缓解花生连作障碍的根际促生菌分离及功能鉴定

[目的] 长期连作障碍严重降低花生生产的产量及品质,根际促生菌可有效降解土壤中自毒化感物质、抑制植物病原菌生长及促进植物生长,从而有效缓解连作障碍问题。筛选优化具有缓解花生连作障碍能力的多功能根际益生微生物,验证其益生作用能力,为根际促生菌株在连作障碍中的应用提供理论依据及技术支持。[方法] 采集连作12年地块花生根际土壤,利用以酚酸为唯一碳源的筛选培养基获得具有酚酸自毒化感物质降解及利用能力的根际促生菌,通过16S rRNA基因测序进行系统发育分析,确定根际促生菌菌株的分类地位,并验证其对植物病原菌生长抑制能力及解磷、解钾、产植物激素吲哚乙酸能力。[结果] 从连作12年的花生发病土壤中获得7株可高效降解酚酸类自毒物质且降解底物多样的根际微生物菌株,经16S rRNA测序比对分别为克雷伯氏菌B02 （Klebsiella sp.B02）、克雷伯氏菌B07 （Klebsiella sp.B07）、克雷伯氏菌B15 （Klebsiella sp.B15）、芽孢杆菌B28 （Bacillus sp.B28）、不动杆菌P09 （Acinetobacter sp.P09）、布鲁氏杆菌VA05 （Brucella sp.VA05）、芽孢杆菌CA04 （Bacillus sp.CA04）。促生实验表明,7株高效降解菌株均可以合成吲哚乙酸,3株具有固氮能力,4株菌具有解有机磷及无机磷的能力,2株菌具有解钾的能力。拮抗实验表明,2株菌可以抑制多种植物病原菌的生长,均为芽孢杆菌属。选取Bacillus sp.B28初步验证对花生种子萌发及幼苗生长的影响,结果表明根际促生菌可显著缓解酚酸对花生种子发芽的抑制,并明显促进花生幼苗的生长。[结论] 获得多株具有降解酚酸类自毒化感物质、抑制植物病原菌生长及促进植物生长的多功能花生根际促生菌,更好地为根际促生菌在连作障碍治理中的有效应用提供菌株及技术支持。     

烟草根际可培养微生物多样性及防病促生菌的筛选

[背景] 根际微生物在植物根部生态系统中扮演着重要角色,影响着植物的营养吸收和健康生长。[目的] 了解常年不发病烟田烤烟品种K326根际可培养微生物的多样性,筛选具有防病促生功能的菌株,为烟草病害绿色防控提供资源。[方法] 采用传统培养方法对烟草根际土壤中的细菌和真菌进行分离鉴定,评价菌株的促生特性及病原菌拮抗能力,并进一步验证典型菌株对盆栽烟苗的促生效果。[结果] 共获得261株微生物菌株,包括160株细菌和101株真菌。经分子鉴定,细菌中以变形菌门（Proteobacteria）和厚壁菌门（Firmicutes）为主要类群;真菌中以子囊菌门（Ascomycota）和毛霉菌门（Mucoromycota）为主要类群。在属水平上,细菌以假单胞菌属（Pseudomonas）和芽孢杆菌属（Bacillus）为主,真菌以曲霉属（Aspergillus）和青霉属（Penicillium）为主。从不同种水平上进一步选择44株细菌为代表菌株,发现它们均具有不同程度的吲哚-3-乙酸（Indole-3-Acetic Acid,IAA）产生能力,9株能够溶解有机磷,16株能够溶解无机磷,13株产生铁载体,14株产生生1-氨基环丙烷-1-羧酸（1-Aminocyclopropane-1-Carboxylate,ACC）脱氨酶。从160株细菌中筛选得到抑制青枯病菌和黑胫病菌的菌株数目分别为25、26株。经盆栽试验发现韩国假单胞菌（P. koreensis） HCH2-3、浅黄绿假单胞菌（P. lurida） FGD5-2和贝莱斯芽孢杆菌（B. velezensis） EM-1对烟苗呈现不同程度的促生作用,其中3株菌联合施加对烟苗的促生效果最明显。[结论] 烟草根际存在着丰富多样的具有防病促生潜力的微生物,并且合成菌群或功能互补的菌株联合施用是未来微生物菌剂研发的重要方向。     

