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Jianmin Qi Hui Chen Aifen Tao Jiantang Xu Lihui Lin Pingping Fan 《Plant Breeding》2014,133(6):777-781
White jute (Corchorus capsularis) and dark jute (Corchorus olitorius) are two important cultivated crops that are used for natural fibre production. Some genetic maps have been developed for dark jute, but the genetic map information for white jute (C. capsularis) is limited. In this study, a linkage map comprising 44 sequence‐related amplified polymorphisms (SRAPs), 57 intersimple sequence repeats (ISSRs) and 18 randomly amplified polymorphic DNA (RAPD) covering 2185.7 cM with a mean density of 18.7 cM per locus was constructed in an F2 population consisting of 185 individuals derived from a cross between two diverse genotypes of ‘Xinxuan No. 1’ and ‘Qiongyueqing’ in white jute. These markers were evenly distributed in the linkage groups without any clustering. This genetic linkage map construction will facilitate the mapping of agronomic traits and marker‐assisted selection breeding in white jute. 相似文献
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本研究主要对杨树RAPD体系进行优化,以银白杨为试材,利用L16(45)正交试验设计和2个单因素试验,研究各主要参数的适宜浓度。结果表明,杨树RAPD优化后的反应体系:20μL反应体系中,含Mg2+ 1.5 mmol/L、Taq酶2.0 U、引物0.2μmol/L、dNTPs 0.2 mmol/L、模板DNA 50 ng。扩增程序为:94℃预变性3 min、94℃变性40 s、38℃退火60 s、72℃延伸90 s,共38个循环;最后在72℃延伸7 min后4℃保温。通过1.5%的琼脂糖凝胶电泳检测其扩增产物,证明该体系稳定可靠,可以用于杨树的遗传学分析。 相似文献
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In marigold, an F2 segregation population of 167 plants was constructed from a cross of a line (M525A) carrying the male sterility trait × an inbred line (f53f). In line M525A, the male sterility trait was controlled by the recessive gene, Tems . The intersimple sequence repeat (ISSR) and sequence-related amplified polymorphism (SRAP) techniques combined with bulked segregant analysis were used to develop markers linked to the trait. From a survey of the 38 ISSR primers and 170 SRAP primer combinations, only one SRAP marker that was closely linked to the target trait was identified and successfully converted into sequence characterized amplified region (SCAR) marker that was located within 2.4 cM from Tems locus. The marker was validated with five other two-type lines and in each case the male fertile plants were reliably identified. This SCAR marker therefore permits the efficient marker-assisted selection of male sterile individuals in breeding programmes of marigold and will greatly facilitate the breeding of F1 cultivars. 相似文献
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目标区域扩增多态性(TRAP):一种新的植物基因型标记技术 总被引:9,自引:0,他引:9
TRAP是一种新的基于PCR的植物基因型标记技术,具有简单、稳定、效率高的特点。借助日益增长的庞大的生物序列信息,TRAP利用生物信息工具和EST数据库信息,产生目标候选基因区多态性标记。TRAP技术采用两个18核苷酸引物产生标记。一个为固定引物,依据EST序列设计;另一个为随机引物,针对外显子和内含子的特点,设计为分别富含GC或AT核心区的任意序列。PCR扩增前5个循环采用35℃的退火温度,后35个循环采用50℃的退火温度。对不同的植物种类,每一个PCR反应可产生多达50个可统计DNA片段。本文在阐述了TRAP的原理与流程后,对该技术的优势和应用情况进行了总结,并对其前景做了展望。 相似文献
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Promoter anchored amplified polymorphism based on random amplified polymorphic DNA (PAAP-RAPD) in cotton 总被引:1,自引:0,他引:1
Non-coding sequences account for a majority of the higher plant genome, some of which have important effects in gene regulation
and plant development. In an effort to develop molecular marker systems to search for polymorphisms associated with high fiber
yield and quality in cotton, we have developed a methodology that could specifically target the regulatory regions of the
cotton genome. In this study we designed 10-nucleotide degenerate promoter primers based on conserved core promoter sequences
and tested their applicability in PCR amplifications in combination with 10-mer random amplified polymorphic DNA (RAPD) primers.
