苦苣苔科植物远缘杂交亲和性初探

该研究对苦苣苔科的蛛毛苣苔属[Paraboea(C.B.Clarke)Ridley]、马铃苣苔属(Oreocharis Bentham)、报春苣苔属(Primulina Hance)、吊石苣苔属(Lysionotus D.Don)、台闽苣苔属(Titanotrichum Solereder)、半蒴苣苔属(Hemiboea C.B.Clarke)、长蒴苣苔属(Didymocarpus Wallich)、光叶苣苔属(Glabrella Mich.MollerW.H.Chen)进行属间远缘杂交,对马铃苣苔属的马铃苣苔组、川滇马铃苣苔组、大叶石上莲组进行组间远缘杂交,统计不同杂交组合的结实情况及播种后的种子萌发情况,以明确不同属(组)间远缘杂交亲和性,为通过远缘杂交进行苦苣苔科种质资源的创新奠定基础。结果表明:(1)不同属作为父本,与同一母本进行杂交,以及同一属内不同种作为父本,与同一母本进行杂交,结实情况差异显著。(2)不同属作为母本,与同一父本进行杂交,以及同一属内不同种作为母本,与同一父本进行杂交,结实情况同样存在显著差异。(3)相同两个种正反交杂交成功率也并不一致。(4)马铃苣苔属与半蒴苣苔属,报春苣苔属与吊石苣苔属,吊石苣苔属与报春苣苔属和半蒴苣苔属,光叶苣苔属与报春苣苔属和吊石苣苔属之间部分种进行远缘杂交能够正常结实,且杂交种子能够正常萌发。(5)马铃苣苔属内组间杂交无明显生殖隔离的现象。     

Molecular phylogenetic relationships of China Seas groupers based on cytochrome <Emphasis Type="Italic">b</Emphasis> gene fragment sequences

The classification and evolutionary relationships are important issues in the study of the groupers. Cytochrome b gene fragment of twenty-eight grouper species within six genera of subfamily Epinephelinae was amplified using PCR techniques and the sequences were analyzed to derive the phylogenetic relationships of the groupers from the China Seas. Genetic information indexes, including Kimura-2 parameter genetic distance and T _S/T _V ratios, were generated by using a variety of biology softwares. With Niphon spinosus, Pagrus major and Pagrus auriga as the designated outgroups, phylogenetic trees, which invoke additional homologous sequences of other Epinephelus fishes from GenBank, were constructed based on the neighbor-joining (NJ), maximum-parsimony (MP), maximum-likelihood (ML) and minimum-evolution (ME) methods. Several conclusions were drawn from the DNA sequences analysis: (1) genus Plectropomus, which was early diverged, is the most primitive group in the subfamily Epinephelinae; (2) genus Variola is more closely related to genus Cephalopolis than the other four genera; (3) genus Cephalopolis is a monophyletic group and more primitive than genus Epinephelus; (4) Promicrops lanceolatus and Cromileptes altivelis should be included in genus Epinephelus; (5) there exist two sister groups in genus Epinephelus.     

桑黄孔菌属的研究进展

历来"桑黄"的种类混淆不清,直至近年来才被确定为桑树桑黄,与其亲缘性相近的物种也一同被划分入广义纤孔菌属的新分支—桑黄孔菌属。本文整理曾被当作"桑黄"的物种,阐述其正名和桑黄孔菌属确立的过程,对目前该属内物种的生物活性和栽培研究进展进行综述,旨在为桑黄孔菌属真菌资源的研究开发提供参考。     

Incidence and diversity of viruses in cowpeas and weeds in the unmanaged farming systems of savanna zones in Nigeria

