Pleurotus eryngii species complex: sequence analysis and phylogeny based on partial EF1α and RPB2 genes

The Pleurotus eryngii species complex comprises at least six varieties (var. eryngii (DC.: Fr) Quel., ferulae Lanzi, elaeoselini Venturella et?al., nebrodensis (Inzenga) Sacc., tingitanus Lewinsohn et?al. and tuoliensis C.J. Mou). This species is unique among the genus Pleurotus because in nature it is found in association with certain species of the Apiaceae (Umbelliferae) and Asteraceae (Compositae) families. Sequences of partial regions of the translation elongation factor (EF1α) and RNA polymerase II (RPB2) genes were analyzed in order to detect nucleotide polymorphisms that might unequivocally distinguish varieties eryngii, ferulae, elaeoselini and nebrodensis. A phylogenetic analysis was also performed with an aim to establish phylogenetic relationships among those. Sequence analysis of the partial EF1α and RPB2 genes contained nucleotide polymorphisms able to unequivocally distinguish variety nebrodensis from the rest. However, distinction among eryngii, elaeoselini and ferulae was achieved only through the RPB2 gene. The phylogenetic analyses from the combined data sets (EF1α and RPB2) indicated that P. eryngii is a monophyletic group and that varieties eryngii, elaeoselini and ferulae are closely related. P. eryngii var. nebrodensis was placed in a distinct clade clearly differentiated from the other varieties but still monophyletic with the P. eryngii complex. The limited nucleotide variation in partial EF1α and RPB2 among varieties eryngii, ferulae and elaeoselini supports the placement of these groups as varieties and not species within the complex.     

Paeonia daurica Andrews or P. mascula ssp. triternata (Pall. ex DC.) Stearn & P. H. Davis (Paeoniaceae)?

The peony in the Crimea of Ukraine and its allied populations have been variously taxonomically treated, as Paeonia daurica Andrews or P. mascula ssp. triternata (Pall. ex DC.) Stearn & P. H. Davis. Supported by the National Geographical Society, we have conducted extensive field observations and population sampling of this group in Turkey. In addition, relevant herbarium specimens from the herbaria B, BEO, BM, BUCA, E, G, GZU, K, P, SA, SOM, UPA, and WU were critically examined. Principal coordinate analysis was performed using MVSP-Version 3.13b analysis software. As a result, P. daurica was shown to be clearly differentiated from P. mascula in the number of leaflets/segments of the lower leaves and the shape of the terminal leaflets. P. daurica is diploid, except for three local tetraploids in the Caucasus, whereas P. mascula is consistently tetraploid. The two units were not found growing together, even in southern Turkey, where they are sympatric. P. daurica is considered to be a good species, which ranges from Croatia to Iran through Turkey and the Caucasus, and comprises six subspecies.  © 2007 The Linnean Society of London, Botanical Journal of the Linnean Society , 2007, 154 , 1–11.     

中国白灵侧耳自然群体的交配型因子分析

为了研究我国新疆白灵侧耳自然种群的遗传多样性,估测自然种群的遗传丰度,对51株新疆野生白灵侧耳的84个原生质体单核体的交配型因子进行分析。在84个单核体中存在54个不同的A因子和59个不同的B因子。x2检测表明交配型因子A和B的系列因子均为等概率分布。据此估算我国白灵侧耳自然群体中有79个A因子,100个B因子,自然群体中的单核交配型总数79×100=7,900个,双核交配型总数31,201,050个。可见,新疆白灵侧耳遗传多样性丰富,新疆是我国白灵侧耳的野生种质资源宝库。     

中国栽培白灵菇学名的订正

采用形态学方法对采集于新疆野生的阿魏蘑标本进行分类鉴定,结果表明其形态学特征符合刺芹侧耳托里变种Pleurotus eryngii var. tuoliensis的特征范围,它与欧洲已报道的Pleurotus nebrodensis有明显的差异。此外本研究还采用分子生物学方法对从该野生样品分离的菌株CCMSSC 02514进行了rDNA ITS序列分析,结果表明它与我国栽培白灵菇菌种CCMSSC 00973、KH5和AFRL 6022完全相同,以此构建系统发育树,将我国栽培白灵菇种质与意大利的Pleurotus nebrodensis、Pleurotus eryngii var. ferulae以及来自荷兰的Pleurotus eryngii等截然分为两组。因此,形态学和rDNA ITS序列分析结果支持我国栽培的白灵菇与欧洲的Pleurotus nebrodensis为不同种,我国的白灵菇是刺芹侧耳独立进化的一个分支,其名称应该为刺芹侧耳托里变种Pleurotus eryngii var. tuoliensis。     

