Three ascomycetes on leaves of evergreen Ilex trees from Japan: Rhytisma ilicis-integrae sp. nov., R. ilicis-latifoliae, and R. ilicis-pedunculosae sp. nov.

Three species belonging to the genus Rhytisma causing tar spot were collected on leaves in evergreen trees of Ilex species from Japan. Rhytisma ilicis-latifoliae, the known species, is found on Ilex latifolia, and R. ilicis-integrae sp. nov. and R. ilicis-pedunculosae sp. nov. are found on I. integra and I. pedunculosa, respectively. Ascomata are formed on the abaxial part of the stromata in all the Rhytisma species studied, and spermogonia are formed on the amphigenous parts in R. ilicis-latifoliae and on the adaxial part in R. ilicis-integrae and R. ilicis-pedunculosae. Shape and size of asci, ascospores, and spermatia are distinctly different among the three species. The morphology of germination tubes from ascospores and appressoria is unique for each Rhytisma species. Yellowish spots arise on the newly developing leaves in mid-May, then abundant spermatia are produced in spermogonia in the three Rhytisma species. In the next year, ascospores are produced in ascomata from early April to late May in R. ilicis-integrae and from early April to early June in R. ilicis-latifoliae and R. ilicis-pedunculosae, and they are considered to be the inocula of disease infection.     

Antimicrobial activity against select food-borne pathogens by phenolic antioxidants enriched in cranberry pomace by solid-state bioprocessing using the food grade fungus Rhizopus oligosporus

Listeria monocytogenes, Vibrio parahaemolyticus and Escherichia coli 0157:H7 are important food-borne pathogens. Control of these pathogens using synthetic antimicrobials such as currently approved antibiotics is challenging due to potential development of resistance. A profile of antimicrobials compared to a single compound could be potentially more effective. Cranberry pomace is a byproduct of the cranberry processing industry. Solid-state bioprocessing of cranberry pomace using the food grade fungus Rhizopus oligosporus improves phenolic and antioxidant profiles. We hypothesize that these phytochemicals mobilized during pomace bioprocessing could improve the antimicrobial functionality. The objective of this research was to use the food grade fungus R. oligosporus to release phenolic aglycones and relate its antioxidant functionality and diphenyl mobilization to antimicrobial activity against L. monocytogenes, V. parahaemolyticus and E. coli 0157:H7. Bioprocessing of pomace was done for 20 days with R. oligosporus. Total phenolics in water extracts were assayed using a Folin-Ciocalteu method. The antioxidant potential was measured using a β-carotene oxidation system (APF) and 1,1-diphenyl-2-picrylhydrazyl-radical (DPPH) system. Changes in phenolic profiles were analyzed using HPLC. Antimicrobial activities of the extracts during growth were tested using standard disk assays. Soluble phenolics, antioxidant activity and ellagic acid concentrations were enriched by bioprocessing and antimicrobial activity of the extracts against L. monocytogenes and V. parahaemolyticus correlated with highest soluble phenolics and APF in the same extracts. For E. coli 0157:H7 inhibition correlated with the extracts corresponding to highest DPPH and ellagic acid concentration. The bioprocessing-based antimicrobial activity depended on different phenolic functional properties of the extracts. Sensitivity towards soluble phenolics reflected in DPPH activity suggested inhibition by the disruption of the membrane by hyperacidification. Whereas, sensitivity to APF and ellagic acid suggests potential antimicrobial activity by membrane-transport disruption. The variation in sensitivity of pathogens has implications for designing new food grade antimicrobials. This bioprocessing strategy can be an innovative approach to produce broad spectrum antimicrobials against important food-borne pathogens.     

Production of phenolic antioxidants by the solid-state bioconversion of pineapple waste mixed with soy flour using Rhizopus oligosporus

The ability of Rhizopus oligosporus to produce enhanced levels of free phenolics from pineapple residue mixed with soy flour as potential nitrogen source was investigated. Concurrently, phenolic-linked β-glucosidase and the antioxidant activity of the extracts were followed. Two treatments were studied: (A) 9 g of pineapple residue and 1 g of soy flour (P9); (B) 5 g of pineapple residue and 5 g of soy flour (P5). The increase of water extractable phenolics was 39.3% for P9 treatment and 79.4% for P5 treatment. During early stages of growth high antioxidant activity, low phenolic content and low β-glucosidase specific activity was observed. High antioxidant activity was likely due to the presence of insoluble polymeric phenolics, know for their high antioxidant activity. A marked decrease of the antioxidant activity of P5 treatment during late stages of growth was observed due to likely formation of free soluble phenolics. The moderate total phenolics content and high β-glucosidase specific activity of P9 treatment in late stages is likely the consequence of low nitrogen content in this treatment. The bioconversion of pineapple residue by R. oligosporus leads to enhanced levels of phenolic compounds, mainly for P5 treatment. This approach offers a novel strategy to enhance the value of pineapple wastes.     

