猪源产细菌素芽孢杆菌的筛选及抑菌特性

【背景】抗生素作为生长促进剂在畜牧业中的滥用,出现了严重的耐药基因富集和扩散问题,发掘新兴的生长促进剂作为饲料添加剂市场潜力巨大,目前益生菌制剂的开发最具潜力。【目的】通过对散养健康育肥猪粪便中芽孢杆菌的分离筛选,获得对典型肠道病原菌具有显著抑菌活性的芽孢杆菌,确定其产生的细菌素特性,以此对芽孢杆菌作为猪养殖业生长促进剂的潜力进行评价。【方法】采用梯度稀释涂平板法分离可培养细菌,利用牛津杯法检测菌株的抑菌活性。通过微生物形态及16S r RNA基因序列分析,确认6株产细菌素菌株的分类地位,并对其抗生素耐药性、细菌素稳定性及生理生化特征进行比较分析。【结果】从116株纯培养物中筛选得到6株对指示病原细菌具有显著抑菌效应的产细菌素芽孢杆菌,其中2株为贝莱斯芽孢杆菌(Bacillus velezensis),3株为枯草芽孢杆菌(Bacillus subtilis),1株为地衣芽孢杆菌(Bacillus licheniformis)。菌株B.licheniformis DY7和B.subtilis FX4对致泻、产肠毒素、出血性Escherichia coli均有显著的抑制效果,对头孢噻肟和红霉素高度敏感,其细菌素在p H 3.0-9.0、50-100°C水浴处理后仍具有明显的抑菌活性。【结论】猪源产细菌素芽孢杆菌DY7和FX4具有高效的病原细菌抑菌能力,所产细菌素稳定性较好,具有作为动物生长促进剂的应用潜力。     

牦牛源产细菌素屎肠球菌的分离鉴定和益生特性

【背景】抗生素的滥用导致牦牛肠道常见病原菌耐药性增加,益生菌作为对抗耐药性细菌的新型武器,应用前景广阔。【目的】获取益生特性优良的牦牛源益生菌。【方法】将20份牦牛粪便样本在含0.5%CaCO3的MRS培养基上分离纯化,以大肠杆菌和金黄色葡萄球菌为指示菌,用牛津杯法筛选有抑菌活性的菌株;排除酸和过氧化氢后,经耐酸耐热试验和蛋白酶敏感试验筛选产细菌素菌株,用形态学和16S rRNA基因序列分析鉴定;通过对大肠杆菌、沙门氏菌等腹泻病原菌体外抑菌试验、耐模拟胃肠液、测定自聚集能力和疏水性及抗生素敏感试验分析益生特性。【结果】从20份牦牛粪便样本中共分离出11株产生溶钙圈的菌株,其中6株对大肠杆菌和金黄色葡萄球菌抑菌效果显著,经复筛得到2株产细菌素的乳酸菌SC6和SC9,经鉴定均为屎肠球菌(Enterococcus faecium)。其中SC9对腹泻病原菌抑菌效果明显,有良好的耐受性和肠道黏附能力,对5种常用抗生素均敏感。【结论】屎肠球菌SC9有一定的抗逆性和潜在的益生能力,具备作为益生菌的潜力。     

