Sample records for aflp molecular markers

  1. Molecular mapping of the Pinus monticola Cr2 gene using AFLP and SCAR markers

    A.K.M. Ekramoddoullah


    Full Text Available White pine blister rust (WPBR, caused by Cronartium ribicola, is a devastating disease in five-needle pines. Genetic resistance is an important component of integrated strategies to control WPBR. The major resistance gene Cr2, discovered by Kinloch etal.(1999, is also effective against British Columbia (BC isolates of WPBR (Hunt et al. 2004. Pyramiding Cr2 gene with other resistancegenes is being pursued as a strategy in BC white pine breeding. To facilitate this strategy, we have recently identified a few RAPD markerslinked to Cr2 at one side (Liu et al. 2006. The objective of the present study was to identify amplified fragment length polymorphism(AFLP markers linked to both sides of Cr2 for its more precise apping. Use of the AFLP technique combined with bulked segregant analysis (BSA and haploid segregation analysis allowed the identification of five AFLP markers. Of these five AFLP markers in the Cr2 linkage, markers EacccMccgat-365, EactgMcccac- 290, and EacagEacag-750 werelinked in coupling and EacagMcccag-160r and EacccMccgat-180r in repulsion. Following cloning and sequencing of the AFLP andRAPD markers, specific PCR primers were designed and used in the amplification of sequence characterized amplified region(SCAR markers at both sides of Cr2. EacccMccgat- 365 and RAPD marker U570-843 reported previously were converted into SCARmarkers. These two SCARs segregated in a 1:1 (presence:absence ratio and the scoring cosegregated with their respective AFLP orRAPD marker. The SCAR marker EacccMccgat- 365-scar was positioned at 3.1 Kosambi cM from one side of Cr2 and U570-843-scarlocalized at 1.4 Kosambi cM from other side. Both SCAR markers can be useful in breeding programs with marker-assisted selection procedureto screen for resistance. This study represents the first report of the development of PCR-based sequence-specific markers linkedto blister rust resistance in five-needle pines. These findings may

  2. Genetic variation of space flight carried rice and mutant analysis by AFLP molecular marker

    Rice seeds were carried by 'Shenzhou No.3' space shuttle, a mutant with golden chaff, stem and leaf was selected and named Golden 1 after the seeds returned to the earth. Except the golden color, other traits of Golden 1 are no obviously different with its original material H9808. Genetic analysis identified that color variation was control by a pair of recessive gene. The DNA fragments of the mutant were compared with its parent by AFLP molecular markers. Five specific bands were found through a serial selection. (authors)

  3. Sesame mutant induced space flight treatment and analysis of polymorphism by AFLP molecular marker

    Seeds of two sesame varieties (Yuzhi 8 and H98) were carried by 'Shijian 8' satellite for space treatment. Variants is SP2 and SP1 were observed and the genetic diversity of mutation generation was analyzed using 30 pair of AFLP markers. The results showed that: (1) variants of leaf, plant height, plant characters, floral organs, capsule, fertility, pre-flowering date were observed in SP2 and SP3 of two varieties, variations frequency and variations type of two sesame varieties were significantly different, but only the small capsule variations and tall plant variations could be inherited from SP2 to SP3 generation; (2) AFLP analysis of primers randomly selected to analyze variant plants in SP2 and SP3 generation, showed that multiple sites in sesame genome were induced by space environment and mutation rate was high, while mutation rate was different in molecular level between varieties; (3) variation sites were diverse between different variations types, and mutation sites were also diverse between different plants of same mutation type, this result indicated that variation of sesame genome DNA could not been expressed totally; (4) The same mutant of botany characteristics and the same locus mutation of molecular level were both detected. (authors)

  4. High Lycopene AFLP- SCAR Molecular Marker of Fresh Tomato%鲜食番茄茄红素基因的AFLP-SCAR分子标记

    侯丽霞; 刘淑梅; 王施慧; 吕鑫


    Lycopene (LYC) is a function of the natural pigment, because of its high antioxidant functions have attracted much attention. For screening high-lycopene varieties of fresh tomatoes to accelerate the process of molecular marker-assisted breeding, AFLP-SCAR marks of high lycopene were studied by using materials high-lycopene selling line F-516F8 and low-lycopene selfing line Nor. Using genetical population of Ft and F2, AFLP and BAS techniques with 64 pairs of E/M primers screening, we obtained one AFLP marker E-AG/M-CAT. Through recovery, and sequencing of idio-fragment, transformation of SCAR, BLAST, marker E-AG/M-CAT was locating on the eighth chromosome. Utilizing SCAR makers can evaluate the content of lycopene at DNA level when the tomato materials are in seedling period, which greatly reducing the workload of field screening. SCAR makers can also be used on molecular marker assisted selection, improving the breeding efficiency.%番茄红素(Lycopene)是功能性天然色素,因其高抗氧化功能而备受关注.为筛选高番茄红素品种,加速鲜食番茄分子标记辅助育种进程,本试验以高茄红素番茄骨干系F-516F8和低茄红素骨干系Nor作为试验材料,研究了鲜食番茄高茄红素基因的AFLP-SCAR分子标记.利用杂交F1代、自交F2代遗传群体,通过64对E/M引物组合的筛选,采用AFLP分析技术和改良BAS法,获得1个与高番茄红素基因连锁的AFLP标记E-AG/M-CAT.通过特异片段回收、测序、SCAR转化、BLAST比对分析,该标记定位在番茄第8染色体.利用SCAR标记可在苗期从DNA水平上对番茄材料的番茄红素含量高低进行鉴定,大大减少了田间筛选工作量,并可用于分子标记辅助育种,提高育种效率.

  5. Genetic diversity analysis of Croton antisyphiliticus Mart. using AFLP molecular markers.

    Oliveira, T G; Pereira, A M S; Coppede, J S; França, S C; Ming, L C; Bertoni, B W


    Croton antisyphiliticus Mart. is a medicinal plant native to Cerrado vegetation in Brazil, and it is popularly used to treat urogenital tract infections. The objective of the present study was to assess the genetic variability of natural C. antisyphiliticus populations using AFLP molecular markers. Accessions were collected in the states of Minas Gerais, São Paulo, and Goiás. The genotyping of individuals was performed using a LI-COR® DNA Analyzer 4300. The variability within populations was found to be greater than the variability between them. The F(ST) value was 0.3830, which indicated that the populations were highly structured. A higher percentage of polymorphic loci (92.16%) and greater genetic diversity were found in the population accessions from Pratinha-MG. Gene flow was considered restricted (N(m) = 1.18), and there was no correlation between genetic and geographic distances. The populations of C. antisyphiliticus exhibited an island-model structure, which demonstrates the vulnerability of the species. PMID:26909989

  6. Diversidade genética em maracujazeiro-amarelo utilizando marcadores moleculares fAFLP Genetic diversity in yellow passion fruit utilizing fAFLP molecular markers

    Rita Maria Devós Ganga; Carlos Ruggiero; Eliana Gertrudes de Macedo Lemos; Gisele Ventura Garcia Grili; Michele Mantovani Gonçalves; Edvan Alves Chagas; Ester Wickert


    Marcadores moleculares fAFLP foram utilizados para estimar a diversidade genética entre 36 acessos de maracujá-amarelo (Passiflora edulis f. flavicarpa Deg.) coletados em 18 estados do Brasil. Os resultados obtidos permitiram concluir que os marcadores fAFLP se mostraram consistentes na avaliação da variabilidade genética, detectando e quantificando a ampla divergência genética entre os 36 acessos analisados, bem como a não-formação de estruturação geográfica. Tais resultados podem auxiliar n...

  7. Genetic diversity and relationship in American and African oil palm as revealed by RFLP and AFLP molecular markers

    Barcelos Edson


    Full Text Available The objective of this work was to evaluate the genetic diversity, its organization and the genetic relationships within oil palm (Elaeis oleifera (Kunth Cortés, from America, and E. guineensis (Jacq., from Africa germplasm using Restriction Fragment Length Polymorphism (RFLP and Amplified Fragment Length Polymorphism (AFLP. In complement to a previous RFLP study on 241 E. oleifera accessions, 38 E. guineensis accessions were analyzed using the same 37 cDNA probes. These accessions covered a large part of the geographical distribution areas of these species in America and Africa. In addition, AFLP analysis was performed on a sub-set of 40 accessions of E. oleifera and 22 of E. guineensis using three pairs of enzyme/primer combinations. Data were subjected to Factorial Analysis of Correspondence (FAC and cluster analysis, with parameters of genetic diversity being also studied. Results appeared congruent between RFLP and AFLP. In the E. oleifera, AFLP confirmed the strong structure of genetic diversity revealed by RFLP, according to geographical origin of the studied material, with the identification of the same four distinct genetic groups: Brazil, French Guyana/Surinam, Peru, north of Colombia/Central America. Both markers revealed that genetic divergence between the two species is of the same magnitude as that among provenances of E. oleifera. This finding is in discrepancy with the supposed early tertiary separation of the two species.

  8. AFLP marker linked to water-stress-tolerant bulks in barley (Hordeum vulgare L.)

    Altinkut A.; Kazan K.; Gozukirmizi N.


    The amplified fragment length polymorphism (AFLP) assay is an efficient method for the identification of molecular markers, useful in the improvement of numerous crop species. Bulked Segregant Analysis (BSA) was used to identify AFLP markers associated with water-stress tolerance in barley, as this would permit rapid selection of water-stress tolerant genotypes in breeding programs. AFLP markers linked to water-stress tolerance was identified in two DNA pools (tolerant and sensitive), which w...

  9. Widespread utility of highly informative AFLP molecular markers across divergent shark species.

    Zenger, Kyall R; Stow, Adam J; Peddemors, Victor; Briscoe, David A; Harcourt, Robert G


    Population numbers of many shark species are declining rapidly around the world. Despite the commercial and conservation significance, little is known on even the most fundamental aspects of their population biology. Data collection that relies on direct observation can be logistically challenging with sharks. Consequently, molecular methods are becoming increasingly important to obtain knowledge that is critical for conservation and management. Here we describe an amplified fragment length polymorphism method that can be applied universally to sharks to identify highly informative genome-wide polymorphisms from 12 primer pairs. We demonstrate the value of our method on 15 divergent shark species within the superorder Galeomorphii, including endangered species which are notorious for low levels of genetic diversity. Both the endangered sand tiger shark (Carcharodon taurus, N = 18) and the great white shark (Carcharodon carcharias, N = 7) displayed relatively high levels of allelic diversity. A total of 59 polymorphic loci (H(e) = 0.373) and 78 polymorphic loci (H(e) = 0.316) were resolved in C. taurus and C. carcharias, respectively. Results from other sharks (e.g., Orectolobus ornatus, Orectolobus sp., and Galeocerdo cuvier) produced remarkably high numbers of polymorphic loci (106, 94, and 86, respectively) from a limited sample size of only 2. A major constraint to obtaining much needed genetic data from sharks is the time-consuming process of developing molecular markers. Here we demonstrate the general utility of a technique that provides large numbers of informative loci in sharks. PMID:17150982

  10. Genetic diversity of Cuban pineapple germplasm assessed by AFLP Markers

    Ermis Yanes Paz; Katia Gil; Laureano Rebolledo; Andrés Rebolledo; Daniel Uriza; Octavio Martínez; Miriam Isidrón; Leyanes Díaz; José Carlos Lorenzo; June Simpson


    The Cuban pineapple germplasm collection represents the genetic diversity of pineapple cultivated in that country and includes other important genotypes obtained from the germplasm collections in Brazil and Martinique. The collection has previously been characterized with morphological descriptors but a molecular characterization has been lacking. With this aim, 56 six genotypes of A. comosus and one of Bromelia pinguin were analyzed with a total of 191 AFLP markers. A dendrogram that represe...

  11. Genetic diversity of Cuban pineapple germplasm assessed by AFLP Markers

    Ermis Yanes Paz


    Full Text Available The Cuban pineapple germplasm collection represents the genetic diversity of pineapple cultivated in that country and includes other important genotypes obtained from the germplasm collections in Brazil and Martinique. The collection has previously been characterized with morphological descriptors but a molecular characterization has been lacking. With this aim, 56 six genotypes of A. comosus and one of Bromelia pinguin were analyzed with a total of 191 AFLP markers. A dendrogram that represents the genetic relationships between these samples based on the AFLP results showed a low level of diversity in the Cuban pineapple collection. All Ananas comosus accessions, being the majority obtained from farmers in different regions in Cuba, are grouped at distances lower than 0.20. Molecular characterization was in line with morphological characterization. These results are useful for breeding and conservation purposes.

  12. Study on the sex-related AFLP marker of the Yangtze finless porpoise


    The sex-related molecular marker of the Yangtze finless porpoise was screened using Amplified Fragment Length Polymorphism (AFLP) technique combined with the bulked segregant analysis. Totally 36 AFLP primer combinations were used to detect the genome DNA bulks of the female and male porpoises, and one sex-related AFLP marker was finally obtained. The marker can be applied to sex identification, and provides a base for further cloning of sex-related genes and analyzing of Y chromosome haplotypes of the Yangtze finless porpoise.

  13. AFLP marker linked to water-stress-tolerant bulks in barley (Hordeum vulgare L.

    A. Altinkut


    Full Text Available The amplified fragment length polymorphism (AFLP assay is an efficient method for the identification of molecular markers, useful in the improvement of numerous crop species. Bulked Segregant Analysis (BSA was used to identify AFLP markers associated with water-stress tolerance in barley, as this would permit rapid selection of water-stress tolerant genotypes in breeding programs. AFLP markers linked to water-stress tolerance was identified in two DNA pools (tolerant and sensitive, which were established using selected F2 individuals resulting from a cross between water-stress-tolerant and sensitive barley parental genotypes, based on their paraquat (PQ tolerance, leaf size, and relative water content (RWC. All these three traits were previously shown to be associated with water-stress tolerance in segregating F2 progeny of the barley cross used in a previous study. AFLP analysis was then performed on these DNA pools, using 40 primer pairs to detect AFLP fragments that are present/absent, respectively, in the two pools and their parental lines. One separate AFLP fragment, which was present in the tolerant parent and in the tolerant bulk, but absent in the sensitive parent and in the sensitive bulk, was identified. Polymorphism of the AFLP marker was tested among tolerant and sensitive F2 individuals. The presence of this marker that is associated with water-stress tolerance will greatly enhance selection for paraquat and water-stress tolerant genotypes in future breeding programs.