耐盐碱微生物菌种的筛选鉴定及其功能性与促生性

【背景】近年来,甘肃省土地盐碱化的日益加剧已对当地农业生产和生态环境产生严重影响。【目的】从甘肃省兰州市兰州新区和张掖市黑河生态保护区盐碱土壤中分离筛选出耐盐碱菌株,并对其进行鉴定和部分功能性评价与促生性分析,从而为改良甘肃省盐碱地提供微生物资源和理论基础。【方法】利用不同盐碱浓度的改良培养基对分离后的菌落进行条件培养,筛选出具有高耐盐碱能力的菌种,通过16S rRNA基因序列鉴定菌种类型,并测试其在不同培养基条件下功能性和促生性的不同作用。【结果】共筛选得到5株高耐盐碱菌株,鉴定结果分别为巨大芽孢杆菌（Bacillus megaterium）、金黄节杆菌（Arthrobacter aurescens）、费氏中华根瘤菌（Sinorhizobium fredii）、地衣芽孢杆菌（Bacillus licheniformis）和不动杆菌属（Acinetobacter sp.）。经鉴定分析5株菌均具有溶磷和分泌吲哚乙酸的能力;金黄节杆菌和地衣芽孢杆菌具有解钾能力;金黄节杆菌、费氏中华根瘤菌和不动杆菌属具有固氮能力;地衣芽孢杆菌、巨大芽孢杆菌和费氏中华根瘤菌分泌的ACC脱氨酶活性分别达到0.272、0.217和0.159 U/mg,金黄节杆菌和不动杆菌属几乎无ACC脱氨酶活性。【结论】五株耐盐碱菌在溶磷、解钾和固氮功能方面各自具有不同的效果且兼具一定的促生作用,这为后期微生物改良盐碱地的应用提供了物种资源和理论基础。     

甘草内生细菌的分离及拮抗菌株鉴定

从乌拉尔甘草健康植株的根茎叶中共分离到内生细菌98株,经初步鉴定芽孢杆菌属（Bacillus sp.）为优势种群,约占30%;从不同生长年份甘草的根、茎、叶组织中分离内生细菌种群密度从5.0×10⁴ cfu/g～2.9×10⁷ cfu/g鲜重不等。采用平板对峙方法筛选出6株对植物病原菌有明显体外拮抗活性的菌株,通过菌落、菌体形态观察、生理生化反应及16S rDNA序列分析,同时结合Biolog细菌自动鉴定系统验证,鉴定这6株拮抗菌分属萎缩芽孢杆菌(Bacillus atrophaeus)、多粘类芽孢杆菌(Paenibacillus polymyxa)、枯草芽孢杆菌(Bacillus subtilis)、Paenibacillus ehimensis。     

低温环境中T-2毒素降解菌的分离鉴定及特性研究

离筛选低温环境中T-2毒素的降解菌并探明其生化特性,探究T-2毒素降解微生物生化特性异同点,为T-2毒素降解微生物的检验提供生化判断参考。针对暴露于－20 ℃低温环境中的低浓度T-2毒素标准品,采用LC-MS/MS定量分析T-2毒素残留量,利用营养琼脂培养基（NA）和马铃薯葡萄糖琼脂培养基（PDA）共分离出5株降解菌,16S rDNA结合生化鉴定结果为死谷芽胞杆菌（Bacillus vallismortis）、蜡状芽胞杆菌（Bacillus cereus）、阴沟肠杆菌（Enterobacter cloacae）、弯曲假单胞菌（Pseudomonas geniculata）和尼泊尔葡萄球菌（Staphylococcus nepalensis）。这些分离株在－20 ℃条件下对低浓度T-2毒素均有不同程度的降解能力,其中蜡状芽胞杆菌（Bacillus cereus）降解能力最强,降解率为91%。它们的理化特性具有很多相似性,主要表现在5株菌均不能利用D 塔格糖(dTAG）,对ELLMAN都表现出阴性;Bacillus vallismortis Bp1234-7、Pseudomonas geniculata Bp24-4、〖WTBX〗Staphylococcus nepalensis Bp124-5和Enterobacter cloacae Bp123-7都能分解D-葡萄糖,而Bacillus cereus Bp1234-8不能。     