沙棘根瘤内生细菌中抑菌促生菌株的筛选和鉴定

【背景】植物内生细菌可产生具有抑菌和促生活性的物质,既能抑制植物病原菌对寄主植物的侵染,也能促进植物的生长。沙棘根瘤内生细菌是根瘤内除共生固氮的弗兰克氏菌外,与沙棘共生的一大类微生物。研究具有抑菌和促生活性的植物内生菌,可为微生物菌肥的研究提供理论基础。【目的】筛选具有优良抑菌和促生活性的沙棘根瘤内生细菌,初步研究其抑菌和促生活性,并对菌株进行鉴定。【方法】采用双层琼脂法、琼脂扩散法、双层平板对峙法、牛津杯法进行沙棘根瘤拮抗性内生细菌的筛选。选取抑菌活性较高的内生细菌,分别采用Salkowski比色法、ChromeAzurolS(CAS)平板检测法和钼锑抗比色法进行产吲哚乙酸、铁载体及溶磷能力的测定。采用发酵液灌根法测定沙棘根瘤内生细菌SR308对黄瓜促生作用的盆栽效果。通过形态和培养特征、生理生化试验及16S rRNA基因序列分析法对菌株TT201和SR308进行鉴定。【结果】从131株沙棘根瘤内生细菌中筛选出9株具有较强抑菌活性的内生细菌,其中菌株TT201抑菌性最佳、抑菌谱广;菌株SR308的促生活性最好,其发酵液对黄瓜的生长具有较强的促进作用。对具有较强抑菌和促生活性的菌株TT201和SR308进行鉴定的结果表明,菌株TT201为侧孢短芽孢杆菌(Brevibacilluslaterosporus),菌株SR308为蕈状芽孢杆菌(Bacillusmycoides)。【结论】获得2株具有优良抑菌和促生活性的沙棘根瘤内生细菌,为进一步开发微生物农药及菌肥提供了资源。     

小龙虾肠道产木聚糖酶细菌的分离与鉴定

【背景】小龙虾肠道微生物是小龙虾降解纤维素和半纤维素的主要驱动力。【目的】研究肠道内细菌的相对丰度,为揭示肠道微生物在小龙虾纤维素降解过程中的作用提供理论支撑。【方法】采用纯培养法从小龙虾肠道筛选产木聚糖酶细菌,并且对小龙虾肠道细菌进行16S高通量测序。【结果】形态学和16SrRNA基因分子鉴定表明,筛选到的4株产木聚糖酶细菌均属于芽孢杆菌科芽孢杆菌属;结合进一步的生理生化特征鉴定,结果为:菌株Z-3为枯草芽孢杆菌(Bacillus subtilis),菌株Z-4为贝莱斯芽孢杆菌(Bacillus velezensis),菌株Z-29为蜡状芽孢杆菌(Bacillus cereus),菌株Z-30为高地芽孢杆菌(Bacillus altitudinis);16S rRNA基因高通量测序结果表明:在属水平上,小龙虾肠道细菌主要是Candidatus Bacilloplasma、拟杆菌属、弧菌属、不动杆菌属、Dysgonomonas、Tyzzerella3、气单胞菌属和希瓦氏菌属细菌。【结论】小龙虾肠道内细菌资源丰富,且芽孢杆菌属细菌在木质纤维素降解过程中发挥一定功能。     

细菌素Paracin 1.7的纯化与性质研究

摘要：【目的】分离纯化（Lactobacillus paracasei）HD1.7所产生的细菌素并分析其特性。【方法】细菌素Paracin 1.7的纯化采用色谱技术,其分子量检测采用十二烷基磺酸钠－聚丙烯酰胺凝胶电泳（SDS-PAGE）,利用琼脂扩散法测定细菌素活力。【结果】Paracin 1.7分离于我国传统发酵食品酸菜发酵液中,其产生菌为副干酪乳杆菌。 Paracin 1.7可以抑制其它微生物的生长,为细菌素。该菌在稳定期可产生大量Paracin 1.7。经过阳离子交换层析、凝胶过滤层析以及高效液相色谱（HPLC）,对该细菌素进行了初步纯化,并经Tricine-SDS-PAGE检测其分子量大约为11 kDa。Paracin 1.7抑菌谱较广,其抑菌范围包括Proteus, Bacillus, Enterobacter, Staphylococcus, Escherichia, Lactobacillus, Microccus, Pseudomonas, Salmonella, Saccharomyces,其中有些为食品源致病菌。该细菌素在酸性及高温下稳定,对几种蛋白质酶敏感。该细菌素对敏感菌株的作用方式为抑菌。在4oC保存4个月后,Paracin 1.7的抑菌活性保持稳定。【结论】基于细菌素Paracin 1.7的性质,该细菌素可用作食品防腐剂。     