  14. Molecular characterization of cultivated species of the genus Pachyrhizus Rich. ex DC. by AFLP markers

    Santayana, Monica; Rossel, Genoveva; Núñez, Jorge;


    Yam beans (Pachyrhizus DC.) are legume root crops that have attracted scientific interest because of high contents of starch, protein, and iron in their roots. In this study, 58 accessions of three cultivated Pachyrhizus species were characterized by amplified fragment length polymorphism (AFLP) ...

  15. A detailed linkage map of lettuce based on SSAP, AFLP and NBS markers

    Syed, N.; Sorensen, A.P.; Antonise, R.; Wiel, van de C.C.M.; Linden, van der C.G.; Westende, van 't W.P.C.; Hooftman, D.A.P.; Nijs, den H.C.M.; Flavell, A.


    Molecular markers based upon a novel lettuce LTR retrotransposon and the nucleotide binding site-leucine-rich repeat (NBS-LRR) family of disease resistance-associated genes have been combined with AFLP markers to generate a 458 locus genetic linkage map for lettuce. A total of 187 retrotransposon-sp

  16. Association of AFLP and SSR markers with agronomic and fibre quality traits in Gossypium hirsutum L.

    Arunita Rakshit; S. Rakshit; J. Singh; S. K. Chopra; H. S. Balyan; P. K. Gupta; Shripad R. Bhat


    Molecular markers linked to QTL contributing to agronomic and fibre quality traits would be useful for cotton improvement. We have attempted to tag yield and fibre quality traits with AFLP and SSR markers using F2 and F3 populations of a cross between two Gossypium hirsutum varieties, PS56-4 and RS2013. Out of 50 AFLP primer combinations and 177 SSR primer pairs tested, 32 AFLP and four SSR primers were chosen for genotyping F2 individuals.Marker-trait associations were studied for eight agronomic and five fibre quality traits through simple and multiple regression analysis (MRA) using a set of 92 AFLP polymorphic loci and four SSR markers. Simple linear regression analysis (SLRA) identified 23 markers for eight different traits whereas multiple regression analysis identified 30 markers for at least one of the 13 traits. SSR marker BNL 3502 was consistently identified to be associated with fibre strength. While all the markers identified in SLRA were also detected in MRA, as many as 16 of the 30 markers were identified to be associated with respective traits in both F2 and F3 generations. The markers explained up to 41 per cent of phenotypic variation for individual traits. A number of markers were found to be associated with multiple traits suggesting clustering of QTLs for fibre quality traits in cotton.

  17. Variabilidade genética de Colletotrichum guaranicola usando marcadores AFLP Variability of Colletotrichum guaranicola using AFLP markers

    Jânia Lilia da Silva Bentes


    Full Text Available Foi detectada a variabilidade genética de vinte isolados de Colletotrichum guaranicola (Albuq. provenientes de diferentes localidades produtoras de guaraná no Amazonas, utilizando-se marcadores moleculares AFLP. Foi possível separar os isolados em dois grupos. O coeficiente de variação genética entre os isolados foi de 0,0216 e a similaridade genética foi de 94,95%, confirmando que os isolados pertencem à mesma espécie, no entanto, foi observada variabilidade intra-específica.The genetic variability of twenty Colletotrichum guaranicola (Albuq. isolates from different fields of guarana in Amazonas, was studied using molecular AFLP markers. The isolates were separated into two groups. The genetic variability coefficient was 0.0216 and the genetic similarity was 94.5%, confirming that the isolates belongs to the same species, however, an intra-specific variability was observed.

  18. A Preliminary Study of Genetic Variation in Populations of Monstera adansonii var. klotzschiana (Araceae) from North-East Brazil, Estimated with AFLP Molecular Markers

    Andrade, I. M.; Mayo, S. J.; van den Berg, C.; Fay, M. F.; Chester, M.; Lexer, C.; Kirkup, D.


    Background and Aims This study sought genetic evidence of long-term isolation in populations of Monstera adansonii var. klotzschiana (Araceae), a herbaceous, probably outbreeding, humid forest hemi-epiphyte, in the brejo forests of Ceará (north-east Brazil), and clarification of their relationships with populations in Amazonia and the Atlantic forest of Brazil. Methods Within-population genetic diversity and between-population dissimilarity were estimated using AFLP molecular markers in 75 individuals from eight populations located in Ceará, the Brazilian Atlantic Forest and Amazonia. Key Results The populations showed a clinal pattern of weak genetic differentiation over a large geographical region (FST = 0·1896). A strong correlation between genetic and geographical distance (Mantel test: r = 0·6903, P = 0·002) suggests a historical pattern of isolation by distance. Genetic structure analysis revealed at least two distinct gene pools in the data. The two isolated Ceará populations are significantly different from each other (pairwise ΦPT = 0·137, P = 0·003) and as diverse (Nei's gene diversity, average He = 0·1832, 0·1706) as those in the Atlantic and Amazon forest regions. The population in southern Brazil is less diverse (Nei's gene diversity, average He = 0·127) than the rest. The Ceará populations are related to those of the Atlantic forest rather than those from Amazonia (AMOVA, among-groups variation = 11·95 %, P = 0·037). Conclusions The gene pools detected within an overall pattern of clinal variation suggest distinct episodes of gene flow, possibly correlated with past humid forest expansions. The Ceará populations show no evidence of erosion of genetic diversity, although this was expected because of their isolation. Their genetic differentiation and relatively high diversity reinforce the importance of conserving the endangered brejo forests. PMID:17823112

  19. Genetic diversity in natural populations of Jacaranda decurrens Cham. determined using RAPD and AFLP markers

    Bianca W. Bertoni


    Full Text Available Jacaranda decurrens (Bignoniaceae is an endemic species of the Cerrado with validated antitumoral activity. The genetic diversity of six populations of J. decurrens located in the State of São Paulo was determined in this study by using molecular markers for randomly amplified polymorphic DNA (RAPD and amplified fragment length polymorphism (AFLP. Following optimization of the amplification reaction, 10 selected primers generated 78 reproducible RAPD fragments that were mostly (69.2% polymorphic. Two hundred and five reproducible AFLP fragments were generated by using four selected primer combinations; 46.3% of these fragments were polymorphic, indicating a considerable level of genetic diversity. Analysis of molecular variance (AMOVA using these two groups of markers indicated that variability was strongly structured amongst populations. The unweighted pair group method with arithmatic mean (UPGMA and Pearson's correlation coefficient (RAPD -0.16, p = 0.2082; AFLP 0.37, p = 0.1006 between genetic matrices and geographic distances suggested that the population structure followed an island model in which a single population of infinite size gave rise to the current populations of J. decurrens, independently of their spatial position. The results of this study indicate that RAPD and AFLP markers were similarly efficient in measuring the genetic variability amongst natural populations of J. decurrens. These data may be useful for developing strategies for the preservation of this medicinal species in the Cerrado.

  20. Genetic Relationships among Prunus mume var. pendula Using AFLP Markers

    Ming Jun; Zhang Qixiang; Ru Guangxin; Mao Qingshan; Yan Xiaolan; Lan Yanping


    Genetic relationships among Prunus mume var. pendula were studied by using AFLP markers. 18 accessions representing 14 cultivars ofPrunus murne var. pendula were selected from the germplasm collection at the Research Center of China Mci Flower. Seven Mse I-EcoR I AFLP primer combinations revealed 450 legible bands, and 269 of which were polymorphic markers. A similarity matrix was prepared using the simple matching coefficient of similarity and Nei's (72) distance coefficient. A UPGMA dendrogram demonstrated the genetic relationships of the cultivars. The information given by AFLP markers was basically consistent with the morphological classification and the evolutionary history of the morphotypes, and roughly supported the new revised classification system for Chinese Mci Cultivars. But there were still several exceptions: 1) the 'Guhong Chuizhi' inserted between the 'Tiaoxue Chuizhi' and the 'Danfen Chuizhi'; 2) the 'Wufu Chuizhi' kept off the Pink Pendant Form, and the 'Moshan Chuizhi' was removed from Viridiflora Pendant Form; 3) the 'Danbi Chuizhi' and the 'Shuangbi Chuizhi' of Viridiflora Pendant Form got together well but fell within the Pink Pendant Form.

  1. Genetic linkage map of Brassica campestris L. Using AFLP and RAPD markers

    卢钢; 曹家树; 陈杭


    A genetic linkage map comprised of 131 loci was constructed with an F2 population derived from an inter-subspecific cross between Brassica 'qisihai'. The genetic map included 93 RAPD loci, 36 AFLP loci and 2 morphological loci organized into 10 main linkage groups (LGs) and 2 small groups, covering 1810.9cM with average distance between adjacent markers being approximately 13.8cM. The map is suitable for identification of molecular markers linked to important agronomic traits, QTL analysis, and even for marker-assisted selection in breeding programs of Chinese cabbage and turnip.

  2. Evolutionary analysis of allotetraploid hybrids of red crucian carp × common carp,based on ISSR,AFLP molecular markers and cloning of cyclins genes

    LIU LiangGuo; YAN JinPeng; LIU ShaoJun; LIU Dong; YOU CuiPing; ZHONG Huan; TAO Min; LIU Yun


    The allotetraploid hybrids of red crucian carp × common carp are the first reported artificially cultured polyploid fish with bisexual fertility and stable inheritance in vertebrate.Using ISSR and AFLP markers and the cyclins genes,the genomes and cyclin gene sequence changes were analyzed between the allotetraploid hybrids and their parents.The results indicated that the allotetraploids inherited many genetic characteristics from their parents and the genetic characteristics were stable after 15 generations.However,the allotetraploids had a closer genetic relationship with their original female parents and represented a bias toward the maternal progenitor.DNA fingerprinting analysis showed that the allotetraploids had undergone sequences deletion from their original parents and that the deleted sequences were mostly from the male parent's genome.Some non-parental bands were found in the allotetraploid hybrids.Sequences analysis of the cyclin A1 and B1 genes showed nonsynonymous substitutions of single nucleotides in codons that were different from their original parents,leading to non-parental amino acid loci.We speculate that the non-additivity in the allotetraploids,compared with their progenitors,could be an adjustment to the genomic shock from heterozygosity and polyploidy, allowing maintenance of genetic stability.

  3. Comparison Between AFLP and RFLP Markers in Detecting the Diversity of Rice (Oryza sativa L.)

    CHEN Liang; LIANG Chun-yang; SUN Chuan-qing; JIN De-min; JIANG Ting-bo; WANG Bin; WANG Xiang-kun


    AFLP and RFLP were used to study the diversity of 20 rice cultivars. 15 primer combinations were used in the AFLP analysis and 47 - 118 bands were amplified in each lane. A total of 107 polymorphic bands were detected in the RFLP analysis using 49 RFLP probes. The cluster analysis based on RFLP data showed that 20 rice cultivars could be divided into an indica group and a japonica group, as did the AFLP data; however AFLP is more suitable in detecting the difference of pedigree and ecotype among the 20 cultivars.The genetic distance based on AFLP and RFLP data showed the same tendency, but AFLP markers increased the measure of genetic distance among intra-subspecific cultivars and decreased the measure of genetic distance among inter-subspecific cultivars relative to RFLP markers. That indicated that AFLP is more suitable than RFLP in the diversity study and RFLP is more suitable to study the indica-japonica differentiation of cultivars.

  4. Confirmation of Clematis hybrids using molecular markers

    The hybrid origin of two progeny from reciprocal crosses of Clematis tubulosa and C. brevicaudata was investigated using molecular markers generated by randomly amplified polymorphic DNA (RAPD), amplified fragment length polymorphism (AFLP), and single nucleotide polymorphisms (SNPs). Morphologi...

  5. Phylogenetic Relationships in Genus Arachis Based on SSR and AFLP Markers

    TANG Rong-hua; ZHUANG Wei-jian; GAO Guo-qing; HE Liang-qiong; HAN Zhu-qiang; SHAN Shi-hua; JIANG Jing; LI Yang-rui


    Fourteen wild species of different sections in the genus Arachis and 24 accessions of the AABB allotetraploid A. hypogaea (cultivated peanut) from several countries which belong to different botanical varieties, were analyzed by SSR and AFLP marker systems. The assay-units per system needed to distinguish among all the tested accessions were at least five for SSR or two for AFLP. The genetic distance detected by the SSR markers ranged from 0.09 to 0.95, and the mean was 0.73; and the genetic distance detected by the AFLP markers ranged from 0.01 to 0.79 with an average of 0.42. All the tested peanut SSR primer pairs were multilocus ones, and the amplified fragments per SSR marker in each peanut genome ranged from 2 to 15 with the mean of 4.77. The peanut cultivars were closely related to each other, and shared a large numbers of SSR and AFLP fragments. In contrast, the species in the genus Arachis shared few fragments. The results indicated that the cultivated peanut (A. hypogaea L.) varieties could be partitioned into two main groups and four subgroups at the molecular level, and that A. duranensis is one of the wild ancestors of A. hypogaea. The lowest genetic variation was detected between A. cardenasii and A. batizocoi, and the highest was detected between A. pintoi and the species in the section Arachis. The relationships among the botanical varieties in the cultivated peanut (A. hypogaea L.) and among wild species accessions in section Arachis and those in other sections in the genus Arachis were discussed.

  6. Genetic diversity among some currants (Ribes spp.) cultivars as assessed by AFLP markers

    Currants cultivation has increased its popularity in Turkey due to the use of more currants in Turkish cuisine. To provide farmers with well adapted currants cultivars, some currants cultivars have been planted in various geographical regions of Turkey. In this study, genetic diversity among some of these currants cultivars has been analyzed using AFLP markers. Our results indicated that red and black currants genotypes are genetically distinct, sharing very small proportion of AFLP markers. Selected currants genotypes from Turkey shared all AFLP markers suggesting that they might be the same genotype. (author)

  7. Genetic linkage map of Brassica campestris L.using AFLP and RAPD markers

    卢钢; 陈杭; 等


    A genetic linkage map comprised of 131 loci was constructed with an F2 population derived from an inter-subspecific cross between Brassica campestris L.ssp.chinensis cv.aijiaohang” and ssp.rapifera cv.,”'isihai”.The genetic map included 93 RAPD loci,36 AFLP loci and 2 morphological loci organized into 10 main linkage groups(LGs) and 2 small groups,covering 1810.9cM with average distance between adjacent markers being approximately 13.8cM.The map is suitable for identification of molecular markers linked to important agronomic traits.QTL analysis,and even for marker-assisted selection in breeding programs of Chinese cabbage and turnip.

