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1.
编码18-5.8-25S核糖体RNA的45S rDNA基因,是1个简单的多基因家族基因,一般以串联方式相连,对应于核仁组织区(NOR).首次利用荧光原位杂交技术(FISH),研究45S rDNA在散生竹类的毛竹和斑竹,混生竹类的茶秆竹、日本矮竹、菲白竹和铺地竹,以及丛生竹类的白绿竹染色体上的分布.结果表明:本研究所涉及的散生竹和混生竹的几个竹种,只观察到1对随体染色体的次缢痕区域有45S rDNA位点,而丛生竹类的白绿竹除随体染色体具有45S rDNA位点外,某些非随体染色体上也有不同拷贝数的45S rDNA位点存在.  相似文献   

2.
The structure of the female sex-determining W chromosome was examined in wild-type females and in three mutant female lines of the codling moth, Cydia pomonella. The mutant lines were isolated from progeny of irradiated females using the abnormal appearance of sex chromatin body in female somatic polyploid nuclei as a cytogenetic marker. A detailed cytogenetic analysis was carried out using three different fluorescence in situ hybridization techniques: genomic in situ hybridization (GISH), comparative genomic hybridization (CGH), and fluorescence in situ hybridization (FISH) with the codling moth W-chromosome painting probe. The FISH techniques enabled us to identify the W chromosome and its fragments in mitotic and meiotic chromosome complements and thus helped us to resolve complicated W chromosome aberrations induced by irradiation.  相似文献   

3.
细胞学在探讨菊花起源与菊属植物进化中的应用   总被引:1,自引:0,他引:1  
论述了菊属植物细胞学研究现状及研究成果对探讨菊花的起源、菊属植物进化的意义;总结了前人采用细胞学方法对野生菊属植物中的某些种如毛华菊、甘菊、野菊等与菊花之间亲缘关系的研究结果,指出前人对于菊花脑的分类地位、菊花染色体数目变化规律等问题仍存有争议并有待进一步的研究;最后讨论了新的研究方法对细胞学发展的应用前景。  相似文献   

4.
Thirty‐eight isolates of Rhizoctonia spp. were isolated from Scots pine (Pinus sylvestris) seedlings with damping‐off symptoms, originating from two forest nurseries in central‐west Poland (Wronczyn and Jarocin) and from diseased seedlings grown in soil from Wronczyn nursery. Majority of these isolates (79%) had multinucleate cells and were identified as Rhizoctonia solani. The remaining isolates were recognized as binucleate Rhizoctonia spp. R. solani isolates were characterized using hyphal anastomosis and were divided into five anastomosis groups (AG). The most prevalent was AG5 (37% of isolates), followed by AG2‐1 (30%) and 27% of the isolates were identified as AG4. Groups AG1‐IB and AG2‐2 were only represented by single isolates. The virulence recorded as mortality (in percentage) was comparatively high for binucleate and multinucleate isolates of Rhizoctonia spp. Sequence analysis of the polymerase chain reaction (PCR)‐amplified internal transcribed spacer (ITS) rDNA region was used for phylogenetic analysis. The dendrogram showed that isolates were distinctly separated based on their AG types and there was no relationship between pathogenicity on Scots pine seedlings and the AG to which the isolates belong to. The results are discussed with respect to pathogenic potential of the various AG groups.  相似文献   

5.
葡萄座腔菌属(B otryosphaeria)系统分类评述   总被引:4,自引:0,他引:4       下载免费PDF全文
本文系统论述了葡萄座腔菌(Botryosphaeria Ces. & De Not.)及其无性型的系统分类现状.首先概述了Botryosphaeria真菌模式种的建立及无性型分类的演化过程,主要介绍了Botryosphaeria真菌系统分类的一个重要观点:即Botryosphaeria代表了两个不同的系统发育单元的集合,其无性阶段可能分别对应于色二孢属(Diplodia)和壳梭孢属(Fusicoccum).同时介绍了真菌多无性型(pleoanamorphy)、同等无性型(synanamorphs)概念和Fusicoccum真菌座腔壳砖隔孢属(Dichomera)同等无性型的主要特征.分析了一些重要有性型种,如茶藨子葡萄座腔菌(B. dothidea),多主葡萄座腔菌(B. ribis),贝伦格葡萄座腔菌(B. berengeriana),B. parva,B. lutea等之间的系统分类关系,介绍了近年发现的20多个有性与无性型新种.其次介绍了主要的分子系统学研究方法并分析了其在Botryosphaeria真菌系统分类中的应用状况.最后通过分析与国外研究水平的差距,讨论了今后我国重点研究的方向.  相似文献   

