Genetic variation and species identification of Thai Boesenbergia (Zingiberaceae) analyzed by chloroplast DNA polymorphism

Genetic variation and molecular phylogeny of 22 taxa representing 14 extant species and 3 unidentified taxa of Boesenbergia in Thailand and four outgroup species (Cornukaempferia aurantiflora, Hedychium biflorum, Kaempferia parviflora, and Scaphochlamys rubescens) were examined by sequencing of 3 chloroplast (cp) DNA regions (matK, psbA-trnH and petA-psbJ). Low interspecific genetic divergence (0.25-1.74%) were observed in these investigated taxa. The 50% majority-rule consensus tree constructed from combined chloroplast DNA sequences allocated Boesenbergia in this study into 3 different groups. Using psbA-1F/psbA-3R primers, an insertion of 491 bp was observed in B. petiolata. Restriction analysis of the amplicon (380-410 bp) from the remaining species with Rsa I further differentiated Boesenbergia to 2 groupings; I (B. basispicata, B. longiflora, B. longipes, B. plicata, B.pulcherrima, B. tenuispicata, B. thorelii, B. xiphostachya, Boesenbergia sp.1 and Boesenbergia sp.3; phylogenetic clade A) that possesses a Rsa I restriction site and II (B.curtisii, B. regalis, B. rotunda and Boesenbergia sp.2; phylogenetic clade B and B. siamensis; phylogenetic clade C) that lacks a restriction site of Rsa I. Single nucleotide polymorphism (SNP) and indels found can be unambiguously applied to authenticate specie-origin of all investigated samples and revealed that Boesenbergia sp.1, Boesenbergia sp.2 and B. pulcherrima (Mahidol University, Kanchanaburi), B. cf. pulcherrima1 (Prachuap Khiri Khan) and B. cf. pulcherrima2 (Thong Pha Phum, Kanchanaburi) are B. plicata, B. rotunda and B. pulcherrima, respectively. In addition, molecular data also suggested that Boesenbergia sp.3 should be further differentiated from B. longiflora and regarded as a newly unidentified Boesenbergia species.     

Sphaerularia vespae sp. nov. (Nematoda, Tylenchomorpha, Sphaerularioidea), an endoparasite of a common Japanese hornet, Vespa simillima Smith (Insecta, Hymenoptera, Vespidae)

Sphaerularia vespae sp. nov., an endoparasite of a common Japanese hornet, Vespa simillima is described from Hokkaido, Japan, and its molecular sequence profiles are given. This newly discovered nematode appears to belong to the genus Sphaerularia, judging from its characteristic parasitic form, the uterium, which looks like a sausage with many verrucae on its surface. Sphaerularia vespae sp. nov. is distinguished from S. bombi, the only other nominal species of the genus, by the morphology of the male bursa, female tail, and anterior end of parasitic juveniles. SSU, ITS, and D2D3 (LSU) DNA sequences were determined for S. vespae sp. nov. and compared with those from related nematodes obtained from the GenBank database. The sequences from S. vespae sp. nov. were close to those of S. bombi and several another tylenchid entomoparasitic nematodes. Sphaerularia vespae sp. nov. parasitizes overwintering gynes of the hornet and practically sterilizes them, as S. bombi does for bumblebee gynes.     

铃刺蛾属在中国的首次发现及七新种记述（鳞翅目：刺蛾科）

首次报道铃刺蛾属Kitanola Matsumura及环铃刺蛾K. uncula (Staudinger)在中国有分布,并记述7新种,即针铃刺蛾K. spina sp. nov.,小针铃刺蛾K. spinula sp. nov.,线铃刺蛾K. linea sp. nov.,灰白铃刺蛾K. albigrisea sp. nov.,蔡氏铃刺蛾K. caii sp. nov.,短颚铃刺蛾K. brachygnatha sp. nov.和宽颚铃刺蛾K. eurygnatha sp. nov.。编制了本属中国已知种的检索表,提供了成虫彩色照片和外生殖器特征图。新种模式标本均保存在中国科学院动物研究所。     

基于DNA序列的白头翁属植物自然杂交证据

对长白山区兴安白头翁(Pulsatilla dahuricaSprengel)、朝鲜白头翁[P.cernua(Thunb.)Berchtold&Presl]以及其自然杂交种(P.dahurica×P.cernua)进行ITS和psbA-trnH序列分析,结果表明,自然杂交种与兴安白头翁遗传距离为0;ITS电泳显示,在6个信息位点的峰值同时具有朝鲜白头翁和兴安白头翁信息的双重峰,进一步证实前者是朝鲜白头翁和兴安白头翁的自然杂交种.     