Fields surveys were conducted to assess the incidence of commonly known legume viruses on cowpeas and weed hosts within and around the cowpeas farms in nine locations across the three agro-ecological zones of Nigeria. Of 315 cowpea leaf samples collected and tested for eight viruses, 69.5% were found to be infected. Bean common mosaic virus-blackeye mosaic (BCMV-BlCM), genus Potyvirus had the highest incidence (70%) and was also the most prevalent (78%). Cowpea aphid-borne mosaic virus (CABMV, genus Potyvirus) had 64% as incidence, incidence of Southern bean mosaic virus (SBMV, genus Sobemovirus) was 21%. Bean pod mosaic virus (BPMV, genus Comovirus) was detected in 1% of the samples tested. Cowpea mosaic virus (CPMV, genus Comovirus) was undetected. Other viruses tested included Cowpea mottle virus (CPMoV, genus Carmovirus), Cucumber mosaic virus (CMV, genus Cucumovirus), and Cowpea mild mottle virus (CPMMV, genus Carlavirus). Multiple virus infections were detected in 68.0% of the infected cowpea leaf samples. The combination of BCMV-BlCM and CABMV was the most common, occurring in 76.4% of all samples. Virus incidence in weeds around the cowpea plots was 2.5% (9 out of 356) whereas 1.5% (5 out of 332) of the weeds collected within the cowpea plots were infected. Some of the weeds infected were Chromoleana odorata, Centrosema sp., Thithonia diversifolia and Talinum triangulare.     

Evaluation of Lespedeza germplasm genetic diversity and its phylogenetic relationship with the genus Kummerowia

The genetic diversity of the genus Lespedeza is not well known and the phylogenetic relationship of Lespedeza with the genus Kummerowia is unclear. We report the first study in which polymorphic expressed sequence tag-simple sequence repeat (EST-SSR) markers derived from Medicago, cowpea and soybean were used to assess the genetic diversity of the USDA Lespedeza germplasm collection and clarify its phylogenetic relationship with the genus Kummerowia. Phylogenetic analysis partitioned 44 Lespedeza accessions into three main groups some of which were species-specific and eight subgroups. This data set revealed some misidentified accessions, and indicated that the two species in the genus Kummerowia are closely related to the genus Lespedeza. Morphological reexamination was used to correct the misidentified accessions within the genus Lespedeza. Our results demonstrated that phylogenetic analysis with morphological reexamination provides a more complete approach to classify accessions in plant germplasm collection and conservation.     

Studies on antagonistic marine actinomycetes from the Bay of Bengal

Screening of 26 marine sediment samples near 9 islands of the Andaman Coast of the Bay of Bengal resulted in the isolation of 88 isolates of actinomycetes. On the basis of sporophore morphology and structure of the spore chain, 64 isolates were assigned to the genus Streptomyces, 8 isolates to the genus Micromonospora, 5 to the genus Nocardia, 7 to the genus Streptoverticilium and 4 to the genus Saccharopolyspora. Among 64 Streptomyces spp., 44 isolates showed antibacterial activity and 17 isolates showed antifungal activity. Three isolates showed very promising antagonistic activities against multi-drug resistant pathogens.     

The Phylogeny of Acetic Acid Bacteria Based on the Partial Sequences of 16S Ribosomal RNA: The Elevation of the Subgenus Gluconoacetobacter to the Generic Level

Thirty-six strains of acetic acid bacteria classified in the genera Acetobacter, Gluconobacter, and Acidomonas were examined for their partial base sequences in positions 1220 through 1375, 156 bases, of 16S rRNA. The strains of the Q₁₀-equipped Gluconobacter species examined were divided into two subgroups, which included the type strains of Gluconobacter oxydans, the type species of the genus Gluconobacter, and of a second species, Gluconobacter cerinus, respectively. The base differences numbered four between the two type strains. The strains of the Q₉-equipped species examined classified in the type subgenus Acetobacter of the genus Acetobacter were not very distant phylogenetically from those of the genus Gluconobacter. The calculated number of base differences was 9–6 between the type strains of G. oxydans and G. cerinus and the type strains of Acetobacter aceti and Acetobacter pasteurianus. In contrast, the strains of the Q₁₀-equipped species examined classified in the subgenus Gluconoacetobacter of the genus Acetobacter were very distant phylogenetically from those of the Acetobacter and Gluconobacter species mentioned above. The number of base differences was calculated to be 14-8. Furthermore, the strains of the methanol-assimilating, Q₁₀-equipped species of the genus Acidomonas examined were located in phylogenetically isolated positions. The type strain of Acidomonas methanolica (≡ Acetobacter methanolicus), the type species of the genus Acidomonas, had 16–9 base differences. The data obtained here indicated that the members of the subgenus Gluconoacetobacter of the genus Acetobacter can be distinguished at the generic level. The new genus Gluconoacetobacter was proposed with the type species, Gluconoacetobacter liquefaciens, in recognition of the genus Acidomonas along with the genera Acetobacter and Gluconobacter in the classification of the acetic acid bacteria.     

