红色毛癣菌的研究进展

红色毛癣菌是最常见的也是世界范围内传播是广的一种皮肤癣菌,属毛癣菌属,本文简述了最近几年在该菌的基因鉴定,基因分类,基因结构的分子生物学研究和免疫学研究方面的主要进展,在基因鉴定中发展了红色毛癣菌特异性探针TR20S和毛癣菌属特异性引物TR1、TR2和NS5,NS6;利用测序方法对毛癣菌属进行了重新分类,并且用探针,限制性片段多态性分析等方法对红色毛癣菌进行了种内分型研究。     

红色毛癣菌致儿童脓癣1例

报道儿童脓癣1例。患儿男,5岁,头部左后侧圆形脓肿破溃结痂伴瘙痒疼痛半个月余。取断发及脓液直接镜检及真菌培养均阳性,做小培养确定菌种为红色毛癣菌,经综合治疗后痊愈。     

红色毛癣菌菌株的特异性PCR鉴定方法研究

目的建立一种快速的红色毛癣菌分子生物学鉴定方法。方法根据红色毛癣菌保守区域-真菌核糖体DNA（rDNA）的转录间隔区（ITS）设计特异性引物,采用上游：ITS19865＇GAC ACC AAG AAA AAA TTC TCT GAA GA3＇,下游：ITS24415＇GTC CTG AGG GCG CTG AA3＇为引物对45株红色毛癣菌、5株须癣毛癣菌和1株紫色毛癣菌菌株的DNA进行PCR扩增,观察产物电泳带型的差异。结果 45株红色毛癣菌均能扩增出目的片段,5株须癣毛癣菌和1株紫色毛癣菌均无目的片段扩增出。结论红色毛癣菌可用特异引物PCR方法快速鉴定。     

香菇品种遗传多样性RAPD分子标记的研究

对32个中国栽培香菇品种和2个野生子实体的遗传多样性进行了RAPD标记研究,9个引物共扩增出113条DNA带,83.19%具有多态性。结果显示,34个香菇菌株间均有遗传上的差异,但遗传相关性极高,表明中国栽培菌株间的遗传背景单一。     

南方红豆杉不同居群遗传多样性的RAPD研究

用随机扩增多态性方法对广东、湖南、江西等3省的12个南方红豆杉自然居群进行了基因组DNA多态性分析,从100条引物中共筛选出10个引物,获得RAPD谱带86条,多态性谱带占51％。聚类分析结果表明：南方红豆杉居群间的遗传距离与这些居群的地理分布相关,即相同或相邻产地的居群间的遗传距离较小,不同产地个体间的遗传距离较大。粤北南方红豆杉的9个居群的遗传多样性较低,可能与近年来资源遭到严重破坏,及其生长缓慢、种子萌发率低、成活率不高等原因有关。     

106株红色毛癣菌基因分型与药敏分析CSCD

目的探索海南地区的红色毛癣菌基因型与感染部位、药敏的关系。方法基于红色毛癣菌核糖体rDNA非转录区(NTS)的基因分型进行种内分型,依据“CLSI-M38-A2”方案进行药敏实验。结果106株红色毛癣菌TRS-1基因型有5种带型,其中TypeⅠ52株(49.06%),TypeⅡ14株(13.21%),TypeⅢ5株(4.72%),TypeⅣ型3株(2.83%),其他带型32株(30.18%)。TRS-2基因型有3种带型,其中TypeⅠ68株(64.15%),TypeⅡ6株(5.66%),其他带型32株(30.19%)。药敏结果MIC几何均数由低至高分别为特比萘芬(0.0092μg/mL)、伏立康唑(0.0181μg/mL)、伊曲康唑(0.1491μg/mL)、酮康唑(0.1630μg/mL)、氟康唑(2.3164μg/mL)。有42株菌表现出对抗真菌药物不敏感,不敏感菌株TRS-1和TRS-2分型均以TypeⅠ为主。结论海南地区流行的红色毛癣菌TRS-1和TRS-2基因型均以TypeⅠ为主。以对特比萘芬(0.0092μg/mL)、伏立康唑(0.0181μg/mL)MIC几何均数最低。本结果发现本地区的红色毛癣菌不敏感菌株与基因带型关系不大,也与感染部位无关,可能与来源有关。     