基于ITS序列分析对我国主要栽培的侧耳品种的鉴定及评价

利用PCR产物克隆测序测定了20个我国主要栽培的侧耳品种的ITS序列,另外从GenBank获得侧耳属15个种25条ITS序列及亚侧耳属2个种的ITS序列。以Hohenbueheliagrisea和H.tremula为外群,运用PAUP软件中的简约分析法(parsimonyanalysis)构建的系统发育树表明:侧耳属Pleurotus是单起源的,20个主要栽培的侧耳品种分别聚在三个组,即Ostreatus-eryngii-populinus复合组、Pulmonarius组、Citrinopileatus-cornucopiae组。Ostreatus-eryngii-populinus组含刺芹侧耳Pleurotuseryngii、白灵侧耳P.nebrodensis、香侧耳Pleurotussp.、阿魏侧耳P.eryngiivar.ferulae、平963-1Pleurotussp.及糙皮侧耳P.ostreatus、日本秀珍Pleurotussp.、平802Pleurotussp.、姬菇Pleurotussp.、灰白侧耳P.spodoleucus、缘刺侧耳Pleurotussp.、凤尾菇P.sajor-caju;Pulmonarius组含肺形侧耳P.pulmonarius、小白平菇Pleurotussp.、平8804Pleurotussp.、平侧5Pleurotussp.、美味侧耳P.sapidus;Citrinopileatus-cornucopiae组含黄白侧耳P.cornucopiae、金顶侧耳P.citrinopileatus、鸡汁菌Pleurotussp.。系统树还显示黄白侧耳与金顶侧耳、白灵侧耳与刺芹侧耳亲缘关系密切,而凤尾菇与肺形侧耳分属于不同的组,属于两个不同的种。基于ITS序列分析,本文还针对目前我国栽培的主要侧耳品种在名称使用上的混淆和混乱进行了初步的评价和讨论。     

Genetic diversity and population structure of the Italian fungi belonging to the taxa Pleurotus eryngii (DC.:Fr.) Quèl and P. ferulae (DC.:Fr.) Quèl

A study using allozymes and PCR fingerprinting was conducted to estimate the genetic diversity of Italian populations of two economically important cultivated fungal taxa, Pleurotus eryngii and P. ferulae. Very little is known about the genetic diversity distribution pattern of these taxa. Heterozygote deficiency was observed at few loci; in fact the inbreeding coefficients were not high, which demonstrates that mechanisms restrain the inbreeding act at the local level. Estimates of genetic differentiation indicated a pattern of greater variation within, rather than between, populations. These results were supported by AMOVA analysis, which attributed a low proportion of the total genetic variation to large geographical scale divergence, and indicated that most of the genetic diversity was because of differences within populations. This distribution pattern of genetic variation of P. eryngii and P. ferulae populations seems to be the result of high gene flow, by efficient basidiospore dispersal, and outcrossing mechanisms, which restrain inbreeding within populations.     

基于DNA拓扑异构酶II基因部分序列的侧耳属系统发育分析

对侧耳属（Pleurotus）14个种的14个菌株,亚侧耳属（Hohenbuehelia）1个种和离褶伞属（Lyophyllum）1个种的DNA拓扑异构酶II（EC 5.99.1.3）部分序列进行扩增分析。以灰盖鬼伞（Coprinus cinereus）、新型隐球酵母（Cryptococcus neoformans）和玉米黑粉菌（Puccinia polysora）等担子菌为外群,采用最大距离法、最大简约法和最大似然法构建侧耳属的系统发育树。结果表明：作为外群的三个物种独立于所有供试材料;侧耳属菌株作为一大类与供试的离褶伞属和亚侧耳属分开。侧耳属14个种又可细分成六支：可能为侧耳属起源最早的P. flabellatus、P. salmoneostramineus和P. djamor组成一组;P. tuberregium和P. pulmonarius均自成一组;P. abalonus和P. cystidiosus均产生束梗孢,聚为同组;P. eryngii var. eryngii、P. eryngii var. ferulae和P. nebrodensis为一组;另一组由P. ostreatus、P. columbinus、P. cornucopiae和P. citrinopileatus组成。按照侧耳属物种菌丝类型的不同,构建的系统发育树将该属14个种进行系群划分,单、二系菌丝分属于各个不同的分支中,说明单、二系菌丝均为多系起源,与用28S rDNA序列进行系统分析的结果一致。     