  8. Genetic diversity revealed by AFLP markers in Albanian goat breeds

    Hoda Anila


    Full Text Available The amplified fragment length polymorphism (AFLP technique with three EcoRI/TaqI primer combinations was used in 185 unrelated individuals, representative of 6 local goat breeds of Albania, and 107 markers were generated. The mean Nei’s expected heterozygosity value for the whole population was 0.199 and the mean Shannon index was 0.249, indicating a high level of within-breed diversity. Wright’s FST index, Nei’s unbiased genetic distance and Reynolds’ genetic distance were calculated. Pairwise Fst values among the populations ranged from 0.019 to 0.047. A highly significant average FST of 0.031 was estimated, showing a low level of breed subdivision. Most of the variation is accounted for by differences among individuals. Cluster analysis based on Reynolds’ genetic distance between breeds and PCA were performed. An individual UPGMA tree based on Jaccard’s similarity index showed clusters with individuals from all goat breeds. Analysis of population structure points to a high level of admixture among breeds.

  9. AFLP markers for the assessment of genetic diversity in european and North American potato varieties cultivated in Iran

    Saeed Tarkesh Esfahani


    Full Text Available Information about the genetic diversity of potato germplasm in Iran is important for variety identification andto enhance the classification of germplasm collections and exploit them in breeding programs and for the development andintroduction of new varieties. AFLP fingerprinting was applied to a group of cultivated potato varieties to find if there is anygeographical differentiation in potato diversity from Europe and North America. The high level of polymorphism within potatovarieties and the high number of variety-specific bands suggest that AFLPs are powerful markers for diversity analysis inpotato varieties. No region-specific AFLP markers were found (present in varieties from the same origin and absent inothers. The UPGMA dendrogram revealed four distinct clusters corresponding almost to the geographical origin of thevarieties. However, the bootstrap support for branches was rather weak. No clusters clearly distinguished varieties fromEurope and North America. Varieties from the same geographical origins however tended to group together within eachcluster. The mean similarity and the UPGMA dendrogram both suggest that North American varieties have nearly identicalgenetic diversity to European varieties. The results of AMOVA revealed large within-region variations which accounted for94.5% of the total molecular variance. The between-region variation, although accounting for only 5.5% of the total variation,was statistically significant. AFLP technology was successfully used to evaluate diversity between different geographicalgroups of potatoes and is recommended for potato genetic studies.

  10. A Genetic Linkage Map of Brassica rapa Based on AFLP Markers

    ZHAO Jian-jun; WANG Xiao-wu; Guusje Bonnema; SUN Ri-fei; XU Ze-yong; Dick Vreugdenhi; Maarten Koornneef


    A F2 mapping population was developed by crossing a Chinese cabbage-pe-tsai variety CC156 and an oil type Rapid cycling RC144 which were different from each other in morphology, maturity, self-compatibility, plant height, etc. Using 244 AFLP markers a map was constructed containing 10 main linkage groups covering a total distance of 857 cM,corresponding to 3.5 cM per marker. Length of linkage groups varied from 43 to 125 cM and the number of AFLP markers linkage to each group ranged from 7 to 41.

  11. Outcrossing rate between 'Haden' and 'Tommy Atkins' mangoes estimated using microsatellite and AFLP markers

    Carlos Antonio Fernandes Santos


    Full Text Available The objective of this work was to estimate outcrossing rates between Haden and Tommy Atkins mango cultivars, using AFLP and microsatellite markers. Progenies of an isolated 'Haden' plant, identified in a 'Tommy Atkins' commercial orchard, in Petrolina, PE, Brazil, were analyzed. Total DNA was isolated from the progeny leaves and used for AFLP and microsatellite reactions. Multilocus outcrossing rates (t m were estimated by direct count of AFLP or microsatellite markers and by the mLTR software. Outcrossing rates ranged from 0.85 to 0.87 with the analysis based on seven AFLP markers, and from 0.83 to 0.91 based on three microsatellite primers. No unexpected band patterns were observed for 'Haden' and 'Tommy Atkins'. The estimates obtained with the mLTR software were close to those obtained by direct AFLP and microsatellite allele counting, which indicates that the multilocus model was appropriate for this kind of study. The microsatellites mMiCIR005, mMiCIR030, and mMiCIR036 can be used to elucidate the origin of 'Haden' and 'Tommy Atkins' seedlings.

  12. Identification of AFLP Markers Linked to Lr19 Resistance to Wheat Leaf Rust

    LI Xing; YANG Wen-xiang; LI Ya-ning; LIU Da-qun; YAN Hong-fei; MENG Qing-fang; ZHANG Ting


    AFLP analyses were carried out on Thatcher, 23 near-isogenic lines and F2 generation of TcLrl9 × Thatcher, to develop molecular markers for gene Lr19 resistance to wheat leaf rust. Seven markers linked to Lr19 resistance trait were obtained,which were P-AGT/M-GAG289 bp (3.3 cM), P-ACA/M-GGT102 bp (4.1 cM), P-ACA/M-GGT106 bp (4.1 cM), P-AAC/M-CAG123 bp(4.9 cM), P-AAC/M-GGT203 bp (5.0 cM), P-ACA/M-GGT290 bp (5.7 cM), and P-ATC/M-GAG293 bp (9.6 cM). All of these specific fragments were isolated from the polyacrylamide gels, reamplified, cloned, and sequenced. The research may facilitate genetic mapping, physical mapping, and the eventual cloning of Lr19.

  13. Genetic diversity of wild and cultivated Rubus species in Colombia using AFLP and SSR markers

    Sandra Bibiana Aguilar


    Full Text Available The Andean blackberry belongs to the genus Rubus, the largest of the Rosaceae family and one of the mostdiverse of the plant kingdom. In Colombia Rubus glaucus Benth, known as the Andean raspberry or blackberry, is one of thenine edible of the genus out of forty-four reported species. In this study wild and cultivated genotypes, collected in the CentralAndes of Colombia were analyzed by AFLP and SSR markers. Sexual reproduction seems to play an important role inmaintaining the genetic variability in R. glaucus, and the viability of using the SSR of Rubus alceifolius to characterizeColombian Rubus species was clearly demonstrated. All species evaluated produced very specific banding patterns,differentiating them from the others. Both AFLP and SSR produced bands exclusive to each of the following species: R.robustus, R. urticifolius, R. glaucus, and R. rosifolius. The SSR markers differentiated diploid and tetraploid genotypes of R.glaucus.

  14. AFLP Marker Linked to Turnip Mosaic Virus Susceptible Gene in Chinese Cabbage (Brassica rapa L.ssp.pekinensis)

    HAN He-ping; SUN Ri-fei; ZHANG Shu-jiang; LI Fei; ZHANG Shi-fan; NIU Xin-ke


    Turnip mosaic virus (TuMV) which has several strains causes the most important virusdisease in Chinese cabbage in terms of crop damage. In China, Chinese cabbage is infectedby a mixture of strains, breeding of cultivar for the TuMV resistance has become themajor aim. Screening the molecular marker linked to the TuMV-resistance gene formolecular assisted selection is the major method to improve the breeding efficiency. Inthis study, we used AFLP technique and the method of bulked segregant analysis(BSA) tostudy the progeny of Brp0058 x Brp0108, and identified two DNA molecular marker linked toTurnip mosaic virus-resistance gene with a recombination frequency 7.5 cM and 8.4 cM.

  15. Analysis of genetic diversity in crocuses with Carpathian Basin origin using AFLP-markers.

    Surányi, G; Máthé, C; Mosolygó, Agnes; Borbély, G; Vasas, G


    Crocus taxonomy has until now been based primarily on morphology, taking chromosome numbers into consideration. The genetics and genome structure of the genus, the relationships and diversity within the genus are not well known. Amplified fragment length polymorphism (AFLP) is a whole genome approach to study genetic variation that is gaining in popularity for lower-level systematics. The present study employed the AFLP technique for analyzing relationships among taxa of the Crocus genus (particularly the Crocus vernus aggregate) with Carpathian Basin origin. The molecular variance obtained was based on amplification, separation and detection of EcoRI and Tru1I double-digested Crocus spp. genomic DNAs. Our results confirm the relatedness of C. tommasinianus, C. vittatus and C. heuffelianus at the Verni series of the Crocus genus. C. banaticus is taxonomically isolated as the sole member of the subgenus Crociris based on unique morphological features, but the difference is not convincing from AFLP data. The second interesting AFLP analysis result is the position of C. scepusiensis which separated it from the Crocus vernus aggregate. PMID:21565773

  16. Genetic similarity among soybean (Glycine max (L Merrill cultivars released in Brazil using AFLP markers

    Ana Lídia V. Bonato


    Full Text Available Genetic similarity among soybean genotypes was studied by applying the amplified fragment length polymorphism (AFLP technique to 317 soybean cultivars released in Brazil from 1962 to 1998. Genetic similarity (GS coefficients were estimated using the coefficient of Nei and Li (Nei and Li 1979, and the cultivars were clustered using the unweighted pair-group method with averages (UPGMA. The parentage coefficients of 100 cultivars released between 1984 and 1998 were calculated and correlated with the genetic similarity obtained by the markers. The genetic similarity coefficients varied from 0.17 to 0.97 (x = 0.61, with 56.8% of the coefficients being above 0.60 and only 9.7% equal to or less than 0.50. The similarity coefficients have remained constant during the last three decades. Dendrogram interpretation was hindered by the large number of cultivars used, but it was possible to detect groups of cultivars formed as expected from their genealogy. Another dendrogram, composed of 63 cultivars, allowed a better interpretation of the groups. Parentage coefficients among the 100 cultivars varied from zero to one (x = 0.21. However, no significant correlation (r = 0.12 was detected among the parentage coefficients and the AFLP genetic similarity. The results show the efficiency of AFLP markers in large scale studies of genetic similarity and are discussed in relation to soybean breeding in Brazil.

  17. Genetic differentiation of Helicoverpa armigera (Hübner) and H.Assulta (Guenée) (Lepidoptera: Noctuidae) based on AFLP markers



    Here we use amplified fragment length polymorphism (AFLP) to assess genetic differentiation of Helicoverpa armigera and H. assulta. The results indicated that both species-specific fingerprints and cluster analysis showed the ability of AFLP technique to discriminate the two sibling species; among a total 1963 AFLP markers amplified from nine primer combinations: 777 (39.6%) were H. armigera-specific,602 (30.7%) were H. assultaspecific,and 584 (29.7%) were common bands. The mean number of H. armigera-specific bands was significantly more than that of H. assulta-specific bands for nine primer combinations (P<0.05); the intraspecific distance of H. armigera and H. assulta was 0.123 0 and 0.110 7 respectively,and the interspecific distance was 0.178 3. In addition,the percentage of polymorphic loci and estimated average heterozygosity were used to estimate genetic diversity of the two species. This study therefore demonstrates that AFLP analysis is a sensitive and reliable technique to study genetic differentiation and genetic relationships between species and provides sufficient molecular markers for future linkage map construction,location and eventual cloning of genes involved in traits differentiation.

  18. Genetic variation in wild populations of the tuber crop Amorphophallus konjac (Araceae) in central China as revealed by AFLP markers.

    Pan, C; Gichira, A W; Chen, J M


    Amorphophallus konjac is an economically important crop. In order to provide baseline information for sustainable development and conservation of the wild plant resources of A. konjac, we studied the genetic diversity and population structure of this species using amplified fragment length polymorphism (AFLP) molecular markers. We sampled 139 individuals from 10 wild populations of A. konjac in central China. Using five AFLP primer combinations, we scored a total of 270 DNA fragments, most of which were polymorphic (98.2%). Percentage of polymorphic loci, Nei's genetic diversity index, and Shannon's information index showed high levels of genetic variation within A. konjac populations. Analysis of molecular variance indicated that most of the variance (68%) resided within populations. The coefficient of genetic differentiation between populations was 0.348 and the estimated gene flow was 0.469, indicating that there was limited gene flow among the populations. Unweighted pair group method with arithmetic mean cluster analysis and principal coordinates analysis indicated that geographically close populations were more likely to cluster together. The Mantel test revealed a significant correlation between geographic and genetic distances (R2 = 0.2521, P konjac and the complex geography of central China are likely to have contributed to the current pattern of genetic variation of this species. In the present study, we provide several suggestions on the future protection of the wild plant genetic resources of A. konjac. PMID:26782525

  19. The application of AFLP fingerprinting in breeding of Brassica napus

    Cuřínová, Petra


    AFLP markers are widely used in breeding in some other crops, but their utilization in breeding of Brassica crops is not so frequent. AFLP markers are used for molecular characterization of particular varieties or genotypes and for evaluation of genetic diversity. The aim of this thesis was the application of this method in breeding of rapeseed and in comparative study of genetic variability of different oil seed rape cultivars of Czech, Czechoslovak and German origin. AFLP is based on select...

