凝胶电泳与共头霉分类

对于共头霉的唯一已知种总状共头霉Syncephalastrum racemosum Cohn ex Schroet.(S 1)和新发现的具单孢孢子囊的新种单孢共头霉下的三个变种,原变种S.monosporum Zheng et al. var. monosporum( S7, S8, S9),冠囊变种var. cristatum Zheng et al. (S10, S11)及多重生变种var. pluriproliferum Zheng et al.(S12) 6株菌的蛋白和酯酶进行了聚丙烯酰胺凝胶电泳的研究.这4个分类群的蛋白图谱和酯酶酶谱互不相同,而同一分类群内各个菌株之间则有很高的相似性.从计算出的相似性百分数来看,3个新变种相互之间有较为密切的关系,而和S. racemosum虽然较疏远,但也有相当程度的亲缘关系.这些结果被认为支持了根据形态鉴定而定这三个新变种属于同一新种内并将其和S. racemosum一并归属于一个属内.由此可见,聚丙烯酰胺凝胶电泳是共头霉分类的有用的辅助手段.     

共头霉的单孢孢子囊变种

我们从江苏、浙江两省采到的一些样品中分离得到六株毛霉菌种并鉴定为分别属于三个变种的同一个新种,即单孢共头霉原变种(Syncephalastrum monosporum Zheng et al.sp. nov. var. monosporum),单孢共头霉冠囊变种(Syncephalastrum monosporum Zheng et al. var. cristatum Zheng et al. var. nov.),以及单孢共头霉多重生变种(Syncephalastrum monosporum Zheng et al. var. pluriproliferum Zheng et al. var. nov.).它们与共头霉属的唯一已知种总状共头霉(Syncephalastrum racemosum Cohn ex Schroeter)的主要区别在于全部孢子囊都是单孢的小型孢子囊(柱孢囊)而不是像后者一样孢子成单行排列的多袍柱孢囊.我们把这些菌归人共头霉属而没有为它们专门成立一个新属的理由除因它们与总状共头霉有明显的亲缘关系外,主要还因我们认为在不具备其它重要区别特征的情况下,在毛霉目的分类中,无论在科级或属级的水平上,都不应过分强调单孢孢子囊的作用.尽管单孢共头霉是毛霉目内唯一具单孢子柱孢囊的种,我们仍然把它们归入共头霉属内.至于我们为什么要把这六株菌鉴定为一个种的三个变种而不是三个各自独立的种,则是由于我们承认共头霉属的唯一已知种总状共头霉是一个变异性很大的种而并没有把它分成许多不同的种,因此,我们对这个属的另外一个种,即单孢共头霉同样采用较大的种概念.     

六种疫霉菌体可溶性蛋白质和酯酶的聚丙烯酰胺凝胶平板电泳研究

本文对槐生疫霉、掘氏疫霉、樟疫霉、烟草疫霉、棕榈疫霉及辣椒疫霉等６种疫霉８１株菌菌体可溶性蛋白质和酯酶进行了聚丙烯酰胺凝胶平板电泳研究。结果指出,种间蛋白质图谱差异明显,种内菌株间基本一致。就酯酶图谱来说,种内菌株间存在一定差异,但种间差异更为显著。因此,本试验进一步支持槐生疫霉新种的建立。同时表明,菌体可溶性蛋白质电泳图谱在疫霉种的鉴定和分类上具有重要意义,而酯酶电泳图谱在一定程度上也有助于疫霉     

疫霉蛋白质的聚丙烯酰胺凝胶盘状电泳

对北京,河北、青海、山东、湖北、广东等地不周寄主上收集的,属于 Phytophthoracapsici,P．Colocasiae,P．Heveac,P．Infestans, P．Melonis P．Nicotianac var. nicotianae 和 P．Nicotianae var. parasitica 7个种和变种的17株菌的蛋白质,进行了聚丙烯酰胺凝胶盘状电泳的分析和比较,经多批次试验,7个种和变种各自恒定地呈现典型的蚤白质图谱,显示出种间的差异。其中 P. infestans 占11株,它们虽采自不同地区和寄主,又分属5个生理小种,经多次电泳,电泳图谱基本相同,但不同菌株存在一些差异。同一株菌,培养条件对电泳图谱影响不大,不论是培养在含苹果酸的培养基上,还是培养在延胡索酸培养基上,电泳图谱基本一致,只是电泳材料必须新鲜。     