红色毛癣菌临床分离株的菌落形态和镜下结构特征再分析

目的探讨红色毛癣菌的菌落形态和镜下结构特征以及与感染部位的相关性。方法采用传统培养方法对192株红色毛癣菌进行表型分型,选取其中39株在28℃、30℃、35℃3种温度孵育6d、10d、14d时观察菌落形态和生长速度。结果192株红色毛癣菌共分离出3种表型：绒毛型、沟纹型、粉末型（或颗粒型）。在相同培养基上,28℃、30℃时菌落生长速度无显著差异（P〈0.05）,均快于35℃（P〈0.05）。在相同温度时,菌落在SDA上的生长速度快于PDA培养基。在28℃、30℃时菌落形态比较稳定,在35℃时变异较大。菌落在PDA培养基上产孢丰富,镜下显示有较多的大、小分生孢子,而在SDA培养基上只有少量的小分生孢子,几乎见不到大分生孢子。各种浅部感染均以绒毛型为主,绒毛型在手足癣中占比例最高,沟纹型在体股癣中所占比例较高,粉末型在甲癣中占的比例较高。结论红色毛癣菌的菌落形态与培养基和培养温度有关,其表型和镜下结构与感染部位均有一定的相关性。     

红色毛癣菌不同生长阶段的基因表达谱分析

红色毛癣菌是最重要的浅表感染真菌.目前对红色毛癣菌分子和遗传学方面的研究还比较有限.应用cDNA芯片技术对红色毛癣菌不同生长阶段的基因表达谱进行了分析,获得了2044个在不同生长阶段差异表达的基因.聚类分析表明,这些基因可以分为3种不同的表达模式.本研究验证了先前的结论,即在红色毛癣菌中存在预储存的mRNA.对不同生长状态下转录谱和基因功能分析表明,糖酵解过程中的一些重要的酶具有相似的表达模式,说明在红色毛癣菌的生长过程中,这些酶可能具有共同的调控模式.同时,在红色毛癣菌生长过程中,一些参与诸如小GTPase调控、cAMP依赖的调控途径以及MAPK信号传导途径的基因表达也发生了变化.尽管这些基因和相关的调控途径在红色毛癣菌中的具体生物学功能还不清楚,但它们可能在其生长过程中起了调控作用.     

长筒石蒜居群遗传多样性RAPD分析

利用RAPD标记对长筒石蒜3个居群的遗传多样性及分化程度进行了研究。12条随机引物扩增出94个可分析位点,多态位点比率(PPB)为65.96%,表明长筒石蒜具有比较高的遗传多样性。经POP-GENE32分析表明:Nei’s基因多样性指数(h)为0.1897,香农多样性指数(Ⅰ)为0.2945,基因分化系数(GST)为0.1191,基因流(Nm)为3.6980。经WINAMOVA分析表明:居群内遗传变异占71.75%,而居群间只占28.25%。遗传多样性分析表明,各居群的遗传多样性水平由高到低为琅琊山居群>宝华山居群>盱眙居群。遗传分化表明:长筒石蒜各居群间遗传分化程度较低;大部分遗传变异存在于居群内部,表明其具有较强的进化潜力,自然情况下不会处于濒危状态,野生种质资源的破坏,主要来自于人为干扰。     