贝莱斯芽孢杆菌(Bacillus velezensis) JJYY防控土传病害效果评价及其全基因组测序分析和抗菌成分鉴定

【背景】土传病害是世界农业可持续发展的关键限制因子,在我国蔬菜和中药材上发生日趋严重,引起严重的连作障碍。生物防治对环境和农产品安全,是目前研究的热点和重点。【目的】明确对土传病原菌具有广谱抗菌活性菌株JJYY的种类及其防控效果和主要抗菌产物,为新型生物农药开发奠定基础。【方法】结合扫描电镜观察、生理生化分析和16S rRNA基因扩增测序技术鉴定菌株JJYY。分别利用比浊法和菌丝生长抑制法测定该菌提取物对4种土传病原细菌和5种土传病原真菌的EC₅₀,利用盆栽试验评价对番茄青枯病等病害的防控效果。利用二代Illumina NovaSeq与三代PacBio Sequel相结合测序技术对菌株JJYY进行全基因组测序,使用PGAP等软件进行基因注释等分析,利用反相制备液相色谱和质谱初步分离鉴定抗菌物质。【结果】菌株JJYY是一株贝莱斯芽孢杆菌（Bacillus velezensis）,该菌提取物对4种土传细菌和5种土传真菌的EC₅₀分别为0.940-1.092 mg/mL和2.733-3.678 mg/mL。对番茄青枯病、菊花根腐病和辣椒枯萎病的最高防效在80.00%-87.74%之间,与化学药剂链霉素或噁霉灵无显著差异（P>0.05）。该菌基因组大小为3 929 792 bp,编码3 895个基因,与B.velezensis JS25R和B.subtilis 168具有3 445个和2 997个同源编码基因。预测该基因组共有12个次级代谢产物合成功能基因簇。从该菌株提取物中共分离出9种抗菌组分,其中2个初步判定为已知大环内酯类抗生素macrolactin D和7-O-malonyl-macrolactin A。【结论】菌株JJYY是一株对土传病原细菌和真菌病害均有较高防效的贝莱斯芽孢杆菌,该菌基因组与已知贝莱斯芽孢杆菌不同,并产生多种已知和未知的抗菌物质,继续深入分析鉴定其抗菌物质和抗菌机制将为开发新型高效的生物农药奠定良好基础。     

芽孢杆菌WL911促低温下小麦生长效应及其功能基因分析

【背景】青藏高原特殊生境孕育着强耐逆性的微生物资源。【目的】探究青藏高原特殊生境的芽孢杆菌对小麦的促生效应。【方法】以分离自青海省海西蒙古族藏族自治州乌兰县的红柳（Tamarix ramosissima）根围菌株WL911为研究对象进行16S rRNA基因及gyrB基因鉴定;测定其低温适生性;以WL911菌悬液（浓度为1×10⁷ cfu/mL）对4℃条件下的小麦（Triticum aestivum）品种“青麦7号”幼苗进行灌根处理,测定其对小麦幼苗的促生效应;采用二代测序平台Illumina HiSeq×10对菌株WL911进行全基因组测序、功能注释及相关功能基因分析。【结果】菌株WL911鉴定为贝莱斯芽孢杆菌（Bacillus velezensis）,可耐受4℃低温;菌株WL911对“青麦7号”幼苗生长具有显著的促生效果,26℃时小麦株高、根长和鲜重分别提高12.22%、50.12%和70.99%;4℃条件下小麦生物量提高的同时,叶绿素含量提高19.72%,H₂O₂、丙二醛（malondialdehyd,MDA）的积累量分别下降23.50%、40.68%,超氧化物歧化酶（superoxide dismutase,SOD）、过氧化物酶（peroxidase,POD）和过氧化氢酶（catalase,CAT）活性分别提高45.82%、17.35%和14.18%;WL911基因组大小为3 915 549 bp,G+C百分含量为46.47%;生物学功能gene ontology （GO）注释到的基因占基因总量的75.74%,包括促生相关功能基因glnB、yclQ和fetB等,可参与合成生长激素、营养元素等途径;存在mnhA、mnhE、pheT、pheS和proV等关键基因参与Na⁺外排机制、编码脯氨酸和酚类化合物等反应;也存在编码冷休克蛋白CspC、CspB和CspD的关键基因cspA。【结论】B.velezensis WL911（登录号为OP874802）是一种优质的生物肥料研发菌株。     

