正交设计优化青蒿SRAP-PCR反应体系及引物筛选     

石紫薇  梁妍  姜婷婷  梁剑平  贾宁  辛志君  陆锡宏  周翔  李雪虎 《分子植物育种》2020,(10):3303-3310

为了建立青蒿的SRAP最佳扩增体系,并筛选出SRAP多态性引物,本研究以青蒿叶片DNA为模板,采用正交试验设计,以Mg^2+、dNTP Mix、Taq DNA聚合酶、引物和DNA模板5种因素5个水平,对青蒿SRAP反应体系进行研究。结果表明,青蒿SRAP-PCR最佳反应体系为:引物0.6μmol/L、Mg^2+2.0 mmol/L、模板DNA 5.1 ng、Taq DNA聚合酶2.0 U、dNTPs 0.25 mmol/L,总体积为25μL。各因素对扩增反应均有不同影响,其中引物浓度的影响最大,dNTPs的影响最小。运用该体系对不同种质资源的青蒿进行验证,证明该体系稳定可靠,并在30个引物组合中筛选出了25对扩增条带清晰,多态性丰富的引物组合。这一结论为今后利用SRAP标记技术进行青蒿分子遗传学研究提供了科学依据。  相似文献   

Development of a core set of KASP markers for assaying genetic diversity in Brassica rapa subsp. chinensis Makino     

Peirong Li  Tongbing Su  Huiping Wang  Xiuyun Zhao  Weihong Wang  Yangjun Yu  Deshuang Zhang  Changlong Wen  Shuancang Yu  Fenglan Zhang 《Plant Breeding》2019,138(3):309-324

Single‐nucleotide polymorphisms (SNPs) are rapid, economical and reliable genotyping tools. Non‐heading Chinese cabbage (Brassica rapa L. subsp. chinensis Makino) is now an economically important vegetable crop worldwide. In this study, 1,167 SNPs were evaluated for 7polymorphism among 70 representative non‐heading Chinese cabbage inbred lines using a Kompetitive Allele Specific PCR (KASP) genotyping assay. On the basis of identified polymorphisms and the results of a principal component analysis, we selected 50 core SNPs that were balanced sufficiently to provide adequate information for genetic identification. The core SNPs were used for construction of a neighbour‐joining dendrogram that separated the 70 inbred lines into four main groups and several subgroups corresponding to Caixin, Heiyebaicai, Huangxinwu, Naibaicai, Taitsai, Pak‐choi, and Wutatsai. This categorization was superior to that achieved using a dataset of 479 polymorphic SNPs. To confirm the utility of the core SNP markers in genetic identification, we tested their stability and resolution using 162 commercial hybrid cultivars. The SNPs, which represent a cost‐effective, accurate marker set for germplasm analysis and cultivar identification, are suitable for molecular marker‐assisted breeding in non‐heading Chinese cabbage.  相似文献   

鸡的微卫星DNA标记与胴体性状的相关分析   总被引：4，自引：1，他引：3  

陈晖  郑丽祯  陈岩锋  郑嫩珠  缪中纬  陈宽维  汤青萍 《中国家禽》2004,8(Z1):137-140

测定了青脚麻鸡与隐性白鸡杂交后自交的F2代个体的全净堂重、全净堂率、脂肪宽带、腹脂重和腹脂率6个胴体性状,检测10个微卫星DNA标记,对标记基因型胴体性状值进行方差分析和多重比较,结果表明:MCW328对于10周龄体重和全净膛重两性状各基因型均有显著的差异(P<0.05);ADL292对于腹脂重性状各基因型有极显著的差异(P<0.01),对于腹脂率有显著的差异(P<0.05)。MCW328标记基因与控制10周龄体重和全净膛重QTL连锁,ADL292标记基因与控制腹脂重和腹脂率QTL连锁。  相似文献   

Genetic diversity of herbicide-resistant and -susceptible Avena fatua populations in North Dakota and Minnesota     

L W MENGISTU  C G MESSERSMITH  & M J CHRISTOFFERS 《Weed Research》2005,45(6):413-423

Genetic diversity within and among 20 herbicide-resistant (HR) and 16 herbicide-susceptible (HS) Avena fatua multi-field populations was determined using 82 polymorphic loci resulting from two intersimple sequence repeat (ISSR) primers and one long-primer random amplified polymorphic DNA (LP-RAPD) primer. Collections from the Red River Valley of North Dakota and Minnesota, sampled in 1964 and 2000, represented A. fatua populations before and after intensive exposure to herbicides. A 1995 collection from south-west North Dakota represented A. fatua exposed to low herbicide selection. Despite differences in years of herbicide exposure among collections, both HR and HS populations from every collection maintained nearly similar levels of ISSR and RAPD diversity. Genetic differentiation among populations (G_ST) varied from 11% to 13% among HR populations and from 9% to 16% among HS populations, indicating that 84–91% of total variation remained within HS or within HR populations. Minimal difference in gene diversity between HR and HS is consistent with multiple origins of resistance, where HR A. fatua most likely evolved from diverse founding individuals.  相似文献   

分子标记用于赤眼蜂分子监测的研究   总被引：5，自引：0，他引：5  

李正西  沈佐锐 《农业生物技术学报》2001,9(1):65-68

通过对松毛虫赤眼蜂（Trichogramma dedrolimi Matsumura）以及玉米螟赤眼蜂（Trichogramma ostriniae Pang et Chen）rDNA-ITS2基因的克隆测序，并联机检索GenBank核酸序列库中其它赤眼蜂的相关序列，利用核酸分析软件找到松毛虫赤眼蜂和玉米螟赤眼蜂ITS2序列中一段有鉴别意义的标志，并据此设计出蜂种的特异PCR引物，实现了松毛虫赤眼蜂和玉米螟赤眼蜂rDNA特异区带的PCR扩增。研究表明该特异区带具有种的特异性。我们认为该分子标记技术可用于目前我国生防中常用的松毛虫赤眼蜂和玉米螟蒌眼蜂的分子监测，如田间种群动态监控和寄生效果评估。  相似文献   