红色毛癣菌的生物学特性研究

目的 观察红色毛癣菌在不同温度、不同培养基上的生长和产孢情况,并对其进行分子生物学鉴定.方法 ①大培养:采用沙堡葡萄糖琼脂(SDA)和马铃薯葡萄糖琼脂(PDA)平皿,27℃、35℃黑暗培养,测量菌落直径,绘成生长曲线.②小培养(钢圈法):采用SDA、PDA、溴甲酚紫乳固体葡萄糖琼脂(BCP-MSG)、乳蜜琼脂(M)和复合维生素B(VitB)培养基,27℃、30℃黑暗培养,观察镜下菌丝生长、孢子产生情况.③进行rDNA 18S和ITS序列测定.结果 在SDA,PDA上,27℃条件下菌落生长速度较35℃快;在5种培养基上,SDA、PDA产孢较快较多,复合维生素B培养基产孢较慢,但产生大分生孢子较多.30℃产孢更丰富.对部分菌株rDNA ITS、18S PCR扩增产物纯化后直接测序,结果在GenBank中比对、分析,相似度为98%～100%,均鉴定为红色毛癣菌.结论 SDA、PDA均为鉴定和分离红色毛癣菌的合适培养基.5种培养基均可用来刺激红色毛癣菌产孢,其中SDA、PDA产孢较早、较丰富.红色毛癣菌rDNA 18S和ITS序列测定是一种快速准确的红色毛癣菌分子生物学鉴定方法.     

Identification and genetic homogeneity of Trichophyton tonsurans isolated from several regions by random amplified polymorphic DNA

Trichophyton tonsurans is an anthropophilic dermatophyte mostly causing tinea capitis and tinea corporis. This study was carried out to identify T. tonsurans and to clarify whether it has any genetic differences depending on the phenotype or region of isolation by random amplified polymorphic DNA (RAPD) analysis with three random primers. The assay was performed in 11 Korean, 2 Japanese, 2 Taiwanese, 5 Brazilian and 1 American isolates of T. tonsurans together with the other 10 anamorphic species of dermatophytes and 3 Arthroderma spp. All tested species of dermatophytes showed distinct bands and T. tonsurans was differentiated from other dermatophytes. It was most clearly ifferentiated from T. mentagrophytes by using primer 5-GAAGGCTCCC-3 (OPAO-15). No difference was found in RAPD band patterns in all strains of T. tonsurans with these random primers. It was considered that T. tonsurans is a genetically homogeneous species regardless of its isolation regions, morphologic or physiologic characteristics.This revised version was published online in October 2005 with corrections to the Cover Date.     

Genotypic diversity in a localized population of Ralstonia solanacearum as revealed by random amplified polymorphic DNA markers

AIMS: To assess genotypic diversity within Ralstonia solanacearum isolates of a single field. METHODS AND RESULTS: A total of 44 field isolates and 22 in vitro generated clones of R. solanacearum were studied for genotypic diversity by random amplified polymorphic DNA (RAPD) technique. Genomic DNA of these isolates and clones was extracted by proteinase-K-SDS lysis mini-prep method. RAPD analysis was done with 30 decamer primers. The data were analysed using NTSYSpc 2.02h software. Forty-two out of 44 field isolates and all the clonal isolates were identified as distinct genotypes at 70% similarity level. CONCLUSION: Very high level of genome variability was observed within the field and clonal isolates of R. solanacearum. This might be a reason for the wide host range of this bacterium and for quick breakdown of wilt resistance in host plants. SIGNIFICANCE AND IMPACT OF THE STUDY: The results suggest that it would be difficult to design specific diagnostic protocol for R. solanacearum even for a localized population and to breed cultivars with broad-spectrum resistance.     

三种摇蚊幼虫RAPD扩增条件的优化及在昆虫系统发育分析中的应用

为了研究摇蚊科昆虫种群遗传的多样性,以促进对其资源的合理保护,以萨摩亚摇蚊Chironomus samoensisEdwards基因组DNA为模板,对摇蚊幼虫的RAPD扩增条件进行优化,建立了摇蚊幼虫RAPD扩增反应的最佳体系:按照利用优化的RAPD扩增条件进行研究,实验有着良好的重现性。用16个随机引物对3种摇蚊幼虫类群各10个个体进行RAPD扩增,其中萨摩亚摇蚊共扩增出78个条带,多态座位率为41.03%,Shannon遗传多样性指数为0.2570,群体内相似度为0.8730;红裸须摇蚊Propsilocerus akamusi(Tokunaga)共75个条带,多态座位率为44.0%,Shannon遗传多样性指数为0.2472,群体内相似度为0.8731;刺铗长足摇蚊Tanypus punctipennis(Fabricius)共67个条带,多态座位率为41.79%,Shannon遗传多样性指数为0.1943,群体内相似度为0.9066。聚类分析结果表明,刺铗长足摇蚊与红裸须摇蚊的亲缘关系较近。     

