平菇不同菌株的同工酶探讨

本文对来源不同的四个侧耳属(Pleurotus)菌株进行了酯酶和过氧化物酶同工酶的分析测定。结果表明,供试菌株的同工酶谱各不相同其酯酶酶谱的差异更为明显。因此初步认为所试四个菌株可作为平茹的不同品种进行深入研究。     

毛木耳种质资源的酯酶同工酶分析

采用垂直平板聚丙烯酰胺凝胶电泳技术对毛木耳56个菌株酯酶同工酶的酶谱多样性进行了研究。结果表明,56份材料有一定的遗传变异,共检测到迁移率不同的10条谱带,各菌株分别具有1~6条酶带,共有26种酶谱类型。菌株被分成了8大群:第一群包括黄耳zh等34个菌株;第二群包括白背木耳等8个菌株;第三、第四群分别为黑木耳和915两个单独的菌株;第五群包括99等4个菌株;第六群包括43等4个菌株;第七群包括50385和AP067两个菌株;第八群包括小上3和杂交34两个菌株。酯酶同工酶分析技术是鉴别种及品种以下菌株的有效手段。     

灵芝栽培菌株酯酶同工酶的酶谱多样性

采用酯酶同工酶聚丙烯酰胺凝胶电泳(PAGE)方法,对灵芝属(GanodermaKarst.)9株灵芝进行品系间鉴定,并采用非加权组平均法(UPGMA)进行聚类分析。试验结果表明:在9个菌株(16 d)中共检测到40条酶带,各个菌株分别具有3至6条酶带,9个菌株共有4种酶谱类型。在相异系数为0.62时所有供试菌株归为1个群,在相异系数为0.81时,把9个菌株分为5个群。酯酶同工酶聚丙烯酰胺凝胶电泳鉴定方法可有效应用于灵芝属真菌品种鉴定及亲缘关系分析。     

黑木耳菌株酯酶同工酶酶谱多样性研究

目的：为了探索黑木耳菌株之间的遗传距离,构建出供试菌株酯酶同工酶鉴别图。方法：采用垂直板聚丙烯酰胺凝胶电泳对21个黑木耳菌株酯酶同工酶的酶谱多样性进行了研究。结果：酯酶同工酶电泳各个菌株分别具有2~7条酶带,其中在Rf值为0.344处,21个菌株都有谱带出现。21个菌株之间的遗传相似系数在0.167-1.000之间,应用NTSYS软件进行聚类分析,当相似水平为0.73时,可将供试的21个黑木耳菌株分为6个不同的类群。结论：酯酶同工酶可以有效快捷的对黑木耳菌株进行菌种鉴定,是黑木耳菌株遗传多样性研究的理想手段。     

木耳栽培菌株酯酶同工酶的酶谱多样性研究

采用聚丙烯酰胺凝胶电泳技术对木耳（Ａｕｒｉｃｕｌａｒｉａａｕｒｉｃｕｌａ（Ｈｏｏｋ）Ｕｎｄｅｒｗ）１０个栽培菌株酯酶同工酶的酶谱多样性进行了研究。１０个供试菌株的幼龄菌丝（７ｄ）中仅检测到２５条酶带,各个菌株分别具有２～３条酶带,１０个菌株仅有２种酶谱类型;老龄菌丝（７２ｄ）中共检测到４４条酶带,各个菌株分别具有３～６条酶带,１０菌株共有８种酶谱类型。研究表明,木耳双核体菌丝中某些酯酶同工酶基因位点在一定的发育时期才开始表达,老龄菌丝酯酶同工酶酶谱在木耳菌株鉴别和遗传育种研究中具有更大的应用价值。     