Apochromic populations of Dactylorhiza incarnata s.l. (Orchidaceae): diversity and systematic significance as revealed by allozyme markers and morphology

Apochromic forms of the Eurasian Dactylorhiza incarnata s.l. were studied in northern Europe to reveal their genetic (allozyme) and morphological diversity and to assess their systematic significance. The study included eight localities with sympatric populations of plants with anthocyanin‐pigmented and apochromic flowers. Parallel samples of the two morphs were taken from each locality. Genetic variation was only found at the allozyme loci pgd, pgi and ugpp. Statistically significant differences in allele frequencies between the two colour morphs were found in two localities and demonstrate that the occurrence of apochromic individuals in D. incarnata s.l. is not always because of spontaneous mutation. At least in some localities the apochromic plants form distinct breeding groups (but local populations of different colour morphs may also be composed of several more or less distinct breeding groups). Based on molecular and morphometric data, it is proposed that the apochromic study populations from calcareous fens should be referred to D. incarnata var. ochroleuca, whereas the apochromic study populations from non‐calcareous fens are better treated as aberrant local populations of var. incarnata s.l. Possible evolutionary patterns and processes are discussed and guidelines for identification of var. ochroleuca from morphological features are given. © 2009 The Linnean Society of London, Botanical Journal of the Linnean Society, 2009, 159 , 396–407.     

Four new species of Pleurosigma (Bacillariophyta) from Alexandria, Egypt

Four new species of Pleurosigma are described from the region of Alexandria in Egypt: P. williamsii sp. nov. , P. mariutensis sp. nov. , P. ramlehensis sp. nov. and P. rushdyensis sp. nov. The taxa had been previously misidentified as P. rigidum W. Sm., P. elongatum W. Sm., P. speciosum var. sumatricum H. Perag. and P. formosum W. Sm., respectively. Due to the close resemblance of P. williamsii, P. mariutensis , P. ramlehensis and P. rushdyensis to P. rigidum , P. elongatum , P. speciosum var. sumatricum and P. formosum and the lack of good illustrations of these previously described species, their valvar features are shown and discussed in relation to the new taxa. © 2002 The Linnean Society of London, Botanical Journal of the Linnean Society , 2002, 140 , 77–92.     

基于DNA拓扑异构酶Ⅱ基因部分序列的侧耳属系统发育分析

对侧耳属（Pleurotus）14个种的14个菌株,亚侧耳属（Hohenbuehelia）1个种和离褶伞属（Lyophyllum）1个种的DNA拓扑异构酶Ⅱ（EC 5．99．1．3）部分序列进行扩增分析。以灰盖鬼伞（Coprinus cinereus）、新型隐球酵母（Cryptococcus neoformans）和玉米黑粉菌（Puccinia polysora）等担子菌为外群,采用最大距离法、最大简约法和最大似然法构建侧耳属的系统发育树。结果表明：作为外群的3个物种独立于所有供试材料;侧耳属菌株作为一大类与供试的离褶伞属和亚侧耳属分开。侧耳属14个种又可细分成6支：可能为侧耳属起源最早的P．flabellatus、P．salmoneostramineus和P．djamor组成一组;P．tuberregium和P．pulmonarius均自成一组;P．abalonus和P．cystidiosus均产生束梗孢,聚为同组;P．eryngii var．eryngii、P．eryngii var．ferulae和P．nebrodensis为一组;另一组由P．ostreatus、P．columbinus、P．cornucopiae和P．citrinopileatus组成。按照侧耳属物种菌丝类型的不同,构建的系统发育树将该属14个种进行系群划分,单、二系菌丝分属于各个不同的分支中,说明单、二系菌丝均为多系起源,与用28S rDNA序列进行系统分析的结果一致。     