1株产细菌素乳酸菌的筛选和鉴定

目的 从植物性材料中筛选产细菌素的乳酸菌。方法 琼脂扩散法。结果 所筛选的产细菌素R260菌株经鉴定为植物乳杆菌。排除有机酸、过氧化氢等干扰因素后,发酵液仍有很强的抑菌作用;用胰蛋白酶和胃蛋白酶处理后,发酵液抑菌活性急剧下降,因而确定产生的抑菌物质具有蛋白质性质,是一种细菌素。抑菌谱试验测定表明,此菌株的发酵液不仅抑制革兰阳性菌,而且对部分革兰阴性菌也有抑制作用,因此产生的是一类广谱细菌素。结论筛选到了1株产广谱细菌素的乳酸菌。     

北黄海沉积物可培养产蛋白酶细菌分离鉴定

【目的】揭示北黄海沉积物中可培养产胞外蛋白酶细菌及蛋白酶多样性,增加人们对北黄海生态系统中产蛋白酶菌多样性的认识,为海洋产蛋白酶微生物的挖掘提供菌群资源。【方法】分别将5个北黄海沉积物样品梯度稀释涂布至酪蛋白明胶筛选平板,选择性分离产蛋白酶细菌;并通过分析基于16S rRNA基因序列的系统发育关系,揭示这些细菌的分类地位和遗传多样性;分别测定胞外蛋白酶活性并对酶活较高的39株菌进行基于苯甲基磺酰氟(PMSF,丝氨酸蛋白酶抑制剂)、邻菲罗啉(o-phenanthroline,O-P,金属蛋白酶抑制剂)、E-64(半胱氨酸蛋白酶抑制剂)和pepstatin A(天冬氨酸蛋白酶抑制剂)4种抑制剂的酶活抑制实验以及所有菌株对3种底物(酪蛋白、明胶、弹性蛋白)的水解能力;分析这些细菌所产胞外蛋白酶的特性及多样性。【结果】从5个北黄海沉积物样品中分离获得66株产蛋白酶细菌,这些菌株隶属于Bacteroidetes、Proteobacteria、Actinobacteria和Firmicutes 4个门的7个属,其中Pseudoalteromonas(69.9%)、Sulfitobacter(12.1%)和Salegentibacter(10.6%)是优势菌群;沉积物中可培养的产蛋白酶细菌的丰度为104 CFU/g;蛋白酶酶活抑制实验表明所有测定菌株产生的胞外蛋白酶属于丝氨酸蛋白酶和/或金属蛋白酶,仅有少数菌株所产蛋白酶具有半胱氨酸蛋白酶或天冬氨酸蛋白酶活性。【结论】北黄海沉积物中可培养产蛋白酶细菌类群较为丰富,Pseudoalteromonas、Sulfitobacter和Salegentibacter菌株是优势菌群,测定菌株所产胞外蛋白酶主要是丝氨酸蛋白酶和/或金属蛋白酶。     

紫菜外生细菌抑菌活性及其多聚酮合酶（PKS I） 基因筛选

摘要：【目的】：基于紫菜外生细菌抑菌活性的研究,本文对具有广谱抑菌活性的菌株进行了多聚酮合酶(Polyketide synthase I, PKS I)基因的筛选,以期获得PKS I 阳性菌株及探讨紫菜藻际微生物区系细菌的拮抗机制与PKS I 途径的关系。【方法】：利用琼脂柱法筛选出具广谱抑菌活性的菌株31株,以其基因组DNA 为模板,设计引物扩增酮基合成酶(Ketosynthase, KS) 片段基因并将其克隆到 pMD19-T Vector,筛选出PKS I 阳性菌,进行16S rDNA 测序分析。【结果】：紫菜外生细菌表现出广谱抑菌性。从温州病烂紫菜外生菌中筛选出3株具强抑菌活性的PKS I 阳性菌,BLAST 比对结果显示：菌株WPhG3、WPySw1和WPySw2 扩增得到PKS I 的KS 结构域核苷酸序列所对应的氨基酸序列与Bacillus subtilis subsp. subtilis str. 168（NP_389602）、Bacillus subtilis (ABR19776)和Aspergillus carbonarius (AAZ99721) 的PKS I 的KS 结构域的同源性分别达到98%、99%和98%;16S rDNA 系统发生学分析显示它们均与Bacillus 的同源性最高。【结论】：紫菜藻际微生物群落组成复杂,通过多条途径调节藻际微生物区系的平衡。PKS I 途径可能是温州病烂紫菜外生菌Bacillus 表现抑菌活性的一种方式。     