香菇双单杂交后代不同发育阶段酯酶同工酶研究

选用香菇野生株分别与栽培株及杂交株进行双单杂交 ,得到 8个杂交后代 ,且具有结实能力 ,分别对液培 2 0d杂交后代菌丝与液培原基酯酶同工酶进行了比较研究。结果表明 ,液培 2 0d菌丝菌株间酯酶同工酶酶谱显示出多样性 ,可以作为鉴定菌株的辅助遗传标记 ,而原基菌株间酯酶同工酶酶谱谱带较少 ,呈现趋同效应 ,不宜作为鉴定香菇菌株的辅助遗传标记。     

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。     

九株根霉可溶性蛋白和三种酶的电泳图谱比较研究

本文了根霉属９个菌株天然态及解态可溶性蛋白、酯酶同工酶、葡萄糖淀粉酶的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。文中首次了根霉的ＳＯＤ同工酶,并对蛋白质和酶,     

培养时间对黑木耳酯酶同工酶谱的影响

为准确地将酯酶同工酶技术应用于食用菌菌株鉴别和遗传育种研究以及进行快速的菌种区分、鉴定,本试验对10个黑木耳菌株不同培养时期的胞内酯酶(EST)同工酶谱进行了研究,并对菌株间亲和性试验结果进行分析。结果表明:同一培养时期的各菌株间同工酶谱存在明显差异,不同培养时期同一菌株同工酶谱也存在一定的差异。培养20d诱导的同工酶谱可有效区分、鉴定各菌株。聚类分析结果表明:培养20d的酶谱聚类分析结果与亲和性试验结果相一致。     

6个杏鲍菇菌株生理生化研究

目的:对收集的6个杏鲍菇菌种菌丝进行生理、生化研究,酯酶同工酶法分析菌株间的差异与关系。方法:利用杏鲍菇菌丝阶段的生长速度测定、诱变剂抗性试验和酯酶同工酶比较,对杏鲍菇菌株营养生长阶段的理化指标进行研究,对酯酶同工酶谱进行相似系数和遗传距离分析,确立各菌株的异同。结果:菌株间菌丝生长速度差异不显著;抗诱变剂实验结果显示,2、5、6号菌株对诱变剂抗性较强;酯酶同工酶分析表明,杏鲍菇6个菌株同工酶酶带存在的差异很大。结论:通过试验表明,所选6株杏鲍菇菌种为各自独立菌株。     

16S rDNA sequence analysis of Xylella fastidiosa strains

The 16S rDNA encoding the small subunit ribosomal RNA were amplified by PCR, cloned, and sequenced from 16 strains of Xylella fastidiosa originating from nine different hosts. In pair-wise comparisons, X. fastidiosa strains showed a maximum variation of 1.0% or 14 nucleotide positions. When all 16 sequences were considered as a set, 54 variable positions were found. Analysis of the sequence data indicated that the X. fastdiosa strains formed three rDNA groups. Group one includes Pierce's disease and mulberry leaf scorch strains; Group two, periwinkle wilt, plum leaf scald, phony peach, oak leaf scorch, and elm leaf scorch strains; and Group three, citrus variegated chlorosis and coffee leaf scorch strains. All X. fastidiosa strains exhibited significantly higher levels of sequence heterogeneity (63 to 83 nucleotide positions) when compared to species from Xanthomonas and Stenotrophomonas. Our data demonstrate that 16S rDNA sequence data could provide valuable information for future classification of X. fastidiosa at the sub-species level.     

Polymerase chain reaction (PCR)-based typing analysis of atypical isolates of the fish pathogen Aeromonas salmonicida.

Two hundred and five isolates of atypical Aeromonas salmonicida, recovered from a wide range of hosts and countries were characterized by polymerase chain reaction (PCR) targeting four genes. The chosen genes were those encoding the extracellular A-layer protein (AP), the serine protease (Sprot), the glycerophospholipid:cholestrol acetyltransferase protein (GCAT), and the 16S rRNA (16S rDNA). All the atypical A. salmonicida isolates could be assigned to 4 PCR groups. Group 1 comprised 45 strains which tested positive for PCR amplification, using the 16S rDNA, GCAT2, Sprot2, and AP primer-sets. Group 2 comprised 88 strains with produced PCR products using the 16S rDNA, GCAT2 and AP primer-sets. Group 3 comprised 21 strains which produced PCR products using 16S rDNA, GCAT2 and Sprot2 primer-sets, and group 4 comprised 51 strains which produced PCR products using the 16S rDNA and GCAT2 primer-sets only. A. salmonicida subsp. salmonicida isolates tested, belonged to group 1. The PCR primer-sets separated A. salmonicida from other reference strains of Aeromonas species and related bacteria with the exception of Aeromonas hydrophila. The results indicated that PCR typing is a useful framework for characterization of the increasing number of isolations of atypical A. salmonicida.     

