用于SSR分析的大豆DNA的快速提取   总被引：40，自引：9，他引：40  

关荣霞  常汝镇  邱丽娟 《大豆科学》2003,22(1):73-74

描述了一种可从大豆种子及叶片中快速提取DNA的方法，既节省时间，又可获得供上百次PCR扩增使用的DNA，并通过实验证明该方法获得的DNA可用于大豆SSR分析。  相似文献   

基于定量PCR方法初步分析西藏开菲尔粒中细菌与酵母菌数量变化     

王兴兴  丁梦帆  潘迎捷  王永杰 《上海海洋大学学报》2015,24(4):625-631

运用实时荧光定量PCR方法分析西藏开菲尔粒中细菌与酵母菌的数量变化规律,需要有效提取其微生物总基因组DNA。本文通过同时添加溶菌酶与溶壁酶优化实验方案,成功获得西藏开菲尔粒中微生物的总基因组DNA。荧光定量PCR分析提取自培养1、4、8和12周的西藏开菲尔粒的微生物总基因组DNA,结果显示:4个时间点细菌16S rRNA与酵母菌26S rRNA基因拷贝数均分别约为每微升107拷贝与105拷贝,表明细菌与酵母菌的数量相对稳定。该结果为进一步研究西藏开菲尔粒中细菌与酵母菌间的共生关系奠定了一定的基础。  相似文献   

Phylogenetic reassessment of Mycosphaerella spp. and their anamorphs occurring on Eucalyptus. II.     

《Studies in Mycology》2006

Species of Eucalyptus are widely planted as exotics in the tropics and Southern Hemisphere and to some extent in southern Europe, for timber and fibre production. Species of Mycosphaerella are commonly associated with leaves and twigs of Eucalyptus and can result in defoliation, dieback, and even tree death. In the present study, numerous isolates of Mycosphaerella species were collected from leaf litter, living leaves exhibiting leaf spot symptoms or severe Mycosphaerella leaf blotch symptoms. Isolates were compared based on DNA sequence data for the internal transcribed spacer region (ITS1 & ITS2) and the 5.8S gene. These data, together with characteristics of the fungal growth on three different media, morphology of the anamorph and teleomorph structures as well as ascospore germination patterns were used to describe 21 new species.Taxonomic novelties: Colletogloeopsis stellenboschiana Crous sp. nov., Mycosphaerella davisoniellae Crous sp. nov. (anamorph Davisoniella eucalypti H.J. Swart), Mycosphaerella eucalyptorum Crous & M.J. Wingf. sp. nov. Mycosphaerella gamsii Crous sp. nov., Mycosphaerella perpendicularis Crous& M.J. Wingf. sp. nov., Mycosphaerella pluritubularis Crous & J.P. Mansilla sp. nov., Mycosphaerella pseudafricana Crous & T. Coutinho sp. nov., Mycosphaerella pseudocryptica Crous sp. nov. (anamorph Colletogloeopsis sp.), Mycosphaerella pseudoendophytica Crous & G. Hunter sp. nov. (anamorph Pseudocercosporella sp.), Mycosphaerella pseudosuberosa Crous & M.J. Wingf. sp. nov. (anamorph Trimmatostroma sp.), Mycosphaerella quasicercospora Crous & T. Coutinho sp. nov., Mycosphaerella scytalidii Crous & M.J. Wingf. sp. nov. (anamorph Stenella sp., synanamorph, Scytalidium-like.), Mycosphaerella secundaria Crous & A.C. Alfenas sp. nov., Mycosphaerella stramenti Crous & A.C. Alfenas sp. nov., Mycosphaerella stramenticola Crous & A.C. Alfenas sp. nov., Mycosphaerella sumatrensis Crous & M.J. Wingf. sp. nov., Mycosphaerella verrucosiafricana Crous & M.J. Wingf. sp. nov., Septoria eucalyptorum Crous sp. nov., Septoria provencialis Crous sp. nov., Stenella pseudoparkii Crous & M.J. Wingf. sp. nov. (teleomorph Mycosphaerella sp.), Stenella xenoparkii Crous & M.J. Wingf., sp. nov. (teleomorph Mycosphaerella sp.).  相似文献   

Re-evaluating the taxonomic status of Phaeoisariopsis griseola,the causal agent of angular leaf spot of bean     