短头熊蜂肠道拮抗菌株的分离筛选鉴定及其生物特性评价

【背景】近年来,由于栖息地减少、农药的大量使用及病原菌侵染等综合因素,导致全世界的熊蜂种类与数量逐年减少,病原菌的侵染可通过微生物在自身生长过程中会产生的抑菌物质进行有效抑制或杀灭。【目的】短头熊蜂(Bombus breviceps)长期生存在野外环境中,其肠道内存在着大量微生物资源。从短头熊蜂肠道内筛选拮抗菌株,并对其抑菌特性进行研究。【方法】采用牛津杯双层法筛选拮抗菌株,测定抑菌活性最佳菌株发酵液的抑菌物质稳定性与抑菌广谱性等抑菌特性,并借助细胞膜通透性、流式细胞仪检测等试验探究其抑菌机制。【结果】得到了5株具有明显抑菌作用的拮抗菌株,其中果杆菌(Fructobacillus tropaeoli)CZ01对金黄色葡萄球菌(Staphylococcus aureus)、沙门氏菌(Salmonella choleraesuis)、大肠杆菌(Escherichia coli)、福氏志贺氏菌(Shigella flexneri)和无乳链球菌(Streptococcus agalactiae)这5种病原指示菌都具有高度抑菌效果。菌株CZ01对金黄色葡萄球菌的抑菌效果最佳,抑菌圈直径可达到(21.21±0.25) mm,在121 ℃处理后仍具有67.36%以上的抑菌活性,调整pH值为10.0时仍具有78.16%的抑菌活性。【结论】短头熊蜂肠道微生物资源较丰富,尤其是果杆菌(F.tropaeoli)CZ01具有抑菌活性高、稳定性好、抑菌谱广等特性,对金黄色葡萄球菌具有良好的杀灭效果,显示出良好的应用潜能。     

土壤产铁载体细菌的筛选及其对铁氧化物的活化与利用

【背景】土壤中铁主要以难溶态铁氧化物的形式存在,有效性较低,产铁载体细菌对铁氧化物的活化是提高铁利用效率的有效途径。【目的】从林木土壤筛选产铁载体细菌,并观察菌株对难溶性铁氧化物的利用效应,可为土壤微生物资源开发及其在养分调控中的作用提供理论依据。【方法】通过CAS检测法从林木根系附近表层土壤中分离产铁载体细菌,借助生物培养实验分析温度和pH对微生物生长和铁载体产生的影响,通过振荡平衡实验,探究细菌产铁载体对铁氧化物的活化效应。【结果】通过CAS检测法从林木根系附近表层土壤中分离得到12株产铁载体细菌,16S rRNA基因扩增子测序初步鉴定结果显示筛选细菌均为假单胞菌属。选取铁载体产生能力和生长活性较高的两株细菌ARSB02和CNRSB01作为重点研究对象,结果显示,不同条件下CNRSB01的生物量和铁载体产量均高于菌株ARSB02,22 h时菌株ARSB02和CNRSB01的铁载体活性单位分别达到67.07%和84.60%。pH 5.0-8.0的范围内两株细菌可以保持较好的铁载体产生能力,菌株ARSB02和CNRSB01在pH7.0时铁载体产生能力最强,铁载体活性单位分别达到38.9...     