古尔班通古特沙漠苔藓结皮土壤中促生菌的分离筛选及促生特性研究

从古尔班通古特沙漠南缘苔藓结皮土壤中分离筛选植物促生菌并阐明其促生特性，对于发掘和应用功能性微生物制剂，促进植被恢复和提升荒漠固沙能力等具有积极意义。采用富集方法从苔藓生物结皮下0～20 cm土壤中分离可培养微生物。利用选择性培养基筛选具溶磷、产铁载体、分泌吲哚乙酸（IAA）特性的菌株。采用钼锑抗比色法、CAS检测法、Salkowski比色法分别测定菌株溶磷量、产生铁载体的浓度、分泌IAA的含量，并对性能优良的菌株进行分类鉴定。共筛选出促生菌31株，其中溶磷菌31株，产铁载体菌13株，分泌IAA菌28株。菌株LB5WH溶解无机磷能力最强，溶磷量为302.24 mg/L；菌株LB15产铁载体能力最强，产生铁载体的浓度 Su值（铁载体活性单位）为85.7%；菌株GA20分泌IAA活性最强，分泌量为15.46 mg/L。菌株LB5WH、LB15同时具有溶磷、产铁载体和分泌IAA活性，菌株GA20具有溶解有机磷、分泌IAA活性。菌株溶解无机磷能力要强于溶解有机磷能力，溶解无机磷菌株的溶磷量与培养液pH值之间呈显著负相关。经鉴定，菌株LB5WH属于芽胞杆菌属（Bacillus），菌株GA20属于寡养单胞菌属（Stenotrophomonas），菌株LB15属于赖氨酸芽胞杆菌属（Lysinibacillus）。这三株细菌的促生功能较多，具有进一步开发为微生物肥料的潜能。本研究为丰富荒漠区促生菌资源，进一步深入研究苔藓结皮下土壤-微生物-植物互作的生态调控机制及荒漠植物促生菌的促生机理提供菌种资源。     

内芽孢杆菌属细菌（Paenibacillus daejeonensis）SS02抗菌蛋白的分离纯化和其性质的研究

内芽孢杆菌属细菌（Paenibacillus daejeonensis）SS02,是一株能强烈抑制玉米纹枯菌（Rhizoctonia cerealis）、白色念珠菌 (Candidal vaginitis)等病害真菌的拮抗菌株。SS02菌株培养液经硫酸铵分级盐析,Sephadex G-75凝胶柱层析和DEAE-32纤维素柱层析分离纯化出一种抗菌蛋白,命名为SD22。蛋白电泳分析结果表明,此蛋白分子量为56000,等电点为6.4。此蛋白对热和紫外线稳定,对蛋白酶部分敏感。纯化后的SD22对玉米纹枯菌(Rhizoctonia cerealis)、油菜菌核病(Sclerotinia sclerotiorum)、苹果轮纹病菌（Physalospora piricala）、绿色木霉(Trichodema viride)、绿色粘帚霉（Gliocladium viride）、弯孢霉菌(Curvularia leaf spot)、镰刀霉菌(Fusarium sp.)、赤霉菌(Fusarium head blight)、球孢白僵菌（Beauveria bassiana）、大肠杆菌（Escherichia coli）、金黄色葡萄球菌（Staphylo-coccus aureus）、枯草芽孢杆菌（Bacillus subtilis）、白色念珠菌(Candidal vaginitis）、冬瓜枯萎菌（Fusarium oxysporum Schl.emend.Snyder & Hanse）等菌有很强的抑制作用。     