The genus Setulipes (Marasmiaceae) in Madagascar and the Mascarenes, including a key to other African taxa

The authors report first records for the genus Setulipes in Madagascar, with the presence of Setulipes cf. hakgalensis and two new species, Setulipes funaliformis and S. moreaui, as well as a new species from Mauritius: S. mauritiensis. A key to these taxa, as well as to other African species, is supplied.     

Morphological variation of Juniperus oxycedrus subsp. oxycedrus (Cupressaceae) in the Mediterranean region

The intra- and inter-population variation of Juniperus oxycedrus subsp. oxycedrus were examined biometrically on the basis of morphological characters of needles, seeds and cones. Cluster analysis of Euclidean distances and discriminant analysis were performed to verify the hypothesis of geographic differentiation and origin from the West- and East-Mediterranean Pleistocene refugial regions. The West-Mediterranean populations of J. oxycedrus subsp. oxycedrus differed from the eastern ones. The intra-populational differentiation of three of the West-Mediterranean populations was significantly higher than within the other West- and all the East-Mediterranean ones.Results confirm the hypothesis of probable different origins of the East- and West-Mediterranean populations of the taxon.     

Song dialects on the Atlantic islands: goldcrests of the Azores ( Regulus regulus azoricus, R. r. sanctae-mariae, R. r. inermis)

Territorial songs in island populations of songbirds are often highly divergent from those of neighbouring continental relatives. This is shown for the three goldcrest subspecies (Regulus regulus azoricus, R. r. sanctae-mariae and R. r. inermis) endemic on six islands of the Azorean archipelago. All investigated populations display a high intra- and inter-individual acoustic variation. On each island, up to six different song types have been found; and a single male sings up to three types. In contrast, all northwestern European populations of R. r. regulus and R. r. anglorum share only a single song type. In playback experiments, none of 18 tested dialect songs of Azorean goldcrests evoked notable territorial reaction in German and Czech goldcrest males (ssp. regulus). Two differing dialect groups of the goldcrest can be distinguished on the Azores. Populations of the eastern islands, São Miguel and Santa Maria, share common song types which are not found on the islands of the central and western groups. Dialect repertoires on the westernmost islands, Flores, Faial and Pico, are dominated by a different song type. In the geologically younger western crater of São Miguel, both western and eastern song types coexist. Acoustic similarities to a population from neighbouring Terceira suggest the western part of São Miguel as the origin for the westward expansion of R. regulus on the Azores.     

中国橐吾属(菊科-千里光族)的分类学研究(四):康定橐吾和Senecio ficariifolius的名实订正(英文)

将康定橐吾(Ligularia kangtingensis S.W.Liu)处理为细茎橐吾[L.hookeri(C.B.Clarke)Hand.-Mazz.]的异名。Senecio ficariifolius H.Lév.Vaniot长期以来被处理为细茎橐吾的异名,但我们检查模式材料后发现其头状花序盘状,与头状花序辐射状的细茎橐吾明显不同,而与头状花序盘状的贵州橐吾[L.leveillei(Vaniot)Hand.-Mazz.]没有区别,故将其处理为贵州橐吾的异名。     

Morphological and genetic study of Prorocentrum donghaiense Lu from the East China Sea, and comparison with some related Prorocentrum species

In this paper, the detailed morphology of Prorocentrum donghaiense Lu from both field samples and cultures was examined, and a taxonomic comparison was made between P. donghaiense and some related Prorocentrum spp. using morphological and molecular data and other published information. There were distinct differences among these species in morphological characteristics that historically have been presented as conservative features. The discrepancies extended beyond that of individual variations within the same species due to environmental factors. Therefore, these morphological features may not be conservative but, rather, polymorphic depending on environmental conditions. Based on this analysis, we suggest that the high-biomass bloom-forming species in the East China Sea, previously reported as Prorocentrum dentatum Stein, is P. donghaiense Lu. The species reported from the East China Sea and Japanese and Korean waters appear to be the same species. Molecular data also suggest that P. dentatum (CCMP1517) and P. donghaiense are genetically identical. Therefore, the geographic distribution of P. donghaiense may be much wider than expected.     

