six species, all in the subclass Octocorallia.     

AFLP analysis of relationships among cassava and other Manihot species

 Despite the worldwide importance of cultivated cassava (M. esculenta Crantz) its origin and taxonomic relationships with other species in the genus have not been clearly established. We evaluated a representative sample of the crop’s diversity and six wild taxa with AFLPs to estimate genetic relationships within the genus. Groupings of accessions of each species by data analysis corresponded largely with their previous taxonomic classifications. A mixed group, consisting of Manihot esculenta subsp. flabellifolia and M. esculenta subsp. peruviana, was most similar to cassava, while M. aesculifolia, M. brachyloba, and M. carthaginensis were more distant. Species-specific markers, which may be useful in germ-plasm classification or introgression studies, were suggested by the unique presence of AFLP products in samples of each of the three wild species. Heterogeneity of similarities among individuals of certain species suggested the existence of intraspecific gene pools, a hypothesis that was supported by morphological or ecogeographic evidence with varying degrees of success. Quantitative assessment of genetic diversity revealed greater homogeneity among cassava accessions than among itsclosest wild relatives. The demonstration of unique genetic diversity in the two M. esculenta subspecies and their genetic similarity to the crop supports the hypothesis that these materials may be the ancestors of cassava. Received: 4 November 1996 / Accepted: 20 December 1996     

夏季南亚热带森林演替中后期优势种幼叶花色素苷的光保护作用

为探讨夏季南亚热带森林演替过程中优势树种幼叶的光保护机制,以演替中期优势树种木荷(Schima superba)、黧蒴(Castanopsis fissa)、锥栗(C.chinensis)和演替后期优势种华润楠(Machilus chinensis)、厚壳桂(Cryptocarya chinensis)、黄果厚壳桂(C.concinna)为材料,分析了2种生长光强(全光照和30%全光照)下6种优势种幼叶和成熟叶的叶片表型、光合色素含量、花色素苷含量、抗氧化能力、类黄酮含量、总酚含量和最大量子产量(Fv/Fm)恢复效率间的差异。结果表明,两个演替阶段幼叶的叶绿素含量(Chl a+b)、Chl a/b比成熟叶低,但光保护物质比成熟叶多;演替中期幼叶的花色素苷含量和总抗氧化能力比演替后期的高,而类黄酮和总酚含量比演替后期的低;全光照下幼叶的总酚、类黄酮、总抗氧化能力及Fv/Fm恢复效率都要比30%全光照的高,并且含有花色素苷的幼叶恢复得更快。因此,植物的光合能力与自身的光保护潜力成反比关系,演替中期优势种幼叶的光保护在很大程度上是因为花色素苷的积累而演替后期优势种是因为自身抗氧化物质(类黄酮、总酚)的共同作用。     

干旱胁迫对喀斯特地区野生茶树幼苗生理特性及根系生长的影响

以贵州喀斯特地区4种野生茶树(Sect Thea(L.)Dyer)无性系幼苗为材料,采用盆栽控水方法研究其对干旱胁迫的生理生长响应,初步评价其抗旱性,并采用田间模拟持续干旱试验进行验证。结果表明:(1)随着干旱胁迫程度的加强,4种野生茶树叶片的相对含水量逐渐降低;细胞质膜透性、丙二醛(MDA)含量和可溶性糖含量均呈上升趋势;游离脯氨酸含量和超氧化物歧化酶(SOD)活性先升后降;过氧化物酶(POD)活性除秃房茶随胁迫程度加强呈上升趋势外,其他种均呈先升后降趋势。(2)随着干旱胁迫程度的加强,4种茶树幼苗总生物量干重均逐渐下降,根冠比先升后降;根系总长除在秃房茶中呈先升后降趋势外,在其他3个种中均逐渐下降;根系总表面积、根系总体积在大理茶和茶中逐渐下降,而在大厂茶和秃房茶中先升后降;根系平均直径在秃房茶中逐渐降低,大厂茶中先升后降,茶中逐渐升高;根系活力在大理茶中逐渐下降,在其他3种中均先升后降;而比叶面积则在大厂茶中呈下降趋势,在其他3种中均先升后降。(3)基于生长生理指标的隶属函数抗旱性评价结果表明,MDA含量和细胞质膜透性与野生茶树抗旱性密切相关,4种野生茶树幼苗的抗旱顺序为秃房茶(Camellia gymnogyna Chang)大厂茶(Camellia tachangensis F.C.Zhang)茶(Camellia Sinensis(L.)-O.Kuntze)大理茶(Camellia taliensis(W.W.Smish)),田间持续干旱试验验证了基于隶属函数抗旱性评价方法的准确性和可靠性。     