Molecular phylogenetic relationships of China Seas groupers based on cytochrome b gene fragment sequences

The classification and evolutionary relationships are important issues in the study of the groupers. Cytochrome b gene fragment of twenty-eight grouper species within six genera of subfamily Epinephelinae was amplified using PCR techniques and the sequences were analyzed to derive the phylogenetic relationships of the groupers from the China Seas. Genetic information indexes, including Kimura-2 parameter genetic distance and T _S/T _V ratios, were generated by using a variety of biology softwares. With Niphon spinosus, Pagrus major and Pagrus auriga as the designated outgroups, phylogenetic trees, which invoke additional homologous sequences of other Epinephelus fishes from GenBank, were constructed based on the neighbor-joining (NJ), maximum-parsimony (MP), maximum-likelihood (ML) and minimum-evolution (ME) methods. Several conclusions were drawn from the DNA sequences analysis: (1) genus Plectropomus, which was early diverged, is the most primitive group in the subfamily Epinephelinae; (2) genus Variola is more closely related to genus Cephalopolis than the other four genera; (3) genus Cephalopolis is a monophyletic group and more primitive than genus Epinephelus; (4) Promicrops lanceolatus and Cromileptes altivelis should be included in genus Epinephelus; (5) there exist two sister groups in genus Epinephelus. These authors contributed equally to this work.     

Phylogenetic Relationships of Species of the Genus Kluyveromyces van der Walt (Saccharomycetaceae) Deduced from the Partial Sequences of 18S and 26S Ribosomal RNAs

In order to clarify the phylogenetic relationships of the species classified in the genus Kluyveromyces (Saccharomycetaceae), three partial base sequences of 18S and 26S rRNAs of eighteen strains were determined. The regions determined of the strains corresponded to positions 1451 through 1618 (168 bases) of 18S rRNA and to positions 1611 through 1835 (225 bases) and 493 through 622 (130 bases) of a strain (IFO 2376) of Saccharomyces cerevisiae. The analyses of the partial base sequences suggested that the genus Kluyveromyces is phylogenetically heterogeneous, ranging from the strains that are quite close to the strain of S. cerevisiae to the strains that are distinct enough to be classified in genera separate from the genus Saccharomyces. From our sequence data, we concluded that the extent of the genus Kluyveromyces should be restricted to only one species, K. polysporus, the type species of the genus. Kluyveromyces phaffii was also distinct enough to deserve another genus. Kluyveromyces cellobiovorus was not close to any of the strains of Kluyveromyces species examined, and should be excluded from the genus. Most of the strains of the species examined were fairly close to the strain of S. cerevisiae.     

纤维单胞菌属放线菌的研究进展

纤维单胞菌属(Cellulomonas)的一些菌株能够产生多种纤维素酶,在纤维素降解方面显示出明显优势。1923年Bergey等以产黄纤维单胞菌为模式菌建立了纤维单胞菌属。1991年Stackebrand和Prauser又以纤维单胞菌属为模式属建立了纤维单胞菌科(Cellulomonadaceae)。目前,纤维单胞菌属包含有从多种环境中分离培养得到的26个有效描述种。纤维单胞菌属菌株在分类学上的典型特征是:细胞壁的肽聚糖成分主要含有Orn和Glu/Asp,以MK-9(H4)为主要的甲基萘醌,主要的脂肪酸成分为anteiso-C15:0和C16:0,极性脂成分主要包括双磷脂酰甘油(DPG)和磷脂酰肌醇甘露糖甙(PIM)。基因组DNA的G+C含量为(68.5–76.0)mol%。最近,本实验室分离到2株纤维单胞菌,应用多相分类研究手段确定了他们的分类学地位。本文将结合我们的研究,对纤维单胞菌属的建立、分类学特征及其在生态和酶资源应用方面进行综述。     