Morphological studies on two marine colepid ciliates from Qingdao, China, Nolandia orientalis spec. nov. and Pinacocoleps similis (Kahl, 1933) comb. nov. (Ciliophora, Colepidae)

Recent studies have revealed that the “lower” marine ciliates are far more diverse than previously suspected. During a survey on the ciliate fauna in coastal waters of Qingdao, northern China, we have isolated about 30 new or poorly known taxa. In the present study two colepid species are investigated, Nolandia orientalis spec. nov. and Pinacocoleps similis (Kahl, 1933) comb. nov. (basionym: Coleps similis Kahl, 1933). Their morphology and infraciliature are documented based on living observations and silver impregnations. The new species Nolandia orientalis differs from the type species N. nolandi mainly in the structure of tier plates. The structure of the tier plates was also the basis for transferring Coleps similis Kahl, 1933 to the genus Pinacocoleps and for three further new combinations: Pinacocoleps heteracanthus (Noland, 1937) comb. nov. (basionym: Coleps heteracanthus Noland, 1937), P. spiralis (Noland, 1937) comb. nov. (basionym Coleps spiralis Noland, 1937) and Pinacocoleps arenarius (Bock, 1952) comb. nov. (basionym: Coleps arenarius Bock, 1952).     

头花杜鹃、陇蜀杜鹃及杜鹃属植物叶绿体基因组比较分析

头花杜鹃(Rhododendron capitatum)和陇蜀杜鹃(R. przewalskii)是极具观赏价值的野生花卉和药用植物。为探讨头花杜鹃和陇蜀杜鹃叶绿体基因组的遗传结构及进化特征,该研究利用 Illumina HiSeq 4000 平台对头花杜鹃和陇蜀杜鹃的叶绿体全基因组进行测序,经组装和注释后,结合 7 个已发表的杜鹃属植物叶绿体全基因组进行比较基因组学分析和系统发育分析。结果表明:(1)头花杜鹃和陇蜀杜鹃叶绿体全基因组呈典型的环状四分体结构,均由一个大单拷贝区(105 990、109 191 bp)、一个小单拷贝区(2 617、2 606 bp)和一对反向重复区(45 825、47 516 bp)构成,全长分别为200 257、206 829 bp。(2)头花杜鹃和陇蜀杜鹃叶绿体基因组中共鉴定出 263 个SSR位点,大部分 SSR 偏好使用 A/T 碱基,密码子偏好使用 A/U 结尾。(3)杜鹃属植物叶绿体全基因组中普遍存在基因丢失以及基因组重排等结构变异现象。该研究丰富了杜鹃属植物的基因组资源,为头花杜鹃、陇蜀杜鹃的资源开发、遗传进化、育种及系统发育相关研究提供了理论参考。     

Cloning and expression of cellulase gene EG1 from Rhizopus stolonifer var. reflexus TP-02 in Escherichia coli

A novel gene, EG encoding enzymes involved in carboxymethyl cellulose (CMC) degradation was isolated, sequenced from the filamentous fungus Rhizopus stolonifer var. reflexus TP-02, and expressed in Escherichia coli BL21. The results showed that the gene amplified from the cDNA of the strain could be classified as the family of endoglucanase. During the fermentation process, the maximum endoglucanase activity (i.e. 0.715 IU/ml) of the recombinant bacteria was obtained at 36 h. The SDS–PAGE analysis on purified samples showed that a band with apparent molecular weight of about 40 kDa was detected after staining with Coomassie brilliant blue.     

中国毛茛科翠雀属的分类学研究(十二):竞生翠雀花的名实订正

通过标本室和野外观察,发现根据云南西北部中甸标本描述的毛茛科竞生翠雀花（Delphinium yangii W.T.Wang）与此前记载分布于该地及四川西南部（康定、小金、汶川）的宝兴翠雀花（D.smithianum Hand.-Mazz.）属于同一分类实体,故将前者处理为后者的异名。     

中国毛茛科翠雀属的分类学研究(四):五花翠雀花的名实订正

通过标本室和野外观察,发现根据云南中甸标本描述的毛茛科五花翠雀花(Delphinium quinqueflorum W.T.Wang)与当地颇为常见的澜沧翠雀花(D.thibeticum Finet&Gagnep.)没有本质区别,仅为其稍矮小的植株,故将前者处理为后者的异名。     

亚沉茶渍亚洲亚种Lecanora subimmersa subsp.asiatica Zahlbr.的订正研究北大核心CSCD

亚沉茶渍亚洲亚种Lecanora subimmersa subsp.asiatica Zahlbr.曾被基于文献研究转隶至平茶渍属(Aspicilia)或处理为亚沉茶渍原亚种L.subimmersa(Fée)Vain.subsp.subimmersa的异名,但模式标本的研究缺乏仍使该变种的概念不清。利用形态学、解剖学和化学等方法对L.subimmersa subsp.asiatica Zahlbr.的模式标本进行了综合研究,发现L.subimmersa subsp.asiatica与滇茶渍[L.oreinoides(Körb.)Hertel&Rambold]的形态特征及次生代谢产物一致,故将L.subimmersa subsp.asiatica作为L.oreinoides的异名处理。依据研究标本,对滇茶渍进行了形态学描述,同时提供了相关形态学图片,澄清了亚沉茶渍亚洲亚种的概念。     

