Extrachromosomal DNA in Bacillus thuringiensis var. kurstaki, var. finitimus, var. sotto, and in Bacillus popilliae

Four entomopathogenic bacteria contained extrachromosomal deoxyribonucleic acid (DNA) molecules of various sizes. Bacillus thuringiensis var. kurstaki contained twelve elements banding on agarose gels that ranged from 0.74 to > 50 × 10⁶ daltons, three of which were giant extrachromosomal DNA elements. B. thuringiensis var. sotto contained one giant extrachromosomal DNA element with a molecular size of about 23.5 × 10⁶ daltons and two lesser elements of 0.80 and 0.62 × 10⁶ daltons. B. thuringiensis var. finitimus harbored two giant DNA elements corresponding to >50 × 10⁶ daltons and two lesser bands with relative small size (0.98 and 0.97 × 10⁶ daltons). B. popilliae contained no giant extrachromosomal DNA elements but did contain two smaller elements corresponding to 4.45 and 0.58 × 10⁶ daltons. The possible use of extrachromosomal DNA elements that prove to be autonomous replicons for recombinant DNA studies is discussed.     

Unveiling Gymnosporangium corniforme, G. unicorne, and G. niitakayamense sp. nov. in Taiwan

Three rust fungi from high mountains and pear-producing areas in Taiwan were described using morphological and molecular data based on 34 specimens. Gymnosporangium corniforme was demonstrated to produce spermogonia and aecia on Sorbus randaiensis based on molecular analyses and inoculation experiments. The pear rust pathogen G. unicorne was discovered in Taiwan for the first time. Gymnosporangium niitakayamense sp. nov. was observed on the leaves of Photinia niitakayamensis. It was distinct from other species in peridial cell wall structures, i.e., smooth outer wall, rugose side wall, and coralloid projections on the inner wall, and in having echinulate aeciospores.     

Three sesquiterpene lactones from Helianthus niveus subsp. canescens and H. argophyllus

Further investigation of the minor terpenoids in Helianthus niveus subsp. canescens and H. argophyllus afforded two known 12,8-fused sesquiterpene lactones, 3-acetylchamissonin and simsiolide as well as one new germacranolide, argophyllin C. Identification and structure elucidation of these lactones by ¹H NMR and chemical transformations are described.     

Pterocarpans of Neorautanenia edulis and N. Amboensis

The root bark of Neorantanenia edulis and N. amboensis yielded six new natural pterocarpanoids, the structures of which have been determined mainly by physical methods and by partial syntheses. They include edulane, edudiol, neoraucarpan, neorautanin, edulenol, and neoraucarpanol.     

Furanosesquiterpenoids of Commiphora erythraea and C. myrrh

The isolation and structure determination of two new furanosesquiterpenoids, 2-O-acetyl-8,12-epoxygermacra-1(10),4,7,11-tetraene and 2-O-methyl-8,12-epoxygermacra-1(10)-4,7,11-tetraene, from gum exudates of Commiphora myrrh are reported. These two compounds, together with the known furanodienone isolated from C. erythraea, were found to have ixodicidal activity against Rhipicephalus appendiculatus tick larvae.     

Unusual iridoid glycosides in Veronica sects. Hebe and Labiatoides

In a chemosystematic investigation of three Southern hemisphere species of Veronica, namely the Australian Veronica derwentiana Andrews and Veronica perfoliata R.Br. (formerly Derwentia species), and the New Zealand Veronica catarractae G. Forster (formerly a species of Parahebe), the water-soluble constituents were isolated and identified by spectroscopic methods. Apart from other iridoid glucosides common to the genus, three unusual substituted benzoyl esters of aucubin (derwentiosides A–C) were obtained from V. derwentiana and a chlorinated iridoid glycoside (catarractoside) from V. catarractae in addition to other iridoids common to the genus. The chemical profile of V. perfoliata is similar to that of Northern hemisphere species of Veronica because of the presence of characteristic 6-O-catalpol esters. The profile of V. derwentiana is unique, since 6-O-esters of aucubin rather than of catalpol dominate, however, the acyl groups are the same as those present in catalpol esters found in some other Veronica sections. V. catarractae also contains one of the catalpol esters characteristic of Veronica, but in addition three 6-O-rhamnopyranosyl substituted iridoid glycosides, one of which is 6-O-rhamnopyranosylcatalpol. Esters of the latter compound are previously only known from the more derived species in recent phylogenetic trees of sect. Hebe to which V. catarractae now also belongs, but as a more basal member.     