  20. Marcadores fAFLP na caracterização de três genótipos de umezeiro selecionados como porta-enxertos para pessegueiro fAFLP markers to characterize three mume genotypes selected as rootstocks for peach tree

    Ester Wickert


    Full Text Available O objetivo deste trabalho foi caracterizar a diversidade genética existente em três genótipos de umezeiro (Clone 05, cv. Rigitano e Clone 15 e identificar marcadores moleculares fAFLP (fluorescent Amplified Fragment Lenght Polymorphism passíveis de serem utilizados na discriminação dos três genótipos de umezeiro selecionados como porta-enxertos para pessegueiro. Foram utilizadas 24 diferentes combinações de primers seletivos fAFLP que geraram 648 marcas, das quais 272 foram diferenciadoras dos três genótipos entre si. As marcas diferenciadoras permitiram o agrupamento dos clones de umezeiro de acordo com sua similaridade através do Método da Distância e algorítmo Neighbour Joining. As mesmas marcas foram utilizadas para calcular a distância genética entre os clones. Com o uso de marcadores fAFLP foi possível discriminar os três genótipos de umezeiro entre si, destacando-se as combinações Fam ACT/CAT, Joe AGG/CTT e Ned AGC/CAA, que permitiram a diferenciação individual de cada um dos clones. A maior distância genética foi encontrada entre a cv. Rigitano e o Clone 15. Os marcadores fAFLP revelaram maior proximidade genética entre o Clone 05 e a cv. Rigitano.The objective of this work was the identification of fAFLP markers to be used in molecular characterization of three mume genotypes selected as rootstocks for peach tree. Twenty-four different fAFLP primer combinations were used and allowed the recognition of 648 markers, comprising 272 markers which were able to discriminate the three clones one from the other. These markers were used to calculate the groupment of the clones according to their similarities with the distance method and neighbour joining algorithm. The same markers were also used to calculate the genetic distance among the clones. The fAFLP markers were efficient to identify the clones, mainly by the combinations of selective primers Fam ACT/CAT, Joe AGG/CTT and Ned AGC/CAA. fAFLP markers allowed the

  1. Genetic diversity in bambara groundnut (Vigna subterranea (L.) Verdc) landraces revealed by AFLP markers.

    Massawe, F J; Dickinson, M; Roberts, J A; Azam-Ali, S N


    Bambara groundnut (Vigna subterranea (L.) Verdc), an African indigenous legume, is popular in most parts of Africa. The present study was undertaken to establish genetic relationships among 16 cultivated bambara groundnut landraces using fluorescence-based amplified fragment length polymorphism (AFLP) markers. Seven selective primer combinations generated 504 amplification products, ranging from 50 to 400 bp. Several landrace-specific products were identified that could be effectively used to produce landrace-specific markers for identification purposes. On average, each primer combination generated 72 amplified products that were detectable by an ABI Prism 310 DNA sequencer. The polymorphisms obtained ranged from 68.0 to 98.0%, with an average of 84.0%. The primer pairs M-ACA + P-GCC and M-ACA + P-GGA produced more polymorphic fragments than any other primer pairs and were better at differentiating landraces. The dendrogram generated by the UPGMA (unweighted pair-group method with arithmetic averaging) grouped 16 landraces into 3 clusters, mainly according to their place of collection or geographic origin. DipC1995 and Malawi5 were the most genetically related landraces. AFLP analysis provided sufficient polymorphism to determine the amount of genetic diversity and to establish genetic relationships in bambara groundnut landraces. The results will help in the formulation of marker-assisted breeding in bambara groundnut. PMID:12502264

  2. Development of a SCAR marker for male gametophyte of Gracilariopsis lemaneiformis based on AFLP technique

    Zhou, Wei; Ding, Hongye; Sui, Zhenghong; Wang, Zhongxia; Wang, Jinguo


    The red alga Gracilariopsis lemaneiformis (Bory) is an economically valuable macroalgae. As a means to identify the sex of immature Gracilariopsis lemaneiformis, the amplified fragment length polymorphism (AFLP) technique was used to search for possible sex- or phase-related markers in male gametophytes, female gametophytes, and tetrasporophytes, respectively. Seven AFLP selective amplification primers were used in this study. The primer combination E-TG/M-CCA detected a specific band linked to male gametophytes. The DNA fragment was recovered and a 402-bp fragment was sequenced. However, no DNA sequence match was found in public databases. Sequence characterized amplified region (SCAR) primers were designed from the sequence to test the repeatability of the relationship to the sex, using 69 male gametophytes, 139 female gametophytes, and 47 tetrasporophytes. The test results demonstrate a good linkage and repeatability of the SCAR marker to sex. The SCAR primers developed in this study could reduce the time required for sex identification of Gracilariopsis lemaneiformis by four to six months. This can reduce both the time investment and number of specimens required in breeding experiments.

  3. First genetic linkage map of Taraxacum koksaghyz Rodin based on AFLP, SSR, COS and EST-SSR markers.

    Arias, Marina; Hernandez, Monica; Remondegui, Naroa; Huvenaars, Koen; van Dijk, Peter; Ritter, Enrique


    Taraxacum koksaghyz Rodin (TKS) has been studied in many occasions as a possible alternative source for natural rubber production of good quality and for inulin production. Some tire companies are already testing TKS tire prototypes. There are also many investigations on the production of bio-fuels from inulin and inulin applications for health improvement and in the food industry. A limited amount of genomic resources exist for TKS and particularly no genetic linkage map is available in this species. We have constructed the first TKS genetic linkage map based on AFLP, COS, SSR and EST-SSR markers. The integrated linkage map with eight linkage groups (LG), representing the eight chromosomes of Russian dandelion, has 185 individual AFLP markers from parent 1, 188 individual AFLP markers from parent 2, 75 common AFLP markers and 6 COS, 1 SSR and 63 EST-SSR loci. Blasting the EST-SSR sequences against known sequences from lettuce allowed a partial alignment of our TKS map with a lettuce map. Blast searches against plant gene databases revealed some homologies with useful genes for downstream applications in the future. PMID:27488242

  4. Identification on commercialized products of AFLP markers able to discriminate slow- from fast-growing chicken strains.

    Fumière, Olivier; Dubois, Marc; Grégoire, Dimitrie; Théwis, André; Berben, Gilbert


    The European chicken meat market is characterized by numerous quality marks: "Label de Qualité Wallon" in Belgium, "Label Rouge" in France, denominations of geographical origin, organic agriculture, etc. Most of those certified productions have specifications requiring the use of slow-growing chicken strains. The amplified fragment length polymorphism (AFLP) technique has been used to search molecular markers able to discriminate slow-growing chicken strains from fast-growing ones and to authenticate certified products. Two pairs of restriction enzymes (EcoRI/MseI and EcoRI/TaqI) and 121 selective primer combinations were tested on individual DNA samples from chicken products essentially in carcass form that were ascribed as belonging to either slow- or fast-growing strains. Within the resulting fingerprints, two fragments were identified as type-strains specific markers. One primer combination gives a band (333 bp) that is specific for slow-growing chickens, and another primer pair generates a band (372 bp) that was found to be characteristic of fast-growing chickens. The two markers were isolated, cloned, and sequenced. The effectiveness and the specificity of the two interesting determinants were assessed on individuals of two well-known strains (ISA 657 and Cobb 500) and on commercialized products coming from various origins. PMID:12590443

  5. Mapping a stripe rust resistance gene YrC591 in wheat variety C591 with SSR and AFLP markers.

    Li, Y; Niu, Y C; Chen, X M


    Stripe rust, caused by Puccinia striiformis Westend. f. sp. tritici (PST), is one of the most destructive diseases of common wheat (Triticum aestivum L.). To determine inheritance of stripe rust resistance and map the resistance gene(s) in wheat variety C591, F(1), F(2,) and F(3) progenies derived from the Taichung 29 x C591 cross were inoculated with Chinese PST race CY32 in the greenhouse. Genetic analysis identified a single dominant gene, temporarily designated YrC591. A total of 178 SSR and 130 AFLP markers were used to test the parents and resistant and susceptible bulks. From the bulk segregant analysis, seven polymorphic SSR and two AFLP markers were selected for genotyping the F(2) population. SSR marker Xcfa2040-7B, and SCAR marker SC-P35M48 derived from AFLP marker P35M48 ( 373 ) were identified to be closely linked to the resistance gene with genetic distances of 8.0 and 11.7 cM, respectively. The SSR markers mapped the resistance gene on chromosome arm 7BL. In the seedling test with five PST races, the reaction patterns of C591 were different from wheat cultivars or lines carrying Yr2 or Yr6 that also are found on chromosome 7B. The results indicate that YrC591 is probably a novel stripe rust resistance gene. PMID:18946654

  6. Genetic diversity in cultivated carioca common beans based on molecular marker analysis

    Juliana Morini Küpper Cardoso Perseguini


    Full Text Available A wide array of molecular markers has been used to investigate the genetic diversity among common bean species. However, the best combination of markers for studying such diversity among common bean cultivars has yet to be determined. Few reports have examined the genetic diversity of the carioca bean, commercially one of the most important common beans in Brazil. In this study, we examined the usefulness of two molecular marker systems (simple sequence repeats - SSRs and amplified fragment length polymorphisms - AFLPs for assessing the genetic diversity of carioca beans. The amount of information provided by Roger's modified genetic distance was used to analyze SSR data and Jaccards similarity coefficient was used for AFLP data. Seventy SSRs were polymorphic and 20 AFLP primer combinations produced 635 polymorphic bands. Molecular analysis showed that carioca genotypes were quite diverse. AFLPs revealed greater genetic differentiation and variation within the carioca genotypes (Gst = 98% and Fst = 0.83, respectively than SSRs and provided better resolution for clustering the carioca genotypes. SSRs and AFLPs were both suitable for assessing the genetic diversity of Brazilian carioca genotypes since the number of markers used in each system provided a low coefficient of variation. However, fingerprint profiles were generated faster with AFLPs, making them a better choice for assessing genetic diversity in the carioca germplasm.

  7. Species phylogeny and diversification process of Northeast Asian Pungitius revealed by AFLP and mtDNA markers

    Takahashi, Hiroshi; Møller, Peter Rask; Shedko, Sergei V.;


    Northeast Asia, although the taxonomy and evolutionary relationships among them remain unclear. We used amplified fragment length polymorphism (AFLP) and mitochondrial DNA (mtDNA) markers to infer phylogenies among individuals collected from sympatric and allopatric populations, including the type....... kaibarae, and P. bussei). The brackish-water, freshwater, and Omono types previously discovered in Japan were reidentified as P. pungitius, P. sinensis, and P. kaibarae, respectively. A marked incongruence was noted between the phylogenies of AFLP and mtDNA markers, suggesting the occasional occurrence of...... hybridization and mtDNA introgression among distinct species. Our results highlight that the marginal seas of Northeast Asia played a key role as barriers to or facilitators of gene flow in the evolution of species diversity of Pungitius concentrated in this region...

  8. Molecular characterization of native potato (Solanum spp. Chungui, Ayacucho, using AFLP

    Juan C. Gonzales Mamani


    Full Text Available Genetic diversity of 25 morphotypes of native potatoes Solanum spp. from Chungui (La Mar, Ayacucho were assess. Morphotypes collected were micropropagated in Murashigue Skoog medium (1962. DNA extraction proceeded using the CTAB method modified from 3 weeks leaves crop, good quality and quantity of DNA was able to use the AFLP. Enzymatic digestion of the DNA was performed using EcoRI and MseI. Three combinations of AFLP primers with three selective nucleotides were used, resulting in a total of 85 clearly discernable bands, of which 63 were polymorphic. The combination E37 – M50 showed the most informative polymorphic index content of 0.43. The presence/absence of polymorphic bands was evaluated using the Simple Matching coefficient similarity and clustering analysis using the UPGMA. The dendrogram produced had a cophenetic correlation coefficient r= 0.7. At the level 0.64 of Simple Matching coefficient similarity, the dendrogram grouped the morphotypes of native potatoes in 4 genetic groups, it not found duplicated morphotypes, despite having some morphotypes very similar. Our results would be showing the highly informative power of AFLP markers for the analysis of genetic diversity of native potatoes.

  9. Highly informative single-copy nuclear microsatellite DNA markers developed using an AFLP-SSR approach in black spruce (Picea mariana and red spruce (P. rubens.

    Yong-Zhong Shi

    Full Text Available Microsatellites or simple sequence repeats (SSRs are highly informative molecular markers for various biological studies in plants. In spruce (Picea and other conifers, the development of single-copy polymorphic genomic microsatellite markers is quite difficult, owing primarily to the large genome size and predominance of repetitive DNA sequences throughout the genome. We have developed highly informative single-locus genomic microsatellite markers in black spruce (Picea mariana and red spruce (Picea rubens using a simple but efficient method based on a combination of AFLP and microsatellite technologies.A microsatellite-enriched library was constructed from genomic AFLP DNA fragments of black spruce. Sequencing of the 108 putative SSR-containing clones provided 94 unique sequences with microsatellites. Twenty-two of the designed 34 primer pairs yielded scorable amplicons, with single-locus patterns. Fourteen of these microsatellite markers were characterized in 30 black spruce and 30 red spruce individuals drawn from many populations. The number of alleles at a polymorphic locus ranged from 2 to 18, with a mean of 9.3 in black spruce, and from 3 to 15, with a mean of 6.2 alleles in red spruce. The polymorphic information content or expected heterozygosity ranged from 0.340 to 0.909 (mean = 0.67 in black spruce and from 0.161 to 0.851 (mean = 0.62 in red spruce. Ten SSR markers showing inter-parental polymorphism inherited in a single-locus Mendelian mode, with two cases of distorted segregation. Primer pairs for almost all polymorphic SSR loci resolved microsatellites of comparable size in Picea glauca, P. engelmannii, P. sitchensis, and P. abies.The AFLP-based microsatellite-enriched library appears to be a rapid, cost-effective approach for isolating and developing single-locus informative genomic microsatellite markers in black spruce. The markers developed should be useful in black spruce, red spruce and other Picea species for

  10. The construction of a linkage map of Alstroemeria aurea by AFLP markers

    Han, T.H.; Eck, van H.J.; Jeu, de M.J.; Jacobsen, E.


    An AFLP based linkage map has been generated for the ornamental cropspecies Alstroemeria aurea. In view of the large genome size of Alstroemeria (25,000 Mb) the number of selective nucleotides for AFLPamplification was increased to EcoRI+4/MseI+4 to generatefingerprints of moderate complexity. In ad

  11. Species phylogeny and diversification process of Northeast Asian Pungitius revealed by AFLP and mtDNA markers.

    Takahashi, Hiroshi; Møller, Peter R; Shedko, Sergei V; Ramatulla, Temirbekov; Joen, Sang-Rin; Zhang, Chun-Guang; Sideleva, Valentina G; Takata, Keisuke; Sakai, Harumi; Goto, Akira; Nishida, Mutsumi


    Pungitius is a highly diversified genus of sticklebacks (Gasterosteidae) occurring widely in northern parts of the Northern Hemisphere. Several ecologically and genetically divergent types that are largely isolated reproductively but occasionally hybridize in sympatry have been discovered in Northeast Asia, although the taxonomy and evolutionary relationships among them remain unclear. We used amplified fragment length polymorphism (AFLP) and mitochondrial DNA (mtDNA) markers to infer phylogenies among individuals collected from sympatric and allopatric populations, including the type localities of the described species. Phylogenetic analyses based on 2683 polymorphic AFLP loci confirmed seven species, each of which (except for one entirely allopatric species P. platygaster) was clearly differentiated from one or two other sympatric species and constituted a highly supported monophyletic clade with conspecific allopatric populations. The phylogeny showed that two lineages arose early; one gave rise to two species (circumpolar species P. pungitius and Paratethys species P. platygaster) and the other to five species endemic to Northeast Asia (P. sinensis, P. tymensis, P. polyakovi, P. kaibarae, and P. bussei). The brackish-water, freshwater, and Omono types previously discovered in Japan were reidentified as P. pungitius, P. sinensis, and P. kaibarae, respectively. A marked incongruence was noted between the phylogenies of AFLP and mtDNA markers, suggesting the occasional occurrence of hybridization and mtDNA introgression among distinct species. Our results highlight that the marginal seas of Northeast Asia played a key role as barriers to or facilitators of gene flow in the evolution of species diversity of Pungitius concentrated in this region. PMID:26997522

  12. Genetic variability of Brazilian isolates of Alternaria alternata detected by AFLP and RAPD techniques Variabilidade genética de isolados Brasileiros de Alternaria alternata por meio de marcadores moleculares de AFLP e RAPD

    Francisco Dini-Andreote


    Full Text Available The Alternaria brown spot (ABS is a disease caused in tangerine plants and its hybrids by the fungus Alternaria alternata f. sp. citri which has been found in Brazil since 2001. Due to the recent occurrence in Brazilian orchards, the epidemiology and genetic variability of this pathogen is still an issue to be addressed. Here it is presented a survey about the genetic variability of this fungus by the characterization of twenty four pathogenic isolates of A. alternata f. sp. citri from citrus plants and four endophytic isolates from mango (one Alternaria tenuissima and three Alternaria arborescens. The application of two molecular markers Random Amplified Polymorphic DNA (RAPD and Amplified Fragment Length Polymorphism (AFLP had revealed the isolates clustering in distinct groups when fingerprintings were analyzed by Principal Components Analysis (PCA. Despite the better assessment of the genetic variability through the AFLP, significant modifications in clusters components were not observed, and only slight shifts in the positioning of isolates LRS 39/3 and 25M were observed in PCA plots. Furthermore, in both analyses, only the isolates from lemon plants revealed to be clustered, differently from the absence of clustering for other hosts or plant tissues. Summarizing, both RAPD and AFLP analyses were both efficient to detect the genetic variability within the population of the pathogenic fungus Alternaria spp., supplying information on the genetic variability of this species as a basis for further studies aiming the disease control.A mancha marrom ou mancha de Alternaria é uma doença causada pelo fungo Alternaria alternata f. sp. citri, encontrada no Brasil desde 2001 em plantas de tangerina e seus híbridos. Por se tratar de uma doença recente no Brasil, a epidemiologia e variabilidade genética deste patógeno compõem importantes pontos a serem estudados. Este trabalho teve como objetivo avaliar a variabilidade genética deste pat

  13. Pattern of ancient goat migration revealed by AFLP molecular markers

    ECONOGENE Consortium; P. Ajmone Marsan; Joost, S; P. Crepaldi; Patrini, M.; Pellecchia, M.; E. Milanesi; R. Negrini


    Domestic goat (Capra hircus) is a very adaptable and geographically spread livestock species. Recent studies on mitochondrial DNA diversity suggest that goats have been the most widely transported and traded livestock species. For this reason it is thought to have played a central role in the demic spread of agriculture during the Neolithic agricultural revolution (Luikart et al. 2001).