内芽孢杆菌属细菌（Paenibacillus daejeonensis）SS02抗菌蛋白的分离纯化和其性质的研究

内芽孢杆菌属细菌（Paenibacillus daejeonensis）SS02,是一株能强烈抑制玉米纹枯菌（Rhizoctonia cerealis）、白色念珠菌 (Candidal vaginitis)等病害真菌的拮抗菌株。SS02菌株培养液经硫酸铵分级盐析,Sephadex G-75凝胶柱层析和DEAE-32纤维素柱层析分离纯化出一种抗菌蛋白,命名为SD22。蛋白电泳分析结果表明,此蛋白分子量为56000,等电点为6.4。此蛋白对热和紫外线稳定,对蛋白酶部分敏感。纯化后的SD22对玉米纹枯菌(Rhizoctonia cerealis)、油菜菌核病(Sclerotinia sclerotiorum)、苹果轮纹病菌（Physalospora piricala）、绿色木霉(Trichodema viride)、绿色粘帚霉（Gliocladium viride）、弯孢霉菌(Curvularia leaf spot)、镰刀霉菌(Fusarium sp.)、赤霉菌(Fusarium head blight)、球孢白僵菌（Beauveria bassiana）、大肠杆菌（Escherichia coli）、金黄色葡萄球菌（Staphylo-coccus aureus）、枯草芽孢杆菌（Bacillus subtilis）、白色念珠菌(Candidal vaginitis）、冬瓜枯萎菌（Fusarium oxysporum Schl.emend.Snyder & Hanse）等菌有很强的抑制作用。     

Y-chromosome Data and Tribal Affiliations of Allenopithecus and Miopithecus

The phylogeny of Old World monkeys has remained unresolved in part because of a lack of resolution in the Cercopithecinae. Competing morphological hypotheses have had Allen's swamp monkey (Allenopithecus nigroviridis) and the talapoins (Miopithecus spp.) as basal branches of either the tribe Cercopithecini or the tribe Papionini. Previous molecular analyses have not adequately addressed the issue. To better understand the evolutionary history of these primates, we sequenced and subjected to phylogenetic analysis 3.1 kb of 2 loci (TSPY and SRY) from the non-recombining portion of the Y-chromosome. Individuals from the genera Allenopithecus, Miopithecus, Erythrocebus, Chlorocebus, and Cercopithecus were surveyed and their sequences compared with those previously published for the Papionini and Colobinae. The results suggest Allenopithecus and Miopithecus are more closely related to the Cercopithecini than Papionini. Our data also support the hypothesis that within the Cercopithecini, Erythrocebus and Chlorocebus share a close evolutionary relationship, distinct from the other members of the tribe.     

Ovule and seed anatomy ofCistaceae and relatedMalvanae

Ovule and especially seed anatomy of eight species ofCochlospermaceae, Bixaceae, Cistaceae, Monotoideae, Pakaraimaeoideae (two subfamilies ofDipterocarpaceae), andSarcolaenaceae were investigated. All representatives show a bixoid chalazal region in the seed as probable exclusive synapomorphy among angiosperms. The palisade layer of the exotegmen is curved inwards at its proximal end and forms a dome-shaped structure. A plug of hypostase tissue with an annulus/core structure fits into this dome. Moreover, two additional tissue types in the hypostase can be found in some representatives of the group. These and other micromorphological, wood anatomical, and floral morphological characters, indicate that the taxa form a monophyletic group close toMalvales s. str. The form of the starch grains in the endosperm is compared and is described for the first time forPakaraimaea (Dipterocarpaceae) andLeptolaena (Sarcolaenaceae). The position ofDiegodendron close toBixa and the presumably more distant positions ofMuntingia andNeuradaceae are discussed.     