Alkaloids of Zanthoxylum monophyllum and Z. punctatum

The known alkaloids N-methylcorydine and magnoflorine were major constituents of stems and branches of Zanthoxylum punctatum (Rutaceae). Berberine was the major alkaloid of Z. monophyllum, which also contained a new pyrano-2-quinolone alkaloid, zanthophylline, and its desmethyl analog. Zanthophylline has an —NCH₂OAc functional group which is readily hydrolyzed by dilute acid. Methyl vanillate and the furocoumarin columbianetin were also found in Z. monophyllum.     

Sesquiterpene lactones from Liatris acidota, L. Aspera and L. Mucronata

Examination of Liatris mucronata gave the known 15-hydroxylated heliangolides liscundin, liscunditrin, punctaliatrin and two new closely related heliangolides. Liatris acidota furnished six new similar but 15-deoxygenated heliangolides as well as euparin and 3β-acetoxytaraxaster-20-en-30-aldehyde. Liatris aspera gave a known glycosidic germacradienolide and a new germacradienolide as well as several known flavones and a known benzofuran. Structures were established by spectroscopic methods and chemical transformations.     

Rhizobium pisi sv. trifolii K3.22 harboring nod genes of the Rhizobium leguminosarum sv. trifolii cluster

The taxonomic status of the Rhizobium sp. K3.22 clover nodule isolate was studied by multilocus sequence analysis (MLSA) of 16S rRNA and six housekeeping chromosomal genes, as well as by a subsequent phylogenic analysis. The results revealed full congruence with the Rhizobium pisi DSM 30132^T core genes, thus supporting the same taxonomic position for both strains. However, the K3.22 plasmid symbiosis nod genes demonstrated high sequence similarity to Rhizobium leguminosarum sv. trifolii, whereas the R. pisi DSM 30132^Tnod genes were most similar to R. leguminosarum sv. viciae. The strains differed in the host range nodulation specificity, since strain K3.22 effectively nodulated red and white clover but not vetch, in contrast to R. pisi DSM 30132^T, which effectively nodulated vetch but was not able to nodulate clover. Both strains had the ability to form nodules on pea and bean but they differed in bean cultivar specificity. The R. pisi K3.22 and DSM 30132^T strains might provide evidence for the transfer of R. leguminosarum sv. trifolii and sv. viciae symbiotic plasmids occurring in natural soil populations.     

Prenylated acetophenones from Melicope obscura and Melicope obtusifolia ssp. obtusifolia var. arborea and their distribution in Rutaceae

Four prenylated acetophenones 2,6-dihydroxy-4-geranyloxyacetophenone (1), 4-geranyloxy-2,6,β-trihydroxyacetophenone (2), 2,6-dihydroxy-4-geranyloxy-3-prenylacetophenone (3), and 4-geranyloxy-3-prenyl-2,6,β-trihydroxyacetophenone (4) have for the first time been isolated from Melicope obscura (1 and 2) and Melicope obtusifolia ssp. obtusifolia var. arborea (3 and 4). The distribution of prenylated acetophenones in Rutaceae is reviewed and the results, including the new records, indicate that prenylated acetophenones are valuable as chemotaxonomic markers for the subfamily Rutoideae, tribe Xanthoxyleae sensu Engler.     

A new wilt and die-back disease of Acacia mangium associated with Ceratocystis manginecans and C. acaciivora sp. nov. in Indonesia

Species of Ceratocystis are well-known wound related pathogens of many tree species, including commercially planted Acacia spp. Recently, several Ceratocystis isolates were collected from wilting A. mangium in plantations in Indonesia. The aim of this study was to identify these Ceratocystis isolates and to investigate their ability to cause disease on two plantation-grown Acacia spp. using greenhouse and field inoculation experiments. For identification, morphological characteristics and comparisons of DNA sequence data for the ITS, β-tubulin and TEF 1-α gene regions, was used. Ceratocystis isolates were identified as C. manginecans, a serious pathogen of mango trees in Oman and Pakistan and a previously undescribed species, described here as C. acaciivora sp. nov. Both fungi produced significant lesions in inoculation experiments on A. mangium and A. crassicarpa, however, C. acaciivora was most pathogenic suggesting that this fungus is the primary cause of the death of trees under natural conditions.     

