共查询到20条相似文献,搜索用时 15 毫秒
1.
F.C.F. Yau K.L. Wong P.C. Shaw P.P.H. But J. Wang 《Biodiversity and Conservation》2002,11(9):1653-1662
Random amplified polymorphic DNA fingerprints characteristic of thethree snakes Zaocys dhumnades, Agkistrodonacutus and Bungarus multicinctus multicinctuswere generated using primer OPF-14. Z. dhumnades is anendangered species included in the Convention on International Trade inEndangered Species of Wild Fauna and Flora, and A. acutus,B. multicinctus multicinctus and Z. dhumnades are listed inthe Chinese Pharmacopoeia. The species-specific polymorphic bands Aa specific toA. acutus, Bmm specific to B. multicinctusmulticinctus and Zd specific to Z. dhumnadeswere identified and the sequences of these bands were used to design polymerasechain reaction primers for sequence characterized amplified region (SCAR)analysis of the three snakes. A multiplex SCAR analysis was established toauthenticate the snakes used in Chinese medicine reliably and efficiently. 相似文献
2.
Cysticercosis is one of the most important zoonosis, not only because of the effects on animal health and its economic consequences, but also due to the serious danger it poses to humans. The two main parasites involved in the taeniasis-cysticercosis complex in Brazil are Taenia saginata and Taenia solium. Differentiating between these two parasites is important both for disease control and for epidemiological studies. The purpose of this work was to identify genetic markers that could be used to differentiate these parasites. Out of 120 oligonucleotide decamers tested in random amplified polymorphic DNA (RAPD) assays, 107 were shown to discriminate between the two species of Taenia. Twenty-one DNA fragments that were specific for each species of Taenia were chosen for DNA cloning and sequencing. Seven RAPD markers were converted into sequence characterized amplified region (SCAR) markers with two specific for T. saginata and five specific for T. solium as shown by agarose gel electrophoresis. These markers were developed as potential tools to differentiate T. solium from T. saginata in epidemiological studies. 相似文献
3.
A method is described for the development of DNA markers for detection of Helicoverpa armigera (Hubner) (Lepidoptera: Noctuidae) in predator gut analysis, based on sequence characterized amplified regions (SCARs) derived from a randomly amplified polymorphic DNA (RAPD) band. A 1200-bp DNA fragment of H. armigera, absent in the predator band pattern and in other closely related prey species, was identified by RAPD analysis. This fragment was cloned and its extremes sequenced to design extended strand-specific 20-mer oligonucleotide primers. Three pairs of SCAR primers, which amplified three different DNA fragments, were used to study the effect of fragment length on detection of prey in the predator gut. Using the pair of primers that amplified the longest fragment of H. armigera DNA, a single band of 1100 bp was obtained, but its detection was not possible in the predator gut. Detection of the ingested prey was possible with the other two pairs of SCAR primers, obtaining bands of 600 and 254 bp, respectively. Detection of H. armigera DNA in the gut of the predator Dicyphus tamaninii was evaluated immediately after ingestion (t = 0) and after 4 h. Detection of H. armigera DNA after 4 h was only possible using the pair of primers that amplified the shortest fragment (254 bp). The test for specificity, using these last pair of primers, showed that H. armigera was the only species detected. The detection threshold was defined at a 1:8192 dilution of a H. armigera whole egg in all samples. 相似文献
4.
Akamatsu HO Grünwald NJ Chilvers MI Porter LD Peever TL 《Journal of microbiological methods》2007,71(1):82-86
Three kinds of genetic markers including simple sequence repeats (SSRs), single nucleotide polymorphisms (SNPs) and sequence characterized amplified regions (SCARs) were developed from Aphanomyces euteiches. Of 69 loci tested, seven SSR, two SNP and two SCAR markers were codominantly polymorphic. These codominant markers and dominant markers described herein will facilitate population genetic and evolutionary studies of this important plant pathogen. 相似文献
5.