  14. Pattern of ancient goat migration revealed by AFLP molecular markers

    ECONOGENE Consortium


    Full Text Available Domestic goat (Capra hircus is a very adaptable and geographically spread livestock species. Recent studies on mitochondrial DNA diversity suggest that goats have been the most widely transported and traded livestock species. For this reason it is thought to have played a central role in the demic spread of agriculture during the Neolithic agricultural revolution (Luikart et al. 2001.

  15. Application of AFLP markers for population genetic study on half-smooth tongue sole Cynoglossus semilaevis

    LIU Yunguo; LI Junfeng; YE Naihao


    The genetic diversity of wild and hatchery populations of half-smooth tongue sole Cynoglossus semilaevis, based on observation of amplified fragment length polymorphism (AFLP) was described. Two hundred individuals from four wild populations, Laizhou (LZ), Weihai (WH), Qingdao (QD), Rizhao (RZ), and one hatchery population, Mingbo (MB), were screened using eight different AFLP primer combinations. A total of 384 loci were screened in the five studied populations. 48.4%, 51.3%,50.7%, 49.3% and 45.8% of these loci were polymorphic among the individuals tested in the LZ, WH,QD, RZ and MB populations, respectively. The number of polymorphic loci detected by single primer combinations ranged from 17 to 35. The average heterozygosity of the LZ, WH, QD, RZ and MB populations was 0.072, 0.093, 0.092, 0.090 and 0.063, respectively. The WH population showed the highest genetic diversity in terms of total number of AFLP bands, total number of polymorphic bands,average heterozygosity and percentage of low frequency (0-0.2) polymorphic loci among all the populations,while the LZ population was the lowest among the wild populations. Compared with the wild populations,the hatchery population showed a low genetic viability.

  16. Construction of two genetic linkage maps in cultivated tetraploid alfalfa (Medicago sativa using microsatellite and AFLP markers

    Santoni Sylvain


    Full Text Available Abstract Background Alfalfa (Medicago sativa is a major forage crop. The genetic progress is slow in this legume species because of its autotetraploidy and allogamy. The genetic structure of this species makes the construction of genetic maps difficult. To reach this objective, and to be able to detect QTLs in segregating populations, we used the available codominant microsatellite markers (SSRs, most of them identified in the model legume Medicago truncatula from EST database. A genetic map was constructed with AFLP and SSR markers using specific mapping procedures for autotetraploids. The tetrasomic inheritance was analysed in an alfalfa mapping population. Results We have demonstrated that 80% of primer pairs defined on each side of SSR motifs in M. truncatula EST database amplify with the alfalfa DNA. Using a F1 mapping population of 168 individuals produced from the cross of 2 heterozygous parental plants from Magali and Mercedes cultivars, we obtained 599 AFLP markers and 107 SSR loci. All but 3 SSR loci showed a clear tetrasomic inheritance. For most of the SSR loci, the double-reduction was not significant. For the other loci no specific genotypes were produced, so the significant double-reduction could arise from segregation distortion. For each parent, the genetic map contained 8 groups of four homologous chromosomes. The lengths of the maps were 2649 and 3045 cM, with an average distance of 7.6 and 9.0 cM between markers, for Magali and Mercedes parents, respectively. Using only the SSR markers, we built a composite map covering 709 cM. Conclusions Compared to diploid alfalfa genetic maps, our maps cover about 88–100% of the genome and are close to saturation. The inheritance of the codominant markers (SSR and the pattern of linkage repulsions between markers within each homology group are consistent with the hypothesis of a tetrasomic meiosis in alfalfa. Except for 2 out of 107 SSR markers, we found a similar order of markers on

  17. Molecular variation of Trypanosoma brucei subspecies as revealed by AFLP fingerprinting

    Agbo, E.E.C.; Majiwa, P.A.O.; Claassen, H.J.H.M.; Pas, te, M.F.W.


    Genetic analysis of Trypanosoma spp. depends on the detection of variation between strains. We have used the amplified fragment length polymorphism (AFLP) technique to develop a convenient and reliable method for genetic characterization of Trypanosome (sub)species. AFLP accesses multiple independent sites within the genome and would allow a better definition of the relatedness of different Trypanosome (sub)species. Nine isolates (3 from each T. brucei subspecies) were tested with 40 AFLP pri...

  18. Assessing genetic diversity of wild populations of Japanese flounder using AFLP markers

    XU Xiaofei; ZHANG Quanqi; WANG Zhigang; QI Jie; ZHANG Zhifeng; BAO Zhenmin; Heisuke Nakagawa


    Amplified fragment length polymorphism (AFLP) analysis was used to evaluate the genetic diversity of four wild geographical populations of Japanese flounder (Paralichthys olivaceus). A total of 775 loci (58.32% of which was polymorphic) in the range between 100 and 1 300 base pairs were detected from 110 individuals using seven primer combinations. The percentage of polymorphic loci detected by single primer combination for each population was calculated, ranging from 19.59% to 53.33%. Genetic similarities within and among the populations were calculated from the binary matrices of presence - absence. Phylogenetic tree of four populations was constructed by using the UPGMA method using PHYLIP Version 3.5. According to intrapopulation genetic similarities, CW population displayed the highest genetic diversity value and KY population had the lowest genetic diversity value.The distance between CW and CF populations was the farthest, which was possibly resulted from the farthest distance of Weihai of Shandong and Fujian of China compared with the geographical distance between other locations of populations. The subpopulation differentiation value ( Gst ) is 0.356 5, showing a certain extent of differentiation among the four geographical populations. AFLP technology was confirmed to be an effective tool to assess within- and among-population genetic diversity of Japanese flounder. The present survey provided significant insights for research in the Japanese flounder breeding program.

  19. A genetic linkage map of marine shrimp Penaeus ( Fenneropenaeus) chinensis based on AFLP, SSR, and RAPD markers

    Liu, Bo; Wang, Qingyin; Li, Jian; Liu, Ping; He, Yuying


    The Chinese shrimp Penaeus ( Fenneropaeneus) chinensis is an important species in marine fishery and aquaculture in China. A female Chinese shrimp Penaeus ( Fenneropaeneus) chinensis was captured from west coast of the Korean peninsula and mated with a “Yellow Sea No. 1” male to produce the first filial generation (F1) 100 F2 full-sib progeny from brother-sister crosses between F1 families was used for the mapping study. A genetic linkage map of the Chinese shrimp was constructed, based on 354 markers, including 300 amplified fragment length polymorphism (AFLP) markers, 42 microsatellite (SSR) markers, and 12 randomly amplified polymorphism (RAPD) markers. Forty-seven linkage groups (LGs) were identified. The total map length was 4 580.5 cM, with an average spacing of 11.3 cM, covering 75.8% of the estimated genome size. The construction of this genetic linkage map was part of a genetic breeding program. This linkage map will contribute to the discovery of genes and quantitative trait loci (QTLs) in Chinese shrimp.




    Molecular markers are defined as the fragments of DNA sequence associated with a genome, which are used to identify a particular DNA sequence. Nowadays, with the explosive growth of genetic research and bacterial classification, molecular marker is an important tool to identify bacterial species. Taking account to its significant roles in clinic, medicine and food industry, in this review article, we summarize the traditional research and new development about molecular markers (also called g...

  1. Application of molecular markers in apple breeding

    Marić Slađana


    Full Text Available Apple is economically the most important species of genus Malus Miller. In respect of production, trade and consumption, it ranks first among deciduous fruit and third on a global scale among all fruit species. Apple breeding is carried out on a large scale in several scientific institutes throughout the world. Due to this activity, apple is a fruit species with the highest number of described monogenic traits; 76 genes, encoding morphological traits, pest and disease resistance, as well as 69 genes encoding enzymes. The development of molecular markers (RFLPs, AFLPs, SCARs and SSRs has allowed the mapping of the apple genome and the development of several saturated genetic maps, to which genes controlling important traits are assigned. Markers flanking these genes not only play an important role in selecting parental combinations and seedlings with positive traits, but they are also particularly important in detecting recessive traits, such as seedless fruit. In addition they enable pre-selection for polygenic quantitative traits. In recent years, particular attention has been paid to biochemical and physiological processes involved in the pathway of important traits e.g., ripening and the storage capability of apple fruit.

  2. Prediction of single-cross hybrid performance for grain yield and grain dry matter content in maize using AFLP markers associated with QTL.

    Schrag, T A; Melchinger, A E; Sørensen, A P; Frisch, M


    Prediction methods to identify single-cross hybrids with superior yield performance have the potential to greatly improve the efficiency of commercial maize (Zea mays L.) hybrid breeding programs. Our objectives were to (1) identify marker loci associated with quantitative trait loci for hybrid performance or specific combining ability (SCA) in maize, (2) compare hybrid performance prediction by genotypic value estimates with that based on general combining ability (GCA) estimates, and (3) investigate a newly proposed combination of the GCA model with SCA predictions from genotypic value estimates. A total of 270 hybrids was evaluated for grain yield and grain dry matter content in four Dent x Flint factorial mating experiments, their parental inbred lines were genotyped with 20 AFLP primer-enzyme combinations. Markers associated significantly with hybrid performance and SCA were identified, genotypic values and SCA effects were estimated, and four hybrid performance prediction approaches were evaluated. For grain yield, between 38 and 98 significant markers were identified for hybrid performance and between zero and five for SCA. Estimates of prediction efficiency (R (2)) ranged from 0.46 to 0.86 for grain yield and from 0.59 to 0.96 for grain dry matter content. Models enhancing the GCA approach with SCA estimates resulted in the highest prediction efficiency if the SCA to GCA ratio was high. We conclude that it is advantageous for prediction of single-cross hybrids to enhance a GCA-based model with SCA effects estimated from molecular marker data, if SCA variances are of similar or larger importance as GCA variances. PMID:16896712

  3. Using AFLP markers and the Geneland program for the inference of population genetic structure

    Guillot, Gilles; Santos, Filipe


    such as single nucleotide polymorphisms (SNP) markers but this difference becomes negligible for data sets of common size (number of individuals n≥100, number of markers L≥200). The latest Geneland version (3.2.1) handling dominant markers is freely available as an R package with a fully clickable graphical...

  4. An AFLP marker linked to the Pm-1 gene that confers resistance to Podosphaera xanthii race 1 in Cucumis melo

    Ana Paula Matoso Teixeira


    Full Text Available Brazil produced 330,000 metric tons of melons in 2005, principally in the Northeast region where one of the most important melon pathogens is the powdery mildew fungus Podosphaera xanthii. The disease is controlled mainly by incorporating single dominant resistance genes into commercial hybrids. We report on linkage analysis of the Pm-1 resistance gene, introgressed from the AF125Pm-1 Cantalupensis Charentais-type breeding line into the yellow-fleshed melon (Group Inodorus breeding line AF426-S by backcrossing to produce the resistant line AF426-R, and the amplified fragment length polymorphism (AFLP marker M75/H35_155 reported to be polymorphic between AF426-S and AF426-R. Segregation analysis of M75/H35_155 using a backcross population of 143 plants derived from [AF426-R x AF426-S] x AF426-S and screened for resistance to P. xanthii race 1 produced a recombination frequency of 4.9%, indicating close linkage between M75/H35_155 and Pm-1. Using the same segregating population, the M75/H35_155 marker had previously been reported to be distantly linked to Prv¹, a gene conferring resistance to papaya ringspot virus-type W. Since M75/H35_155 is linked to Prv¹ at a distance of 40.9 cM it is possible that Pm-1 and Prv¹ are also linked.

  5. AFLP genome scan in the black rat (Rattus rattus) from Madagascar: detecting genetic markers undergoing plague-mediated selection.

