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Manuel TORAL IBAEZ Margarita CARU Miguel A.HERRERA Luis GONZALEZ Luis M.MARTIN Jorge MIRANDA Rafael M.NAVARRO-CERRILLO 《Journal of Zhejiang University. Science. B》2009,10(2)
A protocol of polymerase chain reaction-random amplified polymorphic DNAs (PCR-RAPDs) was established to analyse the gene diversity and genotype identification for clones of Sequoia sempervirens (D.Don) Endl.in Chile.Ten (out of 34) clones from introduction trial located in Voipir-Viilarrica,Chile,were studied.The PCR-RAPDs technique and a modified hexadecyltrimethylammonium bromide (CTAB) protocol were used for genomic DNA extraction.The PCR tests were carried out employing 10-mer random primers.The amplification products were detected by electrophoresis in agarose gels.Forty nine polymorphic bands were obtained with the selected primers (BG04,BF07,BF 12,BF13,and BF 14) and were ordered according to their molecular size.The genetic similarity between samples was calculated by the Jaccard index and a dendrogram was constructed using a cluster analysis of unweighted pair group method using arithmetic averages (UPGMA).Of the primers tested,5 (out of 60) RAPD primers were selected for their reproducibility and high polymorphism.A total of 49 polymorphic RAPD bands were detected out of 252 bands.The genetic similarity analysis demonstrates an extensive genetic variability between the tested clones and the dendrogram depicts the genetic relationships among the clones,suggesting a geographic relationship.The results indicate that the RAPD markers permitted the identification of the assayed clones,although they are derived from the same geographic origin. 相似文献
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为了对本课题组得到的大批优质棉花种质资源进行RAPD分子标记遗传多样性鉴定,我们进行了对棉花有效的引物对的筛选,并完成了24个优质棉花材料的遗传多样性分析.结果从48个随机引物对中筛选出了20对引物,这些引物对这24个优质棉花材料的有效性非常高,共扩增出113条谱带,平均每个引物扩增出5.65条,而且这20个随机引物对这24个材料的多态性谱带扩增效果非常明显,多态率高,能明显分辨出来源不同的材料.将这些多态性标记通过聚类软件分析,可将这24种材料明显的分为六类群,遗传多态性得到明显的体现. 相似文献
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RAPD在蔬菜种子纯度鉴定中应用的反应条件优化 总被引：2，自引：0，他引：2
随着分子生物学的发展,ＲＡＰＤ（随扩增多态性ＤＮＡ）在越来越多的领域得到应用,但在蔬菜纯度鉴定方面,国内外报道较少,为了更好地将ＲＡＰＤ方法应用于蔬菜种子纯度鉴定,就反应条件进行了探索,以几种蔬菜种子为材料,使用进口ＰＣＲ仪在,对几种蔬菜种子进行ＲＡＰＤ分子过程中,从ＤＮＡ提取方法,模板ＤＮＡ量,Ｍｇ＾２＋浓度,Ｔａｑ－ＤＮＡ聚合酶的量,ｄＮＴＰｓ浓度,随机引物浓度及反应缓冲液的量等几个方面进行研 相似文献
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中国桫椤科6种植物的RAPD分析 总被引：8，自引：1，他引：8
应用随机扩增多态性DNA (RAPD)的方法分析中国桫椤科 6种植物 :小羽桫椤(Cyatheatsangii)、粗齿桫椤 (Alsophiladenticulata)、黑桫椤 (Alsophilapodophylla)、白桫椤(Sphaeropterisbrunoniana)、海南白桫椤 (Sphaeropterisbrunoniana)、桫椤 (Alsophilaspinulosa)。经筛选 4 0个引物 ,发现 14个引物的带型清晰并呈多态性。采用UPGMA法对求出的遗传距离进行聚类分析 ,得到的结果显示 :所研究 6种植物明显地聚合为 3个类群 ,支持夏群的分类处理 相似文献
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Assessment of genetic diversity by simple sequence repeat markers among forty elite varieties in the germplasm for malting barley breeding 总被引：1，自引：0，他引：1
Jun-mei Wang Jian-ming Yang Jing-huan Zhu Qiao-jun Jia Yue-zhi Tao 《Journal of Zhejiang University. Science. B》2010,11(10):792-800