6.
利用Giemsa C-分带和荧光原位杂交的方法对龟甲竹根尖细胞有丝分裂中期染色体进行了研究.结果表明:龟甲竹的核型公式为2n=48=26m+14sm+4st(2SAT)+4t,核型分类属ⅡB型;带型公式为2n=48=4CI+2CI+T+2CT++IOCT++4I+2I++6T++6C+8T+2IT+2I+T,每条染色体都显示出了可以明显区别的特征带,带纹强弱差异明显.以45S rDNA为探针对龟甲竹根尖染色体进行了荧光原位杂交,杂交位点定位于1对染色体上.将染色体c-带带型和45S rDNA FISH带型相结合可以将龟甲竹的每条染色体区分开.  相似文献   

7.
【目的】我国经济林树种板栗和杉木上的重要害螨一直被认为是同一种螨——针叶小爪螨,但针叶小爪螨的山东板栗种群和浙江杉木种群间存在生殖隔离,且生殖隔离并非由地理隔离和能调控寄主生殖的内共生菌引起,似乎二者已分化为2个独立的种。本研究目的是明确其成种原因,丰富物种形成理论。【方法】从之前做杂交试验时所采集的针叶小爪螨板栗和杉木种群中各随机取3头雌成螨,提取单头雌成螨的总DNA,使用根据其他小爪螨28S rDNA两端保守序列设计的引物扩增28S rDNA,扩增产物纯化后测序。比较2种群的28S rDNA序列,并基于28S rDNA序列构建小爪螨属系统发育树,结合已有研究分析两种群分化原因。【结果】每个种群的3个个体的28S rDNA序列完全一致,无种群内变异。2种群的28S rDNA序列一致性为98. 3%,遗传距离为1. 7%。在基于28S rDNA序列构建的小爪螨属系统发育树上,浙江杉木种群与采自日本的日本柳杉上的本岛小爪螨亲缘关系最近,山东板栗种群与采自日本的日本栗上的栗小爪螨的亲缘关系最近。【结论】板栗和杉木上的叶螨并非由同一种叶螨因适应不同寄主植物分化而成,很可能是2个独立的物种。  相似文献   

8.
Genetic variation among Armillaria ostoyae isolates was studied by rDNA-restriction fragment length polymorphism (RFLP) and random amplified polymorphic DNA (RAPD) analysis. A total of 20 A. ostoyae isolates, mainly obtained from Picea spp. of different geographical origins, were examined. Southern hybridization of whole-cell DNAs digested with AvaII and probed with biotin-labelled cloned rDNA from Saccharomyces carlsbergensis allowed the differentiation of five RFLP groups. UPGMA cluster analysis of RAPD profiles (138 scorable bands) generated by 10 decamer primers (OPA 01-OPA 10) grouped the isolates in subclusters at similarity levels between 40% and 96%, indicating high intraspecific genetic variability. Some isolates of different geographical origins subgrouped together, suggesting that similar mutational events have occurred independently and that genetic exchange and recombination occurs among the DNAs in natural populations. The potential role of historical and current spread of spruce plants on the genetic variation of A. ostoyae isolates in Europe is discussed. Using the primer pair ARM-1 and ARM-2, an Armillaria-specific ITS-DNA fragment of about 660 bp was obtained. No intraspecific RFLP of this amplicon could be revealed, indicating low genetic variability of this region. The established informative RFLP and RAPD markers and also the Armillaria-specific ITS-DNA fragment may be powerful tools for further epidemiological, phylogenetic and host-pathogen interaction studies with A. ostoyae.  相似文献   