Nystatin and killer toxin sensitivity of free and immobilizedSaccharomyces cerevisiae

The inhibitory effect of nystatin and killer toxin on the growth of free and covalently-immobilizedSaccharomyces cerevisiae cells was studied. The resistance of immobilized cells to both agents was accompanied by increased amounts of phospholipids and sterols. The possible relationship between these changes in the membrane composition and the transduction of a signal across the cytoplasmic membrane is discussed.     

Amino acids and sugars in nectar,and their putative evolutionary significance

Individual amino acids and sugars from flower nectar of 32 plant species with different pollination systems were quantified and compared. Data show that there is no correlation between sugar and amino acid concentration. Furthermore there is no correlation between composition and concentration of amino acids and evolutionary advancement, nor any direct relation with pollination systems. However, higher sugar concentrations are often linked with more advanced morphological characters. Nectars from pierced or damaged flowers or nectars contaminated with pollen exhibit modifications and increases in amino acid composition. The presence of proline probably indicates such pollen contamination. Most pollinating animals depend on flower nectar in their energetic requirements, yet innumerable alternative amino acid and protein sources exist. Future research has to consider the relationship between nutritional requirements of pollinating animals and dependence on flower nectars.Dedicated to Prof. Dr.L. van der Pijl, Den Haag, in honour of his 80th birthday.     

Cyclooxygenase inhibitory and antioxidant cyanidin glycosides in cherries and berries

Anthocyanins from tart cherries, Prunus cerasus L. (Rosaceae) cv. Balaton and Montmorency; sweet cherries, Prunus avium L. (Rosaceae); bilberries, Vaccinum myrtillus L. (Ericaceae); blackberries, Rubus sp. (Rosaceae); blueberries var. Jersey, Vaccinium corymbosum L. (Ericaceae); cranberries var. Early Black, Vaccinium macrocarpon Ait. (Ericaceae); elderberries, Sambucus canadensis (Caprifoliaceae); raspberries, Rubus idaeus (Rosaceae); and strawberries var. Honeoye, Fragaria x ananassa Duch. (Rosaceae), were investigated for cyclooxygenase inhibitory and antioxidant activities. The presence and levels of cyanidin-3-glucosylrutinoside 1 and cyanidin-3-rutinoside 2 were determined in the fruits using HPLC. The antioxidant activity of anthocyanins from cherries was comparable to the commercial antioxidants, tert-butylhydroquinone, butylated hydroxytoluene and butylated hydroxyanisole, and superior to vitamin E, at a test concentration of 125 microg/ml. Anthocyanins from raspberries and sweet cherries demonstrated 45% and 47% cyclooxygenase-I and cyclooxygenase-II inhibitory activities, respectively, when assayed at 125 microg/ml. The cyclooxygenase inhibitory activities of anthocyanins from these fruits were comparable to those of ibuprofen and naproxen at 10 microM concentrations. Anthocyanins 1 and 2 are present in both cherries and raspberry. The yields of pure anthocyanins 1 and 2 in 100 g Balaton and Montmorency tart cherries, sweet cherries and raspberries were 21, 16.5; 11, 5; 4.95, 21; and 4.65, 13.5 mg, respectively. Fresh blackberries and strawberries contained only anthocyanin 2 in yields of 24 and 22.5 mg/100 g, respectively. Anthocyanins 1 and 2 were not found in bilberries, blueberries, cranberries or elderberries.     

Pollen morphology ofSonchus and related genera,and a general discussion

The pollen morphology of the taxa belonging to the generaAetheorhiza Cass.,Launaea Cass.,Reichardia Roth andSonchus L. in the Iberian Peninsula has been studied with light and electron microscopy. The pollen is 3(-4)-zonocolporate and echinolophate (without polar lacunae, but in general with prelacunae), with equatorial ridges and 15–20 lacunae: 3–4 poral, 6–8 abporal and 6–8 paraporal. Small to medium size, P × E = 19–36 × 23–42 µm; sometimes two different sizes have been found. Exine 3–9 µm thick and ornamentation microreticulate and echinate. The results clearly show the relationships between genera. Moreno-Socías, E., Mejías, J. A., Díez, M. J., 1994: Morfología polínica deLactuceae (Asteraceae) en la Península Ibérica, I.Lactuca y géneros relacionados. — Acta Bot. Malacitana.19: 103–113.     

