半夏居群遗传分化的同工酶和可溶性蛋白电泳的聚类分析

用聚丙烯酰胺凝胶电泳（PAGE）技术分析了安徽产7个半夏（PinelliaternataBreit）居群的4种同工酶和可溶性蛋白谱带。根据其中的PER、EST和可溶性蛋白谱带的分布特征,用Forguson和Aquado＆Avise的公式分别计算其谱带的相似性系数和遗传距离,以类平均法（UPGMA）进行了聚类分析。结果表明：半夏居群间表现出一定的遗传分化;鹞落坪半夏与半夏的黄山、九华山、大别山万佛寺居群间的遗传距离较小（D≤0.214）,结合形态变异,笔者认为鹞落坪半夏只能是半夏居群中形态上较为特殊的地方宗,而不是一新种;狭叶半夏与其它6个居群的遗传距离较大（D=0.416）,是半夏居群中种系分化较为强烈的一个居群。文中还讨论了半夏的遗传分化与形态变异、生境等的相关性。     

中国半夏属植物亲缘关系的SRAP分析

利用序列相关扩增多态性(SRAP)标记对中国半夏属植物5个种的亲缘关系进行了研究.38个引物组合在半夏属植物的5个种中共扩增出752条清晰的谱带,其中628条谱带具有多态性,多态性比率为83.51％,显示出较高的多态性比率;各物种间的遗传相似系数在0.6513～0.7312之间,聚类分析和主坐标分析结果表明,5种半夏属植物被聚为两大类:掌叶半夏单独聚为一类(Ⅱ),而其它4个种聚为一类(Ⅰ).第Ⅰ类可再分为A和B两个亚类:A亚类包括半夏和石蜘蛛;B亚类包括盾叶半夏和滴水珠.滴水珠和盾叶半夏的亲缘关系最近,其次是半夏和石蜘蛛,而掌叶半夏和其它4个种的亲缘关系都较远,这说明掌叶半夏与半夏属其它种呈姐妹群关系.本研究结果对我国半夏属植物资源的开发利用与保护具有重要意义.     

大螟不同地理种群COⅡ基因序列分析

本研究通过对我国大螟Sesamia inferens 9个地理种群的线粒体DNACOII基因的测序,运用软件DNAStar的MegAlign程序分析了大螟不同地理种群之间的COII序列的遗传分歧及相似性;同时运用Mega 3.0软件建立其系统发育关系。结果表明,COII基因在大螟中进化速度较快,不同种群之间已产生较大的差异,序列相似性分析显示大螟种群间序列相似性最低仅为90.7（海南和江西种群）,进化分歧矩阵同时也显示差异最大为0.102（云南和江西种群）。此外,以与大螟同属的非洲大螟及螟蛾科的台湾稻螟为外群进行比较显示,COII基因在属内种间差异明显,遗传相似性为87.7-91.8,进化分歧为0.088-0.137;与台湾稻螟的差异最大,相似性仅为82.2-85.8,进化分歧为0.159-0.202。进化树显示,大螟贵州的丹寨种群、沿河种群和福建的福州种群聚为一个分支,江苏扬州种群和江西新余种群聚为一支,安徽的阜阳种群和芜湖种群关系最近,说明不同地理种群的大螟的遗传分化与地理距离之间具有一定的相关性。     

半夏属的染色体数目、倍性与珠芽发生的关系

本文通过对半夏属Pinellia 5个种10个群体的染色体计数和珠芽数量统计,首次报道了5个染色体数目,同时发现珠芽的发生与染色体基数及多倍化程度有关：x=13的类群无珠芽,而x=9的有珠芽;在有珠芽的半夏P．ternata(Thunb．)Breit．中,平均每叶珠芽数随倍性的提高而增大。半夏是一个多倍体复合种,起源于无珠芽、染色体基数为x=7～9的二倍体祖先,可能是在该属的早期进化中由鹞落坪半夏P.yaoluopingensis X．H．Guo et X．L.Liu的x=13经非整倍性跌落而成,在发生上比鹞落坪半夏进化。     