    Tollenaere, C; Duplantier, J-M; Rahalison, L; Ranjalahy, M; Brouat, C


    The black rat (Rattus rattus) is the main reservoir of plague (Yersinia pestis infection) in Madagascar's rural zones. Black rats are highly resistant to plague within the plague focus (central highland), whereas they are susceptible where the disease is absent (low altitude zone). To better understand plague wildlife circulation and host evolution in response to a highly virulent pathogen, we attempted to determine genetic markers associated with plague resistance in this species. To this purpose, we combined a population genomics approach and an association study, both performed on 249 AFLP markers, in Malagasy R. rattus. Simulated distributions of genetic differentiation were compared to observed data in four independent pairs, each consisting of one population from the plague focus and one from the plague-free zone. We found 22 loci (9% of 249) with higher differentiation in at least two independent population pairs or with combining P-values over the four pairs significant. Among the 22 outlier loci, 16 presented significant association with plague zone (plague focus vs. plague-free zone). Population genetic structure inferred from outlier loci was structured by plague zone, whereas the neutral loci dataset revealed structure by geography (eastern vs. western populations). A phenotype association study revealed that two of the 22 loci were significantly associated with differentiation between dying and surviving rats following experimental plague challenge. The 22 outlier loci identified in this study may undergo plague selective pressure either directly or more probably indirectly due to hitchhiking with selected loci. PMID:20444082

  6. Assessment of genetic diversity among Chinese upland cottons with Fusarium and/or Verticillium wilts resistance by AFLP and SSR markers

    WANG Xingfen; MA Jun; YANG Shuo; ZHANG Guiyin; MA Zhiying


    Genetic diversity among 95 Chinese upland cottons with Fusarium and/or Verticillium wilts resistance was estimated using Amplified Fragment Length Polymorphism (AFLP) and Simple Sequence Repeat (SSR) markers.Twenty EcoRI-MseI AFLP and 19 SSR primers with polymorphism were selected to perform the fingerprinting.The results showed that 20 AFLP primer pairs produced a total of 1 480 major bands among 95 genotypes,and 214 were polymorphic bands.The number of total bands per primer pair ranged from 47 to 109,with an average of 74.0.The polymorphism information content (PIC) values for the 20 primer pairs varied from 0.01 (E-ACT/M-CAT) to 0.24 (E-ACA/MCTA),and the average value was 0.09.Nineteen SSR primers generated 89 DNA bands,of which 61 were polymorphic.The total number of alleles per locus varied from 3 to 8,with an average of 4.7.The average PIC value for the SSR amplification products was 0.69.Genetic similarity estimates for the entire data set ranged from 0.978 to 0.998 based on AFLP and SSR bands.It was proved that the close genetic relationship and narrow genetic diversity existed in the tested cultivars.The clustering patterns could not be correlated to the geographic origin,the pedigree and common parentage of the cultivars.

  7. Assessment of genetic diversity in glandless cotton germplasm resources by using agronomic traits and molecular markers

    Zhikun LI; Xingfen WANG; Yan ZHANG; Guiyin ZHANG; Liqiang WU; Jina CHI; Zhiying MA


    Seventy-one glandless cotton germplasm resources were firstly evaluated genetically by using nine agronomic traits,33 simple sequence repeat (SSR) primers and ten amplified fragment length polymorphism (AFLP)primer combinations.Principal component analysis (PCA) of the agronomic traits showed that the first six principal components (PCs) explained a total of 86.352% of the phenotypic variation.A total of 329 alleles were amplified for 33 SSR primers,and 232 polymorphic bands in a total of 389 bands were obtained by using ten AFLP primer combinations.The average polymorphic information content (PIC) value was 0.80 and 0.18 for SSR primers and AFLP primer combinations,respectively.The DIST (average taxonomic distance) and DICE (Nei and Li's pairwise distance) coefficients ranged from 0.373 to 3.164 and 0.786 to 0.948,respectively,for agronomic traits and SSR & AFLP data based on UPGMA analysis.This suggested that there was a higher diversity in the evaluated population for both agronomic traits and molecular markers.The Mantel's test showed that the correlation between the dendrograms based on agronomic traits and SSR & AFLP data was non-significant.

  8. Diversity of garlic (Allium sativum L.) using SSR, EST and AFLP markers

    Germplasm from the center of origin/diversity is important for the breeding and fingerprinting crop plants. In this study we utilized both dominant and co-dominant markers for the characterization of garlic samples from diverse geographic origins to assess the relative utility of these markers to id...

  9. Duck (Anas platyrhynchos linkage mapping by AFLP fingerprinting

    Yang Kuo-Tai


    Full Text Available Abstract Amplified fragment length polymorphism (AFLP with multicolored fluorescent molecular markers was used to analyze duck (Anas platyrhynchos genomic DNA and to construct the first AFLP genetic linkage map. These markers were developed and genotyped in 766 F2 individuals from six families from a cross between two different selected duck lines, brown Tsaiya and Pekin. Two hundred and ninety-six polymorphic bands (64% of all bands were detected using 18 pairs of fluorescent TaqI/EcoRI primer combinations. Each primer set produced a range of 7 to 29 fragments in the reactions, and generated on average 16.4 polymorphic bands. The AFLP linkage map included 260 co-dominant markers distributed in 32 linkage groups. Twenty-one co-dominant markers were not linked with any other marker. Each linkage group contained three to 63 molecular markers and their size ranged between 19.0 cM and 171.9 cM. This AFLP linkage map provides important information for establishing a duck chromosome map, for mapping quantitative trait loci (QTL mapping and for breeding applications.

  10. Divergência genética entre genótipos de alface por meio de marcadores AFLP Genetics divergence among lettuce genotypes by AFLP markers

    Cristina Soares de Sousa


    Full Text Available Considerando a restrita diversidade de espécies disponíveis para nutrir a carência de vitaminas no Brasil, Kerr e colaboradores, desde 1981, vêm desenvolvendo pesquisas para melhoramento genético de hortaliças ricas em vitamina A. Dentre elas, obtiveram uma cultivar de alface, denominada Uberlândia 10.000 com 10.200 UI de vitamina A em 100 gramas de folha fresca. Este trabalho objetivou comparar o grau de divergência genética entre a cultivar Uberlândia 10.000 e seus parentais para avaliar a eficiência da seleção utilizada, por meio da técnica AFLP. Foram utilizados os seguintes genótipos de alface: Maioba, Salad Bowl-Mimosa, Moreninha-de-Uberlândia, Vitória de Santo Antão, Uberlândia 10.000 lisa 8.ª e 9.ª geração e Uberlândia 10.000 crespa 8.ª e 9.ª geração. A técnica AFLP foi eficiente para identificar genótipos muito próximos e para estudos de progênies em alface. O primer PR15 permitiu a separação da forma lisa e crespa com 1,8% de divergência genética e a oitava da nona geração com apenas 0,71%. Com o estudo da filogenia da cultivar pode-se observar que o programa de melhoramento foi desenvolvido com sucesso, pois a cultivar obtida Uberlândia 10.000 possui alto teor de vitamina A e 92% de similaridade com o parental Vitória de Santo Antão. O primer PR11 conseguiu identificar polimorfismo entre cultivares de alta e baixa resistência à septoriose, sugerindo a possibilidade destas bandas estarem relacionadas à resistência.Considering the restricted diversity of species available to counteract vitamin deficiencies in Brazil, Kerr and coworkers have been engaged since 1981, in developing genetic improved garden vegetables rich in vitamin A. One of these vegetables is the lettuce cultivar Uberlândia 10,000, which contains 10,200 UI of vitamin A per 100 grams of fresh leaves. This study compares the genetic diversity between Uberlândia 10,000 and its parental, evaluating selection efficiency through

  11. Identification of Amplified Fragment Length Polymorphism (AFLP Markers Tightly Associated with Drought Stress Gene in Male Sterile and Fertile Salvia miltiorrhiza Bunge

    Hongbo Guo


    Full Text Available Consistent grain yield in drought environment has attracted wide attention due to global climate change. However, the important drought-related traits/genes in crops have been rarely reported. Many near-isogenic lines (NILs of male sterile and fertile Salvia miltiorrhiza have been obtained in our previous work through testcross and backcross in continuous field experiments conducted in 2006–2009. Both segregating sterile and fertile populations were subjected to bulked segregant analysis (BSA and amplified fragment length polymorphism (AFLP with 384 and 170 primer combinations, respectively. One out of 14 AFLP markers (E9/M3246 was identified in treated fertile population as tightly linked to the drought stress gene with a recombination frequency of 6.98% and at a distance of 7.02 cM. One of 15 other markers (E2/M5357 was identified in a treated sterile population that is closely associated with the drought stress gene. It had a recombination frequency of 4.65% and at a distance of 4.66 cM. Interestingly, the E9/M3246 fragment was found to be identical to another AFLP fragment E11/M4208 that was tightly linked to the male sterile gene of S. miltiorrhiza with 95% identity and e-value 4 × 10−93. Blastn analysis suggested that the drought stress gene sequence showed higher identity with nucleotides in Arabidopsis chromosome 1–5.

  12. Genetic diversity analysis of common beans based on molecular markers

    Homar R. Gill-Langarica


    Full Text Available A core collection of the common bean (Phaseolus vulgaris L., representing genetic diversity in the entire Mexican holding, is kept at the INIFAP (Instituto Nacional de Investigaciones Forestales, Agricolas y Pecuarias, Mexico Germplasm Bank. After evaluation, the genetic structure of this collection (200 accessions was compared with that of landraces from the states of Oaxaca, Chiapas and Veracruz (10 genotypes from each, as well as a further 10 cultivars, by means of four amplified fragment length polymorphisms (AFLP +3/+3 primer combinations and seven simple sequence repeats (SSR loci, in order to define genetic diversity, variability and mutual relationships. Data underwent cluster (UPGMA and molecular variance (AMOVA analyses. AFLP analysis produced 530 bands (88.5% polymorphic while SSR primers amplified 174 alleles, all polymorphic (8.2 alleles per locus. AFLP indicated that the highest genetic diversity was to be found in ten commercial-seed classes from two major groups of accessions from Central Mexico and Chiapas, which seems to be an important center of diversity in the south. A third group included genotypes from Nueva Granada, Mesoamerica, Jalisco and Durango races. Here, SSR analysis indicated a reduced number of shared haplotypes among accessions, whereas the highest genetic components of AMOVA variation were found within accessions. Genetic diversity observed in the common-bean core collection represents an important sample of the total Phaseolus genetic variability at the main Germplasm Bank of INIFAP. Molecular marker strategies could contribute to a better understanding of the genetic structure of the core collection as well as to its improvement and validation.

  13. Similaridade genética de acessos de mangueira de diferentes origens geográficas avaliadas por marcadores AFLP Genetic similarity of mango accessions of different geographic origins evaluated with AFLP markers

    Carlos Antonio Fernandes Santos


    Full Text Available As relações genéticas de 105 acessos de diferentes origens geográficas do banco de germoplasma de mangueira da Embrapa foram determinadas com base no marcador AFLP, de forma a orientar trabalhos de melhoramento e manejo de recursos genéticos da espécie para a região Semi-Árida brasileira. Foram ainda incluídos dois acessos de duas espécies do gênero Mangifera, como "outgroup". O DNA dos acessos foi extraído pelo método do CTAB, as reações de AFLP foram realizadas para os iniciadores EcoRI/MseI e as bandas polimórficas foram analisadas para construção de fenograma, baseando-se no coeficiente de similaridade de Jaccard. Foram obtidas 157 e 54 bandas de AFLP polimórficas e monomórficas, respectivamente, em 13 combinações de iniciadores. O valor co-fenético do fenograma foi estimado em 0,81. Foram observados cinco grupos: 1 cultivares como Amrapali, Malika, híbridos Embrapa-Cpac e algumas variedades americanas formando um grupo; 2 grupo formado, predominantemente, por cultivares americanas, com algumas inclusões de híbridos sul-africanos e brasileiros; 3 grande grupo formado por cultivares brasileiras, com algumas inclusões de cultivares australianas, indianas e americanas; 4 grupo formado por algumas variedades tipo Espada, Rosa e acessos de diferentes origens; e 5 grupo formado por M. foetida e M. similis. Os acessos Carabao e Manilla apresentaram a maior similaridade, 97%. Os acessos estudados apresentaram similaridade superior a 51%, evidenciando a alta variabilidade genética da coleção de germoplasma de mangueira estudada.The genetic relationship among 105 mango accessions of different geographic origins of the Embrapa germplasm collection was estimated based on AFLP marker in order to orient breeding and management of genetic resource activities of this species to the Brazilian Semi-Arid region. Two additional accessions of other species of the Mangifera genus were also included as outgroup. The DNA of the

  14. A genetic linkage map of the Durum x Triticum dicoccoides backcross population based on SSRs and AFLP markers, and QTL analysis for milling traits.

    Elouafi, I; Nachit, M M


    Durum wheat ( Triticum turgidum L. var durum) is mainly produced and consumed in the Mediterranean region; it is used to produce several specific end-products; such as local pasta, couscous and burghul. To study the genetics of grain-milling quality traits, chromosomal locations, and interaction with the environment, a genetic linkage map of durum was constructed and the quantitative trait loci QTLs for the milling-related traits, test weight (TW) and thousand-kernel weight (TKW), were identified. The population constituted 114 recombinant inbred lines derived from the cross: Omrabi 5 /Triticum dicoccoides 600545// Omrabi 5. TW and TKW were analyzed over 18 environments (sites x years). Single-sequence-repeat markers (SSRs), Amplified-fragment-length-polymorphism markers (AFLPs), and seed storage proteins (SSPs) showed a high level of polymorphism (>60%). The map was constructed with 124 SSRs, 149 AFLPs and 6 SSPs; its length covered 2,288.8 cM (8.2 cM/marker). The map showed high synteny with previous wheat maps, and both SSRs and AFLPs mapped evenly across the genome, with more markers in the B genome. However, some rearrangements were observed. For TW, a high genotypic effect was detected and two QTLs with epistasic effect were identified on 7AS and 6BS, explaining 30% of the total variation. The TKW showed a significant transgressive inheritance and five QTLs were identified, explaining 32% of the total variation, out of which 25% was of a genetic nature, and showing QTLxE interaction. The major TKW-QTLs were around the centromere region of 6B. For both traits, Omrabi 5 alleles had a significant positive effect. This population will be used to determine other QTLs of interest, as its parents are likely to harbor different genes for diseases and drought tolerance. PMID:14676946

  15. Caracterización molecular de introducciones colombianas de caña flecha utilizando la técnica AFLP Molecular characterization of colombian wild cane accesions with AFLP

    Hernando Rivera Jiménez


    Full Text Available La fibra de la caña flecha Gynerium sagittatum (Aubl. se utiliza como materia prima para fabricar el "sombrero vueltiao", sombrero típico de la costa caribeña colombiana. Se realizó la caracterización molecular con AFLP para estimar variabilidad genética teniendo en cuenta criterios geográficos y morfológicos de 25 introducciones colombianas del banco de germoplasma de la Universidad de Córdoba. El análisis de correspondencia múltiple discriminó las introducciones en cuatro grupos, donde se identificaron características de importancia artesanal (comercial y atributos agronómicos. Se observó escasa correlación entre distancia geográfica y diferenciación genética, lo cual indicó flujos antrópicos por la reproducción asexual del material.Wild cane (Gynerium sagittatum Aubl. fiber is used as raw material for to make "hat vueltiao". A molecular characterization using AFLP was carriet out to estimate genetic variability of 25 accessions planted at the University of Cordoba, associated with geographical and morphological traits. Multiple discrimination correspondence analyses of introductions separated four groups, based on craft handling and agronomic attributes desirable. There was little correlation between geographic distance and genetic differentiation, indicating an anthropic flows by asexual reproduction.