Phylogeny of cardinalfishes (Teleostei: Gobiiformes: Apogonidae) and the evolution of visceral bioluminescence

The cardinalfishes (Apogonidae) are a diverse clade of small, mostly reef-dwelling fishes, for which a variety of morphological data have not yielded a consistent phylogeny. We use DNA sequence to hypothesize phylogenetic relationships within Apogonidae and among apogonids and other acanthomorph families, to examine patterns of evolution including the distribution of a visceral bioluminescence system. In conformance with previous studies, Apogonidae is placed in a clade with Pempheridae, Kurtidae, Leiognathidae, and Gobioidei. The apogonid genus Pseudamia is recovered outside the remainder of the family, not as sister to the superficially similar genus Gymnapogon. Species sampled from the Caribbean and Western Atlantic (Phaeoptyx, Astrapogon, and some Apogon species) form a clade, as do the larger-bodied Glossamia and Cheilodipterus. Incidence of visceral bioluminescence is found scattered throughout the phylogeny, independently for each group in which it is present. Examination of the fine structure of the visceral bioluminescence system through histology shows that light organs exhibit a range of morphologies, with some composed of complex masses of tubules (Siphamia, Pempheris, Parapriacanthus) and others lacking tubules but containing chambers formed by folds of the visceral epithelium (Acropoma, Archamia, Jaydia, and Rhabdamia). Light organs in Siphamia, Acropoma, Pempheris and Parapriacanthus are distinct from but connected to the gut; those in Archamia, Jaydia, and Rhabdamia are simply portions of the intestinal tract, and are little differentiated from the surrounding tissues. The presence or absence of symbiotic luminescent bacteria does not correlate with light organ structure; the tubular light organs of Siphamia and chambered tubes of Acropoma house bacteria, those in Pempheridae and the other Apogonidae do not.     

Tracking down the "head blob": comparative analysis of wingless expression in the developing insect procephalon reveals progressive reduction of embryonic visual system patterning in higher insects

The evolution of larval head morphology in holometabolous insects is characterized by reduction of antennal appendages and the visual system components. Little insight has been gained into molecular developmental changes underlying this morphological diversification. Here we compare the expression of the segment polarity gene wingless (wg) in the pregnathal head of fruit fly, flour beetle and grasshopper embryos. We provide evidence that wg activity contributes to segment border formation, and, subsequently, the separation of the visual system and protocerebrum anlagen in the anterior procephalon. In directly developing insects like grasshopper, seven expression domains are formed during this process. The activation of four of these, which correspond to polar expression pairs in the optic lobe anlagen and the protocerebral ectoderm, has shifted to postembryonic stages in flour beetle and Drosophila. The remaining three domains map to the protocerebral neuroectoderm, and form by disintegration of a large precursor domain in flour beetle and grasshopper. In Drosophila, the precursor domain remains intact, constituting the previously described “head blob”. These data document major changes in the expression of an early patterning gene correlated with the dramatic evolution of embryonic visual system development in the Holometabola.     

Molecular phylogeny of Phebalium (Rutaceae: Boronieae) and related genera based on the nrDNA regions ITS 1+2

Parsimony analyses of the internal transcribed spacer regions of nuclear ribosomal DNA (ITS 1 & ITS 2) for 38 taxa sampled from the Phebalium group (Rutaceae: Boronieae) and two outgroups confirm that, with the exception of Phebalium sensu stricto and Rhadinothamnus, six of the currently recognised genera within the group are monophyletic. The data indicate that Phebaliums. str. is paraphyletic with respect to Microcybe, and Rhadinothamnus is paraphyletic with respect to Chorilaena. Rhadinothamnus and Chorilaena together are the sister group to Nematolepis. Drummondita, included as an outgroup taxon, clustered within the ingroup as sister to Muiriantha and related to Asterolasia.The phylogeny suggests that the evolution of major clades within a number of these genera (e.g. Phebalium) relates to vicariance events between eastern and south-western Australia. Leionema is an eastern genus, with the most basal taxon being the morphologically distinct Leionema ellipticum from northern Queensland. Leionema also includes one species from New Zealand, but this species (as with some others) proved difficult to sequence and its phylogenetic position remains unknown. Taxonomic changes at the generic level are recommended.The authors wish to thank Paul G.Wilson, PERTH, for advice and discussion, and Paul Forster, BRI, for collecting and providing material of Leionema ellipticum. The project was supported by a Melbourne University Postgraduate Award (to BM), the Australian Biological Resources Study (ABRS), Australian Systematic Botany Society and Wolf Den (Australia) Investments.     