大螟不同地理种群COII基因序列分析

本研究通过对我国大螟Sesamia inferens 9个地理种群的线粒体DNA COII基因的测序,运用软件DNAStar的MegAlign程序分析了大螟不同地理种群之间的COII序列的遗传分歧及相似性;同时运用Mega 3.0软件建立其系统发育关系.结果表明,COII基因在大螟中进化速度较快,不同种群之间已产生较大的差异,序列相似性分析显示大螟种群间序列相似性最低仅为90.7(海南和江西种群),进化分歧矩阵同时也显示差异最大为0.102(云南和江西种群).此外,以与大螟同属的非洲大螟及螟蛾科的台湾稻螟为外群进行比较显示,COII基因在属内种间差异明显,遗传相似性为87.7～91.8,进化分歧为0.088～0.137;与台湾稻螟的差异最大,相似性仅为82.2～85.8,进化分歧为0.159～0.202.进化树显示,大螟贵州的丹寨种群、沿河种群和福建的福州种群聚为一个分支,江苏扬州种群和江西新余种群聚为一支,安徽的阜阳种群和芜湖种群关系最近,说明不同地理种群的大螟的遗传分化与地理距离之间具有一定的相关性.     

山楂叶螨线粒体基因组的特征与系统发育与分析

山楂叶螨Amphitetranychus viennensis (Zacher,1920)又名山楂红蜘蛛,主要危害苹果、梨、桃、山楂和李等蔷薇科果树。山楂叶螨的分类归属在叶螨属Tetranychus与双叶螨属Amphitetranychus之间存在争议。因此,获得山楂叶螨的线粒体基因组可在解决其分类问题上提供更多依据。本研究通过长PCR扩增与高通量测序结合的方法得到完整山楂叶螨基因组,其大小为13 085 bp,共注释37个基因,包括13个蛋白编码基因、2个rRNA和22个tRNA。对tRNA的二级结构进行预测后发现有大量长度收缩的tRNA无法折叠成经典三叶草结构。遗传进化分析表明山楂叶螨与叶螨属的物种进化关系较近,但是山楂叶螨与叶螨属物种的遗传距离大于叶螨属内的遗传距离,因此通过对山楂叶螨线粒体基因组的分析支持山楂叶螨独立出叶螨属并归入双叶螨属。     

基于线粒体COⅡ基因的中国蝽科分子系统学研究(半翅目,异翅亚目)

扩增并测定了我国蝽科4亚科8属11种昆虫线粒体COⅡ基因585 bp的序列,对序列的碱基组成、转换颠换、遗传距离等进行分析,探讨了COⅡ基因在该科的分子进化机制.并基于COⅡ基因序列数据,分别采用邻接法(NI)、最大简约法(MP)和贝叶斯推论法(BI)建立蝽科分子系统发育关系.研究结果表明,蝽科昆虫COⅡ基因A T含量平均为71.7%,存在较强的A T含量偏向性,氨基酸的变异率为27.2%;亚科间的遗传距离介于0.168～0.242之间,大于亚科内属种间的遗传距离,蝽科与盾蝽科2外群之间遗传距离最大,两科之间存在明显的间断.分子系统发育树表明,短喙蝽亚科为蝽科中较为原始的类群,分化较早,益蝽亚科与舌盾蝽亚科关系较近,形成一对姐妹群,蝽科中捕食性种类--益蝽亚科是较为特化的类群,它是由植食性种类分化而来.蝽科4亚科间的分子系统发育关系为Phyllocephalinae (Pentatominae (Asopinae Podopinae).     