微卫星DNA与延边黄牛肉用生长性状关系   总被引：12，自引：1，他引：11  

金海国  曹阳  臧延青  黄秉周  严昌国  崔允哉 《中国兽医学报》2004,24(3):285-288

选择与肉牛主要生产性状紧密连锁的 5个微卫星位点 ,以肉牛线性体型评分方法作为衡量肉牛生长发育的指标 ,运用最小二乘法拟合线性模型 ,分析了延边黄牛肉用生产性状与微卫星标记的关系 ,并分析了微卫星 ETH10和IDVGA4 6位点的不同基因型与肉用生产性状的关系。结果表明 ,微卫星 EHT10位点的等位基因 2 0 9对外貌线性评定中的背线性状、肌肉度线性评定中的肩部性状有正面影响 ,等位基因 2 13对外貌线性评定中的系部、肌肉度线性评定中的尻形状和细致度线性评定中的骨骼及皮肤性状有正面影响 ,而等位基因 2 15对外貌线性评定中的背线性状、肌肉度线性评定中的尻形状有负面影响 ;微卫星 IDVGA4 6位点的等位基因 2 4 9对背线、前肢和后肢等外貌线性评定性状 ,肌肉度线性评定的腰宽以及细致度线性评定的骨骼和皮肤性状有正面影响 ,等位基因 2 0 3对外貌评定性状的头部、背线 ,肌肉度评定性状腰宽以及细致度评定性状骨骼和皮肤有负面影响 ,等位基因 2 4 5对腰宽性状也起到负面影响  相似文献   

柑橘抗CTV转基因与分子标记研究进展   总被引：1，自引：0，他引：1  

徐小峰  周常勇 《果树学报》2005,22(4):372-375

综述了柑橘抗衰退病基因工程中两方面的研究进展。介绍多种来源于柑橘衰退病毒(Citrustriztezavirus,CTV)核酸序列的转基因柑橘和抗性种质资源中抗性基因的分子标记,以及所涉及的方法和遇到的问题。目前研究表明,虽然已成功实现对病毒衣壳蛋白(CP)等基因的转化和Ctv等抗性基因的标记,但尚未获得对CTV有高度抗性的转基因柑橘,而抗性基因亦不能实现定点克隆和转化。因此上述两方面研究还有待深入。  相似文献   

Orobanche species and population discrimination using intersimple sequence repeat (ISSR)     

H Benharrat  C Veronesi  C Theodet  P Thalouarn 《Weed Research》2002,42(6):470-475

Summary Orobanche species are commonly identified using morphological characteristics. In many cases, the distinction of closely related species is difficult, and a molecular tool is more suitable to differentiate them. In this study, genomic polymorphism between morphologically distinct species was investigated through amplification by polymerase chain reaction (PCR) of intersimple sequence repeat (ISSR) regions. Five primers were used to study genetic variation in the morphologically distinct species O. hederae and O. amethystea, as well as the closely related species O. cernua and O. cumana. For the first two species, all the primers detected genetic polymorphism. Anchored primers allowed the identification of more specific molecular markers than non‐anchored tri‐ and tetranucleotide primers. Genetic polymorphism was investigated among three O. hederae populations using the two types of primer. One non‐anchored and two anchored primers detected intraspecific variation, which was not correlated with the geographical location of those populations. The primer (GATA)₄ detected polymorphism between five specimens each of O. cernua and O. cumana species collected from different countries, permitting these two closely related species to be clearly differentiated. This study demonstrated that ISSR markers can be highly reliable for precise identification of Orobanche species.  相似文献   

Variation in the response of melon genotypes to Fusarium oxysporum f.sp. melonis race 1 determined by inoculation tests and molecular markers     

Y. Burger  N. Katzir  G. Tzuri  V. Portnoy  U. Saar  S. Shriber  R. Perl-Treves  R. Cohen † 《Plant pathology》2003,52(2):204-211

Screening of genotypes of melon ( Cucumis melo ) for resistance to wilt caused by Fusarium oxysporum f.sp. melonis is often characterized by wide variability in their responses to inoculation, even under carefully controlled conditions. The variability at the seedling stage of 17 genotypes susceptible to race 1 was examined in growth-chamber experiments. Disease incidence varied from 0 to 100% in a genotype-dependent manner. Using four combinations of light (60 and 90  µ E m⁻² s⁻¹) and temperatures of (27 and 31°C), only light intensity showed a statistically significant effect. Marker-assisted selection for fusarium resistance breeding using cleaved amplified polymorphic sequence (CAPS) and sequence-characterized amplified region (SCAR) markers were compared using a single set of genotypes that included 24 melon accessions and breeding lines whose genotype regarding the Fom-2 gene was well characterized. The practical value of the markers for discriminating a range of genotypes and clarifying the scoring of phenotypes was also tested using a segregating breeding population which showed codominant SCAR markers to be useful in marker-assisted selection.  相似文献   

苹果基因组分子生物学研究进展   总被引：4，自引：1，他引：4  

姚玉新  翟衡 《果树学报》2004,21(6):586-591

借助分子标记手段苹果基因组研究取得了巨大成就,涉及苹果的抗性、生长发育、果实品质等各方面基因,综合有关文献,将其研究的主要成果,包括已发现的主要苹果遗传标记、已构建的苹果遗传连锁图谱及苹果的转基因研究进行综述,并对目前苹果基因组研究存在的问题和对策进行了分析,以期为苹果遗传育种提供参考。  相似文献