基于SSR标记探讨三种金花茶植物的遗传多样性和遗传结构

薄叶金花茶、小花金花茶和小瓣金花茶是三种濒危金花茶植物,为了解珍稀濒危植物遗传多样性和遗传结构,该研究利用微卫星标记对他们的7个种群共184个个体进行了遗传多样性和遗传结构分析。结果表明:11个位点共检测到等位基因92个。在物种水平上,小瓣金花茶平均等位基因数(N_A)为3.9、有效等位基因数(N_E)为2.328、观测杂合度(H_o)为0.520、期望杂合度(H_e)为0.501,高于薄叶金花茶和小花金花茶。在种群水平上,有效等位基因数(N_E)在1.788～2.466之间,期望杂合度(H_e)在0.379～0.543之间;种群间遗传分化系数(FST)在0.143 7～0.453 3之间,种群间基因流(N_m)在0.301 5～1.488 9之间。AMOVA分子变异分析显示65.72%的变异存在于种群内。三种金花茶具有较低水平的遗传多样性和高水平的种群间遗传分化。STRUCTURE和PCoA种群遗传结构分析结果将取样种群分为2组,即薄叶金花茶和小花金花茶大部分个体分为一组,小瓣金花茶大部分个体分为一组。现存所有种群应根据实际情况尽快采取就地保护或迁地保护措施。     

Comparative study of the phylogenetic structure in six Apodemus species (Mammalia,Rodentia) inferred from ISSR-PCR data

The utility of the Inter Simple Sequence Repeat-Polymerase Chain Reaction (ISSR-PCR) was explored in order to determine genetic variation in six species of the genus Apodemus (A. flavicollis, A. sylvaticus, A. uralensis, A. agrarius, A. mystacinus and A. epimelas) at the individual level, population level, in separate geographic samples and in the species as a whole. Six optimized primers produced highly reproducible and polymorphic DNA markers with 98.3% polymorphic bands on a total sample of 91 individuals from 32 localities in Europe and Asia. Moreover, each primer allowed for an exact diagnosis of each of the six Apodemus species and thus provides a simple and reliable tool for the hitherto problematic discrimination of species from the subgenus Sylvaemus. Genetic distances between species ranged from 0.079, among the closely related A. flavicollis and A. sylvaticus, to 0.203 between A. mystacinus and A. agrarius. A. flavicollis, A. uralensis and A. sylvaticus display a strong population substructure. The range of genetic distances among geographic samples within last two species reaches the values obtained for closely related species. ISSR markers proved to be a simple and reliable tool for species diagnosis, as well as for estimating genetic diversity below the species level and for closely related species, but they showed questionable reliability for larger genetic distances.     

Higher β‐ and γ‐diversity at species and genetic levels in headwaters than in mid‐order streams in Hydropsyche (Trichoptera)

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Comparison of genetic variation and differentiation using microsatellite markers among three rare threatened and one widespread toad lily species of Tricyrtis section Flavae (Convallariaceae) in Japan

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Understanding the amount and distribution of genetic diversity in natural populations can inform the conservation strategy for the species in question. In this study, genetic variation at eight nuclear microsatellite loci was used to investigate genetic diversity and population structure of wild litchi (Litchi chinensis Sonn. subsp. chinensis). Totally 215 individuals were sampled, representing nine populations of wild litchi. All eight loci were polymorphic, with a total of 51 alleles. The expected heterozygosity in the nine populations ranged from 0.367 to 0.638 with an average value of 0.526. Inbreeding within wild litchi populations was indicated by a strong heterozygote defect. Significant bottleneck events were detected in the populations from Yunnan and Vietnam, which could be responsible for lower levels of genetic diversity in these populations. Measures of genetic differentiation (F _ST = 0.269) indicated strong differentiation among wild litchi populations. Significant correlation was found between genetic differentiation and geographical distance (r = 0.655, P = 0.002), indicating a strong isolation by distance in these populations. Bayesian clustering suggested genetic separation among three regional groups, namely, the western group, the central group and the eastern group. Some conservation strategies for wild litchi populations were also proposed based on our results.     