Radiation, multiple dispersal and parallelism in the skinks, Chalcides and Sphenops (Squamata: Scincidae), with comments on Scincus and Scincopus and the age of the Sahara Desert

Phylogenetic analysis using up to 1325 base pairs of mitochondrial DNA from 179 specimens and 30 species of Chalcides, Sphenops, Eumeces, Scincopus and Scincus indicates that Sphenops arose twice independently within Chalcides. It is consequently synonymized with that genus. Chalcides in this broader sense originated in Morocco, diversifying into four main clades about 10 Ma, after which some of its lineages dispersed widely to cover an area 40 times as large. Two separate lineages invaded the Canary Islands and at least five main lineages colonized southern Europe. At least five more spread across northern Africa, one extending into southwest Asia. Elongate bodies with reduced limbs have evolved at least four times in Chalcides, mesic 'grass-swimmers' being produced in one case and extensive adaptation to life in loose desert sand in two others. In clade, Chalcides striatus colonized SW Europe from NW Africa 2.6 Ma and C. chalcides mainland Italy 1.4 Ma, both invasions being across water, while C. c. vittatus reached Sardinia more recently, perhaps anthropogenically, and C. guentheri spread 1200km further east to Israel. C. minutus is a composite, with individuals from the type locality forming a long independent lineage and the remaining ones investigated being most closely related to C. mertensi. In the Northern clade, C. boulengeri and C. sepsoides spread east through sandy habitats north of the Sahara about 5 Ma, the latter reaching Egypt. C. bedriagai invaded Spain around the same time, perhaps during the Messinian period when the Mediterranean was dry, and shows considerable diversification. Although it is currently recognized as one species, the C. ocellatus clade exhibits as much phylogenetic depth as the other main clades of Chalcides, having at least six main lineages. These have independently invaded Malta and Sardinia from Tunisia and also southwest Arabia C. o. humilis appears to have spread over 4000 km through the Sahel, south of the Sahara quite recently, perhaps in the Pleistocene. In the Western clade of Chalcides, C. delislei appears to have dispersed in a similar way. There were also two invasions of the Canary Islands: one around 5 Ma by C. simonyi, and the other about 7 Ma by the ancestor of C. viridanus+C. sexlineatus. C. montanus was believed to be related to C. lanzai of the Northern clade, but in the mtDNA tree it is placed within C. polylepis of the Western clade, although this may possibly be an artifact of introgression. The Eumeces schneideri group, Scincopus and Scincus form a clade separate from Chalcides. Within this clade, the geographically disjunct E. schneideri group is paraphyletic. One of its members, E. algeriensis is the sister taxon to Scincopus, and Scincus may also be related to these taxa. The phylogeny suggests Scincopus entered desert conditions in Africa, up to 9.6 Ma and the same may have been true of Scincus up to 11.7 Ma. Scincus appears to have diversified and spread into Arabia around 6 Ma. Dates of origin and divergence of these skinks, desert Chalcides and other squamates agree with recent geological evidence that the Sahara is at least 5-7 My old. The subspecies Chalcides viridanus coeruleopunctatus is upgraded to the species level as C. coeruleopunctatus stat nov., on the basis of its large genetic divergence from C. v. viridanus.     

