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黄瓜霜霉病抗病基因的RAPD及SCAR标记 总被引:3,自引:0,他引:3
以感霜霉病黄瓜L18-10-2和抗霜霉病黄瓜129为亲本构建F2代分离群体,以F3代植株霜霉病抗性鉴定表示F2代各单株抗病性并得以区分各单株杂合或纯合感病性,采用RAPD技术和转SCAR的方法筛选黄瓜抗霜霉病基因分子标记.结果显示,在318条RAPD引物中有18条引物表现出两亲本间多态性,其中引物P18的SB-SP18561扩增片段与霜霉病抗病基因之间紧密连锁,根据交换率和Kosambi函数公式计算其遗传距离为7.85 cM.回收SBSP18561片段并克隆和测序,其准确长度为561 bp.将该RAPD标记转换为SCAR标记,长度为494 bp,命名为SSBSP18494. 相似文献
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二十八份玉米自交系的RAPD亲缘关系分析 总被引:4,自引:0,他引:4
采用RAPD技术,对28份玉米自交系的亲缘关系进行分析。旨在DNA水平上揭示玉米自交系之间的亲缘关系,为进一步提高玉米杂种优势利用水平提供有益的信息从100个10bp随机引物中筛选出24个多态性较好的引物,对28份玉米自交系DNA进行扩增,扩增出24张DNA指纹图谱,其中多态性DNA谱带106条,占总扩增带数的64%。利用DNA扩增结果进行聚类分析,建立了28个玉米自交系的亲缘天系树状图,将供试材料划分为五个类群,RAPD分析结果与已知系谱的亲缘关系基本一致。 相似文献
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鸢尾属部分植物种质资源的RAPD分析 总被引:6,自引:0,他引:6
采用RAPD分子标记技术,从100个随机引物中筛选出多态性强、重复性好且稳定性高的引物18个,对38份野生鸢尾属材料进行扩增,共扩增出409条带,其中多态性带405条,多态性比率为99.0%,表明野生鸢尾属植物种间有丰富的遗传多态性;根据DNA谱带计算物种间遗传距离,聚类分析结果将鸢尾属38份材料划分为6组,其结果与传统生物学特性划分的6个亚属的分类结果基本一致;物种特有RAPD标记分析表明,利用18个引物可以较好地将鸢尾属38种植物区分开,其中9个材料得到了单一标记的扩增带,表明运用RAPD分子标记对研究鸢尾属植物特异性基因及标记的筛选等有一定的理论和实际应用价值。 相似文献
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利用RAPD和ISSR分子标记分析地黄种质遗传多样性 总被引:8,自引:0,他引:8
用RAPD与ISSR技术对地黄的8个品种和2个脱毒品系进行了种质遗传多样性分析.分别从80条RAPD引物和44条ISSR引物中筛选出适合地黄种质分析的17条RAPD引物和10条ISSR引物用于RAPD和ISSR分析.17条RAPD引物共扩增出177条带, 多态性位点数为109; 多态性位点比率为61.58%;平均多样性指数(I)为0.3135;每个位点的有效等位基因数(Ne)是1.3641; 10条ISSR引物共扩增出110条带. 多态性位点数为79; 多态性位点比率为71.58%;平均多样性指数(I)为0.3577;每个位点的有效等位基因数(Ne)是1.4037. 基于扩增条带数据库建立了各自的Jaccard遗传相关系数矩阵,构建了相似的分子树状图,将10个供试材料分为2类:一类群含组培85.5、大田85.5、组培9302、大田9302、金状元和金白6个材料;另一类群含北京1号、大红袍、地黄9104和野生地黄4个材料.两种分子标记的分析结果呈极显著正相关(r=0.649).结果表明,RAPD与ISSR标记适合于地黄种质遗传多样性分析,ISSR标记技术是一种多态性和重复性优于RAPD技术的实用技术. 相似文献
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利用RAPD与ISSR分子标记检测手段,分析了哈茨木霉T2-16肽类代谢产物处理豇豆土著根瘤菌,对其遗传性状的影响,同时,比较了RAPD和ISSR两种不同分子标记在检测根瘤菌种间的遗传相似性以及遗传变异性的分辨力.实验中,从100条引物中筛选到具有多态性的ISSR引物5条,从80条引物中筛选到具有多态性的RAPD引物6条,用5条ISSR引物扩增出54条带,多态性条带比率为75.93 %;6条RAPD引物扩增出61条带,多态性条带比率为68.85 %.两种分子标记均能揭示出处理前后根瘤菌间的遗传差异,但ISSR标记比RAPD标记可检测到更大的遗传变异.根据两种标记的结果,对供试的根瘤菌进行聚类分析,结果表明,土著根瘤菌经木霉肽类代谢产物处理后,与出发菌株相比,表现出一定程度的遗传分化和遗传差异性. 相似文献
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In the present study, DNA fingerprinting of eight strains of Flavobacterium columnare was done by random amplification of polymorphic DNA (RAPD) fingerprinting method. The strains were collected from Fish Health Management Division, Central Institute of Freshwater Aquaculture, Bhubaneswar, India. A total number of 160 primers were screened for RAPD-PCR, of which 10 primers yielded amplification with all the strains. The molecular weight of amplified bands varied from 0.29–2.63 Kb. The number of bands varied from 1 to 8. Unique band was seen with primer OPY-15 with molecular weight 0.75 Kb that can be used for epidemiological study. Genetic variability was investigated using NTSYS software. Highest genetic similarity was found between MS1 and MS3 followed by MS5 and MS7. Minimum genetic similarity was found between MS2 and MS8. Phylogenetic tree was constructed using UPGMA and neighbor joining methods. 相似文献
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Dynamics of Fungal Communities in Bulk and Maize Rhizosphere Soil in the Tropics 总被引:9,自引:1,他引:8 下载免费PDF全文
Newton C. Marcial Gomes Olajire Fagbola Rodrigo Costa Norma Gouvea Rumjanek Arno Buchner Leda Mendona-Hagler Kornelia Smalla 《Applied microbiology》2003,69(7):3758-3766
The fungal population dynamics in soil and in the rhizospheres of two maize cultivars grown in tropical soils were studied by a cultivation-independent analysis of directly extracted DNA to provide baseline data. Soil and rhizosphere samples were taken from six plots 20, 40, and 90 days after planting in two consecutive years. A 1.65-kb fragment of the 18S ribosomal DNA (rDNA) amplified from the total community DNA was analyzed by denaturing gradient gel electrophoresis (DGGE) and by cloning and sequencing. A rhizosphere effect was observed for fungal populations at all stages of plant development. In addition, pronounced changes in the composition of fungal communities during plant growth development