我国南北大豆产区慢生大豆根瘤菌的遗传多样性和系统发育研究

利用16S rRNA基因RFLP、16S rRNA基因序列分析以及16S-23S rRNA IGS PCR RFLP技术对分离自我国南北大豆产区的慢生大豆根瘤菌进行了群体遗传多样性和系统发育研究。16S rRNA基因PCR RFLP分析以及16S rRNA基因序列分析结果表明:所有供试慢生大豆根瘤菌可分为B.japonicum和B.elkanii两个类群,其中属于B.japonicum的为优势种群,占供试菌株的91%,属于B.elkanii的仅占9%,多样性水平较低。16S-23S rRNA IGS PCRRFLP研究结果表明:属于B.japonicum的慢生根瘤菌具有较丰富的遗传多样性,在69%的相似性水平上可分为群Ⅰ和群Ⅱ两大类群。群I的菌株以分离自黑龙江和河北等北部区域的菌株为代表,群Ⅱ的菌株以分离自广西和江苏等南部地域的菌株为代表,反映出明显的地域特征。两群菌株在系统发育上均与USDA6、USDA110和USDA122等B.japonicum的模式或代表菌株有差异。     

我国主要生态区域绿豆慢生根瘤菌的遗传多样性和系统发育研究

利用16SrRNAPCR-RFLP、16SrRNA序列分析以及16S-23SrRNAIGS(IntergeneticSpacer)PCR-RFLP技术对分离自中国主要生态区域的44株慢生型绿豆根瘤菌和5株参比菌株进行了遗传多样性和系统发育研究。16SrRNAPCR-RFLP分析表明:在76%的相似水平上,所有供试菌株可分为三大类群:群I由LYG1等13株慢生根瘤菌组成,该群在系统发育上与B.japonicum和B.liaoningense的参比菌株存在一定的差异;群Ⅱ由XJ1等21株供试菌株、B.japonicum和B.liaoningense的代表菌株组成;群Ⅲ由10株来自广东和广西的菌株和B.elkanii的代表菌株组成。16S-23SrRNAIGSPCR-RFLP分析将供试菌株分为A、B两大群。群A由34株供试菌株、B.japonicum和B.liaoningense的代表菌株组成。在85%的相似性水平上,可再分为AⅠ、AⅡ和AⅢ3个亚群。群B由10株分离自广西和广东的菌株和B.elkanii的代表菌株组成。在85%的相似性水平上,可再分为BI和BⅡ两亚群,表现出一定的多样性。与16SrRNAPCR-RFLP相比,16S-23SrRNAIGSPCR-RFLP具有更高的解析度,供试菌株表现出更加丰富的遗传多样性。分离自中国新疆、广东和广西等地的菌株在分群上具有较为明显的地域特征。     

Heterogeneity of strains assigned to Gluconobacter frateurii Mason and Claus 1989 based on restriction analysis of 16S-23S rDNA internal transcribed spacer regions