The amplified markers are called promoter anchored amplified polymorphism based on RAPD (PAAP-RAPD). Forty cotton genotypes
with diverse genetic and geographical backgrounds were used to test the PAAP-RAPD system using polyacrylamide gel electrophoresis.
Based on PAAP-RAPD markers amplified from 12 primer combinations, the 40 genotypes were classified into five distinctive groups:
two Upland cotton (Gossypium hirsutum) groups from China, another two Upland cotton groups from the USA, and one group from American Pima cotton (G. barbadense). The groupings are in general consistent with their genetic and geographical origins. Thirty-six PAAP-RAPD and RAPD fragments
were cloned and four of them were further subjected to sequence analysis. Signal scanning using software PLACE confirmed that
they contained an array of cis-regulatory sequences in addition to the core promoter sequences. The results demonstrate the
potential application of PAAP-RAPD as a new marker system specifically targeting regulatory regions of the plant genome. 相似文献
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Identification of polymorphic DNA markers in cultivated peanut (Arachis hypogaea L.) 总被引:17,自引:0,他引:17
The detection of DNA polymorphism in cultivated peanut (Arachis hypogaea L.) is reported here for the first time. The DNA
amplification fingerprinting (DAF) and amplified fragment length polymorphism (AFLP) approaches were tested for their potential
to detect genetic variation in peanut. The AFLP approach was more efficient as 43% of the primer combinations detected polymorphic
DNA markers in contrast to 3% with the DAF approach. However, the number of polymorphic bands identified using primers selected
in both approaches was comparable. In the DAF study, when 559 primers of varying types were screened, 17 (mostly 10-mer types)
detected polymorphism producing an average of 3.7 polymorphic bands per primer with a total of 63 polymorphic markers. In
the AFLP study, when 64 primer combinations (three selective nucleotides) corresponding to restriction enzymes Eco RI and
Mse I were screened, 28 detected polymorphism. On an average, 6.7% of bands obtained from these 28 primer pairs were polymorphic
resulting in a total of 111 AFLP markers. Our results demonstrate that both AFLP and DAF approaches can be employed to generate
DNA markers in peanut and thus have potential in the marker-assisted genetic improvement and germplasm evaluation of this
economically important crop.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
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绿色棉纤维发育过程中DNA表观遗传变化的甲基化敏感扩增多态性分析 总被引:1,自引:1,他引:0
【目的】分析绿色棉纤维发育过程中DNA甲基化状态差异。【方法】利用2组限制性内切酶Eco RⅠ/HpaⅡ和Eco RⅠ/MspⅠ对绿絮棉1号纤维基因组甲基化位点进行识别和切割,并采用甲基化敏感扩增多态性引物对切割产物进行扩增,分析基因组内5'-CCGG-3'位点的甲基化状态;同时对扩增的差异片段进行测序,分析其生物学功能。【结果】66对甲基化敏感扩增多态性引物共扩增出4112个条带;平均每个样品共扩增出822.4个带型,平均每对引物扩增出12.46个片段。随着绿色棉纤维的发育,甲基化条带总数、甲基化条带比率、全甲基化条带百分比均逐渐升高;其中,开花后10、15、20和25 d时甲基化条带总数分别比开花后5 d时增加11.45%,13.86%,20.10%和33.13%。与开花后5 d相比,开花后10、15、20和25 d时纤维DNA发生甲基化变化位点的比例分别为3.15%,3.43%,3.65%和2.52%;而去甲基化位点的比例分别为12.87%,14.72%,13.31%和48.81%。通过测序和Blast分析表明,17个片段与已知的功能基因同源性较高,包括棉花线粒体基因组、丝氨酸蛋白酶基因和酯酶基因,且这些基因均在开花后25 d发生去甲基化。【结论】绿色棉纤维发育过程中DNA发生了甲基化与去甲基化现象。 相似文献
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New molecular markers derived from expressed sequence tag (EST) sequences were mapped on linkage maps of Italian ryegrass by a two-way pseudo-testcross strategy. cDNA sequences were obtained from various tissues of Italian ryegrass ( Lolium multiflorum ) and converted into cleaved amplified polymorphic sequence (CAPS) markers. Of 260 EST primer pairs that amplified a single band, 74 generated bands that showed clear polymorphisms among individuals of an F1 mapping family. Of the 74 polymorphic marker loci, 69 were mapped on an Italian ryegrass linkage map previously constructed using amplified fragment length polymorphism (AFLP), restriction fragment length polymorphism (RFLP), and simple sequence repeat (SSR) markers. The newly-developed EST-CAPS markers would be useful as an efficient tool to identify genetic markers and to identify candidate genes for quantitative trait loci (QTLs) associated with important traits in Italian ryegrass. 相似文献