were found by DGGE. Similar types of fingerprints were observed in two consecutive growth periods. No major differences were detected in the fungal patterns of the two cultivars. Direct cloning of 18S rDNA fragments amplified from soil or rhizosphere DNA resulted in 75 clones matching 12 dominant DGGE bands. The clones were characterized by their HinfI restriction patterns, and 39 different clones representing each group of restriction patterns were sequenced. The cloning and sequencing approach provided information on the phylogeny of dominant amplifiable fungal populations and allowed us to determine a number of fungal phylotypes that contribute to each of the dominant DGGE bands. Based on the sequence similarity of the 18S rDNA fragment with existing fungal isolates in the database, it was shown that the rhizospheres of young maize plants seemed to select the Ascomycetes order Pleosporales, while different members of the Ascomycetes and basidiomycetic yeast were detected in the rhizospheres of senescent maize plants. 相似文献
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Gomes NC Fagbola O Costa R Rumjanek NG Buchner A Mendona-Hagler L Smalla K 《Applied and environmental microbiology》2003,69(7):3758-3766
The fungal population dynamics in soil and in the rhizospheres of two maize cultivars grown in tropical soils were studied by a cultivation-independent analysis of directly extracted DNA to provide baseline data. Soil and rhizosphere samples were taken from six plots 20, 40, and 90 days after planting in two consecutive years. A 1.65-kb fragment of the 18S ribosomal DNA (rDNA) amplified from the total community DNA was analyzed by denaturing gradient gel electrophoresis (DGGE) and by cloning and sequencing. A rhizosphere effect was observed for fungal populations at all stages of plant development. In addition, pronounced changes in the composition of fungal communities during plant growth development were found by DGGE. Similar types of fingerprints were observed in two consecutive growth periods. No major differences were detected in the fungal patterns of the two cultivars. Direct cloning of 18S rDNA fragments amplified from soil or rhizosphere DNA resulted in 75 clones matching 12 dominant DGGE bands. The clones were characterized by their HinfI restriction patterns, and 39 different clones representing each group of restriction patterns were sequenced. The cloning and sequencing approach provided information on the phylogeny of dominant amplifiable fungal populations and allowed us to determine a number of fungal phylotypes that contribute to each of the dominant DGGE bands. Based on the sequence similarity of the 18S rDNA fragment with existing fungal isolates in the database, it was shown that the rhizospheres of young maize plants seemed to select the Ascomycetes order Pleosporales, while different members of the Ascomycetes and basidiomycetic yeast were detected in the rhizospheres of senescent maize plants. 相似文献
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Screening of molecular markers for NPV resistance in Bombyx mori L. (Lep., Bombycidae) 总被引:3,自引:0,他引:3
Q. Yao M. W. Li Y. Wang W. B. Wang J. Lu Y. Dong K. P. Chen 《Journal of Applied Entomology》2003,127(3):134-136
Abstract: Silkworm ( Bombyx mori L.) is one of the important economic insects. Silkworm rearing and silk industry plays an important role in China, India and other developing countries. In the long history of sericultural practice, introduction of silkworm strains with high resistance to diseases has greatly improved cocoon and silk quality and productivity. However, current silkworm breeding is mainly based on traditional method that involves high input of time and labour. In order to increase the selection efficiency and accuracy for future silkworm breeding, it is necessary to establish a molecular marker-assisted selection system. In our study, three silkworm near isogenic lines that had different resistance to nuclear polyhedrosis virus (NPV) were established by means of different hybridization methods. A total of 150 random amplified polymorphic DNA (RAPD) random primers were used to screen molecular markers. Among them, two molecular markers OPA-18700 and OPY-11400 were found linked to major genes resistant and susceptible to NPV, respectively. Validity of the molecular markers was proved in F2 populations. 相似文献