Species containing E genome of Thinopyrum offered potential to increase the genetic variability and desirable characters for wheat improvement. However, E genome specific marker was rare. The objective of the present report was to develop and identify sequenced characterized amplified region (SCAR) markers that can be used in detecting E chromosome in wheat background for breeding purpose. Total 280 random amplified polymorphic DNA (RAPD) primers were amplified for seeking of E genome specific fragments by using the genomic DNA of Thinopyrum elongatum and wheat controls as templates. As a result, six RAPD fragments specific for E genome were found and cloned, and then were converted to SCAR markers. The usability of these markers was validated using a number of Egenome-containing species and wheat as controls. These markers were subsequently located on E chromosomes using specific PCR and fluorescence in situ hybridization (FISH). SCAR markers developed in this research could be used in molecular marker assisted selection of wheat breeding with Thinopyrum chromatin introgressions. 相似文献
6.
Abstract: Bemisia tabaci (Gennadius) is an important pest of agriculture and horticulture crops that includes many morphologically indistinguishable biotypes. Molecular markers can provide a scientific method to rapidly identify biotypes. In this study, we attempted to develop specific primer sets for rapidly and stably identifying various biotypes of B. tabaci . We developed sequence characterized amplified region (SCAR) markers for each of six B. tabaci (Gennadius) biotypes (A, B, Q, Nauru, An and S). Two primer sets (BaAF/BaAR and BaQF/BaQR) were designed from random amplified polymorphic DNA-specific fragments for the A and Q biotypes. Four forward primers, BaBF, BaNaF, BaANF and BaSF, with a common reverse primer, L2-N-3014, for the other four biotypes of B, Nauru, An and S, respectively, were used based on their individual mitochondrial cytochrome oxidase I sequences. Six of these SCARs were useful in distinguishing each biotype from the others. Application of these SCARs will be helpful in rapidly identifying biotypes for quarantine pest control to prevent the invasion of the various biotypes. 相似文献
7.
Quirin EA Ogundiwin EA Prince JP Mazourek M Briggs MO Chlanda TS Kim KT Falise M Kang BC Jahn MM 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》2005,110(4):605-612
Phytophthora capsici causes devastating disease on many crop species, including Capsicum. Resistance in Capsicum annuum is genetically and physiologically complex. A panel of Capsicum germplasm that included genotypes from both C. annuum and C. chinense showing highly resistant, highly susceptible and intermediate or tolerant responses to the pathogen, respectively, was screened with a series of randomly amplified polymorphic sequence primers to determine which genomic regions contribute to the highest level of resistance. One primer, OpD04, amplified a single band only in those C. annuum and C. chinense genotypes showing the highest level of resistance. The amplified product was cloned, sequenced and used to design longer primers in order to generate a sequence characterized amplified region marker which was then mapped in a reference mapping population and a screened population segregating for resistance to P. capsici. These primers were observed to define a locus on pepper chromosome 5 tightly linked to Phyto.5.2, one of six quantitative trait loci (QTL) previously reported to contribute to P. capsici resistance. These results indicate that the Phyto.5.2 QTL may be widely distributed in highly resistant germplasm and provide improved resolution for this QTL. This work also defines the first breeding tools for this system, allowing for the rapid selection of genotypes likely to be highly resistant to P. capsici. 相似文献
8.
Furtado-Filho OV Polcheira C Machado DP Mourão G Hermes-Lima M 《Comparative biochemistry and physiology. Toxicology & pharmacology : CBP》2007,146(1-2):241-254
Crocodilians and other diving vertebrates experience hypoperfusion and hypoxia of several internal organs during long dives. At the end of a dive, reperfusion of aerated blood may cause a physiologically relevant oxidative stress. In this study, we analyzed selected markers of oxidative stress in eight organs of normoxic Paraguayan caiman (Caiman yacare) captured in the Brazilian Pantanal wetlands during the winter of 2001 (six mature-adult males and eight young-adult males; AD-1 and YA-1 groups, respectively), and during the summer of 2002 (six young-adult males (YA-2 group), ten hatchlings and five embryos). Lipid peroxidation products determined by three different assays were generally highest in brain, liver and kidney (in comparison with all other organs), and lowest in white muscles from the tail and hind legs. Liver and kidney showed the highest levels of carbonyl protein, while brain showed low levels. Intermediate levels of oxidative stress markers were mostly found in the heart ventricles and lung. Differences in oxidative stress markers between AD-1 and YA-1 were organ-specific, showing no age-related correlation. However, most oxidative stress markers in YA-2 organs were either higher than (by 1.4- to 3.7-fold) or not significantly different from respective values in hatchlings organs. This pattern (hatchlings versus young-adults) was confirmed using correlation analysis of individual caiman size versus levels of oxidative damage markers in four organs. The higher level of oxidative stress markers in young-adults possibly relates to the fast growth rate (and thus, increased oxidative metabolic rate) of C. yacare in the first years of life. Differences in oxidative stress markers between YA-1 and YA-2 were also observed and were ascribed to seasonal changes in free radical metabolism. These results in normoxic C. yacare represent the first step towards understanding the age-related physiological oxidative stress of a diving reptile from a seasonally changing wetland environment. 相似文献
9.