9.
We studied the location and distribution of a bacterial isolate, a Mycobacterium sp., in buds of Scots pine (Pinus sylvestris L.). Using a probe specific for the 16S rRNA of the Mycobacterium sp., the bacterium was found by in situ hybridization in the meristematic tissues of 40% of all bud samples examined. Because we had previously found other bacterial and fungal endophytes in the meristematic tissues of Scots pine buds, we studied their occurrence in buds during shoot development and dormancy. Using probes targeted to the 16S or 18S rRNA of the endophytes Mycobacterium sp., Methylobacterium spp., Pseudomonas spp. and Rhodotorula minuta, endophytes were found in association with growing tissues, with Methylobacterium spp. being the dominant species. Endophytes were detected in abundance before elongation or differentiation of a bud, but once a tissue was fully developed, endophytes were no longer detected. Metabolic activity of the endophytes was suppressed at the onset of, and during, dormancy of Scots pine, but recovered before the following growing season.  相似文献   

10.
Phytophthora agathidicida (PTA) causes a root rot and collar rot of New Zealand kauri (Agathis australis). This study developed techniques to visualize early infection of kauri by PTA in deliberately inoculated seedlings. Conventional light microscopy was carried out on cleared and stained roots using trypan blue to observe PTA structures. Additionally, scanning electron microscopy (SEM) was used to study the PTA root structures at a higher resolution. A fluorescent in situ hybridization assay (FISH) was developed using a PTA‐specific probe to label PTA structures in planta. Infection progression in roots of 2‐year‐old kauri inoculated with PTA at 5, 10, 16 and 20 days post‐inoculation (d.p.i.) was compared using these three approaches. Light microscopy identified no Phytophthora‐like structures in the control treatments. In PTA‐inoculated plants, lignitubers were produced 5 d.p.i. in cortical cells. Infection was localized after 5 days, but as the infection progressed (up to 20 d.p.i.), the ‘degree’ of root infection increased, as did the number of replicates in which structures were observed. SEM provided higher resolution images; again, no PTA structures were observed in the negative control material examined. The slide‐based FISH‐specificity assay successfully hybridized with PTA hyphae. Fluorescence was observed using 330–380 nm excitation and an emission filter at 420 nm (DAPI), with PTA nuclei fluorescing a bright greenish‐yellow. Cross‐reactivity was not observed when the assay was applied to six other non‐target Phytophthora species. Successful hybridization reactions occurred between the primer and PTA structures in planta. Applying this FISH assay has allowed clear differentiation of the intracellular and intercellular structures of PTA. The technique can be applied to longer term studies or analysis of ex situ inoculation studies aiming to elucidate differential host‐responses to the pathogen. Additionally, the technique could be applied to study the interactions with other fungal endophytes (e.g. mycorrhizal fungi), which could be assessed for biocontrol potential as part of the integrated management of the disease.  相似文献   

11.
小兴安岭代表性菌物系统进化初步研究   总被引:2,自引:0,他引:2  
为探索小兴安岭菌物系统进化关系并验证可应用于该地菌物的分子钟校正时间标准,采集小兴安岭凉水国家级自然保护区代表性菌物25种,对ITS1、5.8S和ITS2 rDNA区域进行PCR扩增、测序。同源序列多重比对后分析序列特征。以核酸进化的最适模型构建最大似然树、最大简约树和邻接树,自展检验重复1 000次估计可靠性。根据分子钟推测菌物分化发生时间。25种ITS序列平均碱基组成为T=28.66%、C=24.11%、A=26.19%和G=21.03%,总体转换/颠换差R=1.274。Tajima检验支持最大似然树和邻接树的系统发育关系。对于长期进化而言,无ITS核苷酸序列以绝对恒定的速率变化,分子钟不具有通用性。构建不受分子钟约束的最大似然树,根据枝长估计红毛盘菌与其他菌物分化发生在500 Ma,灰光柄菇与其他菌物分化发生在约130 Ma,多孔菌在内的其他菌物起源于约100~120 Ma。  相似文献   