IGS2-RFLP、SCoT和ISSR在白灵侧耳遗传多样性分析中的比较研究

应用IGS2-RFLP、SCoT和ISSR标记分析新疆5个采集地点的28个白灵侧耳野生样本的遗传多样性,比较这3种分子标记在白灵侧耳种质资源遗传多样性研究中的效用。结果显示,IGS2-RFLP的3个内切酶、5个SCoT引物、5个ISSR引物分别检测到42、59和77条多态性条带,多态性比率分别为91.3%、92.4%和92.8%。3种标记检测出的有效等位基因数(ne)和位点平均的预期杂合度(Hep)没有显著性差异。表明3种标记都适宜作为遗传学标记对白灵侧耳野生种质进行多样性分析。多态性检测效率最高的标记为ISSR,E=15.4,Ai=23.4,其次为IGS2-RFLP,E=14.0,Ai=22.4,SCoT则为最低,E=11.8,Ai=18.2。3种标记中,SCoT和ISSR标记的聚类结果极其相似,且均能较为准确地反映样本的地理来源,虽然二者的聚类图不完全相同。IGS2-RFLP的标记效率较高,准确性和可重复性最好,可用于菌株标准图谱的制作,更适用于品种权保护中的菌株鉴定鉴别;SCoT和ISSR标记的多态性高,信息量大,评价范围广,则更适用于白灵侧耳种质资源的遗传多样性研究。     

Euphrasia amurensis, E. hirtella var. ramosa and var. karoiana (Orobanchaceae)

The lectotype for Euphrasia hirtella var. ramosa is selected. This taxon and E. hirtella var. karoiana are compared with E. amurensis and confirmed as synonyms, corroborating Juzepcuk (1955 ).  © 2007 The Linnean Society of London, Botanical Journal of the Linnean Society , 2007, 154 , 215–223.     

A multidisciplinary approach to discriminating different taxa in the species complex Pachycondyla villosa (Formicidae)

A multidisciplinary approach provides new evidence that strongly supports the differentiation of three taxa previously confounded in a single species, Pachycondyla villosa . All specimens studied were collected in the same location at Ilhéus, Bahia, Brazil. Bivariate plots of petiole heights vs. petiole lengths and isozyme patterns clearly differentiate the three taxa. Chain lengths and chemical structures of cuticular hydrocarbons are characteristic of each taxon. This congruent evidence shows that there are three different valid species. Ecological hypotheses are discussed to explain the benefits of these species remaining sympatric. © 2002 The Linnean Society of London, Biological Journal of the Linnean Society , 2002, 75 , 249–259.     

An overview of Syntrichia ruralis complex (Pottiaceae: Musci) in the Mediterranean region and neighbouring areas

The Syntrichia ruralis complex is revised in the Mediterranean region and neighbouring areas. A critical study of six quantitative and eight qualitative gametophytic characters from a total of 232 samples has been carried out. On the basis of this survey five taxa have been recognized. An identification key is provided. S. ruralis var. subpapillosissima is elevated to the rank of species as S. subpapillosissima . A lectotype for S. calcicola is proposed. S. ruralis var. submamillosa and S. ruralis var. glacialis are regarded as synonymous with S. subpapillosissima and S. ruralis , respectively. Also, Tortula densa is included in the variability shown by S. calcicola . Syntrichia ruralis var. substereidosa ( Tortula ruralis var. substereidosa ) is excluded from the Syntrichia ruralis complex and is included in the synonymy of S. virescens . © 2002 The Linnean Society of London, Botanical Journal of the Linnean Society , 138 , 209–224.     

Interaction between gfp-tagged Pseudomonas tolaasii P12 and Pleurotus eryngii

Pseudomonas sp., (formerly reported as strain P12) which produces brown blotch disease symptoms on Pleurotus eryngii, has been identified as P. tolaasii based on its biochemical, physiological properties and 16S rDNA sequence analysis. This pathogen is able to infect basidiocarps when surface-inoculated on mushroom casing soil. However, infected basidiocarps develop the brown blotch disease symptoms when the pathogen concentration in the fruiting body tissues is higher than 10(4) cfu/g d.w. Using gfp-tagged cells and confocal laser scanning microscopy, it was possible to show that the pathogen has the ability to tightly attach to the hyphae of Pleurotus eryngii.     

Morphological and pollen differentiation in Solanum cardiophyllum ssp. cardiophyllum and S. cardiophyllum ssp. ehrenbergii

A multivariate analysis to differentiate morphologically the populations of wild potatoes Solanum cardiophyllum ssp. cardiophyllum and S. cardiophyllum ssp. ehrenbergii was carried out. An analysis of the morphology and the viability of pollen of these potato populations was also made. The results of the morphometric analysis indicate that both subspecies are phenetically different. The pollen grain shape and size in ssp. cardiophyllum are different in northern and southern populations. Pollen diameter is significantly different between subspecies. Based on these results we propose that these taxa should be considered as two separate species.  © 2002 The Linnean Society of London, Botanical Journal of the Linnean Society , 2002, 140 , 415–426.     