六种疫霉菌体可溶性蛋白质和酯酶的聚丙烯酰胺凝胶平板电泳研究*

本文对槐生疫霉(Phytophthora robinicola)、掘氏疫霉(P.drechsleri)、樟疫霉(P.cinnamomi)、烟草疫霉(P.nicotianae)、棕榈疫霉(P.palmivora)及辣椒疫霉(P.capsici)等6种疫霉81株菌菌体可溶性蛋白质和酯酶进行了聚丙烯酰胺凝胶平板电泳研究。结果指出,种间蛋白质图谱差异明显,种内菌株间基本一致。就酯酶图谱来说,种内菌株间存在一定差异,但种间差异更为显著。因此,本试验进一步支持槐生疫霉新种的建立。同时表明,菌体可溶性蛋白质电泳图谱在疫霉种的鉴定和分类上具有重要意义,而酯酶电泳图谱在一定程度上也有助于疫霉种的研究。     

六种疫霉菌体可溶性蛋白质和酯酶的聚丙烯酰胺凝胶平板电泳研究

本文对槐生疫霉(Phytophthora robinicola)、掘氏疫霉(P.drechsleri)、樟疫霉(P.cinnamomi)、烟草疫霉(P.nicotianae)、棕榈疫霉(P.palmivora)及辣椒疫霉(P.capsici)等6种疫霉81株菌菌体可溶性蛋白质和酯酶进行了聚丙烯酰胺凝胶平板电泳研究。结果指出,种间蛋白质图谱差异明显,种内菌株间基本一致。就酯酶图谱来说,种内菌株间存在一定差异,但种间差异更为显著。因此,本试验进一步支持槐生疫霉新种的建立。同时表明,菌体可溶性蛋白质电泳图谱在疫霉种的鉴定和分类上具有重要意义,而酯酶电泳图谱在一定程度上也有助于疫霉种的研究。     

水玉霉(Pilobolus)属的种的重新划分

我们在过去的工作中承认水玉霉(Pilobolus)属的9个种(郑、胡,见戴,1979)。近年来我们重新分离得到了这些分类群并对它们进行了再研究。研究结果表明,尽管它们是彼此可以互相区分的分类群,但是,包括我们过去的概念在内,目前被普遍接受的用于这个属的分类的种概念太小.为了与整个毛霉目的其它属的分类系统相一致,我们把这9个分类群重新划分为由9个变种组成的5个种:晶澈水玉霉原变种[Pilobolus crystallinu (Wigg.) Tode var. crystallinus],晶澈水玉霉透孢变种新组合[P. crystallinus var.hyalosporus (Boedijn) Hu &Zheng, comb. nov.],晶澈水玉霉克莱因变种新组合[P. crystallinus var.kleinii (van Tieghem) Zheng &G.-q. Chen, comb. nov.],豆状水玉霉原变种(P. lentiger Corda var. lentiger),豆状水玉霉小型变种新组合[P. lentiger var. minutus (Speg.) Zheng &G.-q. Chen, comb. nov.],长型水玉霉(P. longipes van Tieghem),厚壁水玉霉(P. Oedipus Mont.),露水玉霉原变种[P. roridus (Bolt.) Pers. var. roridus],露水玉霉突囊变种新组合[P. roridus var. umbonatus (Butler) Hu &Zheng, comb. nov.]。水玉霉属先后报道过的种或种下分类群名称共计50个左右,其中一些异名往往被不同的作者归到不同的正名下面。为了解决它们的正确归属问题,我们对全部原始描述作了细致的文献考证然后决定其位置。对那些找不到原始描述或从原始描述中得不出结论的则作为可疑名称处理。可疑名称共计12个:Mucor obliquus Scop., M. urceolatus Dicks.;Pilobolus urceolatus Purt., P. pestis-bovinae Hallier(=P. hallierii Rivolta), P. nanus van Tieghem, P. intermedius, (Coem.) P. A. Karsten(=P. Oedipus Mont. var,intermedius Coem.), P. pullus Massee, P.proliferens McVickar, P. ramosus McVickar, P. simplex McVickar, P. lentiger forma leinii Reyn. &Laysa, P. lentiger forma minutus Reyn. &Laysa.     