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离子束介导大豆DNA转化小麦后代高蛋白株的RAPD标记分析 总被引:9,自引:0,他引:9
利用离子束介导法将大豆DNA导入小麦,经过连续4代田间筛选和蛋白含量测定,获得高蛋白变异株系.采用RAPD分析技术,用34条随机引物对供体大豆、受体小麦和3个高蛋白小麦变异株的基因组DNA进行扩增.有29个引物扩增出清晰稳定的条带,其中18个引物扩增出的条带有不同程度的差异.高蛋白小麦突变株与受体小麦(对照)相比出现了条带的增加、缺失、扩增带深浅等变化,也出现了与受体小麦不同而与供体大豆相同的扩增带.实验结果表明,外源大豆DNA导入受体小麦可以引起后代基因组DNA序列变化,扩大小麦遗传基础. 相似文献
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Yang DE Zhang CL Zhang DS Jin DM Weng ML Chen SJ Nguyen H Wang B 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》2004,108(4):706-711
One single pathogen Fusarium graminearum Schw. was inoculated to maize inbred lines 1,145 (Resistant) and Y331 (Susceptive), and their progenies of F1, F2 and BC1F1 populations. Field statistical data revealed that all of the F1 individuals were resistant to the disease and that the ratio of resistant plants to susceptive plants was 3:1 in the F2 population, and 1:1 in the BC1F1 population. The results revealed that a single dominant gene controls the resistance to F. graminearum Schw.. The resistant gene to F. graminearum Schw. was denominated as Rfg1 according to the standard principle of the nomenclature of the plant disease resistant genes. RAPD (randomly amplified polymorphic DNA) combined with BSA (bulked segregant analysis) analysis was carried out in the developed F2 and BC1F1 populations, respectively. Three RAPD products screened from the RAPD analysis with 820 Operon 10-mer primers showed the linkage relation with the resistant gene Rfg1. The three RAPD amplification products (OPD-201000, OPA-041100 and OPY-04900) were cloned and their copy numbers were determined. The results indicated that only OPY-04900 was a single-copy sequence. Then, OPY-04900 was used as a probe to map the Rfg1 gene with a RIL F7 mapping population provided by Henry Nguyen, which was developed from the cross S3×Mo17. Rfg1 was primarily mapped on chromosome 6 between the two linked markers OPY-04900 and umc21 (Bin 6.04–6.05). In order to confirm the primary mapping result, 25 SSR (simple sequence repeat) markers and six RFLP (restriction fragment length polymorphism) markers in the Rfg1 gene-encompassing region were selected, and their linkage relation with Rfg1 was analyzed in our F2 population. Results indicated that SSR marker mmc0241 and RFLP marker bnl3.03 are flanking the Rfg1 gene with a genetic distance of 3.0 cM and 2.0 cM, respectively. This is the first time to name and to map a single resistant gene of maize stalk rot through a single pathogen inoculation and molecular marker analysis.Communicated by H.F. Linskens 相似文献
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D. Ladhalakshmi A. Vijayasamundeeswari V. Paranidharan R. Samiyappan R. Velazhahan 《World journal of microbiology & biotechnology》2009,25(12):2129-2135
The fungus Peronosclerospora sorghi [Weston and Uppal (Shaw)] infects both sorghum and maize and incites downy mildew disease. Pathogenic and molecular variability
among isolates of P. sorghi from sorghum and maize has been reported. In the present study we developed a DNA sequence characterized amplified region
(SCAR) marker for identification of isolates of P. sorghi from maize by using polymerase chain reaction (PCR). The random amplified polymorphic DNA (RAPD) primer OPB15 consistently
amplified a 1,000 base pairs (bp) product in PCR only from DNA of P. sorghi isolates from maize and not from isolates of sorghum. The PCR-amplified 1,000-bp product was cloned and sequenced. The sequence
of the SCAR marker was used for designing specific primers for identification of maize isolates of P. sorghi. The SCAR primers amplified a 800 bp fragment only from genomic DNA of maize isolates of P. sorghi. The SCAR primers developed in this study are highly specific and reproducible, and proved to be powerful tool for identification
of P. sorghi isolates from maize. 相似文献