Phylogeography of two sympatric giant flying squirrel subspecies,Petaurista alborufus lena and P. philippensis grandis (Rodentia: Sciuridae), in Taiwan

To test the association between Pleistocene forest dynamics relative to elevation and the population dynamics of arboreal small mammals, we examined the phylogeographical predictions for the genetic structure of the red and white giant flying squirrel (Petaurista alborufus lena) and the Indian giant flying squirrel (P. philippensis grandis) using complete mitochondrial control region sequences. Both giant flying squirrels are endemic subspecies to Taiwan and are sympatric in much of their range. In the phylogenetic analyses, we included 35 specimens of P. alborufus lena collected from 20 localities and five specimens with unknown sampling localities. Also, we phylogenetically examined 32 specimens of P. philippensis grandis from 18 localities and three specimens with unknown sampling localities. We identified 36 haplotypes of P. alborufus lena and 33 haplotypes of P. philippensis grandis. Although we did not recognize major phylogroups, we found several minor phylogroups in both subspecies, suggesting similar evolutionary histories. Phylogeographical and demographic tests showed distributions of these two subspecies expanded into coniferous and mixed forests that developed during glaciation in Taiwan's lowlands and middle lands. This suggests that these two Petaurista subspecies shifted elevation from mountainous areas to lowlands during glaciation. © 2011 The Linnean Society of London, Biological Journal of the Linnean Society, 2011, 102 , 404–419.     

Hybridization between Ischnura graellsii (Vander Linder) and 2. elegans (Rambur) (Odonata: Coenagrionidae): are they different species?

Two closely related damselflies, Ischnura graellsii and I. elegans , were analysed for morphological differences and reproductive isolation in the north coast of Galicia (NW Spain). We compared animals from sympatric and allopatric localities, including I. elegans from Belgium and I. graellsii from southern Spain as pure allopatric populations. A set of morphometric characters were studied by means of multivariate discriminant analysis to determine if these two species can be unambiguously distinguished. Discriminant analysis revealed that I. graellsii and I. elegans are well differentiated on the first two axis (86% and 11%, respectively). I. graellsii individuals are distinguished from I. elegans by their smaller size and, specifically, by their narrower and shorter wings and shorter tibiae. In addition, I. elegans has a narrower space between the branches of each cercus, and greater distance between the branches of each paraproct. Sympatric individuals are morphologically intermediate, suggesting hybridization. When the species were put together in the laboratory, they showed partial temporal separation in mating behaviour, but males of I. elegans readily mated with females of I. graellsii , and hybrid individuals were obtained. The opposite heterospecific cross was almost impossible, apparently because of mechanical problems with the tandem linkage. Laboratory-reared hybrids (from male I. elegans × female I. graellsii ) are morphologically intermediate, mainly resembling the maternal phenotype. Although hybridization between both taxa is common, we suggest maintaining the specific status for both phenotypes because they show incipient reproductive isolation, as it is reported in the literature. © 2002 The Linnean Society of London, Biological Journal of the Linnean Society , 2002, 76 , 225–235.     

Hybrid queen butterflies from the cross Danaus chrysippus (L.) × D. gilippus (Cramer):confirmation of species status for the parents and further support for Haldane's Rule

A cross between queen butterflies of the Palaeotropical species Danaus chrysippus and the Neotropical D. gilippus was achieved with difficulty in both directions. Only one progeny ( N  = 70) was reared comprising sterile males and inviable females in a precisely 1 : 1 ratio. Both prezygotic and postzygotic barriers to gene flow are strong. The result supports Haldane's Rule, to which we propose a minor amendment. The F₁ hybrids were intermediate for background colour between the brown (genotype BB ) of gilippus and orange (genotype bb ) of chrysippus . Most F₁ pattern characters were also intermediate. In polymorphic chrysippus populations, because Bb heterozygotes are brown, or nearly so, we suggest the B allele may have evolved towards dominance in sympatry. Hybrid males show positive heterosis for body size. The close similarity of male genitalia between the allopatric, genetically distant species chrysippus  and gilippus , compared to their divergence between gilippus and its largely sympatric sister species eresimus , suggest that reinforcement of sexual isolation or reproductive character displacement have evolved in sympatry.  © 2002 The Linnean Societyof London, Biological Journal of the Linnean Society , 2002, 76 , 535–544.