昌化江河谷隔离对海南岛特有植物盾叶苣苔遗传多样性的影响

为评估盾叶苣苔的遗传多样性与遗传分化格局,探索影响盾叶苣苔遗传变异地理分布的因素,该研究采集盾叶苣苔(Metapetrocosmea peltata)11个种群172份材料,通过PCR扩增和测序分析核糖体转录间隔区(ITS)序列的变异式样。结果表明:(1)盾叶苣苔物种水平的遗传多样性很高(H_T=0.998, π=0.023 5),种群间基因流很弱(N_m=0.04)且存在强烈的遗传分化(G_ST=0.375)。(2)单倍型分析显示,盾叶苣苔的单倍型大多是种群特异的,仅白马岭与南茂岭种群有共享单倍型。(3)Mantel test表明,遗传距离和地理距离存在一定相关性(相关系数r=0.322,P=0.010)。(4)Structure聚类分析将盾叶苣苔划分为6种遗传成分,其地理分布与昌化江河谷导致的隔离样式基本一致,基于Nei遗传距离的种群聚类分析支持这一结果,显示盾叶苣苔遗传多样性的分布受到昌化江河谷的隔离作用。(5)AMOVA分析确定67%的变异来自地区间,表明地理隔离是盾叶苣苔种群分化的重要因素。这表明昌化江及其支流所引起的海南岛山地内部隔离是盾叶苣苔种群发生强烈遗传分化的重要原因,从而导致盾叶苣苔在物种水平具有较高的遗传多样性。上述研究结果为海南特有苦苣苔资源盾叶苣苔的保护和可持续利用提供了理论指导,将有助于理解海南岛特有植物和其他海岛植物遗传变异的地理分布及其影响因素。     

印度-西太平洋胡椒鲷亚科鱼类DNA条形码及分子系统进化研究

为探讨COⅠ基因作为条形码在胡椒鲷亚科鱼类物种鉴定的可行性, 研究测定了胡椒鲷亚科8种鱼类51个个体线粒体COⅠ基因长度为651 bp的序列, 利用MEGA 5.0计算胡椒鲷亚科种内与种间的遗传距离, 基于最大似然法与贝叶斯法构建了系统进化树。结果显示: 胡椒鲷亚科鱼类种间平均遗传距离(0.142)显著大于种内平均遗传距离(0.003), 物种间遗传距离均大于Hebert推荐的物种鉴定最小种间遗传距离0.020(2%)。系统进化树上, 同一物种不同个体间均能形成独立的单系分支, 表明COⅠ基因可作为胡椒鲷亚科物种鉴定的有效条形码基因。研究同时揭示, 在形态分类上被认为是同种异名的两种胡椒鲷(条纹胡椒鲷与东方胡椒鲷)的COⅠ基因序列差异达到0.070, 有可能是两个独立的物种。此外, 在属级水平, 少棘胡椒鲷属与胡椒鲷属种类之间的平均遗传距离小于胡椒鲷属内部种间的遗传距离, 支持少棘胡椒鲷归属于胡椒鲷属的观点。       

基于髭鲷属鱼类Cyt b基因全序列探讨髭鲷属在鲈总科的分类地位

为探讨髭鲷属鱼类的分类地位,克隆了两种髭鲷属鱼类(横带髭鲷和斜带髭鲷)的线粒体Cyt 6基因全长序列,并利用线粒体Cyt b基因序列分析了鲈总科6个科29种的分子系统进化关系.结果表明:髭鲷属与石鲈科的其他5个属的遗传距离超过了科问水平的遗传距离,存在较远的遗传分化.29种鲈总科鱼类分成了两大类群,石鲈科的矶鲈属、仿石鲈属、厚唇椒鲷属、异孔石鲈属和石鲈属的物种聚为一起形成石鲈科的一个独立分支,与亲缘关系较近的鲷科、梅鲷科和笛鲷科形成一个类群;而隶属于石鲈科的髭鲷属与石鲷科及雀鲷科形成另一个类群.髭鲷属与其他石鲈科鱼类的亲缘关系已经超过了科问的遗传分化水平,而在遗传距离及系统进化分析上髭鲷属仍然隶属于鲈总科.本研究支持将髭鲷属提升为髭鲷科的观点.     