Twenty-three strains, which were assigned to Gluconobacter frateurii and maintained at Culture Collection NBRC, were re-identified at the species level on the basis of restriction analysis of 16S-23S rDNA ITS regions by digestion with six restriction endonucleases: Bsp1286I, MboII, AvaII, TaqI, BsoBI, and BstNI. The strains examined were divided into six groups, Group III-1, Group III-2, Group III-3, Group III-4, Group III-5, and Group IV. Group III-1 and Group III-4 respectively were divided into two subgroups, Subgroup III-1a, Subgroup III-1b and Subgroup III-4a, Subgroup III-4b. Gluconobacter frateurii NBRC 3264(T) was included in Group III-2, along with strains NBRC 3265 and NBRC 3270, and G. thailandicus BCC 14116(T) was included in Group III-3, along with strains NBRC 3254, NBRC 3256, NBRC 3258, NBRC 3255, and NBRC 3257. These groupings were supported by a phylogenetic tree based on 16S-23S rDNA ITS sequences. Strains of group III-2 and Group IV were unequivocally re-identified as G. frateurii, but strains of Group III-3, Group III-4, and Group III-5 were not necessarily re-identified as G. frateurii. The results obtained indicate that the 23 strains have a taxonomically heterogeneous nature, and they are referred to as the G. frateurii complex.     

16S rDNA restriction fragment length polymorphism analysis of psychrotrophic vibrios from Japanese coastal water

Restriction fragment length polymorphism (RFLP) analysis was carried out for 136 natural isolates belonging to the family Vibrionaceae. These were collected from inshore areas of Japan, mainly in winter. Twenty-eight 16S rDNA genotypes were obtained by digestion with four restriction endonucleases (HhaI, DdeI, RsaI, and Sau3AI). To estimate the genetic relationships, 53 informative fragments were scored by their presence or absence. A dendrogram was constructed using the unweighted pair group method with the arithmetic averages algorithm. Five RFLP groups (groups I to V) were obtained. Group I corresponded to Vibrio splendidus-like strains. It was confirmed that this group was not only found in Otsuchi Bay, but also in broad coastal areas of Japan. Group II strains were not identified as previously known Vibrio species. Group III strains were regarded as members of the Vibrio main group, which is a major phylogenetic group deduced from 16S rRNA gene analysis in the family Vibrionaceae. The RFLP profile indicated that Group IV strains were closely related to V. hollisae. Group V strains showed RFLP patterns which have not been observed previously. From the clustering analysis, it was concluded that group V strains were not Vibrio species. Most of the isolates studied were not identified as previously described species. It suggests that many psychrotrophic vibrios in cold marine environments remain as unknown species.     

16S rRNA gene and 16S-23S rRNA gene internal transcribed spacer sequences analysis of the genus Myxococcus.

Phylogenetic relationships of the species belonging to the genus Myxococcus were elucidated based on the sequences of 16S rRNA genes and 16S-23S rRNA gene internal transcribed spacer (ITS) regions. The Myxococcus species were consequently classified into four distinct groups. The type strain of Myxococcus coralloides occupied an independent position (Group 1); it has been recently reclassified as Corallococcus coralloides. Group 2 comprised the type strains of both Myxococcus virescens and Myxococcus xanthus, and some strains assigned to Myxococcus flavescens. The type strain of M. flavescens was contained in Group 3 along with the strains of Myxococcus fulvus. Group 4 included the strains belonging to C. coralloides, M. fulvus, and M. stipitatus. The type strain of M. fulvus that was allocated outside Group 4 in the 16S rRNA gene tree belonged to Group 3 in the ITS tree. These results strongly suggest that the morphological characteristics of Myxococcus species are not consistent with the phylogenetic relationships. The Myxococcus species must therefore be redefined according to the phylogenetic relationships revealed in this study.     