Sequence characterized amplified region markers tightly linked to the mating factors of Lentinula edodes. 总被引:1,自引:0,他引:1
Detecting the mating types in shiitake, Lentinula edodes (Berk.) Pegler, is important for making progress in the breeding of this mushroom and determining the compatibility of the pair to cross. Shiitake is a tetrapolar fungus with two unlinking mating factors, A factor and B factor. We screened molecular markers linked to the mating factors using the randomly amplified polymorphic DNA (RAPD) method to develop the mating type identification procedure. Using 147 oligonucleotide primers, a total of 6 linkage markers for the shiitake mating factors, 4 markers for the A factor and 2 markers for the B factor, were discovered with a logarithm of the odds threshold of 3.0 for linkage. Two RAPDs that perfectly segregated with each mating factor among 72 basidiospore strains were detected. Both of these RAPDs were cloned and sequenced to convert them to the sequence characterized amplified region (SCAR) markers. Four primers, two sets of primers, were designed according to the internal sequences of two RAPDs tightly linking to the A factor or B factor. Consequently, we determined the polymerase chain reaction condition for multiplex analyses of these SCAR markers. 相似文献
10.
MAIKEL M. AVESKAMP JOYCE H.C. WOUDENBERG JOHANNES DE GRUYTER ELENA TURCO JOHANNES Z. GROENEWALD PEDRO W. CROUS 《Molecular Plant Pathology》2009,10(3):403-414
Phoma exigua is considered to be an assemblage of at least nine varieties that are mainly distinguished on the basis of host specificity and pathogenicity. However, these varieties are also reported to be weak pathogens and secondary invaders on non‐host tissue. In practice, it is difficult to distinguish P. exigua from its close relatives and to correctly identify isolates up to the variety level, because of their low genetic variation and high morphological similarity. Because of quarantine issues and phytosanitary measures, a robust DNA‐based tool is required for accurate and rapid identification of the separate taxa in this species complex. The present study therefore aims to develop such a tool based on unique nucleotide sequence identifiers. More than 60 strains of P. exigua and related species were compared in terms of partial actin gene sequences, or analysed using DNA amplification fingerprinting (DAF) with short, arbitrary, mini‐hairpin primers. Fragments in the fingerprint unique to a single taxon were identified, purified and sequenced. Alignment of the sequence data and subsequent primer trials led to the identification of taxon‐specific sequence characterized amplified regions (SCARs), and to a set of specific oligonucleotide combinations that can be used to identify these organisms in plant quarantine inspections. 相似文献
11.
Development of a sequence‐characterized amplified region marker using inter‐simple sequence repeats for detection of Puccinia striiformis f. sp. tritici
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Stripe rust, caused by Puccinia striiformis f. sp. tritici (Pst), is the most devastating wheat disease in China. Early and accurate detection of the pathogens would facilitate effective control of the diseases. DNA‐based methods now provide essential tools for accurate plant disease diagnosis. In this study, inter‐simple sequence repeats (ISSR) technique has been successfully applied to develop a sequence‐characterized amplified region (SCAR) marker for diagnosis of stripe rust of wheat and detection of Pst. In this study, one fragment unique to Pst was identified by ISSR and then sequenced. Based on the specific fragment, a pair of SCAR primers (616AF/616AR) was designed to amplify a 299‐bp DNA fragment within the sequenced region. The primers can amplify a unique DNA fragment for all tested isolates of Pst but not for the other pathogens of wheat leaves and the uninfected leaves. The polymerase chain reaction (PCR) assay could detect as low as 0.1 ng of genomic DNA in a 25.0 μl PCR reaction mixture and detect the pathogen from asymptomatic wheat leaves inoculated with Pst under glasshouse conditions. 相似文献
12.