Molecular phylogeny of Phalaenopsis Blume (Orchidaceae) based on the internal transcribed spacer of the nuclear ribosomal DNA

The internal transcribed spacer (ITS1, 5.8S rDNA, and ITS2) region of nuclear ribosomal DNA (nrDNA) was sequenced from 53 species, which represent most of the living species diversity in the genus Phalaenopsis (Orchidaceae). A phylogeny was developed for the genus based on the neighbor-joining and maximum parsimony analyses of molecular data. Results of these analyses provided support for the monophyly of the genus Phalaenopsis and concurred in that the genera Doritis and Kingidium should be treated as being parts of the genus Phalaenopsis as suggested by Christenson (2001). Within the genus Phalaenopsis, neither subgenera Aphyllae nor Parishianae were monophyletic, and they were highly clustered with subgenus Proboscidioides plus sections Esmeralda and Deliciosae of subgenus Phalaenopsis based on ITS data. Those species also have the same characters of morphology of four pollinia and similar biogeographies. Furthermore, neither subgenus Phalaenopsis nor Polychilos was monophyletic. Within the subgenus Phalaenopsis, only section Phalaenopsis was highly supported as being monophyletic. As for the subgenus Polychilos, only section Polychilos was moderately supported as being monophyletic. In conclusion, the present molecular data obtained from the ITS sequence of nrDNA of the genus Phalaenopsis provide valuable information for elucidating the phylogeny of this genus.     

小月菌属放线菌的研究进展

小月菌属(Microlunatus)菌株在治理磷元素造成的环境污染方面呈现优势,具有广阔的研究与开发前景。1995年Nakamura等以积磷小月菌(Microlunatus phosphovorus)为模式种建立了小月菌属。目前,该属包含7个有效描述种,分离自多种生态环境。小月菌属在分类学上的典型特征是:细胞壁含有LL-二氨基庚二酸,优势甲基萘醌成分为MK-9(H_4),磷酸类脂主要包括双磷脂酰甘油和磷脂酰甘油。本文结合我们对新近分离得到的2株小月菌属菌株的分类研究结果和相关文献资料,就小月菌属的建立、分类学特征、属内各成员的分布、及其在化工和医药业上的应用研究进展和开发前景进行了综述。     

NEW CHROMOSOME COUNTS FOR CHUSQUEA AND AULONEMIA (POACEAE: BAMBUSOIDEAE)

Tetraploid chromosome counts of n = 20 are reported for two previously cytologically unknown South American species of Chusquea, C. capitulifora and C. scandens. Diploid counts were previously unknown in this genus, but C. talamancensis of Costa Rica has n = 10. No previous chromosome counts were known for the genus Aulonemia nor any other genus of the subtribe Arthrostylidiinae. Aulonemia pumila and A. amplissima from South America are tetraploid with n = 20.     

Phylogeny of <Emphasis Type="Italic">Litsea</Emphasis> and related genera (Laureae-Lauraceae) based on analysis of <Emphasis Type="Italic">rpb2</Emphasis> gene sequences

The relationship between Litsea and related genera is currently unclear. Previous molecular studies on these taxa using cpDNA and nrITS were unable to produce well-resolved phylogenetic trees. In this study, we explored the potential of the rpb2 gene as a source of molecular information to better resolve the phylogenetic analysis. Although rpb2 was believed to be a single-copy gene, our cloning results showed that most species examined possessed several copies of these sequences. However, the genetic distance among copies from any one species was low, and these copies always formed monophyletic groups in our molecular trees. Our phylogenetic analyses of rpb2 data resulted in better resolved tree topologies compared to those based on cpDNA or nrITS data. Our results show that monophyly of the genus Litsea is supported only for section Litsea. As a genus, Litsea was shown to be polyphyletic. The genera Actinodaphne and Neolitsea were resolved as monophyletic groups in all analyses. They were also shown to be sisters and closer to the genus Lindera than to the genus Litsea. Our results also revealed that the genus Lindera is not a monophyletic group.     

