Makful, S. Purnomo, and Sunyoto. Analysis of Genetic Diversity of Mangosteen Basedon the Amplified Fragment Length Polymorphism (AFLP) Technique. For estimating genetic distance the AFLP and RFLP markers gave the most The dominant markers (AFLP and RAPD) had small CV values indicating a. Key Words: Aegilops, AFLP, DNA fingerprinting, durum wheat. Bu¤dayda ve Aegilops’ta çok teknik deneyim gerektirmektedir. Bununla birlikte ÇPUP.
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Because the mean is not a good indicator of central tendency for skewed data we calculated the minimum number of loci necessary for an accurate representation tekniii the genetic distances by fitting an exponential function based on the mean, median and maximum CV values of the genetic distances obtained by bootstrap sampling to the data for each marker, the results of this analysis being given in the Boxplots shown in Figure 3.
For dominant markers, where the distribution is skewed towards lower genetic distance values, the use of mean or median CV values may lead to errors because some of the genetic distance values will not fall within the required level of precision.
Although the AFLP markers gave the lowest mean PIC value they provided a similar degree of polymorphism information content to that provided by the RAPD markers, which agrees with the results published by Becker et al. However, these molecular markers have technical differences in terms of cost, speed, amount of DNA needed, technical labor, degrees of polymorphism, precision of genetic distance estimates and the statistical power of tests.
In order to compare their relative efficiencies as markers and to find the most suitable marker for maize diversity studies we evaluated 18 inbred tropical maize lines using a number of different loci as markers. As expected, the PIC distributions revealed that, in terms of genetic distance, dominant markers had lower levels of polymorphism as compared to codominant markers.
Differences in the distribution profiles also occurred between dominant and codominant markers, with dominant markers having higher standard deviations than codominant markers. Am J Hum Genet As expected, the magnitude of the coefficient of variation CV values decreased as the number of polymorphic loci bands evaluated increased.
Analisis Keragaman Genetik Manggis Menggunakan Teknik Amplified Fragment Length Polymorphism (AFLP)
The AFLP technique is more laborious and time consuming than RAPD methods but is also more reliable, AFLP being able to detect a large number of polymorphic bands in a single lane rather than high levels of polymorphism at each locus such as is the case for SSR methods.
Bull Soc Vaud Nat Correspondence to Antonio A.
Theor Appl Genet We used the median and maximum coefficient of variation values to evaluate the accuracy of the genetic distance tekbii because although the mean coefficient of variation is often used in the literature caution is needed when dealing with molecular marker data for which there is no assurance that the CVs values are distributed symmetrically.
Each band visualized on the gel was tekii to be the re-sampling unit for dominant markers because for these markers each band is related to one locus. Detailed descriptions of these populations are given in Lanza et al. For the AFLP method 20 primer combinations were used and binary scored 1 or 0 with each band being considered a locus while for the SSR method 68 polymorphic primers were used with the binary data being converted into a genotypic matrix which was used to identify alleles and their respective loci.
Comparison of RAPD, RFLP, AFLP and SSR markers for diversity studies in tropical maize inbred lines
The genetic distances for the codominant markers RFLP and SSR were calculated using the modified Roger’s distance MRD; Goodman and Stuber, based on the allele frequency of each locus which considers the amount of genetic diversity and expresses the quantity of diversity present in each locus or allele, calculations being made using version 1.
Plant material and DNA isolation. Jaccard similarities were calculated using version 2. Indeed, we found that the RFLP and AFLP markers produced sufficient numbers of polymorphic bands to produce reliable genetic distance estimates with high correlations between these two marker systems, the similarity between the results being explainable by the fact that they are similar techniques based on restriction site changes.
From the analysis of our data it appears that the maximum CV value appears to be, in most cases, the best guarantee for producing reliable estimates of genetic distance.
Comparison of the genetic distances generated by different molecular markers in diversity studies have been reported by several authors Hahn et al. How to cite this article. Heterotic group assignment and hybrid performance determined by RFLP marker. The choice of the appropriate number of polymorphic loci required for a reliable estimation of genetic distance is influenced by the criteria used, and it appears that the maximum and median CV values are the best choice for evaluating the precision of the genetic distance estimates based on molecular marker data sets.
SAS language guide for personal computers. The principles and practice of statistics in biological research, 3 rd edn. AFLP also correlates highly with results obtained using the RFLP system and is a fast and reliable system capable of supporting a multiplex approach not requiring previous knowledge of DNA sequencing. Correlations between genetic distances measured with different markers.
Theor Appl Genet 94, Plant Breeding 6: A new technique for DNA fingerprinting. The results also suggest that the number of loci tekniii should be increased.
Eighteen S 3 selected inbred lines from two divergent tropical maize populations eight from BR and ten from BR previously had their genetic distances surveyed using four different marker systems Lanza et al. We found that estimates of polymorphism information content PIC based on RFLP measures had the lowest standard deviations and were the most informative. Codominant markers relate each band to an allele, and therefore the boostrap was applied among locus.
Materials and methods Plant material and DNA isolation Eighteen S 3 selected inbred lines from two divergent tropical maize populations eight from BR and ten from BR previously had their genetic distances surveyed using four different marker systems Lanza et al.
Total genomic DNA trknii isolated from a bulk of five-week-old leaf tissue taken from 16 plants of each line, then being isolated and purified by the method of Hoisington et al.
The box-plots Figure 3 show what happens when the genetic distance CV values, which are different for dominant and codominant markers, are high. Within each sample i.
Briefly, a total of clone-enzyme combinations were analyzed, the maize genome being saturated sflp cM intervals with wflp least one RFLP probe selected by its map location on each chromosome. Our results indicate that, apart from the RAPD markers, the other DNA marker systems provided consistent information for diversity studies on tropical maize populations and produced genetic distance estimates which were in good agreement.
Pearson’s correlation coefficient was calculated for the genetic distances, single cross performance and heterosis as previously described by Benchimol et al.
Our results indicate that AFLP seemed to be the best-suited molecular assay for fingerprinting and assessing genetic relationships among tropical maize inbred lines with high accuracy. VI Isozyme variation among races of maize in Bolivia. Nucl Acids Res Due to this uniqueness, molecular markers have been very useful in genetic evaluations and assignment of tropical maize inbred lines to heterotic groups.
The past limitations associated with pedigree tekmii and morphological, physiological and cytological markers for assessing genetic diversity in cultivated and wild plant species have largely been circumvented by the development of DNA markers such teknki restriction fragment length polymorphisms RFLPs; Botstein et al.
Our results points to the need to adopt different strategies for selecting markers and choosing an upper number of SSR and RAPD markers.