《Studies in Mycology》2006

Angular leaf spot of Phaseolus vulgaris is a serious disease caused by Phaeoisariopsis griseola, in which two major gene pools occur, namely Andean and Middle-American. Sequence analysis of the SSU region of nrDNA revealed the genus Phaeoisariopsis to be indistinguishable from other hyphomycete anamorph genera associated with Mycosphaerella, namely Pseudocercospora and Stigmina. A new combination is therefore proposed in the genus Pseudocercospora, a name to be conserved over Phaeoisariopsis and Stigmina. Further comparisons by means of morphology, cultural characteristics, and DNA sequence analysis of the ITS, calmodulin, and actin gene regions delineated two groups within P. griseola, which are recognised as two formae, namely f. griseola and f. mesoamericana.Taxonomic novelties: Pseudocercospora griseola (Sacc.) Crous & U. Braun comb. nov., P. griseola f. mesoamericana Crous & U. Braun f. nov.  相似文献   

The USDA trypsin inhibitor study. II. Timed related biochemical changes in the pancreas of rats     

I. E. Liener  Z. Nitsan  C. Srisangnam  J. J. Rackis  M. R. Gumbmann 《Plant foods for human nutrition (Dordrecht, Netherlands)》1985,35(3):243-257

A total of 600 rats was distributed equally among 15 diets formulated from mixtures of raw and toasted soy flour and casein so as to provide 5 different levels of trypsin inhibitor (TI) at 3 levels of protein (10%, 20%, and 30%). Animals were sacrificed at the end of 6 months and at 3-month intervals thereafter over a period of 22 months. Increases in the secretory activity of the pancreas, as measured by trypsin and chymotrypsin activities in the pancreas and small intestines, were associated with increasingly higher levels of TI and protein but did not change appreciably with time. Hypertrophy and hyperplasia of the pancreas, as reflected by changes in pancreas weight and RNA and DNA content, developed at an early stage (6 months) and were likewise positively correlated with the levels of TI and protein in the diet. Although the hypertrophic response remained unchanged with time, hyperplasia became more pronounced as the period of exposure to TI was prolonged. The peptide test confirmed the stimulatory effect of TI on the pancreas, but some functional impairment was noted in those animals which had been fed raw soy flour for 18 months of or longer. The appearance of nodules on the pancreas was observed by gross inspection after 15 months, and these became larger and more numerous with time. The incidence of nodules was highly correlated with the level of TI in the diet and was highest when the diet contained 20% protein.  相似文献   

瑞氏木霉碳源阻遏相关基因Cre1的分子改造     

陈小玲  陈东  张穗生  陈英 《广西农业生物科学》2014,(2):288-292

为了提高瑞氏木霉（Trichoderma reesei）纤维素酶的活力,用类似基因改组的方法改造其碳源阻遏相关基因cre1。以瑞氏木霉基因组DNA为模板PCR扩增cre1基因,用DNaseⅠ消化cre1基因后,回收50-100 bp的DNA片段,用T4 DNA连接酶连接,以连接产物作为模板进行无引物PCR,并将PCR产物转入瑞氏木霉原生质体,通过测定滤纸酶活的方法筛选突变菌株,并在NCBI上比对分析突变菌株的cre1基因。结果表明,筛选获得1株纤维素滤纸酶活比出发菌株提高0.7倍的突变菌株cre2-3。cre2-3菌株在液体培养基中呈棉花状,而出发菌株呈小颗粒状,菌株cre2-3发酵液的颜色比出发菌株的更黄亮。推测cre1基因与瑞氏木霉菌株的生长代谢有关。  相似文献   

Genetic Structure of Severe Combined Immunodeficiency Carrier Horses in Morocco Inferred by Microsatellite Data     

Mohammed Piro  Abdelaziz Benjouad  Abdelghafor Karom  Ahmed NabichNoureddine Benbihi  Khalid El Allali  Mohammed Machmoum  Lahoussine Ouragh 《Journal of Equine Veterinary Science》2011,31(11):618-624

A total of 17 microsatellite deoxyribonucleic acid loci used routinely for horse parentage control were used to evaluate genetic diversity among normal Arabian horses and severe combined immunodeficiency (SCID) carrier Arabian horses (ArS) and normal Arab-Barb horses and SCID carrier Arab-Barb horses (ArbeS). On the basis of the genotype of 186 horses, mean allelic diversity was estimated as 6.82, 5.53, and 6.7059 in normal Arabian horses, ArS, and for both groups of Arab-Barb horses, respectively. Five specific alleles were observed in ArS and ArbeS, with one common with ArS at HMS6, whereas five alleles common between ArS and ArbeS had a high frequency. Expected and observed heterozygosity showed great heterogeneity in the population studied and were similar or higher when compared with other studies on Arabian horses. Coefficient of gene differentiation G_st of Nei associated with Nei’s genetic distance and multivariate correspondence analysis indicated a possible differentiation between the studied populations when analyzed separately according to breed. Probability of assignment of a horse to a specific group was assessed using a full and partial Bayesian approach. In all, 80.6% of Arab horses and 78.2% of Arab-Barb horses were assigned properly with a partial Bayesian test, which provided better results than the full one. These findings will be useful for identification of SCID carrier horses by using the microsatellite deoxyribonucleic acid loci used routinely for horse parentage control in our laboratory.  相似文献   