Phylogenetic relationships of the anamorphic genus <Emphasis Type="Italic">Fusicladium</Emphasis> s. lat. as inferred by ITS nrDNA data

The genus Fusicladium s. lat. (incl. Pollaccia and Spilocaea) was phylogenetically analysed using ITS nrDNA sequences. Pollaccia and Spilocaea did not form monophyletic groups of their own, but were intermingled between Fusicladium species, together with which they formed a monophyletic clade. Thus, Pollaccia and Spilocaea should be included in a wider genus concept of Fusicladium, constituting a morphologically variable genus. Furthermore, all Venturia and Fusicladium isolates clustered together on the bases of available ITS data, providing support for the monophyly of the anamorphic genus Fusicladium and the teleomorphic genus Venturia. Within this clade several subclades can be recognized. All taxa on the host family Salicaceae were found in one subclade. Three other subclades comprised taxa on Rosaceae whereas taxa on other host families all clustered separately. Geographic specializations were not observed. Two examples of host switching could be demonstrated, but these were confined to instances involving host species belonging to the same family. Fusicladium convolvularum and F. effusum, two species with unknown teleomorphs, clustered within the Fusicladium/Venturia clade, supporting the correct placement of these taxa in Fusicladium. The placement of Pseudocladosporium hachijoense within the family Venturiaceae was also supported.     

Isolation and characterization of salmonid telomeric and centromeric satellite DNA sequences

Satellite DNA clones with a 37 bp repeat unit were obtained from BglII-digested genomic DNA of Masu salmon (Oncorhynchus masou) and Chum salmon (O. keta). Fluorescence in situ hybridization (FISH) analysis with the isolated clones as a probe showed that these repetitive sequences were localized in the telomeric regions of chromosomes in both species. Southern and dot blot analyses suggested conservation of homologous sequences with similar repeat unit in other salmonids including the species of the genus Oncorhynchus and Salvelinus, but lack or scarcity of such sequences in the genus Hucho and Salmo. Similarly, polymerase chain reaction (PCR)-based cloning of satellite DNA referring to a reported Rainbow trout (O. mykiss) centromeric sequence was successful for the Oncorhynchus, Salvelinus and Hucho species. The obtained satellite DNA clones were localized with FISH in the centromeric regions of chromosomes of the species from these three genera. Although PCR cloning of the centromeric satellite DNA had failed in the Salmo species due to some base changes in the priming sites, dot blot hybridization analysis suggested conservation of homologous satellite DNA in the genus Salmo as in the other three genera. In the neighbor-joining tree of cloned centromeric satellite DNA sequences, the genus Oncorhynchus and Salvelinus formed adjacent clades, and the clade of the genus Hucho included the reported centromeric sequence of the genus Salmo. Conservation pattern and molecular phylogeny of the telomeric and centromeric satellite DNA sequences isolated herein support a close phylogenetic relationship between the genus Oncorhynchus and Salvelinus and between the Salmo and Hucho.     

The Phylogenetic Relationships of the Saturn-shaped Ascospore-forming Species of the Genus Williopsis Zender and Related Genera Based on the Partial Sequences of 18S and 26S Ribosomal RNAs (Saccharomycetaceae): The Proposal of Komagataea Gen. Nov.