Phylogenetic relations of Rhizoplaca Zopf. from Anatolia inferred from ITS sequence data

Like many lichen-forming fungi, species of the genus Rhizoplaca have wide geographical distributions, but studies of their genetic variability are limited. The information about the ITS rDNA sequences of three species of Rhizoplaca from Anatolia was generated and aligned with other species from other countries and also with the data belonging to Lecanora species. The examined species were collected from the volcanic rocks of Mount Erciyes which is located in the middle of Anatolia (Turkey). The sequence data aligned with eight other samples of Rhizoplaca and six different species of Lecanora were obtained from GenBank. The results support the concept maintained by Arup and Grube (2000) that Rhizoplaca may not be a genus separate from Lecanora. According to the phylogenetic tree, Rhizoplaca melanopthalma from Turkey with two different samples of R. melanopthalma from Arizona (AF159929, AF159934) and a sample from Austria formed a group under the same branch. R. peltata and R. chrysoleuca samples from Anatolia located in two other branches of the tree formed sister groups with the samples of the same species from different countries. Although R. peltata remained on the same branch with other samples of the same species from other countries it was placed in a different branch within the group. When the three species from Anatolia were considered alone, it was noticed that Rhizoplaca melanopthalma and Rhizoplaca peltata are phylogenetically closer to each other than Rhizoplaca chrysoleuca; the morphological characteristics also support this result.     

The genus Podocrella and its nematode-killing anamorph Harposporium

Several genera are described in the literature as having morphology similar to the clavicipitaceous genus Podocrella, viz. Atricordyceps, Ophiocordyceps, Wakefieldiomyces and "Cordyceps" peltata. These genera have capitate-stipitate stromata that gradually expand into a horizontally flattened fertile head that is dark, has strongly protruding perithecia and asci containing eight multiseptate filiform ascospores. These ascospores disarticulate at the middle septum to form two lanceolate multiseptate part-as-cospores. In this study several specimens of the above-mentioned genera, including the types, were examined to determine whether they are congeneric with Podocrella. This study also reveals the connection of Podocrella to its anamorph genus, Harposporium, and its relationship to several other clavicipitaceous genera, based on cultural data and large subunit nuclear ribosomal DNA (LSU) sequences. Nematode predation of the Harposporium anamorph of P. peltata is demonstrated. The results show Podocrella and selected Harposporium LSU sequences form a monophyletic group and that this clade is closely related to Aschersonia. A new species of Podocrella from Costa Rica, P fusca, is described, new combinations made for P. peltata and P. harposporifera, and a key to the known species is presented.     

Genetic comparison of Bacillus anthracis and its close relatives using amplified fragment length polymorphism and polymerase chain reaction analysis

