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本文报道了用识别四个GC对的限制性内切酶HaeⅢ、CofI、MsPI酶切高等担子菌伞菌目(Agaricales)、非褶菌目(Aphyllophorales)12个菌株的总DNA,在琼脂糖凝胶上显现线粒体DNA带谱的结果。通过对带型、累加分子量、酶切片段数和同源性的分析表明,不同种类高等担子菌的线粒体DNA变化较大.研究还显示,以多种识别四个GC对的限制酶分析高等担子菌的线粒体DNA及其它们的RFLP,比只用HaeⅢ更灵敏有效。 相似文献
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节丛孢属rDNA ITS区RFLPs分析 总被引:2,自引:0,他引:2
利用PCR朢FLP方法对捕食线虫真菌节丛孢属进行了系统发育研究。以ITS1和ITS4为引物对该属10种12个菌株的核糖体DNA转录间区(ITS)进行了PCR扩增,4种内切酶(AluI、HaeIII、HpaII、TaqI)酶切。结果表明该属的ITS区长度没有明显差异,其长度范围在658~705之间,酶切图谱能体现不同种间的多态性,根据4种内切酶酶切结果,利用UPGMA法构建的节丛孢属分子系统树,基本体现了属内不同种间的亲缘关系,从分子水平证明了节丛孢属形态分类上的合理性,同时对该属属征的扩大提出疑议。 相似文献
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人工栽培蛹虫草退化现象的分子分析* 总被引:4,自引:0,他引:4
利用PCR-RFLP和RAPD方法对野生驯化蛹虫草及其退化菌种进行了基因水平的分析。PCR-RFLP实验,采用真菌通用引物ITS1和ITS4,扩增出5.8S和其两端的两个转录间隔区(ITS),选用5种识别四个碱基的内切酶(HaeIII、AfaI、TagI、AluI和XspI),其中只XspI酶切结果在两个菌种中存在差异,进一步测序表明,整个片段全长534bp,共有13个位点发生碱基突变,且都为c转换为t。RAPD实验,选用4组计80个随机引物进行PCR扩增,共筛选出9个对所有供试个体均存在明显差异的引物。结果表明了退化菌株较野生驯化菌株在DNA水平上发生了频率较高的突变,这些突变可能直接与导致菌种退化的基因相关联。 相似文献
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捕食线虫真菌rDNA ITS区间RFLPs分析 总被引:3,自引:1,他引:2
利用PCR-RFLP方法对捕食线虫真菌进行了系统发育研究。以ITS1和ITS4为引物对3属14种16个菌株的核糖体DNA转录间区(ITS)进行了PCR扩增,4种内切酶(AluI、HaeIII、HpaII、TaqI)酶切,结果表明不同属的ITS区长度没有明显差异,其长度范围在585~695之间。酶切图谱种间差别明显,种内基本一致,同属菌株图谱没有特异性,暗示传统的分属可能过细,某些属的成立还有待商榷,PCR-RFLP对确定疑难种的地位有重要意义,但不适用于种下水平的系统学研究。 相似文献
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水稻线粒体DNA酶切带型研究 总被引:10,自引:0,他引:10
水稻IR36线粒体DNA经6种限制酶酶切,用脉冲电泳和长距离琼脂糖凝胶电泳分离酶切片段,获得高分辨率的清晰带型。每组酶切片段加和测得水稻IR36线粒体基因组大小分别为227kb(HindⅢ)、253kb(EcoRⅠ)、253kb(XhoⅠ)、294kb(BamHⅠ)、239kb(SalⅠ)和283kb(xbal)采用9个来自水稻和玉米线粒体基因组的基因探针与酶切条带杂交发现,水稻线粒体基因组含有包括编码基因在内的重复顺序。 相似文献
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本文采用限制内切酶HindⅢ切割经Sepharose 4 B柱纯化的北京鸭肝线粒体DNA,得到五个片段,其大小分别为:A——6.56kb,B——3.12kb,C——3.12kb,D——2.40kb,E——1.40kb。以质粒pWR33为载体,在HindⅢ切点处插入线粒体DNA的HindⅢ酶切片段,将体外重组质粒转化到大肠杆菌HB101内,经筛选、分析:如菌落原位杂交,限制性内切分析和Southern吸印法分析,首次得到了线粒体DNA的HindⅢB、C、D、E四个片段的克隆子。另外还得到了一个与片段A同源的1.60kb大小的片段。 相似文献
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对印度酸桔(Citrus reticulata) 飞龙枳(Poncirus trifoliata)属间体细胞杂种的3棵8年生植株及其融合亲本的胞质基因组进行了CAPS(Cleaved Amplified Polymorphic Sequences)和RFLP分析。用5对叶绿体和5对线粒体通用引物对(universal primer pairs)对杂种及亲本的总DNA进行PCR扩增,都没有检测到多态性,但扩增产物分别用11种限制性内切酶酶切后,发现3个有多态性的叶绿体CAPS标记和1个线粒体CAPS标记。结果表明杂种的叶绿体都来源于飞龙枳,而线粒体都来源于印度酸桔。为了证实CAPS分析结果的可靠性,用5种限制性内切酶对总DNA进行单酶切,分别与1个叶绿体探针和5个线粒体探针杂交,结果与CAPS分析一致。初步证实该组合体细胞杂种的胞质遗传组成为“印度酸桔的线粒体 飞龙枳的叶绿体”。结果表明细胞融合确实能导致细胞核、线粒体和叶绿体的重新组合,为柑桔体细胞杂种中线粒体偏向来源于悬浮亲本而叶绿体偏向来源于叶肉亲本的胞质分配现象提供了新的证据,并为通过体细胞融合技术定向转移柑桔胞质基因的品种改良思路提供了重要理论依据。 相似文献
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木耳属真菌rDNA特异性扩增片段的RFLP研究 总被引:3,自引:0,他引:3
对木耳属8个种25个菌株的ITS和28SrDNA5’端两个区域分别进行了PCR扩增和限制酶切片段长度多态性(RFLP)研究。ITS-RFMP研究结果表明,HaeⅢ可将黑木耳与其它种区分开,MspⅠ可将盾形木耳、角质木耳、琥珀木耳和黑木耳4个种区分开,而供试的HaeⅢ、TaqⅠ、HinfⅠ和MaPⅠ这四种限制酶均不能将皱木耳、大木耳、网脉木耳及毛木耳4个种区分开,表明它们之间的亲缘关系较近;结果还表明,ITS—rDNA拷贝在毛木耳和琥珀木耳种内是异质性的,而在黑木耳种内是同质性的。285rDNA-RFLP研究结果表明,供试的4种限制酶中,仅MspⅠ可将盾形木耳和角质木耳区分开,而不能将其它种区分开,这显示了28SrDNA序列在木耳属不同种间的保守性。 相似文献
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Organelle DNA inheritance of four 10-year-old somatic hybrid trees between Valencia orange [Citrus sinensis (L.) Osbeck] and Meiwa kumquat (Fortunella crassifolia Swingle) was analyzed by cleaved amplified polymorphic sequence (CAPS) and restriction fragment length polymorphisms (RFLPs). Five chloroplast (cp) and three mitochondrial (mt) universal primer pairs were amplified, but no polymorphisms were detected. When the polymerase chain reaction products were digested by 15 restriction