Riaz S Sadia B Awan FS Khan IA Sadaqat HA Khan IA 《Genetics and molecular research : GMR》2012,11(1):440-447
DNA fingerprints of four rose species, Rosa centifolia, R. Gruss-an-Teplitz, R. bourboniana, and R. damascena, were developed using RAPD-PCR. We identified a unique polymorphic band in R. centifolia. This 762-bp fragment was produced by the random primer GLI-2. The fragment was eluted and directly cloned in a TA cloning vector, pTZ57R/T. Digestion of the plasmid with EcoRI confirmed the cloning of GLI-2(762) in pTZ57R/T. A second enzyme, PstI, used in combination with EcoRI, gave complete digestion of the plasmid, and the 762-bp fragment was confirmed on the gel. Subsequently, the polymorphic amplicon was sequenced with an AB1 373 DNA sequencer system using the PRISM(TM) Ready Reaction DyeDeoxy(TM) Terminator Cycle Sequencing kit. After sequencing, specific primers (23 bp long) were designed based on the sequence of the flanking regions of the original RAPD fragment. These primers will effectively allow fingerprinting for the identification of R. centifolia species. In essence, we developed an SCAR marker to authenticate the identity of R. centifolia species and to distinguish it from its substitutes. Such techniques are required not only to complement conventional parameters in creating the passport data of commercial and medicinal products of rose, but also for routine quality control in commercial and government rosaries and rose nurseries. 相似文献
13.
We describe an unusual repetitive DNA region located in the 3′ end of the light (L)-strand in the mitochondrial control region
of two elephant seal species. The array of tandem repeats shows both VNTR (variable-number tandem repeat) and sequence variation
and is absent from 12 compared mammalian species, except for the occurrence in the same location of a distinct repetitive
region in rabbit mtDNA and a similar repeat in the harbor seal. The sequence composition and arrangement of the repeats differ
considerably between the northern elephant seal (Mirounga angustirostris) and the southern species (M. leonina) despite an estimated divergence time of 1 MY (based on an mtDNA-RNA gene and the nonrepetitive control region). Analysis
of repeat sequence relationships within and between species indicate that divergence in sequence and structure of repeats
has involved both slippagelike and unequal crossingover processes of turnover, generating very high levels of divergence and
heteroplasmy.
Presented at the NATO Advanced Research Workshop onGenome Organization and Evolution, Spetsai, Greece, 16–22 September 1992 相似文献
14.
H. Li S. G. Zhang J. M. Gao C. G. Wang Y. Zhang L. W. Qi L. Chen W. Q. Song 《Biologia Plantarum》2008,52(3):525-528
In this study, bulked segregant analysis (BSA) was used on Larix leptolepis × Larix olgensis hybrids to identify a random amplified polymorphic DNA (RAPD) marker associated with high rooting ability in larch. Two DNA
bulks: H (high rooting ability) bulk and L (low rooting ability) bulk were constructed according to the rooting percentages
of the stock plants. Among the 328 primers, only S356 could amplify a specific band, named S356445, which only existed in the H bulk and was further confirmed following selective genotyping of individual hybrids. Grounded
on the border sequences, S356445 was converted to a sequence characterized amplified region (SCAR) marker, HRL445, which can be useful in marker-assisted
selection (MAS) to screen for larch with high rooting ability. All the results strongly indicated that S356445 and HRL445 were closely associated with high rooting ability in larch. 相似文献
15.