The genetic diversity and relationship among 40 elite barley varieties were analyzed based on simple sequence repeat (SSR)
genotyping data. The amplified fragments from SSR primers were highly polymorphic in the barley accessions investigated. A
total of 85 alleles were detected at 35 SSR loci, and allelic variations existed at 29 SSR loci. The allele number per locus
ranged from 1 to 5 with an average of 2.4 alleles per locus detected from the 40 barley accessions. A cluster analysis based
on the genetic similarity coefficients was conducted and the 40 varieties were classified into two groups. Seven malting barley
varieties from China fell into the same subgroup. It was found that the genetic diversity within the Chinese malting barley
varieties was narrower than that in other barley germplasm sources, suggesting the importance and feasibility of introducing
elite genotypes from different origins for malting barley breeding in China. 相似文献
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Fabiola MORENO-OLIVAS ;Vincent U.GANT Jr ;Kyle L.JOHNSON ;Jose R.PERALTA-VIDEA ;Jorge L.GARDEA-TORRESDEY 《浙江大学学报(A卷英文版)》2014,15(8):618-623
研究目的：二氧化钛（TiO2）纳米颗粒已经广泛应用于化妆品、防晒霜、涂料和牙膏等。这些纳米颗粒性质非常稳定,能在废水和生物固体中转移和分散。现有研究表明,TiO2纳米颗粒对动物正常生理活动具有毒性等负面作用。但是,它们对植物是否具有毒性特别是是否会产生植物基因毒性至今尚不清楚。因此,本文使用随机扩增多态性DNA技术研究TiO2纳米颗粒是否对西葫芦具有基因毒性,为TiO2纳米颗粒排放进入环境后的潜在植物毒性风险评价提供依据。 创新要点：首次发现了TiO2纳米颗粒对西葫芦具有基因毒性。 重要结论：采用随机扩增多态性DNA技术,发现TiO2纳米颗粒污染处理的西葫芦样品与未处理样品的基因组DNA图谱相比,不仅在谱带强度有明显差异,而且存在谱带消失和新谱带产生现象,表明TiO2纳米颗粒对西葫芦具有基因毒性。 相似文献
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生物分子系统学研究中的RAPD技术 总被引：1，自引：0，他引：1
RAPD技术在Williams等人于1990年首先提出后,在检测DNA多态性方面受到了广泛的应用.RAPD技术以其简单、快速、信息量大等特点渗透在基因诊断,法医检测,生物药品的检验,生物种群关系的研究各个领域.以核酸为指标构建的生物系统进化树,不受主观因素的影响,避免了经典植物系统学的人为影响因素,因而受到广泛应用,在分子系统研究中出现了RFLP,RAPDDNA和RNA测序等分子生物学技术,其中RAPD技术不需要经过DNA分子克隆、分子杂交等繁锁步骤,也不需用放射性同位素显示,并且多态性检出率高,因而受到普遍关注. 相似文献
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以南美水蛭抗凝血Xa因子mRNA为模板设计引物,利用RT-PCR随机克隆云南牛蛭（P.yunnanensis.sp.nov）相关基因,结果得到多条片段,经严格筛选和鉴定后,确定大于200 bp ESTs序列,通过对这些ESTs序列进行聚类、拼接,结果得到10条片段,Genebank注册号分别为HQ285410,HQ285411,HQ285412,HQ285413,HQ285414,HQ285415,HQ285416,HQ285417,HQ285418,HQ285419.以上获得的云南水蛭头部和身体细胞部分转录的ESTs片段均与南美水蛭有较大差异,表明云南水蛭与南美水蛭可能使用完全不同的抗凝血机制. 相似文献
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A genetic linkage map comprised of 131 loci was constructed with an F2 population derived from an inter-subspecific cross betweenBrassica campestris L. ssp.chinensis cv. ‘aijiaohuang’ and ssp.rapifera cv. ‘qisihai’. The genetic map included 93 RAPD loci, 36 AFLP loci and 2 morphological loci organized into 10 main linkage
groups (LGs) and 2 small groups, covering 1810.9cM with average distance between adjacent markers being approximately 13.8cM.
The map is suitable for identification of molecular markers linked to important agronomic traits, QTL analysis, and even for
marker-assisted selection in breeding programs of Chinese cabbage and turnip.