九株根霉可溶性蛋白和三种酶的电泳图谱比较研究

本文报导了根霉属(Rhizopus) 9个菌株天然态及解聚态可溶性蛋白、酯酶同工酶、葡萄糖淀粉酶和SOD电泳图谱的比较研究。结果表明：可溶性蛋白图谱和酯酶同工酶谱能显示五种已知供试菌种间的差异,尤其酯酶同工酶谱还能显示米根霉两个供试菌株之间的微小差异。经综合分析全部试验结果后得出的系统树图显示了9个供试菌株间的亲缘关系,并为未知菌株F1(BR12)和Q303提供了鉴定和命名依据。文中首次报导了根霉的SOD同工酶,并对蛋白质和酶电泳图谱用于根霉分类研究进行了讨论。     

刺孢小克银汉霉原变种和轮生刺孢小克银汉霉变种(新组合)

刺孢小克银汉霉[Cunninghamella echinulata(Thaxt.) Thaxt. ex Blakeslce]是小克银汉霉属的模式种,多年来因不同作者是否将贝尼尔小克银汉霉(C. bainieri Naum.)包含在内而对此种持广义(sensu lato)或狭义(scnsu stricto)的不同理解。我们在国内分离到51株刺孢小克银汉霉(广义)菌种,经与获自css的7株菌(包括刺孢小克银汉霉的模式菌种在内)和IMI的1株菌(原定名称均为刺孢小克银汉霉)一起研究后,根据它们的无性型形态特征有明显不同但在配合试验中又可互相配合形成可育的接合孢子的结果,认为将它们处理为同一个种的两个不同变种较为合理.其中,13株中国的菌和3株CBS及IMI的菌被鉴定为刺孢小克银汉霉原变种(C. echinulata var. echinulata);其余的38株中国的菌和5株CBS的菌被鉴定为轮生刺孢小克银汉霉变种(新组合)[C. echinulata var. verticillata (F. S. Paine.) comb. nov.j.以‘轮生(verticillata)’作为新组合变种的加词是根据与贝尼尔小克银汉霉同物异名的‘轮生布拉小克银汉霉(C. blakesleeana var. verticillata (F. S. Paine) Baijal & B. S. Mchrotra)’而来的.在前人的工作中,对刺孢小克银汉霉持狭义理解的作者往往根据孢子枝的分枝方式、巨型暗色孢子的有无、以及40℃能否生长来区分这两个分类群.对刺孢小克银汉霉持广义理解的作者则认为这些特性毫无意义.我们同意后面两个特性意义不大,因为二者的许多菌系都能形成巨型暗色袍子并在40℃生长,但认为孢子枝分枝方式确实是区别这两个分类群的重要依据.此外,根据产孢构造的种类、孢子枝主枝的直径、孢子枝第一次分枝的数目、分枝长度、孢子枝主枝的顶生泡囊形状以及其直径等性状综合考虑,这两个分类群是不难明确区分的,对12株刺孢小克银汉霉原变种和13株轮生刺孢小克银汉霉相互之间所进行的配合试验结果表明,在具一定性亲和力的(+)、(一)菌系同时存在时,两个变种内和变种之间均可形成可育的接合孢子。对上述三种配合的各六对菌所形成的有性型形态的研究结果则表明,除轮生刺孢小克银汉霉变种内形成的接合孢子的壁往往较厚外,各种配合形成的有性型形态基本上是一致的.我们研究组的其他成员最近曾对小克银汉霉的22株菌进行了rDNA的ITS区的PCR产物长度测定,其中包括了刺孢小克银汉霉原变种和轮生刺孢小克银汉霉变种的各3株菌,它们的平均长度(bp)分别为890±7.5和915±7.6.这个结果也支持了我们在这两个分类群可以互相配合的情况下仍将它们作为两个不同变种的处理.这两个分类群虽然已被发表多次,但它们的描述和绘图均过于简单而未能充分显示出它们的区别性特征,因此我们在本文中再次作了描述并再次提供了详尽的线条图和显微照相.     

桔青霉变异株分类地位的研究

桔青霉(Penicillium cirlrinunr Thom)是青霉属中的一个明确的种,但变异范围很大.自然界存在其白色株和不能利用硝酸盐为氮源的所谓营养缺陷型。在分类上对上述变异有两种不同处理方式:一是作为新种或变种,二是认为是突变型而不承认其分类地位。选择何种处理在一定程度上取决于研究者的主观看法。作者结合某些生理生化特性和可溶性蛋白电泳图形以及同工酶酶谱(酯酶,苹果酸脱氢酶,淀粉酶,核糖核酸酶)对桔青霉及其变异株以及邻近种作了比较研究以便提供依据,确认哪一种处理更为合理。研究结果表明桔青霉与其变异株的各种主要特征都极为相似,而与其邻近种则有明显不同。因此作者认为对这类菌株不应作为新种或变种而应作为种内变异株处理,不承认其分类地位.这一结论支持了Raper和Thom以及Pitt的观点。     

Protein enrichment of sago starch by solid-state fermentation with Rhizopus spp.