Amplified fragment length polymorphism (AFLP) analysis allows a rapid, relatively simple analysis of a large portion of a microbial genome, providing information about the species and its phylogenetic relationship to other microbes (Vos et al. 1995). The method simply surveys the genome for length and sequence polymorphisms. The AFLP pattern identified can be used for comparison to the genomes of other species. Unlike other methods, it does not rely on analysis of a single genetic locus that may bias the interpretation of results and does not require any prior knowledge of the targeted organism. Moreover, a standard set of reagents can be applied to any species without using species-specific information or molecular probes. We are using AFLP analysis to rapidly identify different bacterial species. A comparison of AFLP profiles generated from a large battery of Bacillus anthracis strains shows very little variability among different isolates (Keim et al. 1997). By contrast, there is a significant difference between AFLP profiles generated for any B. anthracis strain and even the most closely related Bacillus species. Sufficient variability is apparent among all known microbial species to allow phylogenetic analysis based on large numbers of genetically unlinked loci. These striking differences among AFLP profiles allow unambiguous identification of previously identified species and phylogenetic placement of newly characterized isolates relative to known species based on a large number of independent genetic loci. Data generated thus far show that the method provides phylogenetic analyses that are consistent with other widely accepted phylogenetic methods. However, AFLP analysis provides a more detailed analysis of the targets and samples a much larger portion of the genome. Consequently, it provides an inexpensive, rapid means of characterizing microbial isolates to further differentiate among strains and closely related microbial species. Such information cannot be rapidly generated by other means. AFLP sample analysis quickly generates a very large amount of molecular information about microbial genomes. However, this information cannot be analysed rapidly using manual methods. We are developing a large archive of electronic AFLP signatures that is being used to identify isolates collected from medical, veterinary, forensic and environmental samples. We are also developing the computational packages necessary to rapidly and unambiguously analyse the AFLP profiles and conduct a phylogenetic comparison of these data relative to information already in our database. We will use this archive and the associated algorithms to determine the species identity of previously uncharacterized isolates and place them phylogenetically relative to other microbes based on their AFLP signatures. This study provides significant new information about microbes with environmental, veterinary and medical significance. This information can be used in further studies to understand the relationships among these species and the factors that distinguish them from one another. It should also allow the identification of unique factors that contribute to important microbial traits, including pathogenicity and virulence. We are also using AFLP data to identify, isolate and sequence DNA fragments that are unique to particular microbial species and strains. The fragment patterns and sequence information provide insights into the complexity and organization of bacterial genomes relative to one another. They also provide the information necessary for the development of species-specific polymerase chain reaction primers that can be used to interrogate complex samples for the presence of B. anthracis, other microbial pathogens or their remnants.     

Ochratoxin A producing Penicillium verrucosum isolates from cereals reveal large AFLP fingerprinting variability

AIMS: To examine if molecular amplified fragment length polymorphism (AFLP) fingerprinting of the only ochratoxin A-producing species in European cereals, Penicillium verrucosum, can be used as a method in hazard analysis using critical control points (HACCP). METHODS AND RESULTS: A total of 321 isolates of P. verrucosum were isolated from ochratoxin A-contaminated cereals from Denmark (oats), UK (wheat and barley) and Sweden (wheat). Of these, 236 produced ochratoxin A as determined by thin layer chromatography; 185 ochratoxin A-producing isolates were selected for AFLP fingerprinting. A total of 138 isolates had unique AFLP patterns, whereas 52 isolates could be allocated to small groups containing from two to four isolates with similar AFLP patterns. A total of 155 clones were found among the 185 P. verrucosum isolates, thus 84% of the isolates may represent different genets of P. verrucosum. As the few isolates that were grouped often came from the same farm, and those groups that contained AFLP-identical isolates from different countries were morphotypically different. On single farms up to 35 clones were found. The few groups of ramets from the same genet indicated that a HACCP approach based on clones may require a very large number of AFLP analysis to work in practice, we recommend basing the HACCP approach on the actual species P. verrucosum. A more detailed characterization should rather be based on the profile of species present at different control points, or analysis of the mycotoxins ochratoxin A and citrinin in the isolates. Examination of 86 isolates with HPLC and diode array detection of P. verrucosum showed that 66% produced ochratoxin A, 87% produced citrinin, 92% produced verrucin and 100% produced verrucolone. CONCLUSIONS: Among 184 ochratoxin A-producing Penicillium verrucosum, 155 clonal lineages were indicated by AFLP fingerprinting, indicating a high genetical diversity, yet the species P. verrucosum is phenotypically distinct and valid. SIGNIFICANCE AND IMPACT OF THE STUDY: AFLP fingerprinting of Penicillium verrucosum indicates that genetic recombination takes place in this fungus.     