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Rapid and convenient gel‐free screening of SCAR markers in wheat using SYBR green‐based melt‐profiling 下载免费PDF全文
Gautam Vishwakarma Ajay Saini Bikram Kishore Das Suresh Gopal Bhagwat Narendra Jawali 《Plant Breeding》2016,135(6):643-653
Sequence characterized amplified region (SCAR) markers that are highly desirable in crop breeding for marker‐assisted selection (MAS) are routinely analysed by gel‐based methods that are low‐throughput, time‐consuming and laborious. In this study, we showed a rapid and convenient method for analysis of SCAR markers in a gel‐free manner. Seven SCAR markers, linked to rust resistance genes (Sr24, Sr26 and Sr31) and seed quality traits (Pina, Pinb and Glu‐D1) in wheat (Triticum aestivum), were amplified on a real‐time PCR machine using custom reaction mixture. Subsequently, melting curve analysis was performed, to assess the specificity of amplicons. Using the amplicon‐specific melt‐profiles, the presence/absence of SCAR markers was analysed in fifteen genotypes and five F2 populations. Unlike the fluorescence‐based in‐tube detection methods, the present method used the amplicon‐specific melt‐profiles to evaluate the status of the SCAR markers, thus eliminating the need for gel‐based analysis. Results also showed feasibility of multiplex analysis of two markers with well‐separated melting profiles. Overall, the approach is a rapid, convenient and cost‐effective method for high‐throughput screening of SCAR markers. 相似文献
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Shuancang Yu Fenglan Zhang Xiuyun Zhao Yangjun Yu Deshuang Zhang 《Plant Breeding》2011,130(5):580-583
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The AFLP (amplified fragment length polymorphism) technique has been applied in establishing an extended linkage map of sugar beet. A total of 120 AFLPs were integrated into an existing linkage map based on RFLP markers. Four primer combinations yielded between 19 and 40 polymorphic bands in an F2 population consisting of 94 plants. The AFLP loci were evenly distributed over the nine linkage groups, with the exception of linkage group V where the number of AFLPs was significantly low. The AFLPs were found to be reproducible even against the background of different combinations of Taq DNA polymerases and buffers. However, the quantity of higher molecular weight fragments (>400 bp) was reduced when using plant DNA of poor quality as a template. The results of these experiments are discussed, together with possible applications of AFLPs in sugar beet breeding. 相似文献
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A yellow leaf mutant at a locus named YEL was selected in a population of the cultivated carrot. Genetic analysis of segregating F2 progenies and corresponding F3 families, indicates that the phenotype expressed is controlled by a single recessive nuclear gene. The mutant is stably inherited and is associated with a reduced leaf‐biomass of approximately 30% compared with the wild‐type. Amplified fragment length polymorphism markers were developed and used in bulked segregant analysis. Seventeen marker candidates were detected by using 45 primer pairs. Ten of these could be linked with the YEL locus and mapped in a linkage group with a total length of 33.2 cM. Application of the yellow leaf mutant in carrot research is discussed. 相似文献
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Levels of genetic similarity characterizing 20 grasspea (Lathyrus sativus L.) populations collected in central Italy (17 populations in the Marche region and three populations in the Abruzzo region) were analysed with amplified fragment length polymorphism (AFLP) molecular markers. Two main clusters were found: one included large‐seeded populations from farms that were not market‐oriented (named Household populations) and the second, small‐seeded populations, cultivated in market‐oriented farms (named Commercial populations). Relationships among populations collected in different regions were found, although one population of the Abruzzo region was placed between the two main clusters, suggesting a possible further genetic differentiation within this grasspea germplasm collection. Principal component analysis based on AFLP marker frequency was effective in identifying polymorphic markers showing high discriminating ability between clusters H and C. In particular, seven markers showing high positive and three markers with low negative PC1 scores showed an almost cluster‐specific distribution. These results will be useful for enhancing Italian grasspea germplasm use in plant‐breeding programmes and for extending grasspea cultivation within the sustainable agricultural systems of central Italy. 相似文献