An AFLP-based survey of genetic diversity and relationships of major farmed cultivars and geographically isolated wild populations of <Emphasis Type="Italic">Saccharina japonica</Emphasis> (Phaeophyta) along the northwest coasts of the Pacific

The genetic diversity and relationships of six representative cultivars and six geographically isolated wild populations of Saccharina japonica along the northwest coasts of the Pacific Ocean were investigated using AFLP markers. A total of 547 bands were generated across all samples by ten primer combinations. At the cultivar or population level, the percentage of polymorphic loci (P), gene diversity (H), and Shannon’s information index (I) was highest in Dalian population (P 59.05%; H 0.2057; I 0.3062) and lowest in Lianjiang cultivar (P 9.87%; H 0.0331; I 0.0497). At the species level, P, H, and I were 85.01%, 0.1948, and 0.3096, respectively. Unique bands were detected in all the six wild populations, with Dalian being the most. In comparison, only Yanza cultivar possessed one unique band. The G _ST value was 0.6226 and the gene flow (N _m ) was 0.1515, indicating strong genetic differentiation among cultivars and populations. Two UMPGA dendrograms were constructed based on the Dice similarity coefficients among individuals and on genetic distances among cultivars and populations, which generally revealed three major clades corresponding to three countries. Analysis of molecular variance revealed that a larger proportion (60.21%) of the total genetic variation was attributable to differences among cultivars and populations. The Mantel test suggested that genetic differentiation was positively correlated with geographic distance (r = 0.7962, P = 0.011) in the six wild populations, agreeing with the isolation by distance model. On the whole, low to moderate genetic diversity within cultivars and populations (except Dalian population) and high genetic differentiation among cultivars and populations were detected.     

Revisiting protein heterozygosity in plants��nucleotide diversity in allozyme coding genes of conifer Pinus sylvestris

Allozyme variation has been and continues to be a major source of information on the level of genetic variation among plant species. Deciphering the molecular basis of electrophoretic variation is essential for understanding the forces affecting the protein level variation. In this study, the relationship between allozyme heterozygosity and nucleotide diversity in plants is investigated among and within species. Allozyme and nucleotide diversity in 27 plant species was reviewed. At the multilocus level, the two methods are congruent: a clear correlation between the two measures of genetic diversity among plant species was observed, strengthening the view that effective population size is the major determinant of genome-wide diversity. Nucleotide diversity at six allozyme coding genes (6pgdB, aco, gdh, gotC, mdhA, and mdhB) in conifer Pinus sylvestris was investigated jointly with electrophoretic data. Single non-synonymous charge-changing mutations were found together with electrophoretic alleles that consequently were mutationally unique. Synonymous site nucleotide diversity (point estimate of θ _W—0.009 per bp) and silent site divergence from Pinus pinaster at allozyme coding loci were at comparable levels with other loci in the species. Linkage disequilibrium was extensive compared to earlier estimates from P. sylvestris and other trees, spanning several kilobases. Allozyme coding genes had an excess of closely related haplotypes whose frequency has recently increased possibly as a result of partial selective sweeps or balancing selection, but complex demographic effects cannot be excluded.     

Population structure,gene diversity,and differentiation in natural populations of Cedar pines (Pinus subsect.Cembrae,Pinaceae) in the USSR

Twelve natural populations of four cedar pine species,Pinus sibirica, P. cembra, P. pumila, andP. koraiensis, occurring in the Soviet Union were investigated by starch-gel electrophoresis. Frequencies of 55 alleles at 19 loci were determined. Interpopulation genetic diversity inP. sibirica andP. pumila was only 2–4 per cent of the total genetic diversity. Nei's distance coefficient (Dn) was used to estimate the level of genetic differentiation among conspecific populations and among species. Dn values among populations ranged from 0.006 to 0.038. A dendrogram constructed using Dn values divided cedar pines species into 2 clusters:sibirica-cembra (Dn = 0.030) andpumila-koraiensis (Dn = 0.143). Nei's distance between these clusters was 0.232. On the basis of the data obtained it was possible to draw the following conclusion:P. sibirica, P. pumila, andP. koraiensis are distinct species, whileP. cembra should apparently be regarded as geographicalP. sibirica race.