  16. Sequence-Related Amplified Polymorphism (SRAP Markers: A Potential Resource for Studies in Plant Molecular Biology

    Daniel W. H. Robarts


    Full Text Available In the past few decades, many investigations in the field of plant biology have employed selectively neutral, multilocus, dominant markers such as inter-simple sequence repeat (ISSR, random-amplified polymorphic DNA (RAPD, and amplified fragment length polymorphism (AFLP to address hypotheses at lower taxonomic levels. More recently, sequence-related amplified polymorphism (SRAP markers have been developed, which are used to amplify coding regions of DNA with primers targeting open reading frames. These markers have proven to be robust and highly variable, on par with AFLP, and are attained through a significantly less technically demanding process. SRAP markers have been used primarily for agronomic and horticultural purposes, developing quantitative trait loci in advanced hybrids and assessing genetic diversity of large germplasm collections. Here, we suggest that SRAP markers should be employed for research addressing hypotheses in plant systematics, biogeography, conservation, ecology, and beyond. We provide an overview of the SRAP literature to date, review descriptive statistics of SRAP markers in a subset of 171 publications, and present relevant case studies to demonstrate the applicability of SRAP markers to the diverse field of plant biology. Results of these selected works indicate that SRAP markers have the potential to enhance the current suite of molecular tools in a diversity of fields by providing an easy-to-use. highly variable marker with inherent biological significance.

  17. Chinese Cabbage-pak-choi Transcriptome Map Construction with cDNA-AFLP Techniques

    FAN Shu-ying; LE Jian-gang; CHENG Guang-jie; WU Cai-jun


    Chinese cabbage-pak-choi (Brassica campestris L. ssp. chinensis Makino) transcriptome map with cDNA-APLP techniques was constructed. The inbred line Aijiaohuang 97-3-2, the inbred line Baimanjing 001-24 of turnip [B. campestris ssp. rapifera (Matzg.) Sinsk] and 183 F6 (recombinant inbred population) plants were used as experimental materials. cDNAs were synthesized from total RNA extracted from young leaves at rosette stage. 256 pairs of cDNA-AFLP primers were used to detect the polymorphisms between parents Aijiaohuang 97-3-2 and Baimanjing 001-24. 56 pairs of cDNA-AFLP primers with high polymorphisms were screened from 256 pairs of primer by DNA-AFLP techniques. The genetic diversity of parents and 183 F6 progenies was detected by 56 pairs of cDNA-AFLP primers. The segregation and distribution of cDNA-AFLPs molecular marker were analyzed to construct transcriptome map amongst parents and F6 plants. A total of 164 cDNA-AFLPs marker loci were mapped into 13 linkage groups which covered 1 401.2 cM with an average distance of 9.7 cM. It was the first transcriptome map of Chinese cabbage using cDNA-AFLP technique.

  18. Application of fluorescence-based semi-automated AFLP analysis in barley and wheat

    Schwarz, G.; Herz, M.; Huang, X.Q.; Michalek, W.; Jahoor, A.; Wenzel, G.; Mohler, V.

    Genetic mapping and the selection of closely linked molecular markers for important agronomic traits require efficient, large-scale genotyping methods. A semi-automated multifluorophore technique was applied for genotyping AFLP marker loci in barley and wheat. In comparison to conventional P-33-b...

  19. High-density Linkage Map of Cultivated Allotetraploid Cotton Based on SSR, TRAP, SRAP and AFLP Markers

    Jiwen Yu; Shuxun Yu; Cairui Lu; Wu Wang; Shuli Fan; Meizhen Song; Zhongxu Lin; Xianlong Zhang; Jinfa Zhang


    A high-density linkage map was constructed for an F2 population derived from an interspecific cross of cultivated allotetraploid species between Gossyplum hirsutum L. and G. barbadense L. A total of 186 F2 individuals from the interspecific cross of "CRI 36 × Hai 7124" were genotyped at 1 252 polymorphic loci including a novel marker system,target region amplification polymorphism (TRAP). The map consists of 1 097 markers, including 697 simple sequence repeats (SSRs), 171 TRAPs, 129 sequence-related amplified polymorphisms, 98 amplified fragment length polymorphisms, and two morphological markers, and spanned 4 536.7 cM with an average genetic distance of 4.1 cM per marker. Using 45 duplicated SSR loci among chromosomes, 11 of the 13 pairs of homologous chromosomes were identified in tetraplold cotton. This map will provide an essential resource for high resolution mapping of quantitative trait loci and molecular breeding in cotton.

  20. Molecular markers for thyroid cancer

    The importance of the study of the thyroid nodule lies in excluding the possibility of a malignant lesion because the majority of lesions are benign but there is a malignancy risk of 5 to 10%. Most of them are well differentiated carcinomas originating in the follicular epithelium. In spite of the fact that the majority are benign lesions, distinguishing them from carcinomas is crucial to treatment and adequate follow-up. Fine-needle biopsy allows making the diagnosis in most of cases. However, this method is restricted, particularly when diagnosing follicular lesions. In an effort to improve the diagnostic accuracy of biopsy and to provide new diagnosing criteria, a number of molecular markers have been put forward, some of which has wide range of approval whereas others still awaits to be validated for further implementation. This article presented an updated review of molecular markers with higher number of evidence, more accessible and potentially usable from a methodological viewpoint for diagnosis of the thyroid nodule before surgery. The importance of the study of the thyroid nodule lies in excluding the possibility of a malignant lesion because the majority of lesions are benign but there is a malignancy risk of 5 to 10%. Most of them are well differentiated carcinomas originating in the follicular epithelium. In spite of the fact that the majority are benign lesions, distinguishing them from carcinomas is crucial to treatment and adequate follow-up. Fine-needle biopsy allows making the diagnosis in most of cases. However, this method is restricted, particularly when diagnosing follicular lesions. In an effort to improve the diagnostic accuracy of biopsy and to provide new diagnosing criteria, a number of molecular markers have been put forward, some of which has wide range of approval whereas others still awaits to be validated for further implementation. This article presented an updated review of molecular markers with higher number of evidence, more

  1. Screening of AFLP markers related to growth traits in Tegillarca granosa%泥蚶生长性状相关AFLP分子标记的筛选

    董迎辉; 姚韩韩; 林志华; 肖国强; 柴雪良


    The bloody clam, Tegillarca granosa, is an important commercial bivalve which is extensively cultured from South Korea to Malaysia. Due to its economic value, it is important to cultivate rapid growth strains for the sustainable development of aquaculture industry of T. granosa. The emergence of molecular markers provides a rich theoretical basis for the rapid cultivation varieties. By the method of group selection, the rapid growth strains of T. granosa were bred though two generations. According to the comparison test, the rapid growth strains showed significant advantages in shell length, shell height, shell width and total weight than control group under the same environmental conditions. AFLP marker was applied in this study to evaluate the genetic variation of the rapid growth strains, and to screen the molecular markers associated with the traits of growth. A total of 2180 bands were amplified from 64 individuals of the rapid growth strain and control group by 40 primer combinations. The analysis of Nei's genetic diversity index and Shannon's genetic information index of two groups indicated that the genetic diversity of breeding group was slightly higher than the control group. The genetic distance between two groups was about 0.011 3, the value of GSt was 0.022 4 and Nm was 22.281 1, from which we concluded that there is a little genetic differentiation after selection. Of all amplified bands, 7 bands showed significant differences in frequency between two groups. 2 bands of all were only found in rapid growth strains, with the frequency of 0.812 5 and 0.343 8, respectively. And 2 bands showed significantly higher frequencies in rapid growth strains, while 3 bands showed significantly higher frequencies in control group. The bands with higher frequencies in rapid growth strains may be associated with the traits of growth. The unique AFLP markers associated with growth traits would be useful for growth-related gene cloning, quantitative trait locus (QTL

  2. Genetic Diversity in Musa acuminata Colla and Musa balbisiana Colla and some of their natural hybrids using AFLP Markers.

    Ude, G.; Pillay, M.; Nwakanma, D.; Tenkouano, A.


    Genetic diversity and relationships were assessed in 28 accessions of Musa acuminata (AA) Colla and Musa balbisiana (BB) Colla, and some of their natural hybrids, using the amplified fragment length polymorphisms (AFLP) technique. Fifteen AFLP +3 primer pairs produced 527 polymorphic bands among the accessions. Neighbor-joining and principal co-ordinate (PCO) analyses using Jaccard's similarity coefficient produced four major clusters that closely corresponded with the genome composition of the accessions (AA, BB, AAB and ABB). The AFLP data distinguished between the wild diploid accessions and suggested new subspecies relationships in the M. acuminata complex that are different from those based on morphological data. The data suggested that there are three subspecies within the M. acuminata complex (ssp. burmannica Simmonds, malaccensis Simmonds, and microcarpa Simmonds). 'Tjau Lagada' (ssp. microcarpa), 'Truncata' [ssp truncata (Ridl.) Shepherd] and 'SF247' [ssp. banksii (F.Muell) Simmonds] clustered very closely with 'Gros Michel' and 'Km 5', indicating that more than one M. acuminata subspecies may be involved in the origin of triploid AAA bananas. 'Calcutta 4' (ssp. burmannicoides De Langhe & Devreux) and 'Long Tavoy' (ssp. burmannica) were closely related and could be together in the same subspecies. This study also showed that there is much more genetic diversity within M. balbisiana that was split into two groups: (1) 'I-63' and 'HND' and (2) 'Los Banos', 'MPL' (Montpellier), '10852', 'Singapuri', 'Etikehel', and 'Butohan 1' as the other. PMID:12582577

  3. Veronaea botryosa: molecular identification with amplified fragment length polymorphism (AFLP) and in vitro antifungal susceptibility.

    Badali, Hamid; Yazdanparast, Seyed Amir; Bonifaz, Alexandro; Mousavi, Bita; de Hoog, G Sybren; Klaassen, Corné H W; Meis, Jacques F


    Inter- and intraspecific genomic variability of 18 isolates of Veronaea botryosa originating from clinical and environmental sources was studied using amplified fragment length polymorphism (AFLP). The species was originally described from the environment, but several severe cases of disseminated infection in apparently healthy individuals have been reported worldwide. All tested strains of V. botryosa, identified on the basis of sequencing and phenotypic and physiological criteria prior to our study, were confirmed by AFLP analysis, yielding a clear separation of V. botryosa as a rather homogeneous group from related species. In vitro antifungal susceptibility testing resulted in MIC90s across all strains in increasing order posaconazole (0.25 μg/ml), itraconazole (1 μg/ml), voriconazole (4 μg/ml), terbinafine (4 μg/ml), caspofungin (8 μg/ml), anidulafungin (8 μg/ml), isavuconazole (16 μg/ml), amphotericin B (16 μg/ml), and fluconazole (32 μg/ml). Overall, the isolates showed a uniform pattern of low MICs of itraconazole and posaconazole, but high MICs for remaining agents. The echinocandins (caspofungin and anidulafungin) had no activity against V. botryosa. There was no statistically significant difference between susceptibilities of environmental (n = 11) and clinical (n = 7) isolates of V. botryosa (P > 0.05). PMID:23463524

  4. Molecular Linkage Mapping and Marker-Trait Associations with NlRPT, a Downy Mildew Resistance Gene in Nicotiana langsdorffii.

    Zhang, Shouan; Gao, Muqiang; Zaitlin, David


    Nicotiana langsdorffii is one of two species of Nicotiana known to express an incompatible interaction with the oomycete Peronospora tabacina, the causal agent of tobacco blue mold disease. We previously showed that incompatibility is due to the hypersensitive response (HR), and plants expressing the HR are resistant to P. tabacina at all stages of growth. Resistance is due to a single dominant gene in N. langsdorffii accession S-4-4 that we have named NlRPT. In further characterizing this unique host-pathogen interaction, NlRPT has been placed on a preliminary genetic map of the N. langsdorffii genome. Allelic scores for five classes of DNA markers were determined for 90 progeny of a "modified backcross" involving two N. langsdorffii inbred lines and the related species N. forgetiana. All markers had an expected segregation ratio of 1:1, and were scored in a common format. The map was constructed with JoinMap 3.0, and loci showing excessive transmission distortion were removed. The linkage map consists of 266 molecular marker loci defined by 217 amplified fragment length polymorphisms (AFLPs), 26 simple-sequence repeats (SSRs), 10 conserved orthologous sequence markers, nine inter-simple sequence repeat markers, and four target region amplification polymorphism markers arranged in 12 linkage groups with a combined length of 1062 cM. NlRPT is located on linkage group three, flanked by four AFLP markers and one SSR. Regions of skewed segregation were detected on LGs 1, 5, and 9. Markers developed for N. langsdorffii are potentially useful genetic tools for other species in Nicotiana section Alatae, as well as in N. benthamiana. We also investigated whether AFLPs could be used to infer genetic relationships within N. langsdorffii and related species from section Alatae. A phenetic analysis of the AFLP data showed that there are two main lineages within N. langsdorffii, and that both contain populations expressing dominant resistance to P. tabacina. PMID:22936937

  5. AFLP and AMP Fingerprints as Markers to Evaluate Genetic Differences between Medicago truncatula Line Jemalong and 2HA, a New Line Produced by in vitro Culture Selection

    R.R. Irwanto


    Full Text Available A new line, Medicago truncatula cv. Jemalong 2HA (herein known as 2HA has been developed via repetitive regeneration and selection of M. truncatula cv. Jemalong. During somatic embryogenesis, 2HA produces 500 times more embryos than its progenitor, Jemalong. It is interesting to study if those two lines are isogenic or has genetic differences. The main objectives of the study was to evaluate the genotypic differences between Jemalong and 2HA also to evaluate the methylation event in 2HA utilized two DNA fingerprinting techniques, i.e AFLP fingerprints (Amplified Length of Polymorphism and AMP (Amplified Methylation Polymorphism. The results showed that AFLP analysis using eight primers combinations could not detect any differences between Jemalong and 2HA. However, using AMP methylation sensitive primers it could detect 15 polymorphisms out of 840 markers. These results lead to a conclusion that Jemalong and 2HA are isogenic lines. 2HA may have higher regeneration capacities due to methylation process which occurs during the production of 2HA through repetitive regeneration cycles.