中国毛茛科翠雀属的分类学研究(七):岩瓦翠雀花的名实订正

通过标本室和野外观察,发现根据云南西北部维西标本描述的毛茛科岩瓦翠雀花(Delphinium yanwaense W. T. Wang)与云南西北部颇为常见的中甸翠雀花(D. yuanum Chen)没有本质区别,故将前者处理为后者的异名。中甸翠雀花在叶的分裂程度上变异颇大。     

中国毛茛科翠雀属的分类学研究(十四):光序翠雀花二新异名

广布于中国-喜马拉雅地区的毛茛科光序翠雀花（Delphinium kamaonense Huth）在我国甘肃、青海、四川和西藏具有相当连续的分布区,在分布区内颇为常见,几为杂草。通过标本室和野外观察,发现根据西藏波密、察隅标本描述的展毛光序翠雀花[D. kamaonense var. glabrescens（W. T. Wang）W. T. Wang]和根据西藏类乌齐标本描述的倒心形翠雀花（D. obcordatilimbum W. T. Wang）与光序翠雀花没有本质区别,故将二者均处理为光序翠雀花的异名。     

中国毛茛科翠雀属的分类学研究(五):秋翠雀花二新异名

通过标本室和野外观察,发现根据四川西南部越西标本描述的毛茛科凉山翠雀花(Delphinium liangshanense W. T. Wang)和根据四川西南部木里标本描述的光轴翠雀花(D. leiostachyum W. T. Wang)只是此前记载分布于四川木里、盐源和云南西北部宁蒗一带的秋翠雀花(D. autumnale Hand.-Mazz.)植株较高的类型,故将二者均处理为秋翠雀花的异名.     

中国毛茛科翠雀属的分类学研究(六):拟川西翠雀花二新异名

通过标本室和野外观察, 发现根据四川丹巴标本描述的毛茛科光果拟螺距翠雀花(Delphinium bulleyanum Forrest ex Dielsvar. leiogynum W. T. Wang)和根据四川汶川标本描述的汶川翠雀花(D. wenchuanense W. T. Wang)与此前发现分布于四川宝兴、都江堰、汶川一带的拟川西翠雀花(D. pseudotongolense W. T. Wang)没有区别, 故将二者均处理为拟川西翠雀花的异名。     

中国毛茛科翠雀属的分类学研究(十一): 巴唐翠雀花二新异名

毛茛科巴塘翠雀花（Delphinium batangense Finet & Gagnep.）是我国四川西部和云南西北部一种较广布的高山植物,形态变异颇大。标本室和野外观察表明白缘翠雀花（D.chenii W.T.Wang）和雅江翠雀花（D.yajiangense W.T.Wang）在形态上处于该种的变异范围之内,故将二者均处理为巴塘翠雀花的异名。     

匍枝根霉纤维二糖合成酶胞内糖基供体的初探及结构功能研究

纤维二糖可有效诱导丝状真菌产纤维素酶,前期研究表明匍枝根霉Rhizopus stolonifer TP-02具有纤维二糖合成酶(CBS),可以尿苷二磷酸葡萄糖(UDPG)为糖基供体合成纤维二糖,从而开启纤维素酶的自诱导合成途径。为研究R. stolonifer中纤维二糖的胞内合成途径,通过重叠PCR在GDP-葡糖焦磷酸化酶基因ggp中引入硫胺吡啶抗性基因ptrA,分别转化原菌TP-02和△ugp突变株,构建△ggp和△ugp/△ggp突变株。利用液质联用(LC-MS)检测突变株的胞内糖组分,发现ggp的缺失对胞内纤维二糖合成的影响较弱,但同时缺失ugp则将直接导致二糖合成受阻。RT-qPCR结果显示△ggp突变株中纤维素酶基因转录水平较原株TP-02下调20%左右,而△ugp/△ggp突变株中被测基因的转录水平则出现了高达80%左右的下调。同时对突变株纤维素酶表达水平进行研究,发现△ugp/△ggp突变株中几乎检测不到纤维素酶活力。结果显示,UDPG为R. stolonifer胞内合成纤维二糖的主要糖基供体,而GDPG可能是UDPG的替代物,在UDPG不足时维持胞内二糖合成。此外,利用生物信息学方法对CBS结构功能深入分析,经丙氨酸扫描确定其合成纤维二糖的关键作用残基为Asp210和Asp300,为后续进一步研究及理性改造提供方向和理论依据。