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主要棉花种质在形态学特征和ＲＡＰＤ标记上的遗传多样性②我们用两个主要的棉花栽培种陆地棉和海岛棉进行了有限的种间渐渗育种，然后，分别用ＲＡＰＤ标记从ＤＮＡ水平，用遗传率高且稳定的形态学特征从表型水平，对得到的１６个近纯合的种间杂交系进行了多样性分析。用... 相似文献
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“中国春”小麦不同缺体的RAPD分析 总被引：1，自引：0，他引：1
利用ＰＣＲ－９０Ａ型ＤＮＡ扩增仪，对７个中国春小麦缺体核ＤＮＡ进行了扩增。结果显示，中国春小麦缺体核ＤＮＡ的ＲＡＰＤ产物可分为二种类型：（１）所有供试材料均能扩增出的片段，代表了核ＤＮＡ在进化上的保守性。有一个引物检测到这类片段。（２）全部供试材料间存在的差异片段，这类片段是用来基因定位的主要依据。另外，建立中国春小麦缺体系统的ＲＡＰＤ指纹图谱对研究小麦的系统进化也将有重要意义 相似文献
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改良CTAB法快速提取棉花DNA 总被引：74，自引：43，他引：74
针对棉花（Ｇｏｓｓｐｉｕｍ）富含棉酚、多糖等其它次生干扰物质这一特点而设计的一种快速分离和纯化棉花总ＤＮＡ（ｇＤＮＡ）的方法。该方法是对ＣＴＡＢ（ｃｅｔｙｌｔｒｉｍｅｔｈｙｌａｍｍｏｎｉｕｍｂｒｏｍｉｄｅ）法的改良，它在ＣＴＡＢ法的基础上，首先用ＤＩＥＣＡ（ｄｉｅｔｈｙｌｄｉｔｈｉｏｃａｒｂａｍｉｃａｃｉｄ）抑制酚氧化酶的活性，然后用活性炭和ＰＶＰ４０（ｐｏｌｙｖｉｎｙｌｐｙｒｒｏｌｉｄｏｎｅ）排除棉酚等其它次生干扰物质。试验证明，该方法比较简便、快速、经济、有效，得到的ｇＤＮＡ完全可以满足ＰＣＲ等分子生物学分析。 相似文献
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用RAPD方法鉴定杂交辣椒种子纯度 总被引：6，自引：0，他引：6
本文用ＲＡＰＤ（随机扩增多态性ＤＮＡ）方法对辣椒种子的父本、母本及杂种一代进行分析研究,筛选出适合辣椒杂种一代种子纯度鉴定的随机引物Ｓ５,并对并进了种子纯度的初步鉴定。ＲＡＰＤ结果和田间鉴定的相比,在误差允许范围内,进一步证明了ＲＡＰＤ方法在蔬菜种子纯度鉴定中应用的可行性。 相似文献
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玉米RAPD程序优化研究及其初步探讨 总被引：32，自引：1，他引：32
西方琉黄早四、汶黄、Ｍｏ１７等我国主要玉米自交系为使用国产ＰＣＲ仪，对玉米ＲＡＰＤ分析中ＰＣＲ过程的ＤＮＡ模板浓度，扩以应的循环数进行了研究，并针对国产ＰＣＲ扩增仪（ＰＣＲ－９０ＡＤ）的特点，确定了ＤＮＡ变性，引物和模板ＤＮＡ结合，引物洞模板延伸３个步骤的时间，实验结果表明玉米ＰＣＲ过程使用１０－１５ｎｇ的模板ＤＮＡ，在９４℃模板变性时间１分３０秒，３６℃引物退火２分钟，７２℃引物延伸２分钟３０秒 相似文献
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DNA分子标记技术在品种鉴定和纯度分析上的应用 总被引：21，自引：2，他引：19
本文就ＲＦＬＰ、ＲＡＰＤ、ＳＳＲ、ＡＦＬＰ、等４种ＤＮＡ分子标记技术的原理及其在作物品种鉴定和纯度分析上的应用作了概述，并对它们的应用前景作了探讨。 相似文献
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Somatic hybridization can be used to induce genetic variability in plastidial and mitochondrial genomes, and transfer multiple
uncloned genes across sexual barriers. Somatic hybrids were produced between a dihaploid clone of the common potato, S. tuberosum subsp. tuberosum, and the wild sexually incongruent diploid species S. commersonii. Fusion products were selected on the basis of callus growth and regeneration in vitro. Genome composition of putative somatic hybrids was determined by flow cytometric analysis of nuclear DNA content, RAPD analysis,
and Southern analysis with probes specific to organellar DNA. All regenerated fusion products proved to be hybrids based on
RAPD analysis. Seventy per cent of somatic hybrids were (near) tetraploids, 22% (near) hexaploids and 8%(near) octoploids.