12.
Powdery mildew symptoms and signs were observed on seedlings of Lagerstroemia speciosa within an ornamental plant nursery in Viçosa, State of Minas Gerais, Brazil. Signs were most prominent as intense mycelial growth and sporulation on the shoots, new branches and leaves, which led to tissue necrosis. Based on morphological characteristics and phylogenetic analyses of ITS and 28S rDNA sequences, the powdery mildew pathogen of L. speciosa was identified as Erysiphe australiana, a well‐known powdery mildew pathogen infecting Lagerstroemia spp. worldwide. To our knowledge, this is the first report of E. australiana causing powdery mildew on L. speciosa in Brazil.  相似文献   

13.
根据根癌土壤杆菌Ti质粒的保守序列设计了2对引物,对我国不同木本植物上分离鉴定的10株致病性根癌土壤杆菌进行PCR,致瘤性根癌土壤杆菌(Agrobacterium tumefaciens)扩增出427 bp和338 bp的特异性DNA片段,而发根土壤杆菌(A.rhizogenes)仅扩增出338 bp片段,放线土壤杆菌(A.radiobacter)、丁香假单胞菌(Pseudomonas syringae)和泡桐丛枝病植原体(phytoplasma)皆未有特异扩增带.用CYT/CYT'引物对也可鉴定T-DNA遗传转化瘤组织,而VirD2A/VirD2E可用于工程土壤杆菌株侵染能力鉴定.从北京通州杨树苗圃和河北廊坊桃园的病树冠瘿组织和土壤中经选择培养基分离菌株,然后PCR筛选出6个致病性根癌土壤杆菌菌株,而未能从施用过抗根癌生防制剂的北京玉渊潭公园樱花根癌病园中分离和鉴定出典型的致病根癌土壤杆菌株.将杨树根癌土壤杆菌菌株(CFCC1001)异戊烯基转移酶基因(ipt)片段克隆和序列测定结果显示与A.tumefaciens Ti15955菌株的ipt基因序列同源性为83.64%.用此片段制备的ipt cRNA基因探针进行斑点杂交和Northern blot,结果显示此探针与杨树根癌土壤杆菌以及玫瑰、樱花、海棠、桃树等木本植物上分离鉴定的致病土壤杆菌菌株所扩增出的427 bp片段都有明显杂交信号,但与引起泡桐丛枝病植原体染色质和染色质外DNA均未有明显杂交信号.  相似文献   

14.
广宁红花油茶在广东、广西等原生地种植容易获得高产,具有良好的遗传改良潜力。文章以广宁红花油茶为对象,探索了适用于荧光原位杂交技术中的染色体制片技术,并与传统的染色体压片制作技术进行比较,总结了一套节省时间、技术要求较低和效果较好的染色体根尖压片制作技术,可用于广宁红花油茶的细胞学研究,为开展分子染色体工程育种以及基因组分析奠定技术基础。  相似文献   

15.
利用ITS rDNA序列对我国杨树溃疡病的葡萄座腔菌科病原真菌进行系统发育学分析,共得到4种葡萄座腔菌科病原真菌:Botryosphaeria dothidea,Neofusicoccum parvum,Diplodia seriata和D.mutila。它们与相应的无性型或有性型一一对应。其中,D.seriata在我国是第一次报道为杨树的病原。研究了试验菌株与国内外已报道菌株间的系统发育学关系,分析了它们与环境间的联系。结果表明B.dothidea具有高度的种内遗传多样性。这些结果对葡萄座腔菌科真菌在全球的分布作了必要的补充,也为杨树溃疡病的研究和防治提供进一步的理论基础。  相似文献   