Gilliamella intestini sp. nov., Gilliamella bombicola sp. nov., Gilliamella bombi sp. nov. and Gilliamella mensalis sp. nov.: Four novel Gilliamella species isolated from the bumblebee gut

Spectra of five isolates (LMG 28358^T, LMG 29879^T, LMG 29880^T, LMG 28359^T and R-53705) obtained from gut samples of wild bumblebees of Bombus pascuorum, Bombus lapidarius and Bombus terrestris were grouped into four MALDI-TOF MS clusters. RAPD analysis revealed an identical DNA fingerprint for LMG 28359^T and R-53705 which also grouped in the same MALDI-TOF MS cluster, while different DNA fingerprints were obtained for the other isolates.Comparative 16S rRNA gene sequence analysis of the four different strains identified Gilliamella apicola NCIMB 14804^T as nearest neighbour species. Average nucleotide identity values of draft genome sequences of the four isolates and of G. apicola NCIMB 14804^T were below the 96% threshold value for species delineation and all four strains and G. apicola NCIMB 14804^T were phenotypically distinct. Together, the draft genome sequences and phylogenetic and phenotypic data indicate that the four strains represent four novel Gilliamella species for which we propose the names Gilliamella intestini sp. nov., with LMG 28358^T as the type strain, Gilliamella bombicola sp. nov., with LMG 28359^T as the type strain, Gilliamella bombi sp. nov., with LMG 29879^T as the type strain and Gilliamella mensalis sp. nov., with LMG 29880^T as the type strain.     

中国冠果蝇属一新种和二新记录种及DNA条形码信息(双翅目:果蝇科)

基于形态学和分子遗传学证据,描述了冠果蝇属1新种Stegana(Stegana)helvippecta sp.nov.和2中国新记录种:S.(S.)furta(Linnaeus,1767)和 S.(S.)taba Okada,1971;利用41 条 DNA 条形码信息,分析了18种冠果蝇的遗传距离,并提供了中国产全部1...     

Evidence for a new lineage of primary ambrosia fungi in Geosmithia Pitt (Ascomycota: Hypocreales)

Geosmithia is a genus of mitosporic filamentous fungi typically associated with phloeophagous bark beetles world-wide. During this study, the fungal associates of ambrosia beetles Cnesinus lecontei, Eupagiocerus dentipes, and Microcorthylus sp. from Costa Rica, were studied using morphology and DNA sequences. Fungal associates belonged to four undescribed Geosmithia species. Geosmithia eupagioceri sp. nov. and G. microcorthyli sp. nov. are evidently primary ambrosia fungi of their respective vectors E. dentipes and Microcorthylus species. They both have convergently evolved distinct morphological adaptations including the production of large, solitary and globose conidia, and yeast-like cells. Tunnels of C. lecontei contained an undescribed Geosmithia species, but its nutritional importance for its vector is unclear. An auxiliary ambrosia fungus, Geosmithia rufescens sp. nov., was found associated with both G. eupagioceri and the Geosmithia species associated with C. lecontei. G. microcorthyli is genetically quite similar to the phloem-associated Geosmithia sp. 8 from Europe. Large differences in morphology between these two species suggest the rapid co-evolution resulting from the close symbiosis of the former with its beetle host. The ITS rDNA sequences of G. microcorthyli and Geosmithia sp. 8 were not diagnostic, suggesting that alternative markers such as EF-1α, IGS rDNA or β-tubulin should be used, together with morphological and ecological data, for species delimitation in this genus. The primary ambrosia fungi described here are derived from phloem-associated ancestors, and represent two independent lineages of ambrosia fungi in the Hypocreales and a new ecological strategy within Geosmithia.     