The partial base sequences of 18S and 26S rRNAs of strains of Williopsis and Saturnospora species were analyzed. In the three regions partially sequenced, the higher base differences were observed in the strains examined of the three species, W. californica, W. mucosa, and W. pratensis, compared with those of W. saturnus var. saturnus (type species of genus Williopsis), W. beijerinckii, W. mrakii, W. saturnus var. subsufficiens, W. suaveolens, P. membranaefaciens (type species of genus Pichia), C. matritensis (type species of genus Citeromyces), and S’spora dispora (type species of genus Saturnospora): the percent similarities were 52–82 in positions 493–622, 130 bases, of 26S rRNA, and the number of base differences was 28–6 in positions 1611–1835, 225 bases, of 26S rRNA, and the number of base differences was 25–4 in positions 1451–1618, 168 bases, of 18S rRNA. In the 18S rRNA partial base sequencings, W. mucosa had an identical base sequence with P. anomala (≡ H. anomala, type species of genus Hansenula). Based on the sequence data obtained, the taxonomic positions of the three Williopsis species mentioned above are discussed. The genus Zygowilliopsis Kudriavzev was postulated to be retained and emended, and a new genus, Komagataea was proposed for W. pratensis with a new combination, Komagataea pratensis.     

Oligonucleotide Probes for the Detection of Representatives of the Genus Thermoanaerobacter

Based on the analysis of 16S rRNA nucleotide sequences, oligonucleotide probes were designed for the detection of representatives of the genus Thermoanaerobacter. To increase the specificity of detection, the genus Thermoanaerobacter was divided into three groups. The probe Tab 827 (5"-GCTTCCGCDYCCCACACCTA-3") detected all known representatives of the genus Thermoanaerobacter; the probe Tab_1 844 (5"-TTAACTACGGCACGRAATGCTTC-3") was specific for the first group of species of the genus (T. wiegelii, T. siderophilus, T. sulfurophilus, T. brockii, T. kivui, T. ethanolicus, T. acetoethylicus, and T. thermohydrosulfuricus); the probe Tab_2 424 (5"-CACTAMYGGGGTTTACAACC-3") targeted the second group (T. thermocopriae, T. mathranii, and T. italicus); and the probe Tab_3 184 (5"-TCCTCCATCAGGATGCCCTA-3") was specific for the third group (T. tengcongensis, T. yonseiensis, T. subterraneus, and Carboxydibrachium pacificum, an organism related to the genus Thermoanaerobacter according to its 16S rRNA sequence). The oligonucleotide probes were labeled with Dig-11-dUTP. Hybridization with the probes showed the affiliation with Thermoanaerobacter of several pure cultures that were morphologically similar to representatives of this genus but possessed metabolic features unusual for it (capacity for agarose hydrolysis, anaerobic oxidation of CO, growth at low pH values) or were isolated from habitats previously unknown for Thermoanaerobacter (deep-sea hydrothermal vents).     

南海东海岛近海固氮细菌多样性及对菜心的促生作用

海洋固氮细菌在自然界氮循环中发挥着重要作用,筛选和开发海洋固氮促生的菌种资源,对于生物菌肥的开发应用和农业生产具有重要意义。[目的]研究海洋固氮细菌的生物多样性及对陆地作物的促生作用,筛选优良的植物根际促生菌株。[方法]通过形态特征、生理生化试验和16S rRNA基因序列比对进行菌属鉴定;将解磷、解钾、产蛋白酶和纤维素酶等性能优良的菌株作为菜心盆栽试验的组合菌液,探究对菜心能否起促生作用。[结果]本研究从南海东海岛的海岛沉积物中筛选出18株固氮菌,分布在6个属9种,不动杆菌属4株,假交替单胞菌属1株,芽孢杆菌属8株,嗜冷杆菌属1株,海单胞菌属1株,交替单胞菌属3株。菜心幼苗经过组合菌剂的浇灌,在茎高、茎粗、最大叶宽和最大叶长4个指标均表明对菜心有显著的促生作用。其中,以芽孢杆菌属和不动杆菌属的菌株在菜心的生长过程中起关键的促进作用,对菜心的促生性能最佳。[结论]南海近海具有种类丰富多样的固氮细菌,以芽孢杆菌属和不动杆菌属的菌株促生作用最为显著,具有开发成微生物菌肥的潜力,为优良的海洋促生微生物菌种资源的定向利用及蔬菜的无公害生产提供重要依据。