enzymes, four polymorphic cpDNA-CAPS and two mtDNA-CAPS markers were found. Both the cpDNA and mtDNA in the somatic hybrids were derived from Valencia orange (the embryogenic suspension parent). Genomic DNA of the somatic hybrids and corresponding parents was digested by five restriction endonucleases and hybridized with one chloroplast probe (RbcL- RbcL) and nine mitochondrial probes (coxI, coxII, c oxIII, c ob, atpA, tyr, proI, atp6 and atp9). The results indicated that three hybrid plants shared one strong cpDNA band with both parents and that the remaining one plant had two additional novel bands besides the shared band, while their mtDNA was identical to that of Valencia orange plus non-parental bands. When data on the mtDNA banding patterns were combined with observations on phenotypic performance in the field, it was found that the more complex mtDNA banding pattern coincided with increased vigor of the plant. The stability of the organelle genomes was studied by extracting the genomic DNA of one hybrid plant at monthly intervals for 1 year and then analyzing it using RFLPs. Before the dieback of the shoots, two fragments of the mtDNA were lost while the cpDNAs remained stable. Ploidy analysis by flow cytometry showed that all of the hybrids were stable tetraploids. Four simple sequence repeat primer pairs were applied to detect microsatellite alleles of the four hybrid plants, both parents and the 12 DNA samples from one plant. The results showed that all hybrids had biparental bands uniformly, which indicated that they had the same nuclear background. These results suggest that the mtDNA pattern is correlated with the phenotypic abnormality of Valencia and kumquat somatic hybrid plants and that nuclear-cytoplasm incompatibility may be the cause of dieback. 相似文献
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对木耳属8个种25个菌株的ITS和28SrDNA5’端两个区域分别进行了PCR扩增和限制酶切片段长度多态性(RFLP)研究。ITS-RFMP研究结果表明,HaeⅢ可将黑木耳与其它种区分开,MspⅠ可将盾形木耳、角质木耳、琥珀木耳和黑木耳4个种区分开,而供试的HaeⅢ、TaqⅠ、HinfⅠ和MaPⅠ这四种限制酶均不能将皱木耳、大木耳、网脉木耳及毛木耳4个种区分开,表明它们之间的亲缘关系较近;结果还表明,ITS—rDNA拷贝在毛木耳和琥珀木耳种内是异质性的,而在黑木耳种内是同质性的。285rDNA-RFLP研究结果表明,供试的4种限制酶中,仅MspⅠ可将盾形木耳和角质木耳区分开,而不能将其它种区分开,这显示了28SrDNA序列在木耳属不同种间的保守性。 相似文献
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B. Wang W. Cheng Y. -N. Li D. -D. Li 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1989,77(4):581-586
Summary Certain physicochemical properties of rice mitochondrial DNA (mtDNA) were determined. Certain low-molecular-weight mtDNA bands were found in addition to the major mtDNA band. Rice mtDNA appeared in the electron microscope as a collection of linear molecules with heterogeneous length in the range of 1–156 kb. The major distribution area was 60–105 kb. A small fraction (less than 5%) of rice mtDNA was found in the form of a circular molecule. Some molecules had the appearance of being supercoiled. Replication fork structures were found in both circular and linear mtDNA molecules. In one rice species, Jin Nante, 15 different circular molecules were found. Rice mtDNA was digested with different restriction enzymes. The total molecular weight of rice mtDNA was calculated to be about 300 kb according to the data of restriction enzyme digestion and electron microscopy. 相似文献
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Differentiation of mitochondrial subgroups in the Verticillium lecanii species complex 总被引:2,自引:0,他引:2