Williams CG Reyes-Valdés MH Huber DA 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》2007,116(1):87-94
Validation of a quantitative trait locus (QTL) for outcrossing perennial plants is rarely reported due to complexity of plausible
genetic models and reliance on field designs already available. Here, a particular marker-QTL haplotype exerted a large, positive
effect on height for Pinus taeda and its origin could be traced to a founder, GP3, in a three-generation QTL pedigree. To validate this QTL effect, we used an extended GP3-based pedigree. In the validation cross, each of the 46 offspring was clonally propagated from developing seeds using somatic
embryogenesis technology. Subsequent analyses were conducted separately for seedlings and for other somatic emblings. For
seedlings, the original QTL effect could not be fully validated. For somatic emblings, a strong negative QTL effect was detected
in the validation cross; some evidence from another cross-supported the original positive QTL effect. From this part of the
analysis, three distinct marker-QTL haplotypes at a single locus could be inferred. Validating QTL haplotypes in readily available
field tests was feasible despite the genetic model complexity inherent to outcrossing long-lived perennials. 相似文献
16.
尼罗鳄线粒体基因组全序列分析及鳄类系统发生关系的探讨 总被引:1,自引:0,他引:1
大多数脊椎动物的线粒体基因组(约16—18kb)的组成是相对较稳定的,但在不同类群中,线粒体基因组在基因结构和基因排列方式等方面均显示了极大的多样性,这种多样性可能反映了真核细胞不同的进化路线(Saccone et al.,1999)。就目前的研究而言,线粒体基因组是惟一一个能够从基因组水平上来分析动物系统发生的分子标记,可以从线粒体基因组序列信息、基因组成及基因排列方式等进行多方位的分子进化研究,因而线粒体基因组全序列将成为动物分子系统发生最有力的证据(Saccone et al.,1999)。 相似文献
17.
Molecular authentication of Panax species 总被引:16,自引:0,他引:16
Using conserved plant sequences as primers, the DNA sequences in the ribosomal ITS1-5.8S-ITS2 region have been amplified and determined for six Panax species, P. ginseng C. A. Mey. (Oriental ginseng), P. quinquefolius L. (American ginseng), P. notoginseng (Burkill) F. H. Chen (Sanchi), P. japonicus C. A. Mey. (Japanese ginseng), P. trifolius L. and P. major Ting, as well as two common adulterants of ginseng, Mirabilis jalapa L. and Phytolacca acinosa Roxb. An authentication procedure based upon the restriction fragment length polymorphism (RFLP) in the region is able to differentiate between P. ginseng and P. quinquefolius, and to discriminate the ginsengs from the two common poisonous adulterants. Broader application of this approach to authenticate other morphologically similar Chinese medicinal materials is rationalised. 相似文献
18.
19.
H. Britz B. D. Wingfield T. A. Coutinho M. J. Wingfield 《Molecular ecology resources》2002,2(4):577-580
Fusarium circinatum is the causal agent of pitch canker disease of pines. This pathogen is thought to have originated in Central America and currently poses a serious threat to commercial pine plantations in many areas of the world. In this study, polymorphic molecular markers were developed for F. circinatum using the internal short sequence repeats‐polymerase chain reaction (ISSR‐PCR) technique. Nine sequence characterized amplified polymorphic markers were developed. Thirty‐two putative alleles were observed among 103 F. circinatum isolates from different geographical areas using the nine polymorphic markers. These sequence characterized amplified polymorphic markers can be used as genetic tools in populations studies of F. circinatum. 相似文献
20.
The 5S RNA genes inPinus radiata occur in two size classes of about 850 and 525 bp in length. Representatives from both the long and short size classes were cloned and sequenced. The primary difference in the two size classes was shown to be a 330 bp insertion in the spacer region of the long units. Within an individual breeding clone ofP. radiata there was some sequence heterogeneity between representatives of the short class. The 5S RNA genes in thirty pine species were characterised using either a clone of the short size class or a subclone of the 330 bp insertion characterizing the long size class. Eleven species of subg.Strobus were examined and all lacked the long type of unit of radiata pine. The New World species of subg.Pinus all had both short and long units whereas the Old World species had long units. The implications of these results for the evolution of the 5S DNA sequences and the phylogenetic relationships withinPinus are discussed. 相似文献