An AFLP clock for the absolute dating of shallow-time evolutionary history based on the intraspecific divergence of southwestern European alpine plant species

The dating of recent events in the history of organisms needs divergence rates based on molecular fingerprint markers. Here, we used amplified fragment length polymorphisms (AFLPs) of three distantly related alpine plant species co-occurring in the Spanish Sierra Nevada, the Pyrenees and the southwestern Alps/Massif Central to establish divergence rates. Within each of these species ( Gentiana alpina , Kernera saxatilis and Silene rupestris ), we found that the degree of AFLP divergence ( D _N72) between mountain phylogroups was significantly correlated with their time of divergence (as inferred from palaeoclimatic/palynological data), indicating constant AFLP divergence rates. As these rates did not differ significantly among species, a regression analysis based on the pooled data was utilized to generate a general AFLP rate. The application of this latter rate to AFLP data from other herbaceous plant species ( Minuartia biflora : Schönswetter et al . 2006 ; Nigella degenii : Comes et al . 2008 ) resulted in a plausible timing of the recolonization of the Svalbard Islands and the separation of populations from the Alps and Scandinavia ( Minuartia ), and of island population separation in the Aegean Archipelago ( Nigella ). Furthermore, the AFLP mutation rate obtained in our study is of the same magnitude as AFLP mutation rates published previously. The temporal limits of our AFLP rate, which is based on intraspecific vicariance events at shallow (i.e. late glacial/Early Holocene) time scales, remains to be tested.     

Reconstructing the species phylogeny of Pseudopanax (Araliaceae), a genus of hybridising trees

Pseudopanax (Araliaceae) comprises 12 tree species of diverse morphology and ecology, and is endemic to New Zealand. It is notable for the hybridisation that occurs between P. crassifolius and P. lessonii, which have very different leaves and habits. To provide context for the study of this hybridisation and other investigations, we examined the phylogeny of Pseudopanax using chloroplast DNA sequences (c.5900 base-pairs) and AFLP DNA-fingerprinting. Both approaches resolve two principal groups within Pseudopanax – the Arboreus group and the Crassifolius + Lessonii union – but how they are related to other genera remains unclear. There is, nevertheless, little compelling evidence against the monophyly of Pseudopanax, making unnecessary the recent re-segregation of the Arboreus group as Neopanax. The chloroplast data provided minimal additional resolution, although the position of P. linearis was discordant compared to other data. Analyses of the AFLP data strongly recovered each species, aside from the morphologically heterogeneous P. colensoi, and the two mainland species (P. arboreus and P. crassifolius) that each contained a nested island-endemic (P. kermadecensis and P. chathamicus, respectively). However, relationships amongst species within the two principal groups were poorly resolved. An example was the uncertainty of whether P. crassifolius grouped with P. lessonii and its allies, or the morphologically similar species it had been previously placed with. This in turn raises the issue of how hybridisation might affect phylogenies and the ability to reconstruct them, even when using multiple, independent markers.     

Phylogeny, biogeography and taxonomy of the African wattle-eyes (Aves: Passeriformes: Platysteiridae)

The African wattle-eyes (genera Platysteira and Dyaphorophyia) comprise 10 species endemic to Africa. We analyzed both mitochondrial and nuclear DNA sequence data to test the monophyly of this group and its two genera, provide a preliminary assessment of species limits, and gain insight into the phylogeographic history of the wattle-eye radiation. Analyses based on mitochondrial ND2 sequences failed to recover wattle-eye monophyly, but the alternatives were not well-supported. In contrast, analyses of two nuclear introns (myoglobin intron-2 and beta-fibrinogen intron-5) recovered wattle-eye monophyly, as did combined analyses of mitochondrial and nuclear data. These analyses, however, did not support reciprocal monophyly of the two wattle-eye genera typically recognized, suggesting instead that Platysteira is nested within a paraphyletic Dyaphorophyia. The diversification of most wattle-eye species and many subspecies occurred through the divergence of allopatric populations well before the Pleistocene. Species and subspecies with disjunct distributions are typically characterized by deep genetic divergences, suggesting that many of these populations are evolutionary independent and could be recognized as additional phylogenetic species. In D. castanea and D. chalybea, for example, divergent haplotypes from geographically disjunct populations were paraphyletic with respect to those of D. tonsa and D. jamesoni, respectively. Similarly, Platysteira laticincta is highly divergent from its sister taxon P. peltata ( approximately 9.5% ND2 sequence divergence), consistent with species level recognition of this endangered species. In contrast, more broadly distributed taxa inhabiting a greater diversity of habitats (e.g., P. peltata and P. cyannea) show evidence of gene flow and connectivity among regions, suggesting that previously isolated populations expanded and fused into one another. Our study provides a framework for additional analyses of intraspecific phylogeography and species limits in these colorful birds.     