A high correlation was found between the nuclear DNA content determined by flow cytometry and the number of chloroplasts in
stomata guard cell pairs. Somatic hybrids inherited the parental plastids in a random manner. On the contrary, they preferentially
inherited the mitochondrial DNA fragments of S. tuberosum. The majority of them had a rearranged mitochondrial genome with fragments from both parents. Hybrids were highly vigorous
and morphologically more similar to the cultivated than to the wild parent, produced tubers on long stolons under long photoperiod
conditions, showed a high degree of flowering, but did not produce pollen. In addition, somatic hybrids were generally more
resistant to frost and Verticillium wilt than the cultivated parent, indicating the introgression of relevant resistance genes
from the wild species into the genetic background of S. tuberosum.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
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Metin Tuna Deepak K. Khadka Madan K. Shrestha K. Arumuganathan Avi Golan-Goldhirsh 《Euphytica》2004,135(1):39-46
Determining the ploidy and geneticdiversity of a germplasm is necessarybefore initiating breeding or geneticstudies. This study was conducted tocharacterize the ploidy level of 57 naturalpopulations of orchardgrass (Dactylisglomerata L.) collected from the ranges ofThrace region of Turkey and the diversityamong populations based on RAPD (RandomAmplified Polymorphic DNA) markers. Flowcytometry was used to determine nuclear DNAcontent (pg 2C-1 = DNA content of adiploid somatic nucleus) of 6 plants foreach population. Nuclear DNA contents werecorrelated to ploidy level with root tipchromosome counts on selected plants. Onthe basis of this study, mean nuclear DNAcontent of orchardgrass was determined as9.57 ± 0.33 (with 95% confidenceinterval) while all the plants used inchromosome counting were determined to betetraploid, with 2n = 28 mitoticchromosomes, suggesting that diploidorchardgrass plants are likely very rare orabsent in ranges of Thrace region ofTurkey. In the RAPD assay, over 40polymorphic fragments were generated whichallowed some populations to bedistinguished from the rest by uniquemarkers. A cluster analysis was performedusing Nei's (1972) genetic distance indexwith an unweighted pair group method witharitmetic mean (UPGMA). The clusteranalysis indicated that there is a highlevel of gene flow among naturalorchardgrass populations and thereforegenes distributed quite homogeneouslythrough out the region. The results of thisstudy can be useful in the development ofDactylis germ plasm collectionstrategies in Thrace region for breedingpurpose. 相似文献
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种质是利用和改良动植物、微生物的物质基础,更是实施各个育种途径的原材料,因此优良种质鉴定是一个很关键的问题。种质鉴定方法已由形态水平发展到蛋白质和DNA分子水平。RAPD技术具有敏感、快速、简便、产量高、重复性好及检测容易等突出优点,广泛应用于种质鉴定中,但也有不足之处。基于RAPD标记技术建立起来的SCAR（Sequence Characterized Amplified Region）标记克服了RAPD标记的不足,操作简捷,特异性和重复性较高,是一种有效的分子标记。RAPD-SCAR分子标记技术的利用使种质鉴定更为快速准确,提高了育种速度,缩短了育种周期,为相关的研究工作提供分子遗传学依据。 相似文献
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DNA polymorphism among nine cultivars of Asparagus officinalis L. was measured using random amplified polymorphic DNA (RAPD). Of 69 reproducible amplification products from 12 arbitrary decamer primers, 49 RAPD markers were polymorphic and could be used to distinguish six German and three Dutch asparagus cultivars. Even with very small sample sizes, genetic similarity measurements based on the RAPD data allowed accurate grouping of the nine cultivars into distinct clusters, with the exception of two individuals which clustered to closely related varieties. Two German cultivars showed high genetic similarity and were distinct from the remaining German varieties. The German and Dutch cultivars were clearly separated by a relatively large genetic distance. 相似文献