Identification of lactic acid bacteria from fermented tea leaves (miang) in Thailand and proposals of Lactobacillus thailandensis sp. nov., Lactobacillus camelliae sp. nov., and Pediococcus siamensis sp. nov

Eighteen rod-shaped homofermentatives, six heterofermentatives, and a coccal homofermentative lactic acid bacteria were isolated from fermented tea leaves (miang) produced in the northern part of Thailand. The isolates were placed in a monophyletic cluster consisting of Lactobacillus and Pediococcus species. They were divided into seven groups by phenotypic and chemotaxonomic characteristics, DNA-DNA similarity, and 16S rRNA gene sequences. Groups I to VI belonged to Lactobacillus and Group VII to Pediococcus. All of the strains tested produced DL-lactic acid but those in Group IV produced L-lactic acid. The strains tested in Groups I, II and V had meso-diaminopimelic acid in the cell wall. Six strains in Group I were identified as Lactobacillus pantheris; five strains in Group II as Lactobacillus pentosus; and four strains in Group V as Lactobacillus suebicus. Two strains in Group VI showed high DNA-DNA similarity for each other and MCH4-2 was closest to Lactobacillus fermentum CECT 562(T) with 99.5% of 16S rRNA gene sequence similarity. Five strains in Group III are proposed as Lactobacillus thailandensis sp. nov., and MCH5-2(T) (BCC 21235(T)=JCM 13996(T)=NRIC 0671(T)=PCU 272(T)) is the type strain which has 49 mol% G+C of DNA. Two strains in Group IV are proposed as Lactobacillus camelliae sp. nov., and the type strain is MCH3-1(T) (BCC 21233(T)=JCM 13995(T)=NRIC 0672(T)=PCU 273(T)) which has 51.9 mol% G+C of DNA. One strain in Group VII is proposed as Pediococcus siamensis sp. nov., and MCH3-2(T) (BCC 21234(T)=JCM 13997(T)=NRIC 0675(T)=PCU 274(T)) is the type strain which has 42 mol% G+C of DNA.     

中国太宇谷蛾属分类研究(鳞翅目,谷蛾科)(英文)

记述中国太宇谷蛾属Gerontha Walker10种,其中有4新种:拟华太宇谷蛾G.similihoenei sp.nov.,喙太宇谷蛾G.rostriformis sp.nov.,梯缘太宇谷蛾G.trapezia sp.nov.,褶太宇谷蛾G.rugulosa sp.nov.和3新纪录种:暹罗太宇谷蛾G.siamensis Moriuti,1989(图7,17～18)、清迈太宇谷蛾G.nawapuriensis Moriuti,1989(图8,19)、钻太宇谷蛾G.borea Moriuti,1977(图9,20).首次发现和报道了弯茎太宇谷蛾Gerontha flexura Huang et al .(图6,15-16)和暹罗太宇谷蛾G.siamensis Moriuti的雌性个体.文中给出了所有中国种的检索表.研究标本保存于南开大学生命科学学院昆虫标本室.     

中国水螟亚科二新种记述(鳞翅目:草螟科)

记述了采自天津及河北、福建等地的水螟亚科Nymphulinae 2新种, 即 环波水螟Paracymoriza convallata, sp. nov.和白筒水螟Parapoynx candida, sp. nov..模式标本保存于南开大学生物系, 天津.1. 环波水螟Paracymoriza convallata,新种(图1,3)前翅长♂9.0 mm.正模♂, 福建永泰县青云山 (25.52°N,118.57°E),550 m,2002-Ⅸ-18,王新谱采.新种与华南波水螟Paracymoriza laminalis (Hampson) 相似,主要区别在新种前翅中线外白区前缘封闭;雄性外生殖器抱器瓣基部较窄,端部较圆.2. 白筒水螟Parapoynx candida,新种(图2,4～5)前翅长♂11.5～12.0 mm;♀11.5 mm.正模♂,天津武清县 (39.31°N,116.51°E),1976-Ⅵ-22, 王印来采.副模1♂,天津市板桥农场(38.52°N,117.28°E),1976-Ⅵ(采集人不明);1♀,河北大城县 (38.40°N,116.39°E),1976-Ⅵ-12 (采集人不明).此新种与重筒水螟Parapoynx stratiotata (Linnaeus) 相近,区别在于新种外横线至翅外缘白色;雌性外生殖器无明显囊突.     