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Huei-Mei Chen Chien-An Liu C. George Kuo Ching-Mei Chien Horng-Chi Sun Chung-Chu Huang Yu-Chung Lin Hsin-Mei Ku 《Euphytica》2007,157(1-2):113-122
Bruchid, Callosobruchus spp. (Coleoptera: Bruchidae), is a serious pest during storage of seeds of mungbean (Vigna radiata (L.) Wilczek) and other Vigna species. A source of resistance to this pest has been identified in Vigna
sublobata (Roxb.) Bairig. accession TC1966. Two hundred recombinant inbred lines at the F12 generation have been developed for molecular mapping of bruchid resistance (Br) gene in TC1966. Through bulked segregant analysis (BSA), ten randomly amplified polymorphic DNA (RAPD) markers associated
with the bruchid resistance gene were successfully identified. A total of four closely linked RAPDs were cloned and transformed
into sequence characterized amplified region (SCAR) and cleaved amplified polymorphism (CAP) markers. Seven CAPs developed
from the identified RAPD markers showed tighter linkage with the Br gene than the original RAPD. Through transformation of RAPDs into CAPs, codominant markers for bruchid resistance were successfully
obtained. Homozygous genotypes of these PCR-based markers were estimated to contribute 85% of the variance for seed damage
when the insect assay was performed under favorable growth conditions for bruchid. 相似文献
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Gladys Romero Luisa M. Vásquez Philippe Lashermes Juan C. Herrera 《Plant Breeding》2014,133(1):121-129
Most of the commercial varieties of coffee (Coffea arabica L.) derived from the Timor hybrid (TH) have been shown to contain major genes for coffee leaf rust (CLR) resistance. To identify markers tightly linked to such genes, an F2 mapping population derived from a cross between ‘Caturra’ (susceptible variety) and the TH‐derived DI.200 line (highly resistant) was generated. Using expressed sequence information and a bioinformatics approach, both targeted region amplified polymorphism (TRAPs) markers and simple sequence repeat (SSR) markers were identified. Phenotypic evaluations in the field and under controlled conditions confirmed the existence of one quantitative trait locus for CLR resistance. Four candidate SSR markers were associated with high CLR resistance. They spanning a region of 2.5 cM designated QCLR_4 located within chromosome 4 of the international C. canephora map. The presence of this region was confirmed in a set of elite lines and commercial varieties. The QCLR_4 region corresponds to a new and genetically independent SH locus that could potentially be useful in gene pyramiding with other genes to enhance rust resistance in TH derivatives. 相似文献
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Common bunt caused by Tilletia tritici and T. laevis has occurred worldwide and reduces yield and quality in common and durum wheats. The development of DNA markers linked to bunt resistance to race T1 in the cross, ‘Laura’(S) בRL5407’ (R), was carried out in this study based on the single head derived F4:5 and single seed derived F4:6 populations. Bulked segregant analysis was used to identify two random amplified polymorphic DNA (RAPD) markers linked to the gene for resistance to race T1 in the spelt wheat ‘RL5407′. The two markers identified, UBC548590 and UBC274988, flanked the resistance gene with a map distance of 9.1 and 18.2 cM, respectively. The former was linked in repulsion phase to bunt resistance while the later was in coupling phase. The two RAPD markers and the common bunt‐resistance gene all segregated in Mendelian fashion. Use of these two RAPD markers together could assist in incorporating the bunt‐resistance gene from spelt wheat into common wheat cultivars by means of marker‐assisted selection. 相似文献