Evaluation of antiprotozoal and antimycobacterial activities of the resin glycosides and the other metabolites of Scrophularia cryptophila

Resin glycosides are secondary metabolites exclusive to the convolvulaceous plants. In this study, crypthophilic acids A–C (1–3), the first resin glycosides occurring in another family (Scrophulariaceae), and the other constituents of Scrophularia cryptophila were examined for in vitro antiprotozoal and antimycobacterial potentials. Except for crypthophilic acid B (2), all tested compounds exhibited growth-inhibitory effect against Trypanosoma brucei rhodesiense, with l-tryptophan (6) and buddlejasaponin III (7) being the most potent ones (IC₅₀'s 4.1 and 9.7 μg/ml). In contrast, the activity towards Trypanosoma cruzi was poor, and only crypthophilic acid C (3), 6 and 7 were trypanocidal at concentrations above 40 μg/ml. With the exception of 2 and 6, all compounds were active against Leishmania donovani. Harpagide (4) and 3 emerged as the best leishmanicidal agents (IC₅₀'s 2.0 and 5.8 μg/ml). Only compounds 3, 6 and 7 showed antimalarial activity against Plasmodium falciparum with IC₅₀ values of 4.2, 16.6 and 22.4 μg/ml. Overall the best and broadest spectrum activity was presented by compounds 3 and 7, as they inhibited all four parasitic protozoa. None of the isolates had significant activity against Mycobacterium tuberculosis (MICs >100 μg/ml) or were toxic towards mammalian (L6) cells. This is the first report of antiprotozoal activity for natural resin glycosides, as well as for harpagide (4), acetylharpagide (5), tryptophan (6) and buddlejasaponin III (7).     

Ultrastructure and Development of the Ooecial Walls in Some Calloporid Bryozoans (Gymnolaemata: Cheilostomata)

In the brood chambers (ovicells) of six calloporid cheilostomes studied each skeletal wall consists of four calcified layers: (1) a very thin superficial layer of planar spherulitic crystallites, (2) an upper (outer) layer with wall-perpendicular prismatic ultrastructure, (3) an intermediate lamellar layer, and (4) a lower (inner) wall-perpendicular prismatic layer. Comparative studies of both the ovicell wall ultrastructure and early ovicell formation showed a hypothetical opportunity for evolving complex (multilayered) skeletal walls by fusion of the initially separated gymnocystal and cryptocystal calcifications in Cheilostomata. In two species studied, a bilobate pattern in the final stage of the formation of the ooecial roof was encountered in specimens with the cuticle preserved. A possible explanation to this finding is discussed – the bilobate pattern is suggestive of the hypothetical origin of the brood chamber from (1) two flattened spines, or (2) reduction in spine number of an originally multispinous ovicell.     

Phylogeographic patterns in Leccinum sect. Scabra and the status of the arctic-alpine species L. rotundifoliae

We investigated inter- and intraspecific phylogenetic relationships in the ectomycorrhizal fungal genus Leccinum section Scabra. Species of this section are exclusively associated with Betula and occur throughout the Northern Hemisphere. We compared the phylogenetic relationships of arctic, alpine, boreal and temperate accessions of section Scabra based on DNA sequences of the single-copy nuclear gene Gapdh and the multiple-copy nuclear region 5.8S-ITS2. Exclusively arctic lineages were not detected in species that occur both in arctic-alpine or boreal regions, except in L. rotundifoliae that was restricted to cold climates. L. scabrum and L. holopus showed an intercontinental phylogeographic pattern, and L. variicolor showed a pattern unrelated to geographical distribution. Molecular clock estimates indicated that L. rotundifoliae is as old as other species in section Scabra. Individual gene trees suggest that interspecific hybridisation occurred several times in the evolution of section Scabra.     