Transfer of Kaempferia candida to Curcuma (Zingiberaceae) based on morphological and molecular data

Results of morphological studies supported by molecular analyses of chloroplast psbA‐trnH and partial petA‐psbJ DNA sequences indicate that Kaempferia candida cannot be retained in this genus. The species is here formally transferred to Curcuma and the name is typified. Notes on the species and colour plates are given.     

鸡爪槭种下分类群的DNA条形码筛选

槭属鸡爪槭(Acer palmatum Thunb.)是北温带广泛分布的一类园艺观赏树种,由于频繁的种内杂交渐渗导致其种下分类群的形态性状特征趋同,致使传统的形态学分类难以准确鉴定,新兴的DNA条形码技术为快速、准确的鉴定鸡爪槭种下分类群提供了新的思路。本研究采用5个叶绿体DNA片段(rpl16、psbA-trnH、trnL-trnF、rbcL、matK)和核基因组ITS片段,运用PWG-distance和Tree-Building两种方法对鸡爪槭的8个分类群共32个个体进行DNA条形码分析。结果显示,单个叶绿体基因组片段(分辨率为0%~25%)或核rDNA ITS片段(12.5%)的分辨率较低,不同组合的叶绿体DNA片段(0%~62.5%)、叶绿体片段与核rDNA ITS片段(12.5%~50%)的分辨率则相对较高。其中,rpl16+psbA-trnH+trnL-trnF片段组合的分辨率最高(62.5%),较为符合DNA条形码快速、准确鉴定的要求,因此建议将其作为鸡爪槭种下分类群鉴定的DNA条形码。     

凹唇姜属(Boesenbergia O.Kuntze)和山柰属(Kaempferia L.)(姜科)叶的解剖学

通过凹唇姜属（Boesenbergia curtisii,B.Prainana,B.rotunda和B.plicata)和山柰属（Kaempferia pulchra,D.galanga.K.gilbertii,K.rotunda,K.parviflora和K.angustifolia)种间叶解剖学变化的研究,寻找能用于鉴别种的解剖学特征,结果显示变化表现在气孔的类型,中脉的结构,叶缘和叶柄横切面轮廓,叶片的远 轴面或近轴面下皮层和毛状体的出现或缺如,研究表明这些特征的联合对已研究的种的鉴别是有用的。     

DNA barcoding and phylogenetic analysis of Pectinidae (Mollusca: Bivalvia) based on mitochondrial COI and 16S rRNA genes

DNA sequence data enable not only the inference of phylogenetic relationships but also provide an efficient method for species-level identifications under the terms DNA barcoding or DNA taxonomy. In this study, we have sequenced partial sequences of mitochondrial COI and 16S rRNA genes from 63 specimens of 8 species of Pectinidae to assess whether DNA barcodes can efficiently distinguish these species. Sequences from homologous regions of four other species of this family were gathered from GenBank. Comparisons of within and between species levels of sequence divergence showed that genetic variation between species exceeds variation within species. When using neighbour-joining clustering based on COI and 16S genes, all species fell into reciprocally monophyletic clades with high bootstrap values. These evidenced that these scallop species can be efficiently identified by DNA barcoding. Evolutionary relationships of Pectinidae were also examined using the two mitochondrial genes. The results are almost consistent with Waller’s classification, which was proposed on the basis of shell microstructure and the morphological characteristics of juveniles.     