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黄花矶松基因组DNA的提取以及RAPD反应体系的优化 总被引：1，自引：0，他引：1
RAPD-PCR是从核酸水平鉴定DNA变异的一种快速、有效的方法,在植物的分子水平检测中得到广泛的应用。本文分别采用SDS法和CTAB法对黄花矶松基因组DNA进行了提取,筛选了适合黄花矶松的基因组DNA提取方法,研究了RAPD反应体系中不同Mg2+浓度,不同的退火温度等条件对PCR扩增的影响,建立了适用于黄花矶松基因组分析的一种简便、有效的RAPD方法。 相似文献
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Heterogeneity of random amplified polymorphic DNA sequences in individual seedlings and bulked samples of four cultivars of Brassica napus 总被引：1，自引：0，他引：1
Using four different random amplified polymorphic DNA (RAPD) primers, a qualitative and quantitative assessment was made of the level of DNA sequence heterogeneity present in the seedlings of four representative Australian rapeseed cultivars. It was found that, depending upon the primer/cultivar combination, the seedlings diverged from total homogeneity to almost complete heterogeneity. The increase or decrease of sample-specific RAPD sequences was evaluated in proportional mixtures of DNA from individual seedlings. These results were then compared with those obtained from bulked DNA samples containing DNA from all the seedlings of a cultivar. From these comparisons, it was found that for a specific RAPD to be detectable in a bulked sample, the particular polymorphism had to be present in at least 15% of the individual seedlings. Even so, the bulked samples produced cultivar-specific RAPD banding patterns with all four primers, showing that any of these primers could be used to identify the different rapeseed cultivars. In contrast to the cultivars ‘Oscar’, ‘Dunkeld’ and ‘Narendra’, the cultivar ‘Rainbow’ was found to be highly heterogeneous—as shown by a diversity of RAPD combinations rather than the presence of differing length RAPDs—and it is suggested that this heterogeneity may be related to the improved tolerance of this cultivar to blackleg infection. 相似文献
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冬虫夏草RAPD反应体系的建立及优化 总被引：1，自引：0，他引：1
为建立并优化冬虫夏草RAPD反应体系,通过改变RAPD反应体系中Mg2+、dNTP、引物、模板等主要成分的浓度,结合RAPD扩增效果,建立冬虫夏草RAPD反应体系,然后通过改变主要热循环参数,优化冬虫夏草RAPD反应体系。结果表明：适合冬虫夏草的RAPD反应体系为25 μL体系中内含1×PCR缓冲液、1.5 μmol/μL Mg2+、320 μmol/L dNTP、24 ng引物、20 ng模板、1 U Taq酶;扩增程序的优化结果为：95℃预变性5 min,然后35个循环（94℃变性45 s,36℃复性1 min,72℃延伸2 min）,循环结束后72℃延伸7 min。综上,RAPD技术可用于冬虫夏草的鉴定、评价分析。 相似文献
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RAPD and SCAR markers linked to the sex expression locus M in asparagus 总被引：13，自引：0，他引：13
Bulk segregant analysis (BSA), random amplified polymorphic DNA (RAPD) and sequence characterized amplified region (SCAR)
methods were used to map molecular markers to the sex locus M of asparagus. Two parents, A19 (male, Mm) and MW25 (female,
mm), and 63 progeny were used for the study. Two DNA bulks, one male and one female, were made by pooling equal amounts of
DNA from 10 randomly selected progeny of each sex type. A total of 760 arbitrary decamer oligonucleotide primers were used
for RAPD analysis. Primer OPC15 produced two RAPD markers, OPC15-98 and OPC15-30, both of which were linked to the M locus
at a distance of 1.6 cM. Subsequently, amplified RAPD fragment OPC15-98 was cloned and sequenced. The sequence was then used
to design flanking 24-mer oligonucleotide SCAR primers SCC15-1 and SCC15-2. Both of these SCAR primers amplified a single
980 bp fragment; the same size as the cloned RAPD fragment. However, the SCAR marker was dominant as was the original OPC15-98
band from which it was derived. These RAPD and SCAR markers could be used for scoring male and female progeny in the mapping
population, but were not found to be applicable to other asparagus germplasm studied.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献