Utilized and potentially utilizable seaweeds on the Namibian coast: biogeography and accessibility

Of the 1500 km coast of Namibia, only 80 km is predominantly rocky, the remaining 1420 km being sandy with only minor rocky outcrops. At present two species are utilized, Gracilaria verrucosa for agar and Laminaria schinzii for human consumption. Other potentially utilizable seaweeds occurring on the coast are Ecklonia maxima for alginates, Suhria vittata for agar, Gigartina radula, G. stiriata and Aeodes orbitosa for carrageenans and Porphyra capensis for human consumption. Laminaria schinzii also can be used for alginate production. Due to the diamond-mining areas around the 80 km rocky area at Luderitz (26° 39 S), only 20 km are accessible; hence, at present the seaweeds are being exploited almost to their full potential. The species currently not being utilized could be used, however, to support the industry that already exists at Luderitz. To increase production, access to the diamond areas would have to be gained or a cultivation program initiated.     

Molecular identification and differentiation of Brevibacterium species and strains

Amplified ribosomal DNA restriction enzyme analysis (ARDRA), pulsed field gel electrophoresis (PFGE) and ribotyping were used to differentiate among 24 strains of Brevibacterium linens, Brevibacterium casei and Brevibacterium epidermidis obtained from type culture collections or isolated from various smear ripened cheeses. ARDRA was applied to the 16S rDNA. B. linens was shown to be a quite heterogenic group with 2 to at least 4 copies of rrn operons per strain with aberrant nucleotide sequences. AccI gave genus specific restriction patterns and was used to separate Brevibacterium from Corynebacterium species. The expected species specificity of TaqI applied to B. linens type culture strains, but not to all strains isolated from cheese. By AvaI restriction, B. casei and B. linens were differentiated from B. epidermidis and the orange pigmented Arthrobacter casei, a new species of coryneform bacteria; by XmnI restriction, B. linens and B. epidermidis were differentiated from B. casei. One of 4 B. linens genotypes could not be distinguished from B. casei by this method. Here, the typical orange B. linens pigments were used for classification, which was confirmed by partial sequencing of the 16S rDNA.     

New species and new combinations in Peruvian Orchidaceae

seventeen new species and combinations are proposed in the generaChondrorhyncha, Cischweinfia, Cochlioda, Eloyella, Encyclia, Kefersteinia, Koellensteinia, Macroclinium, Rodriguezia, Solenidiopsis, andStenia. All new species are illustrated. A key is provided for 2-flowered species ofMacroclinium, PeruvianSigmatostalix, and PeruvianStenia. Solenidium (Solenidiopsis) peruvianum Schltr. is lectotypified.     

Phylogeny and biosystematics ofPseudomonotes (Dipterocarpaceae) based on molecular and morphological data

The placement of a recently discovered South American monotypic genus,Pseudomonotes tropenbosii, in subfam.Monotoideae (Dipterocarpaceae) extends the geographical range of the subfamily from Africa to the Neotropics. Although morphological and anatomical evidence suggest similarities betweenPseudomonotes andMonotes, the close alliance of these two genera was questionable due to their disjunct distribution and a lack of phylogenetic analysis. In the present study, we reconstructed the phylogeny ofPseudomonotes and other putatively related taxa usingrbcL sequence data. The analysis ofrbcL sequences of 20 taxa belonging to 15 genera and eight families recovered a single most parsimonious tree. The genusSarcolaena (Sarcolaenaceae) formed a clade sister to the monophyleticDipterocarpaceae clade.Monotes andPseudomonotes formed a strongly supported group, sister to the monophyletic clade withPakaraimaea and the remaining Asiatic dipterocarp species studied. The study strongly supports the placement ofPseudomonotes within subfam.Monotoideae of theDipterocarpaceae.