First Steps Towards an Understanding of a Mode ofCarcinogenic Action for Vanadium Pentoxide     

Schuler D  Chevalier HJ  Merker M  Morgenthal K  Ravanat JL  Sagelsdorff P  Walter M  Weber K  McGregor D 《Journal of toxicologic pathology》2011,24(3):149-162

Inhalation of vanadium pentoxide clearly increases the incidence of alveolar/bronchiolar neoplasms in male and female B6C3F1 mice at all concentrations tested (1, 2 or 4 mg/m³), whereas responses in F344/N rats was, at most, ambiguous. While vanadium pentoxide is mutagenic in vitro and possibly in vivo in mice, this does not explain the species or site specificity of the neoplastic response. A nose-only inhalation study was conducted in female B6C3F1 mice (0, 0.25, 1 and 4 mg/m³, 6 h/day for 16 days) to explore histopathological, biochemical (α-tocopherol, glutathione and F2-isoprostane) and genetic (comet assays and 9 specific DNA-oxo-adducts) changes in the lungs. No treatment related histopathology was observed at 0.25 mg/m³. At 1 and 4 mg/m³, exposure-dependent increases were observed in lung weight, alveolar histiocytosis, sub-acute alveolitis and/or granulocytic infiltration and a generally time-dependent increased cell proliferation rate of histiocytes. Glutathione was slightly increased, whereas there were no consistent changes in α-tocopherol or 8-isoprostane F2α. There was no evidence for DNA strand breakage in lung or BAL cells, but there was an increase in 8-oxodGuo DNA lesions that could have been due to vanadium pentoxide induction of the lesions or inhibition of repair of spontaneous lesions. Thus, earlier reports of histopathological changes in the lungs after inhalation of vanadium pentoxide were confirmed, but no evidence has yet emerged for a genotoxic mode of action. Evidence is weak for oxidative stress playing any role in lung carcinogenesis at the lowest effective concentrations of vanadium pentoxide.  相似文献   

中国部分地区绵羊肺炎支原体的基因多态性分析     

许健  储岳峰  赵萍  高鹏程  贺英  剡根强  逯忠新 《畜牧兽医学报》2011,42(9)

为了解绵羊肺炎支原体(Mycoplasma ovi pneumoniae,Mo)在我国部分地区的流行病学特征,采用随机扩增多态性DNA(RAPD)及限制性片段长度多态性(RFLP)方法对26株Mo基因多态性进行了研究,并利用NTsys2.10e软件对获得的多态性图谱进行了聚类分析.结果在相似系数为0.70时,26株Mo可分成6个RAPD群或6个RFLP群;相似系数为0.90时,分成18个RAPD群或18个RFLP群;相似系数为1.00时,分成25个RAPD群或26个RFLP群.结果表明,我国Mo存在高度的基因多态性,这种多态性与地域差异相关,与宿主来源也具有一定的相关性.研究结果为了解我国Mo的分子流行病学特征打下了基础,也为建立有效的Mo诊断方法和疫苗研制提供了参考依据.  相似文献   

湖南省山羊脑多头蚴的线粒体nad1和nad4基因的序列测定及种系发育分析     

何德肆  徐平源  彭运潮 《畜牧兽医学报》2011,42(12)

本研究旨在阐明脑多头蚴湖南分离株线粒体烟酰胺腺嘌呤二核苷酸( NADH)脱氢酶亚单位1基因(nad1)部分序列(pnad1)和烟酰胺腺嘌呤二核苷酸(NADH)脱氢酶亚单位4基因(nad4)部分序列(pnad4)的遗传变异情况,并用pnad1和pnad4序列重构脑多头蚴与其它带科绦虫的种群遗传关系.利用聚合酶链反应(PCR)扩增脑多头蚴的pnad1和pnad4,应用ClustalX 1.81程序对序列进行比对,再用Phylip3.67程序MP法和Mage4.0程序NJ法绘制种系发育树,并用Puzzle5.2程序构建最大似然树,同时利用DNAstar5.0中的Megalign程序进行同源性分析.结果显示所获得的pnad1和pnad4序列长度分别均为666和887 bp,湖南分离株与已知多头带绦虫位于同一分枝.由于脑多头蚴pnad1和pnad4序列种内相对保守,种间差异较大,故均可作为种间遗传变异研究的标记,从而为脑多头蚴的分子流行病学和其相关疾病的诊断奠定基础.  相似文献