Protein enrichment of sago starch of three different diameters was investigated both in flask culture and under forced aeration in a packed-bed fermenter using two strains of Rhizopus. Protein production by R. oligosporus UQM 145F was superior to Rhizopus sp. UQM 186F in the flask culture without aeration, with both preferring larger diameter (3 to 4 mm) spherical sago-beads. In the packed-bed fermenter with forced aeration, Rhizopus sp. UQM 186F led to more rapid protein production compared to R. ollgosporus UQM 145F and produced equivalent final yields (about 10% protein on a dry wt basis).E. Gumbira-Sa'id, P.F. Greenfield and D.A. Mitchell are with the Department of Chemical Engineering, and H.W. Doelle is with the Department of Microbiology, University of Queensland, Queensland 4072, Australia.     

Protein and Esterase Patterns of Pathogenic, Fusarium oxysporum f. sp. elaeidis and F. oxysporum var. redolens from Africa, and Non-Pathogenic K oxysporum from Malaysia

Protein and esterase patterns of eleven isolates of F. oxysporum f. sp. elaeidis, one isolate of F. oxysporum var. redolens pathogenic to oil palm from Africa and six non-pathogenic isolates of F. oxysporum from oil palm soils in Malaysia were studied by vertical disc-electrophoresis and isoelectric focusing, to determine whether the pathogenic and saprophytic forms of F. oxysporum could be distinguished using these two methods. The protein patterns of all the isolates studied by the two methods were almost identical qualitatively and it was impossible to distinguish between the pathogenic isolates of F. oxysporum f. sp. elaeidis and F. oxysporum var. redolens from Africa and saprophytic isolates of F. oxysporum from Malaysia. Esterase zymograms of the isolates produced by the two methods were different. Esterase zymograms produced by vertical disc-electrophoresis showed great variations between and within the African and Malaysian isolates, but the esterase patterns produced by isoelectric focusing were almost identical qualitatively.     

Analysis of esterase zymograms ofFusarium sambucinum and related species

Esterase zymograms were obtained following polyacrylamide slab gel electrophoresis of protein extractsFusarium sambucinum and related species originating from different geographic locations and different matrices. The sites of esterase activity were recorded, and the R_fs were calculated. The data were used for the construction of phenograms by cluster analysis and nonlinear mapping by computerized classification techniques. The fifteen isolates ofF. sambucinum, the eight isolates ofF. torulosum and the six isolates ofF. spec. nov. each had identical profiles, and are therefore electrophoretically distinct species. The isolates ofF. sarcochroum, one ofF. sambucinum sensu lato (BBA 64280) and fifteen isolates ofF. sambucinum were electrophoretically indistinguishable from each other. We assume they are synonymous. The isolate ofF. bactridioides, one ofF. sambucinum sensu lato (BBA 64993) and eight isolates ofF. torulosum had uniform EST patterns, therefore the two species are electrophoretically identical. We assume they are also synonymous. The remaining three isolates ofF. sambucinum sensu lato are somewhat closely related toF. sambucinum isolates on the basis of our investigations.     

A flax-retting endopolygalacturonase-encoding gene from Rhizopus oryzae

A polygalacturonase from the filamentous fungus Rhizopus oryzae strain sb (NRRL 29086), previously shown to be effective in the retting of flax fibers, was shown by the analysis of its reaction products on polygalacturonic acid to be an endo-type. By zymogram analysis, the enzyme in the crude culture filtrate appeared as two active species of 37 and 40 kD. The endopolygalacturonase-encoding gene was cloned in Escherichia coli and its translated 383-amino acid sequence found to be identical to that of a presumed exopolygalacturonase found in R. oryzae strain YM9901 and 96% identical to a hypothetical protein (RO3G_04731.1) in the sequenced genome of R. oryzae strain 99–880. Phylogenetic analysis revealed the presence of an unique cluster of Rhizopus polygalacturonase sequences that are separate from other fungal polygalacturonases. Conservation of 12 cysteines appears to be a special feature of this family of Rhizopus polygalacturonase sequences. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Differential Patterns of Constitutive Intracellular Laccases of the Vegetative Phase of Pleurotus Species

The constitutive intracellular laccase activity of ten strains of Pleurotus spp. was determined in vitro and by zymograms, using different substrates. Differences in the in vitro activities were observed between all the strains; however, zymogram patterns were only similar for strains within same species, independently of any of the three substrate (2,6-dimethoxyphenol, p-anisidine or o-tolidine) used. The differences observed in the number and positions of the isoforms in the gel suggest that laccase zymograms can be used to differentiate species of this organism.     