The effects of regeneration by fragmentation upon clonal diversity in the tropical forest shrub Poikilacanthus macranthus: random amplified polymorphic DNA (RAPD) results

Poikilacanthus macranthus (Acanthaceae), like many shrubs in the neotropical cloud forest of Monteverde, Costa Rica, is capable of regeneration through stem and twig fragments. Our field sampling indicated that reproduction in P. macranthus is primarily through fragmentation in the Monteverde region. Random amplified polymorphic DNAs (RAPDs) were employed to examine clonal diversity in four populations of P. macranthus , and two primers yielded 11 variable markers that were used to identify genets. Sixty-eight multilocus genotypes were identified out of 277 plants sampled. Clonal diversity values were higher than the average for clonal plant species, as the number of genets per ramet was 0.25, while the mean D value was 0.87 and the mean E value was 0.84.     

The use of random amplified polymorphic DNA (RAPD) for hybrid detection in Scirpus from the river Schelde (Belgium)

Along the unique freshwater tidal zone of the river Schelde (Belgium), plants of Scirpus species occur primarily in small and fragmented populations. The majority of these are native Scirpus lacustris, S. pungens, S. triqueter, S. tabernaemontani and intermediate morphological forms. The distribution area of S. triqueter is even restricted to this tidal habitat. However, several cultivated S. tabernaemontani strains have recently been introduced. The latter species is often used to stabilize riverbanks. To determine the existing genetic diversity among these species, stems from plants from 44 different locations were subjected to random amplified polymorphic DNA (RAPD) analysis, using 22 decanucleotide primers. Data analysis of the amplified DNA fragments enabled us to unambiguously differentiate among these Scirpus taxa. Hybridization between S. triqueter and S. tabernaemontani was documented, and the studied hybrids were always genetically more similar to S. triqueter than to S. tabernaemontani . Among the introduced clones, at least two different origins could be distinguished. Several of the introduced S. tabernaemontani clones were clearly different from individuals of the same species that were native to this region. As there were different hybrid genotypes owing to different hybrid events or to introgression, this complex should be safeguarded from genetic pollution. New S. tabernaemontani genotypes with markers not previously observed in that hybrid complex of the unique tidal freshwater habitat should be avoided in replantation projects.     

Genetic relationship of 32 cell lines of the euplotes octocarinatus species complex revealed by random amplified polymorphic DNA (RAPD) fingerprinting

Random amplified polymorphic DNA (RAPD) fingerprinting was used in this study to determine the genetic relationship of different cell lines of the hypotrichous ciliate Euplotes octocarinatus. Stocks isolated from different habitats in the USA, and from a group of genetically recombined laboratory strains, were characterized. Band-sharing indices (D) for all possible pairwise comparisons revealed a remarkable genetic diversity between the different cell lines. Investigation of the genetic structure in natural populations found diversity--although to a different extent--in all populations investigated. No clonal structure could be observed, as proposed for several protozoa and recently shown for E. daidaleos. These findings suggest frequent conjugation in the population of E. octocarinatus. No correlation between the genetic relationship of cell lines from different habitats and the distance between the corresponding sampling locations was found. Once separated geographically, the exchange of genetic material between populations appears to be nearly impossible. Therefore, these groups tend to separate into sibling species. The data generally support the occurrence of different syngens in the E. octocarinatus species complex. This finding is in accordance with our observation that the morphological 'species' of E. octocarinatus consists of several syngens or sibling species, similar to findings for the Paramecium aurelia-, Tetrahymena pyriformis- and E. vannus- species complexes.     