Chloroplast inheritance patterns in Actinidia hybrids determined by single stranded conformation polymorphism analysis

The inheritance patterns of the chloroplast genomes of the Actinidia hybrids A. eriantha (male parent) x A. chinensis (female parent) and A. chinensis (male parent) x A. melanandra (female parent) were analyzed using single-strand conformation polymorphism (SSCP) analysis of the trnL-trnF and psbA-trnH intergenic spacers. This showed that the artificial hybrids between A. eriantha and A. chinensis all had the haplotype of their male parent. Alignment of the sequences of A. eriantha and A. chinensis revealed four substitutions and one insertion (GATTC) in trnL-trnF and two substitutions in psbA-trnH. In contrast, the haplotypes of the artificial hybrids between A. chinensis and A. melanandra had the same patterns as their female parent. Alignment of the entire region of A. chinensis and A. melanandra revealed 12 substitutions: 1 in trnL-trnF and 11 in psbA-trnH. However, no sequence variation in the trnL-trnF and psbA-trnH intergenic spacers was found. We have developed a simple screening method for detecting the inheritance patterns of Actinidia chloroplast DNA haplotypes using SSCP analysis of the trnL-trnF and psbA-trnH intergenic spacers. Our findings indicate that the inheritance of the chloroplast genome in Actinidia hybrids differs according to the species selected.     

Four new species of the genusSporobolomyces isolated from leaves in Thailand

Thirteen undescribed strains of ballistoconidium-forming yeasts, isolated from leaves collected in the suburbs and along the southeast seacoast of Bangkok, Thailand, were divided into four different groups in the genusSporobolomyces on the basis of morphological, physiological, biochemical, and chemotaxonomical characteristics, and analyses of the sequences of 18S rDNA and internal transcribed spacer regions. DNA-DNA reassociation experiments with related species revealed that these four groups were four new distinct species.Sporobolomyces nylandii sp. nov.,S. poonsookiae sp. nov.,S. blumeae sp. nov. andS. vermiculatus sp. nov. are proposed for these strains.     

帘蛤科贝类rDNA内转录间隔区序列的研究

根据18SrDNA、5．8SrDNA和28SrDNA保守序列设计引物,应用聚合酶链式反应（PCR）扩增了文蛤（Meretrix meretrix L．）、青蛤（Cyclina sinensis G）、硬壳蛤（Mercenaria mercenaria L．）和江户布目蛤（Protothaca jedoensis L．）4种帘蛤科贝类的第一内转录间隔区（ITS1）和第二内转录间隔区（ITS2）序列,并进行了测序。结果表明,文蛤、青蛤、硬壳蛤和江户布目蛤的ITS1扩增产物大小分别为978bp、663bp、757bp和942bp,GC含量分别为61．55％、60．78％、62．48％和64．86％～64．97％,其中ITS1序列长度分别为900bp、585bp、679bp和864bp,是迄今已报道双壳贝类中变化范围最大的,GC含量分别为61．67％、61．03％、63．03％和65．51％～65．62％,江户布目蛤种内ITS1序列有个体差异;ITS2扩增产物大小分别为644bp、618～620bp、593bp和513～514bp,GC含量分别为61．18％、61．29％～61．81％、62．73％和61．48％61．60％,其中ITS2序列长度分别为412bp、386～388bp、361bp和281～282bp,GC含量分别为65．29％、65．21％～66．06％、67．87％和67．38％～67．62％,青蛤和江户布目蛤种内ITS2序列有个体差异。4种蛤ITS1和ITS2序列种间差异很大,有明显的长度多态性,ITS2种间序列相似度73．0％～89．1％,与ITS1的种间序列相似度48．7％～81．5％相比略高。此外,在4种蛤ITS1和ITS2序列中各发现2个与rRNA加工有关的保守区。通过对ITS1和ITS2序列的组装获得了4种蛤5．8SrRNA基因完整序列,序列长度都是157bp,GC含量57．96％～58．60％,4种蛤5．8SrRNA基因相对保守,种间序列差异度0-6．0％,共有10个变异位点,其中转换4处,颠换6处,硬壳蛤和江户布目蛤5．8SrRNA基因序列完全相同。以ITS2序列（包含5．8SrRNA和28SrRNA基因部分序列）为标记,调用北极蛤科的Arctica islandica相应序列数据作外群,构建了帘蛤科贝类的系统发育树,其拓扑结构显示江户布目蛤与硬壳蛤亲缘关系最近,青蛤与其他3物种的亲缘关系最远。