Artemisone inhibits in vitro and in vivo propagation of Babesia bovis and B. bigemina parasites

Artemisone was evaluated, in in vitro and in vivo, for control of bovine babesiosis caused by Babesia bigemina and Babesiabovis parasites. In vitro, artemisone reduced parasitemia in a dose-dependent manner: the inhibitory effects increased gradually, reaching a maximum inhibition of 99.6% and 86.4% for B. bigemina and B. bovis, respectively 72 h after initiation of treatment with initial parasitemia of 0.5%. In calves infected with either B. bigemina or B. bovis artemisone treatment was well tolerated and prevented development of acute babesiosis in all animals except for one B. bovis-infected calf. The treatment did not eliminate all blood parasites, and recovered animals carried a persistent low-level infection. Treatment with artemisone may be useful as an alternative drug for preventing the pathology that results from babesiosis, without interfering with acquired immune protection following recovery from an acute babesiosis infection or vaccination.     

基于28S, COI和Cytb基因序列的薜荔和爱玉子传粉小蜂分子遗传关系研究

薜荔和爱玉子均属于桑科榕属植物,二者为同一物种的原变种与变种的关系,早期研究认为这两种榕树与同一种传粉榕小蜂(Wiebesia pumilae (Hill))建立了稳定的互利共生关系,但近期在形态学、生态学、传粉生物学等方面对二者的研究结果表明,薜荔传粉小蜂和爱玉子传粉小蜂之间可能发生了遗传分化。实验用核糖体28SrDNAD1-D3区、线粒体Cytb及COI基因部分序列,对采自福建3个不同样地的薜荔传粉小蜂和3个不同品系的栽培爱玉子的传粉小蜂进行分析,结果表明:(1)薜荔传粉小蜂和爱玉子传粉小蜂的核糖体28S序列的碱基组成中A,T,G,C 4种含量较平均,C+G的平均含量(56%)稍高于A+T的含量(44%)。线粒体Cytb序列中A+T的含量(76.1%)明显高于C+G的含量(23.9%),COI序列中A+T的含量(71.9%)也明显高于G+C的含量(28.1%),这是膜翅目昆虫线粒体基因的普遍特征。在薜荔和爱玉子传粉小蜂的线粒体Cytb及COI基因中,密码子第三位点A+T的含量最高。(2)比较薜荔和爱玉子传粉小蜂的3种分子标记的变异范围显示,28S进化速度较Cytb及COI序列慢,比较保守,更适合科、亚科等较高分类单元的研究。薜荔传粉小蜂与爱玉子传粉小蜂之间的亲缘关系较近,采用Cytb与COI序列进行分析更为精确。(3)用Cytb及COI序列对薜荔传粉小蜂与爱玉子传粉小蜂之间的遗传距离进行分析显示,薜荔传粉榕小蜂个体间Cytb序列平均遗传距离为0.0054,爱玉子传粉小蜂个体间的Cytb遗传距离为0.0164;薜荔传粉小蜂与爱玉子传粉小蜂群体之间的Cytb序列平均遗传距离为0.1385;COI序列的薜荔传粉榕小蜂个体间遗传距离为0.0048,爱玉子传粉小蜂各样本间平均遗传距离为0.0102;薜荔传粉小蜂与爱玉子传粉小蜂群体间COI序列平均遗传距离为0.1896,两群体间的遗传距离(差异大于10%以上)明显大于群体内各样本之间的遗传距离,表明薜荔传粉小蜂与爱玉子传粉小蜂之间已经发生了很大的遗传分化,其变异水平达到了种间分化水平,即薜荔传粉小蜂与爱玉子传粉小蜂为两个不同的种。     