Identification of Acetobacter strains isolated from Indonesian sources,and proposals of Acetobacter syzygii sp. nov., Acetobacter cibinongensis sp. nov., and Acetobacter orientalis sp. nov

Forty-six strains of acetic acid bacteria newly isolated from flowers, fruits, and fermented foods collected in Indonesia were taxonomically studied. They were Gram-negative rods, produced acetic acid from ethanol, oxidized acetate and lactate to CO(2) and H(2)O, and had Q-9 as the major ubiquinone system. On the basis of DNA-DNA similarity, all strains studied, including type strains and reference strains of the genus Acetobacter, were separated into eleven groups (Groups I to XI). Of the 46 isolates, two isolates were included in Group II and identified as Acetobacter pasteurianus, five in Group IV as A. orleanensis, 16 in Group V as A. lovaniensis, five in Group VII as A. indonesiensis, and three in Group VIII as A. tropicalis. The remaining 15 isolates constituted three new groups based on DNA-DNA similarity; four isolates were included in Group IX, two in Group X, and nine in Group XI. No isolates were identified as A. aceti (Group I), A. peroxydans (Group III), and A. estunensis (Group VI). Phylogenetic analysis based on 16S rDNA sequences of representative strains of the Groups indicated belonging to the strains of the genus Acetobacter. On the basis of DNA base composition, DNA-DNA similarity, and 16S rDNA sequences, three new species of the genus Acetobacter are proposed: Acetobacter syzygii sp. nov. for Group IX, Acetobacter cibinongensis sp. nov. for Group X, and Acetobacter orientalis sp. nov. for Group XI. The distribution of Acetobacter strains in Indonesia is discussed in light of isolation sources.     

Heterogeneity of Strains Assigned to Gluconobacter frateurii Mason and Claus 1989 Based on Restriction Analysis of 16S-23S rDNA Internal Transcribed Spacer Regions

Twenty-three strains, which were assigned to Gluconobacter frateurii and maintained at Culture Collection NBRC, were re-identified at the species level on the basis of restriction analysis of 16S-23S rDNA ITS regions by digestion with six restriction endonucleases: Bsp1286I, MboII, AvaII, TaqI, BsoBI, and BstNI. The strains examined were divided into six groups, Group III-1, Group III-2, Group III-3, Group III-4, Group III-5, and Group IV. Group III-1 and Group III-4 respectively were divided into two subgroups, Subgroup III-1a, Subgroup III-1b and Subgroup III-4a, Subgroup III-4b. Gluconobacter frateurii NBRC 3264^T was included in Group III-2, along with strains NBRC 3265 and NBRC 3270, and G. thailandicus BCC 14116^T was included in Group III-3, along with strains NBRC 3254, NBRC 3256, NBRC 3258, NBRC 3255, and NBRC 3257. These groupings were supported by a phylogenetic tree based on 16S-23S rDNA ITS sequences. Strains of group III-2 and Group IV were unequivocally re-identified as G. frateurii, but strains of Group III-3, Group III-4, and Group III-5 were not necessarily re-identified as G. frateurii. The results obtained indicate that the 23 strains have a taxonomically heterogeneous nature, and they are referred to as the G. frateurii complex.     

两个根瘤菌新群的系统发育学分析

在数值分类、ＳＤＳＰＡＧＥ 全细胞蛋白分析、ＤＮＡＤＮＡ 杂交、１６ＳｒＤＮＡＰＣＲＲＦＬＰ 的基础上,测定了两个分离自干旱地区苜蓿、草木樨根瘤菌新群１ 、２ 的中心株ＸＪ９６０６０ 、ＸＪ９６４０８ 的１６ＳｒＤＮＡ 全序列,并进一步将中心株和３１ 株已知菌、３ 株分自黄土高原的根瘤菌进行了系统发育学分析。结果表明,供试菌株在系统发育树中基本分成Ｓｉｎｏｒｈｉｚｏｂｉｕｍ 、Ｍｅｓｏｒｈｉｚｏｂｉｕｍ 、ＡｇｒｏｂａｃｔｅｒｉｕｍＲｈｉｚｏｂｉｕｍ 、Ｒｈｉｚｏｂｉｕ ｍ 、Ｂｒａｄｙｒｈｉｚｏｂｉｕｍ 、Ａｚｏｒｈｉｚｏｂｉｕｍ 六个分枝。群１ ,２ 落入Ｓｉｎｏｒｈｉｚｏｂｉｕ ｍ 分枝。