Phylogeny and taxonomic revision of plastid-containing euglenophytes based on SSU rDNA sequence comparisons and synapomorphic signatures in the SSU rRNA secondary structure

Sequence comparisons and a revised classification of the Euglenophyceae were based on 92 new SSU rDNA sequences obtained from strains of Euglena, Astasia, Phacus, Trachelomonas, Colacium, Cryptoglena, Lepocinclis, Eutreptia, Eutreptiella and Tetreutreptia. Sequence data also provided molecular signatures for taxa from genus to class level in the SSU rRNA secondary structure, revealed by a novel approach (search for non-homoplasious synapomorphies) and used for taxonomic diagnoses. Photosynthetic euglenoids and secondary heterotrophs formed a clade, designated as Euglenophyceae (emend.) with two orders: Euglenales and Eutreptiales. The mostly marine Eutreptiales (Eutreptia, Eutreptiella; not Distigma) comprised taxa with two or four emergent flagella (the quadriflagellate Tetreutreptia was integrated within Eutreptiella). The Euglenales (freshwater genera with < or = one emergent flagellum) formed nine clades and two individual branches (single strains); however, only two clades were congruent with traditional genera: Trachelomonas (incl. Strombomonas) and Colacium. Euglena was polyphyletic and diverged into four independent clades (intermixed with Astasia, Khawkinea and Lepocinclis) and two individual branches (e.g. E. polymorpha). Phacus was also subdivided into Phacus s. str. and two combined lineages (mixed with Lepocinclis spp. or Cryptoglena). In consequence, Euglena (s. str.), Phacus and other genera were emended and one lineage (mixed Phacus/Lepocinclis-clade) was recognized as the previously neglected genus Monomorphina Mereschkowsky (1877). The sister clade of Phacus s. str. (mixed Euglena/Lepocinclis-clade) was identified as Lepocinclis Perty (emended).     

Planting densities and bird and rodent absence affect size distributions of four dicots in synthetic tallgrass communities

Variability in the size distributions of populations is usually studied in monocultures or in mixed plantings of two species. Variability of size distributions of populations in more complex communities has been neglected. The effects of seeding density (35 or 350 seeds/species/m²) and presence of small vertebrates on the variability of size distributions were studied for a total of 1,920 individuals of 4 species in replicated synthetic communities of 18 species in northern Illinois. End-of season height and above-ground biomass were measured for prairie perennials Dalea purpurea (purple prairie clover), Echinacea purpurea (purple coneflower), Desmanthus illinoensis (Illinois bundleflower) and Heliopsis helianthoides (early sunflower). Variability in biomass distribution of the four target species was twice as great at low than at high densities when small vertebrates were excluded. Our results suggest that inter- and intraspecific competition may affect all individuals more under high-density conditions, thereby reducing the variability in their biomass distributions within this community. This result, a consequence of plant-plant interaction, is obscured when small birds or mammals are present, presumably because either or both add variance that overwhelms the pattern.     

The phylogenetic placement of the Leucogastrales, including Mycolevis siccigleba (Cribbeaceae), in the Albatrellaceae using morphological and molecular data

The morphology of many hypogeous fungi converges on a homogeneous reduced form, suggesting that disparate lineages are subject to a uniform selection pressure. The primary goal of this study was to evaluate the morphology and infer the phylogeny of the Leucogastrales with Mycolevis siccigleba using a Bayesian methodology. A comprehensive morphological assessment was used for an a priori phylogenetic inference to guide the sequencing effort. All structures except spore ornamentation pointed to the Albatrellaceae as the most likely sister taxon. Polyporoletus sublividus, a close relative of Albatrellus, produces ornamented basidiospores with a similar structure to M. siccigleba basidiospores. The ITS from 30 taxa was used for the molecular phylogenetic analysis. P. sublividus was found sister to Mycolevis. Leucophleps spinispora and L. magnata formed a group sister to the Polyporoletus/Mycolevis group, whereas Leucogaster was polyphyletic with respect to the core of the Leucogastrales and sister to A. caeruleoporus. This relationship was expected as previously undescribed chlamydospores produced by members of Albatrellus had a similar morphology to the basidiospores of L. rubescens.