Project (No. 39870509) supported by the National Natural Science Foundation of China 相似文献
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Umezuruike Linus OPARA Dan JACOBSON Nadiya Abubakar AL-SAADY 《Journal of Zhejiang University. Science. B》2010,11(5):332-341
Banana is an important crop grown in Oman and there is a dearth of information on its genetic diversity to assist in crop breeding and improvement programs.This study employed amplified fragment length polymorphism(AFLP) to investigate the genetic variation in local banana cultivars from the southern region of Oman.Using 12 primer combinations,a total of 1094 bands were scored,of which 1012 were polymorphic.Eighty-two unique markers were identified,which revealed the distinct separation of the seven cultiva... 相似文献
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采用改进的CTAB法提取中国特有濒危裸子植物白豆杉(Pseudotaxus chienii)嫩叶总DNA,进行RAPD分析.分别研究了模板DNA、引物、dNTPs、Mg2 和TaqDNA聚合酶用量对RAPDPCR扩增反应结果的影响.筛选并建立了适合于白豆杉RAPD扩增的反应体系:总体积25μL,其中10×PCR缓冲液(500mmol/L KCl,100mmol/L Tris-HCl,1.0%Triton X-100,20mmol/L MgCl2)2.0μL,引物(1.5μmol/L)0.5μL,dNTPs(10mmol/L)0.4μL,模板1μL(35ng),Taq酶0.8μL(0.5U),水20.3μL. 相似文献
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Xiu-ping Lu Bing-guang Xiao Yong-ping Li Yi-jie Gui Yu Wang Long-jiang Fan 《Journal of Zhejiang University. Science. B》2013,14(7):570-577
Diversity arrays technology (DArT) is a microarray-based marker system that achieves high throughput by reducing the complexity of the genome. A DArT chip has recently been developed for tobacco. In this study, we genotyped 267 flue-cured cultivars/landraces, including 121 Chinese accessions over five decades from widespread geographic regions in China, 103 from the Americas, and 43 other foreign cultivars, using the newly developed chip. Three hundred and thirty polymorphic DArT makers were selected and used for a phylogenetic analysis, which suggested that the 267 accessions could be classified into two subgroups, which could each be further divided into 2–4 sections. Eight elite cultivars, which account for 83% of the area of Chinese tobacco production, were all found in one subgroup. Two high-quality cultivars, HHDJY and Cuibi1, were grouped together in one section, while six other high-yield cultivars were grouped into another section. The 330 DArT marker clones were sequenced and close to 95% of them are within non-repetitive regions. Finally, the implications of this study for Chinese flue-cured tobacco breeding and production programs were discussed. 相似文献
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王子迎 《安徽教育学院学报》2005,23(6):94-96,105
采用随机扩增多态性DNA(RAPD)和简单序列重复区间扩增多态性(Inter-simple sequence repeat,ISSR)技术,利用12条RAPD引物和10条ISSR引物对10个安徽香菇菌株进行了DNA标记比较分析.结果表明,ISSR-PCR较RAPD-PCR稳定,更具可操作性;供试菌株存在比较显著的遗传变异,具有较丰富的遗传多样性. 相似文献
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采用改进的CTAB法提取中国特有濒危裸子植物白豆杉(Pseudotaxus chienii)嫩叶总DNA,进行RAPD分析.分别研究了模板DNA、引物、dNTPs、Mg2 和TaqDNA聚合酶用量对RAPD PCR扩增反应结果的影响.筛选并建立了适合于白豆杉RAPD扩增的反应体系:总体积25μL,其中10×PCR缓冲液(500 mmol/L KCl,100 mmol/L Tris-HCl,1.0%Triton X-100,20 mmol/L MgCl2)2.0μL,引物(1.5μmol/L)0.5μL,dNTPs(10 mmol/L)0.4μL,模板1μL(35 ng),Taq酶0.8μL(0.5 U),水20.3 μL. 相似文献
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A genetic linkage map comprised of 131 loci was constructed with an F2 population derived from an inter-subspecific cross between Brassica 'qisihai'. The genetic map included 93 RAPD loci, 36 AFLP loci and 2 morphological loci organized into 10 main linkage groups (LGs) and 2 small groups, covering 1810.9cM with average distance between adjacent markers being approximately 13.8cM. The map is suitable for identification of molecular markers linked to important agronomic traits, QTL analysis, and even for marker-assisted selection in breeding programs of Chinese cabbage and turnip. 相似文献
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Gabriel PEREZ Valentina VERDEJO Clarissa GONDIM-PORTO Julieta ORLANDO Margarita CARU 《Journal of Zhejiang University. Science. B》2014,15(11):966-978
研究目的：开发具有种属特异性序列特征性扩增区域（SCAR）标记物来监测哈茨木霉在其入侵的试验菌群中的定殖和生长,为哈茨木霉应用于生物防治等生态和生物技术中提供支撑。
创新要点：多种木霉属真菌能与各种微观和宏观的生物有机体建立相互作用。利用这些相互作用,木霉可做为原生种群的入侵物种而用于生物防治。本文通过使用试验菌群为研究模型,利用随机扩增多态性DNA（RAPD）技术和序列特征性扩增区域（SCAR）标记物来监测菌群中哈茨木霉的生长状态。
研究方法：利用随机扩增多态性DNA（RAPD）技术,从16个10聚体引物进行多态性筛选,其中1个引物扩增出对应哈茨木霉的条带。对该条带进行克隆测序,并设计5个20-23聚体引物。成功利用引物对2F2／2R2和2F2／2R3278分别特异性地扩增出哈茨木霉BpTloa菌株278bp和448bp的DNA片段。同时,用这两个引物对14个哈茨木霉菌株和几种不同的真菌菌株进行特异性对照试验,也只成功扩增出哈茨木霉菌株。此外,使用真菌DNA混合物和试验真菌群的DNA为模板,采用实时聚合酶链式反应（PCR）对引物对2F2／2R2进行评估。当仅使用100份SCAR标记物或哈茨木霉仅占整个菌群的0．1％时,仍能检测出哈茨木霉。
重要结论：本研究所建立的SCAR分子标记能有效监测菌群中的哈茨木霉的定殖和生长,具有较高特异性、灵敏度和准确度。 相似文献