Presumptive mitochondrial DNA (mtDNA) restriction fragment length polymorphisms (RFLPs) were obtained from 54 fungal isolates identified as Verticillium lecanii, V. psalliotae or ' V. lecanii -like'. Analysis of the mtDNA RFLPs showed 20 different patterns, indicating considerable genetic variation within the V. lecanii species complex. There was no direct correlation between host and mtDNA pattern, or between mtDNA patterns and previously described isoenzyme-defined specific groups. Isolates from tropical and subtropical areas showed considerable variation in genotypes, while isolates from temperate regions appeared less variable. 相似文献
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《Fungal Ecology》2017
The increasing availability of DNA sequence data enables exciting new opportunities for fungal ecology. However, it amplifies the challenge of how to objectively classify the diversity of fungal sequences into meaningful units, often in the absence of morphological characters. Here, we test the utility of modern multilocus Bayesian coalescent-based methods for delimiting cryptic fungal diversity in the orchid mycorrhiza morphospecies Serendipita vermifera. We obtained 147 fungal isolates from Caladenia, a speciose clade of Australian orchids known to associate with Serendipita fungi. DNA sequence data for 7 nuclear and mtDNA loci were used to erect competing species hypotheses by clustering isolates based on: (a) ITS sequence divergence, (b) Bayesian admixture analysis, and (c) mtDNA variation. We implemented two coalescent-based Bayesian methods to determine which species hypothesis best fitted our data. Both methods found strong support for eight species of Serendipita among our isolates, supporting species boundaries reflected in ITS divergence. Patterns of host plant association showed evidence for both generalist and specialist associations within the host genus Caladenia. Our findings demonstrate the utility of Bayesian species delimitation methods and suggest that wider application of these techniques will readily uncover new species in other cryptic fungal lineages. 相似文献
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Shigeyuki Kawano Takahito Suzuki Tsuneyoshi Kuroiwa 《Biochimica et Biophysica Acta (BBA) - Gene Structure and Expression》1982,696(3):290-298
Mitochondrial DNA (mtDNA) of Physarum polycephalum was isolated gently by CsCl centrifugation. The mtDNA was linear with molecular weights ranging from 25·106 to 45·106 and heterogeneous in size. Nevertheless, thermal transition profiles of the mtDNA suggested that this DNA fraction was more homogeneous than nuclear DNA. Exhaustive digestions of this DNA with restriction endonucleases yielded unique fragments, and then the total of their molecular weights of each digest was around 45·106. This value is equivalent to the maximum molecular weight estimated using electron microscopy and electrophoresis. Moreover, EcoRI digests of the mtDNA fractionated by the sucrose gradient showed unequimolar quantities of large fragments and a high background between bands. These results suggest that the mtDNA of Physarum has a homogeneous base sequence, and that the size heterogeneity of the mtDNA is attributable to degradation of the DNA under isolation procedures. The mtDNA was cleaved by EcoRI and XhoI to yield 16 and 7 fragments, respectively. A physical map of these fragments was constructed using the routine mapping procedures. The physical map showed that the mitochondrial genome of Physarum was linear with a molecular weight of 45·106. We concluded therefore that the mitochondrial nucleoid is a structure in which the homogeneous mtDNA is highly amplified. 相似文献