16.
17.
Identification of European Armillaria species using ribosomal DNA (rDNA) restriction-fragment-length polymorphisms (RFLPs) was studied. A total of 44 Armillaria isolates representing five European biological species were examined. Whole-cell DNAs were digested with one or two (double digest) restriction endonucleases and probed with a cloned plasmid carrying one complete rDNA repeat copy of Saccharomyces carlsbergensis. Applying the restriction endonuclease Bgl II in combination with either Eco RI, Bam Hl or Hin dIII, it was possible to detect rDNA RFLPs allowing differentiation between all five European Armillaria species investigated in the study. The most conclusive results were obtained in the rDNA/Ava II RFLP pattern. All biological species showed unique Ava II banding patterns. Generally speaking, the interspecific similarities were around 42% and lower, indicating a distinct species separation. The analysis of rDNA RFLP patterns by a restriction-enzyme-rDNA-probe combination (for instance, Ava II or Bgl II/Hin dIII; probe pMY 60) is a practical means of identifying European Armillaria species for further taxonomic, phylogenetic and host-pathogen interaction studies.  相似文献   

18.
红树林内生真菌1893是从中国南海秋茄(Kandelia candel)上分离的一株内生真菌,在液体发酵条件下能够产生包括1893A和1893B在内的多种新物质。为了正确的鉴定该菌株并为进一步的工业应用做准备,本文结合形态学和分子学两方面对它进行了鉴定,首先,在纯培养条件下,将它和具有以下形态特征的相似菌株进行比较:产生受精丝或像受精丝一样的菌丝,这些菌丝又和像雄器一样的菌丝发生明显的融合现象;环状产囊体与大量菌丝缠绕形成菌核前体。进一步通过该菌株在寄主秋茄和非寄主芒果叶子上的培养性状与其它相似真菌比较,结果将该菌株归于拟茎点霉属(Phomopsis)。其次,在ITS序列水平上,将它和具有形态学或生态学相似性的拟茎点霉进行系统发育分析。最后,综合形态学和分子学,红树林内生真菌1893被定为Diaporthe phaseolorum var.sojae,本文是首次报道Diaporthe phaseolorum var.sojae做为内生真菌寄生于秋茄。图5参43。  相似文献   

19.
以In(NO_3)_3·4.5H_2O和六亚甲基四胺(HMT)为起始物,以活性炭(AC)为载体,采用一步水热原位合成法制备In(OH)_3/AC协同吸附-光催化复合材料,采用XRD、FT-IR、DRS、SEM以及TEM等方法对复合材料的晶相结构、表面结构等进行了表征,并测试了复合材料对甲苯的光催化降解性能。结果表明,AC与In(OH)_3是采用化学键结合;AC的加入不仅影响了In(OH)_3的成核与熟化过程,改变了In(OH)_3的晶胞结构,使晶面间距变小、粒径增加,而且降低了其结晶度;但是AC仅影响复合材料对光的吸收强度,对In(OH)_3能阈结构和粒子形貌基本没有影响。复合材料对甲苯的光催化降解测试结果显示,加入适量的AC(3%)所产生的吸附-催化协同效应可以有效地提高材料的光催化性能,光照60 min时,甲苯的降解率可达98.25%。  相似文献   

20.
采集巨桉林下马勃子实体,培养其菌丝体,提取子实体及菌丝体基因组DNA,进行rDNA-ITS区序列的扩增、克隆和序列测定,并对rDNA-ITS不同区域作序列分析,首次构建马勃的系统发育树.结果表明:野外采集获得马勃子实体l0种,其中成功培养6种,测序结果表明马勃rDNA-ITS区长度在607~766 bp之间,系统发育分析表明硬皮马勃属(Scleroderma)与豆包菌(Pisolithus)亲缘关系较近,秃马勃属(Calvatia)、马勃属(Lycoperdon)及横膜马勃属(Vascellum)之间亲缘关系较近,ITSl-5.8S rDNA-ITS2区可建立马勃类真菌属间系统发育树,ITS2区可用于建立马勃类真菌属内系统发育树,3个待定种硬皮马勃Scleroderma sp.11-1,Scleroderma sp.2-2和Scleroderma sp.5-2为金黄硬皮马勃(S.aurantium)的可能性较大.此研究可为探讨巨桉人工林下外生菌根种类与作用机制、马勃分类系统学及菌丝体的开发研究奠定基础.  相似文献   

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