Nuclear and chloroplast DNA sequences data support the origin of Potamogeton intortusifolius J.B. He in China as a hybrid between P. perfoliatus Linn. and P. wrightii Morong

Internal transcribed spacers (ITS) of nuclear ribosomal DNA and chloroplast rbcL gene sequence data were used to test the hypothesis that natural populations of Potamogeton intortusifolius J.B. He in China originated from hybridization between P. perfoliatus Linn. and P. wrightii Morong. Based on ITS sequences data, P. intortusifolius possessed heterozygous rDNA genotypes which confirmed the hybrid origin of P. intortusifolius. Chloroplast rbcL gene sequences of P. intortusifolius from Yichang population revealed the same chloroplast haplotype as P. perfoliatus and the samples of P. intortusifolius from Weinan population had the same chloroplast haplotype as P. wrightii, which indicated that both putative parental species had been the maternal parent and that the two populations of P. intortusifolius had independent origins. This study confirms P. intortusifolius as a reciprocal hybrid. Because P. × intortusifolius in China has the same hybrid origin as P. × anguillanus Koidz. in Japan, it is suggested that P. × intortusifolius should be a synonym of P. × anguillanus.     

Description of Brenneria roseae sp. nov. and two subspecies, Brenneria roseae subspecies roseae ssp. nov and Brenneria roseae subspecies americana ssp. nov. isolated from symptomatic oak

Gram-negative, facultatively anaerobic bacteria were isolated from symptomatic oak tissue in the UK and USA. Partial gyrB sequencing placed ten strains in the genus Brenneria, with B. goodwinii as the closest phylogenetic relative. The strains were investigated further using a polyphasic approach including MLSA (based on partial gyrB, rpoB, infB and atpD gene sequences), 16S rRNA gene sequencing, DNA–DNA relatedness studies and both phenotypic and chemotaxonomic assays. The MLSA and 16S rRNA gene analyses separated the strains into two groups based on origin, suggesting that they belong to Brenneria as two novel species. However, the DNA–DNA relatedness values revealed a closer relationship between the groups and indicated that they should belong to the same species. As the two groups of strains from the UK and USA can be differentiated from each other phenotypically and by ERIC PCR fingerprints, it is proposed to classify them as novel subspecies of a novel Brenneria species. The name Brenneria roseae sp. nov. (FRB 222^T = LMG 27714^T = NCPPB 4581^T) is proposed, with Brenneria roseae subsp. roseae ssp. nov. (FRB 222^T = LMG 27714^T = NCPPB 4581^T) for the strains from the UK and Brenneria roseae subsp. americana ssp. nov. (FRB 223^T = LMG 27715^T = NCPPB 4582^T) for the strains from the USA.     

Staphylococcus petrasii sp. nov. including S. petrasii subsp. petrasii subsp. nov. and S. petrasii subsp. croceilyticus subsp. nov., isolated from human clinical specimens and human ear infections

Thirteen coagulase-negative, oxidase-negative, and novobiocin-susceptible staphylococci were isolated from human clinical specimens. The isolates were differentiated from known staphylococcal species on the basis of 16S rRNA, hsp60, rpoB, dnaJ, tuf, and gap gene sequencing, automated ribotyping, (GTG)₅-PCR fingerprinting, and MALDI-TOF MS analysis. Phylogenetic analysis based on the 16S rRNA gene sequence indicated phylogenetic relatedness of the analyzed strains to Staphylococcus haemolyticus, Staphylococcus hominis, Staphylococcus devriesei, and Staphylococcus lugdunensis. DNA–DNA hybridization experiments between representative strains CCM 8418^T, CCM 8421^T, and the closest phylogenetic neighbors confirmed that the isolates represent novel Staphylococcus species, for which the name Staphylococcus petrasii sp. nov. is proposed. Genotypic and phenotypic analyses unambiguously split the strains into two closely related subclusters. Based on the results, two novel subspecies S. petrasii subsp. petrasii subsp. nov. and S. petrasii subsp. croceilyticus subsp. nov. are proposed, with type strains CCM 8418^T (=CCUG 62727^T) and CCM 8421^T (=CCUG 62728^T), respectively.