华山新麦草自然居群沿海拔梯度的遗传分化

华山新麦草为我国特有种 ,仅分布于陕西华山 ,是国家一级珍稀濒危植物和急需保护的农作物野生亲缘种。应用 RAPD技术 ,选取 8条引物对华山新麦草自然分布区的 3个山峪(种群 ) 1 1个样方 (亚居群 ) 79个华山新麦草个体的总 DNA进行随机扩增 ,共得到 65个RAPD位点。统计分析表明 ,在华山的 3个山峪 (居群 )中 ,黄埔峪 (居群 )与其它 2个山峪 (居群 )发生较显著的遗传分化 ;华山峪 6个亚居群的个体平均 RAPD位点数有随海拔的升高而下降的趋势 ;6个亚居群间的相似性系数也有随海拔的升高而下降的趋势 ;华山峪的高海拔亚居群和低海拔亚居群间表现出明显差异。主成分分析结果进一步证明了黄埔峪居群与华山峪居群和仙峪居群间、以及华山峪的高海拔亚居群与低海拔亚居群间已发生一定程度的分化。研究结果暗示海拔差异是导致华山新麦草自然居群遗传分化的主要因素 ,海拔差异造成的有限的基因流可能才是影响居群和亚居群遗传分化的主要因子 ,而亚居群内遗传变异程度则与该亚居群的所处的特定生境有关     

Assessment of genetic variation within Indian mustard (Brassica juncea) germplasm using random amplified polymorphic DNA markers

Genetic diversity among 45 Indian mustard (Brassica Juncea L.) genotypes comprising 37 germplasm collections, five advance breeding lines and three improved cultivars was investigated at the DNA level using the random amplified polymorphic DNA (RAPD) technique. Fifteen primers used generated a total of 92 RAPD fragments, of which 81 (88%) were polymorphic. Of these, 13 were unique to accession 'Pak85559'. Each primer produced four to nine amplified products with an average of 6.13 bands per primer. Based on pairwise comparisons of RAPD amplification products, Nei and Li's similarity coefficients were calculated to evaluate the relationships among the accessions. Pairwise similarity indices were higher among the oilseed accessions and cultivars showing narrow ranges of 0.77-0.99. An unweighted pair-group method with arithmetic averages cluster analysis based on these genetic similarities placed most of the collections and oilseed cultivars close to each other, showing a low level of polymorphism between the accessions used. However, the clusters formed by oilseed collections and cultivars were comparatively distinct from that of advanced breeding lines. Genetically, all of the accessions were classified into a few major groups and a number of individual accessions. Advanced breeding lines were relatively divergent from the rest of the accessions and formed independent clusters. Clustering of the accessions did not show any pattern of association between the RAPD markers and the collection sites. A low level of genetic variability of oilseed mustard was attributed to the selection for similar traits and horticultural uses. Perhaps close parentage of these accessions further contributed towards their little diversity. The study demonstrated that RAPD is a simple and fast technique to compare the genetic relationship and pattern of variation among the gene pool of this crop.     

Strain typing of Lentinula edodes by random amplified polymorphic DNA assay

Abstract Single 10-base primers were used to generate randomly amplified polymorphic DNA (RAPD) markers in the shiitake mushroom, Lentinula edodes . Seven primers produced polymorphisms in all 15 strains tested, producing 12–19 bands ranging from 0.34 to 2.52 kb. Thirteen of the 15 strains had unique DNA fingerprints, whereas L. edodes ATCC 28759 and ATCC 28760 exhibited identical RAPD profiles for all the primers. Molecular-genetic markers obtained with the RAPD assay can be used to differentiate strains of L. edodes and have potential applications in mushroom breeding and strain improvement programs.     

Characterization of Zebu cattle breeds in Tanzania using random amplified polymorphic DNA markers

A total of 141 short primers, of arbitrary nucleotide sequence, were used singly in poly-merase chain reactions to amplify DNA fingerprints in pools of DNA representing three Zebu cattle breeds. Two primers, which discriminated between the breed-specific DNA pools were used further to amplify individual pool components in order to establish band frequencies of the amplified fingerprints. One of the primers (ILO 1127) amplified a RAPD fingerprint in 61%of TSZ animals but less than 6% in the other breeds, while another primer (ILO 1065) revealed a DNA sequence common to 89% of the Boran animals and less than 30% in the other two breeds. Bandsharing and mean average percentage difference calculated within and between the three breeds using RAPD fingerprint data showed a higher degree of homogeneity within than across the breeds and indicated measurable divergence between the three breeds. It is concluded that RAPD polymorphisms are useful as genetic markers for cattle breed differentiation.