Parasites, primates, and ant-plants: clues to the life cycle of Controrchis spp. in black howler monkeys (Alouatta pigra) in southern Belize

The purpose of this study was to investigate the prevalence, intensity, and possible transmission routes of the trematode, Controrchis spp. (Dicrocoeliidae), in a population of black howler monkeys (Alouatta pigra) near Monkey River, Belize. Alouatta pigra are arboreal primates that are typically folivorous and frugivorous. Controrchis is a dicrocoeliid trematode and, as such, should require a gastropod and an ant (Formicidae) intermediate host for transmission. From January to July 2005 and February 2006 to June 2007, we collected fecal samples and focal animal data from 18 individual primates in four social groups. Feces were stored in either 10% buffered formalin or 95% ethanol and examined using a double-centrifugation sugar-flotation technique. The prevalence of Controrchis spp. was 89%, and the mean intensity was 2.29 eggs per gram (epg), with a range of 1.00-6.57 epg. A general linear mixed model to examine the effect of sex, rainfall, and time spent feeding on various plant species, while controlling for individual and group identity, revealed that both group and the amount of time spent feeding on trumpet trees (Cecropia peltata) by individuals predicted the prevalence and intensity of Controrchis spp. infestation. Cecropia peltata has a mutualistic relationship with Azteca spp. ants and constitutes an important food source for A. pigra. Our findings provide strong, circumstantial evidence that A. pigra are infected with Controrchis spp. through the ingestion of metacercariae in Azteca spp. when feeding on C. peltata. Because C. peltata is a pioneer tree species and typically occurs in disturbed forests, results from this study suggest an important link between habitat disturbance and parasitism in a wild mammal.     

Comparing EST-based genetic maps between <Emphasis Type="BoldItalic">Pinus sylvestris</Emphasis> and <Emphasis Type="BoldItalic">Pinus taeda</Emphasis>

A genetic map of Pinus sylvestris was constructed using ESTP (expressed sequence tag polymorphism) markers and other gene-based markers, AFLP markers and microsatellites. Part of the ESTP markers (40) were developed and mapped earlier in Pinus taeda, and additional markers were generated based on P. sylvestris sequences or sequences from other pine species. The mapping in P. sylvestris was based on 94 F₁ progeny from a cross between plus-tree parents E635C and E1101. AFLP framework maps for the parent trees were first constructed. The ESTP and other gene sequence-based markers were added to the framework maps, as well as five published microsatellite loci. The separate maps were then integrated with the aid of AFLPs segregating in both trees (dominant segregation ratios 3:1) as well as gene markers and microsatellites segregating in both parent trees (segregation ratios 1:1:1:1 or 1:2:1). The integrated map consisted of 12 groups corresponding to the P. taeda linkage groups, and additionally three and six smaller groups for E1101 and E635C, respectively. The number of framework AFLP markers in the integrated map is altogether 194 and the number of gene markers 61. The total length of the integrated map was 1,314 cM. The set of markers developed for P. sylvestris was also added to existing maps of two P. taeda pedigrees. Starting with a mapped marker from one pedigree in the source species resulted in a mapped marker in a pedigree of the other species in more than 40% of the cases, with about equal success in both directions. The maps of the two species are largely colinear, even if the species have diverged more than 70 MYA. Most cases of different locations were probably due to problems in identifying the orthologous members of gene families. These data provide a first ESTP-containing map of P. sylvestris, which can also be used for comparing this species to additional species mapped with the same markers.Communicated by C. Möllers