Analysis of SDS-dissociated proteins of pathogenic and nonpathogenicFusarium species

SDS — dissociated proteins from eight isolates belonging to threeFusarium species were assessed and compared using polyacrylamide gel electrophoresis. Intra and inter specific variation in banding patterns were analyzed both qualitatively and quantitatively. Special emphasis was placed on variability of protein banding pattern between two nonpathogenic strains ofF. oxysporum (C5 and C14). Protein profiles from host-associated isolates were distinct, and each isolate showed a uniquely characteristic profile. Attempts were made to provide information on the genetic system controlling pathogenicity, compared to nonpathogenicity, at the protein level. The data obtained from electrophoresis support the potential use of this experimental approach to help distinguish between differentFusarium isolates.     

An original habitat of tempeh molds

Tempeh is a traditional Indonesian food made from soybeans fermented with Rhizopus species. Some researchers believe the original habitat of the tempeh molds may be closely related to fresh leaves of Hibiscus species because these leaves artificially infected with the tempeh molds are used to start tempeh fermentation in cottage-scale factories. To verify this hypothesis, we investigated the occurrence of Rhizopus species in Hibiscus leaves and identified the isolated Rhizopus strains precisely. Rhizopus oryzae, one of the tempeh molds, occurred in sample leaves of some Hibiscus species with considerable frequency. This result implies that tempeh molds that lived in Hibiscus leaves might have fermented soybeans accidentally when used to wrap the cooked soybeans. The original habitat of the tempeh mold could be fresh leaves of Hibiscus species.     

Differences in the control of bacteriochlorophyll formation by light and oxygen

Control of bacteriochlorophyll formation was studied with continuous cultures of Rhodospirillum rubrum, Rhodopseudomonas sphaeroides, and Rhodopseudomonas capsulata. Oxygen controlled specific bacteriochlorophyll contents of the three species in a hyperbolical fashion irrespective of the presence of light. In Rps. sphaeroides, this applied to oxygen concentrations above 16% air saturation of the medium while at lower oxygen concentrations control followed a kinetics with negative cooperativity. Cell protein formation of R. rubrum and Rsp. sphaeroides was independent of oxygen concentrations while protein formation of Rps. capsulata increased at lower concentrations. Light controlled bacteriochlorphyll contents of R. rubrum and Rps. sphaeroides in a sigmoidal fashion. When growing at a constant low oxygen concentration cell protein formation increased with light energy flux in Rps. sphaeroides but remained unaffected in R. rubrum. Protein formation of R. rubrum increased with light energy flux only under anaerobic conditions. Two factor analyses were performed with R. rubrum and Rps. sphaeroides to study the combined effects of light and oxygen on bacteriochlorophyll formation. The results showed that both factors act independent of each other.Abbreviations ALA 5-aminolevulinic acid - R Rhodospirillum - Rsp. Rhodopseudomonas     

Variations in amylase and protease activities amongRhizopus isolates

Summary Variations in amylase and protease activities among 16 local isolates ofRhizopus spp. were assessed.Rhizopus microsporus and theR. arrhizus isolates were consistently good producers of amylase. Good producers of protease wereR. microsporus and theR. oligosporus isolates. No amylase or protease activity was detected inR. stolonifer. The results also showed that

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Resumen Se evaluaron las variaciones en las actividades amilasa y proteasa de 16 aislados locales deRhizopus spp. Los aislados deR. microsporus y deR. arrhizus fueron buenos productores de amilasa en tanto que los aislados deR. microsporus y deR. oligosporus lo fueron de proteasa. No se detectó actividad amilasa ni proteasa enR. stolonifer. Los resultados mostraron además que la producción enzimática variaba entre las deferentes especies deRhizopus spp. aislados y tambien entre aislados de la misma especie. UnRhizopus spp. aislado de pan seco fue tan activo como varias cepas aisladas de tempeh.

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Résumé Les variations en activités amylolytique et protéolytique chez 16 souches isolées localement deRhizopus spp. ont été mesurées. Les souches deRhizopus microsporus et deR. arrhizus se sont avérées systématiquement bonnes productrices d'amylase. Les souches deR. microsporus et deR. oligosporus se sont avérées bonnes productrices de protéase. Ni amylase ni protéinase n'ont été détectées chezR. stolonifer. Les résultats ont également montré que la productivité enzymatique variait d'espèce chezRhizopus, et même parmi différentes souches de la même espèce. La souche deRhizopus isolée de pain bis était aussi active en production d'enzyme que certaines souches isolées de tempeh.