  6. AFLP Fingerprint and SCAR Marker of Watermelon Core Collection%西瓜核心种质的AFLP指纹图谱和SCAR标记

    车克鹏; 许勇; 梁春阳; 宫国义; 翁曼丽; 张海英; 金德敏; 王斌


    西瓜(Citrullus lanatus (Thunb.) Mansf.)种质资源的鉴定与评价是对其有效利用的基础.以往的研究表明, 西瓜是一种遗传资源特别狭窄的作物,在用同工酶、RAPD及SSR技术对西瓜种质资源进行鉴定时,发现很难将品种完全区分开来.本研究利用高效可靠的AFLP技术,对30个西瓜核心种质材料进行了遗传分析,最终建立了这30个材料的DNA指纹图谱.在该图谱中,每个材料均有其独特的"指纹",材料之间可以相互区分开来.为了进一步利用AFLP分子标记,将重要抗病种质材料"PI296341"的AFLP特异带转化成了生产上可以直接利用的SCAR标记.%The identification of germplasm is an important step for effective utilization of the available germplasm. In previous studies, isoenzyme, RAPD and SSR techniques had been used to conduct the genetic identification of watermelon (Citrullus lanatus (Thunb.) Mansf.), but their effectiveness was limited due to the extremely narrow genetic background among watermelon genotypes. In this research, amplified fragment length polymorphism (AFLP), which was reported as a reliable technique with high efficiency in detecting polymorphism, was used to conduct genetic analysis and variety identification of thirty genotypes of watermelon core collection that represent a wide range of breeding and commercially available germplasm. As a result, a DNA fingerprint based on 15 bands amplified with four primer combinations was developed. In this fingerprint, each genotype has its unique fingerprint pattern and can be distinguished from each other. Furthermore, in order to facilitate the utilization of AFLP marker in practice, one specific AFLP band of genotype "PI296341" coming from fragment amplified by primer combination E-AT/M-CAT was successfully converted into a sequence characterized amplified region (SCAR) marker.

  7. Assessment of Genetic Stability Among In Vitro Plants of Arachis retusa Using RAPD and AFLP Markers for Germplasm Preservation

    Rachel Fatima Gagliardi; Luiz Ricardo Hanai; Georgia Pacheco; Carlos Alberto Oliveira; Leonardo Alves Carneiro; José Francisco Montenegro Valls; Elisabeth Mansur; Maria Lucia Carneiro Vieira


    Arachis retusa Krapov. et W. C. Gregory et Valls is endemic in the West-central region of Brazil, occurring in areas endangered by human actions. The establishment of in vitro preservation methods for wild species of Arachis isan alternative to seed banks for germplasm storage, multiplication and distribution. The risk of genetic changesinduced by tissue culture and the monitoring of the genetic stability of the biological material before, during andafter storage must be considered in the context of conservation. Random amplified polymorphic DNA (RAPD) andamplified fragment length polymorphism (AFLP) fingerprinting were used to evaluate the genetic stability of invitro plants originated from cotyledons and embryo axes of A. retusa. Cotyledons originated shoots through directorganogenesis and embryo axes displayed multishoot formation induced by 110 mmol/L and 8.8 mmol/L BAP,respectively. Ninety genomlc regions (loci) generated from RAPD and 372 from AFLP analyses were evaluated. Allamplified fragments detected by both techniques in plants derived from the two explant types were monomorphic.The results indicate that the recovered shoots are genetically stable at the assessed genomic regions.

  8. Molecular Markers: an Introduction and Applications

    Firas Rashad Al-Samarai


    Full Text Available The dramatic development of molecular genetics has laid the groundwork for genomics. It has introduced new generations of molecular markers for use in the genetic improvement of farm animals. These markers provide more accurate genetic information and better understanding of the animal genetic resources. Scientists, unfamiliar with the different molecular techniques tend to get lost as each has its own advantages and disadvantages. This review represents a trail to shade alight on the different types of molecular markers by introducing a brief summary on the development of genetic markers including both the classical genetic markers and more advanced DNA-based molecular markers. This review could be helpful to better understand the characteristics of different genetic markers and the genetic diversity of animal genetic resources.

  9. Prediction of hybrid performance in maize using molecular markers and joint analyses of hybrids and parental inbreds.

    Schrag, Tobias A; Möhring, Jens; Melchinger, Albrecht E; Kusterer, Barbara; Dhillon, Baldev S; Piepho, Hans-Peter; Frisch, Matthias


    The identification of superior hybrids is important for the success of a hybrid breeding program. However, field evaluation of all possible crosses among inbred lines requires extremely large resources. Therefore, efforts have been made to predict hybrid performance (HP) by using field data of related genotypes and molecular markers. In the present study, the main objective was to assess the usefulness of pedigree information in combination with the covariance between general combining ability (GCA) and per se performance of parental lines for HP prediction. In addition, we compared the prediction efficiency of AFLP and SSR marker data, estimated marker effects separately for reciprocal allelic configurations (among heterotic groups) of heterozygous marker loci in hybrids, and imputed missing AFLP marker data for marker-based HP prediction. Unbalanced field data of 400 maize dent x flint hybrids from 9 factorials and of 79 inbred parents were subjected to joint analyses with mixed linear models. The inbreds were genotyped with 910 AFLP and 256 SSR markers. Efficiency of prediction (R (2)) was estimated by cross-validation for hybrids having no or one parent evaluated in testcrosses. Best linear unbiased prediction of GCA and specific combining ability resulted in the highest efficiencies for HP prediction for both traits (R (2) = 0.6-0.9), if pedigree and line per se data were used. However, without such data, HP for grain yield was more efficiently predicted using molecular markers. The additional modifications of the marker-based approaches had no clear effect. Our study showed the high potential of joint analyses of hybrids and parental inbred lines for the prediction of performance of untested hybrids. PMID:19916002

  10. Diversidade genética entre híbridos de laranja-doce e tangor 'Murcott' avaliada por fAFLP e RAPD Genetic diversity among hybrids of sweet orange and 'Murcott' tangor evaluated by fAFLP and RAPD markers

    Marinês Bastianel


    Full Text Available O objetivo deste trabalho foi avaliar a diversidade genética em uma população de 148 híbridos de tangor 'Murcott' (Citrus reticulata Blanco x C. sinensis L. Osbeck e laranja 'Pêra' (C. sinensis L. Osbeck obtidos por polinização controlada, pelo uso de marcadores fAFLP e RAPD. Marcadores polimórficos (416 marcadores fAFLP e 33 RAPD foram utilizados para avaliar a similaridade genética entre os híbridos, calculada com o coeficiente Jaccard pelo método UPGMA. A consistência de cada agrupamento foi determinada pelo programa BOOD. Houve alta similaridade genética entre os parentais. A laranja 'Pêra' apresentou maior número (132 de loci em heterozigose em relação ao tangor 'Murcott' (105, corroborando a teoria de origem híbrida para a laranja-doce. Observaram-se dois grupos distintos de plantas, e um deles abrangeu 80% dos híbridos com maior similaridade com a laranja 'Pêra'. A análise bootstrap não revelou consistência estatística entre esses grupos. Marcadores fAFLP são mais eficientes na avaliação do polimorfismo, sendo indicados para seleção de indivíduos híbridos mais próximos a um dos parentais.The objective of this work was to evaluate the genetic diversity in a population of 148 hybrids of 'Murcott' tangor (Citrus reticulata Blanco x C. sinensis L. Osbeck and 'Pêra' sweet orange (C. sinensis L. Osbeck, obtained by controlled polination, using fAFLP and RAPD markers. Polymorphic markers (416 fAFLP and 33 RAPD markers were used to evaluate genetic similarity among the hybrids, calculated by the coefficient of Jaccard, using the UPGMA method. The consistency of each group was determined by software BOOD. There was high genetic similarity within the parents. 'Pêra' sweet orange had a higher number of loci in heterozygosis (132 compared to 'Murcott' tangor (105, supporting the theory of hybrid origin for sweet oranges. Two distinct groups of plants were observed: one group had 80% of the hybrids that displayed

  11. Molecular markers: tools to improve genebank efficiency

    Hintum, van T.J.L.; Treuren, van R.


    Possibilities for using molecular markers to improve genebank efficiency are increasingly present thanks to developments in genebanks and developments in molecular genetics. These possibilities relate to all aspects of genebank management: acquisition, maintenance, characterisation and utilisation.

  12. Identification of an AFLP marker linked to the stripe rust resistance gene Yr1O in wheat


    AFLP analysis of near-isogenic lines of the stripe rust resistance gene Yr10 was carried out with 6 Pst Ⅰprimers and 10 Taq Ⅰ -primers with the donor parent of Yr10 gene as the check. A total of about 4200 distinguishable bands were amplified, of which 5 were stable. The genetic linkage of the 5 polymorphic DNA fragments with the target gene were tested preliminarily on a segregating F2 population derived from a cross between the gene donor parent "Moro" and susceptible cultivar "Mingxian 169". The DNA fragment PT0502 was found closely linked to the Yr10 gene and cloned and sequenced. Based on the sequence specific primers for PCR were designed and synthesized. Genetic linkage analysis with 195 segregating F2 plants indicated that the genetic distance was 05 cM between the main product SC200 fragment produced by PCR with the primers and the Yr10 gene. The primers can be used to detect the Yr10 gene quickly, effectively and exactly.``

  13. Genetic variation and differentiation in wide ranging populations of razor clam ( Sinonovacula constricta) inferred from AFLP markers

    Wang, Jingbo; Li, Qi; Kong, Lingfeng


    The genetic variation and differentiation of the razor clam Sinonovacula constricta distributed along the coast of China were studied through amplified fragment length polymorphism (AFLP) analysis. Six primer combinations generated 193 fragments. The H e values varied from 0.322 to 0.463 and the percentage of polymorphic loci ranged from 74.1% to 98.4%, which indicates a high level of genetic diversity. Cluster analysis by Nei’s pairwise distance grouped all specimens by geographical origins. AMOVA consistently showed that genetic variation among populations was 8.71%, and most of the variation came from the genetic variation within populations (91.29%). Genetic differentiation among the six populations was moderate; pairwise F ST ranged from 0.0282 to 0.1480, which indicated that S. constricta populations along the coast of China are genetically connected. Among all the six populations, the Beihai population is the mostly differentiated from the others, suggesting that Hainan Island and Leizhou Peninsula act as barriers to gene flow. All populations abide isolation by distance model as indicated by Mantel test, except for ZS (Zhoushan) and YQ (Yueqing) populations. Information obtained in this study will provide guidelines for conservation and fishery management of this species in the future.

  14. AFLP and MS-AFLP analysis of the variation within saffron crocus (Crocus sativus L. germplasm.

    Matteo Busconi

    Full Text Available The presence and extent of genetic variation in saffron crocus are still debated, as testified by several contradictory articles providing contrasting results about the monomorphism or less of the species. Remarkably, phenotypic variations have been frequently observed in the field, such variations are usually unstable and can change from one growing season to another. Considering that gene expression can be influenced both by genetic and epigenetic changes, epigenetics could be a plausible cause of the alternative phenotypes. In order to obtain new insights into this issue, we carried out a molecular marker analysis of 112 accessions from the World Saffron and Crocus Collection. The accessions were grown for at least three years in the same open field conditions. The same samples were analysed using Amplified Fragment Length Polymorphism (AFLP and Methyl Sensitive AFLP in order to search for variation at the genetic (DNA sequence and epigenetic (cytosine methylation level. While the genetic variability was low (4.23% polymorphic peaks and twelve (12 effective different genotypes, the methyl sensitive analysis showed the presence of high epigenetic variability (33.57% polymorphic peaks and twenty eight (28 different effective epigenotypes. The pattern obtained by Factorial Correspondence Analysis of AFLP and, in particular, of MS-AFLP data was consistent with the geographical provenance of the accessions. Very interestingly, by focusing on Spanish accessions, it was observed that the distribution of the accessions in the Factorial Correspondence Analysis is not random but tends to reflect the geographical origin. Two clearly defined clusters grouping accessions from the West (Toledo and Ciudad Real and accessions from the East (Cuenca and Teruel were clearly recognised.

  15. Determination of borojo sex (Borojoa patinoi, Cuatrecasas through molecular markers

    Clara Inés Giraldo


    Full Text Available Borojó (Borojoa patinoi, Cuatrecasas, a rubiacea indigenous to the Colombian Pacific Region, produces a fleshy fruit having interesting nutritious characteristics making it a promising plant resource. It is used for fresh beverages; preserves and wine can also be obtained. It is an especially important source of income for some of the native population who sell it in local food markets and the main Colombian cities. Borojó is a dioecious plant; the two genders are not phenotypically distinguishable before flowering (3-4 years after planting. A non-selected plantation's productivity thus becomes substantially reduced. This project was aimed at determining gender-linked amplified fragment length polymorphism (AFLP markers. 20 and 22 bp primers were designed by sequencing one of these gender-linked markers abd used for determining the plants' gender via polymerase chain reaction (PCR. Samples examined by PCR and AFLP were compared; the techniques presented no differences when determining plant gender. Determining gender in borojó is an important tool for agriculture because it allows adequate selection and distribution of female and male plants for establishing greater productivity plantations. Key words: Borojoa patinoi, dioecious plant, AFLP, gender-linked marker, gender diagnosis.

  16. 黄瓜黄色子叶突变体遗传及相关AFLP标记%Genetic Analysis and AFLP Markers Associated to the Yellow Cotyledon Trait in Cucumber

    王惠哲; 李淑菊; 杨瑞环; 管炜


    With a F2and BC1 population derived from F1 of B180H( yellow cotyledon female) and Q12( green cotyledon male) was used as materials to study the inheritance of yellow cotyledon color and molecular marker associated to the cotyledon yellow trait. The results indicated that the yellow cotyledon trait was controlled by a pair of recessive gene,and the green cotyledon trait was in the mode of incomplete dominant inheritance. The polymorphism between yellow and green cotyledon parents of cucumber were studied using BSA method and AFLP technology. 1 024 AFLP markers were screened, and a co-dominant marker E-AG/M-AAG was obtained. Sequencing of the polymorphic fragment indicated that their lengths were 258 bp and 257 bp, respectively, and they were orthologous sequences with an insertion /deletion or mutant of only one nucleotide. The yellow cotyledon F2 plant possessed the 258 bp fragments, and the green cotyledon F2 plants possessed the 257 bp fragments or possessed both two fragments. The marker was closely linked to the cucumber cotyledon yellow color trait-related gene,and the marker was confirmed to be 96. 74% accurate by F2 individuals. The trait was a useful marker for hybridization and purity identification of F, hybrids.%以黄色子叶突变体B180H为母本,颜色正常子叶黄瓜Q12为父本,配制F1、F2、BC1,并通过对6世代子叶叶色特征的观察和统计分析,研究该黄色子叶性状的遗传规律,并利用BSA法、AFLP技术筛选与黄色子叶性状基因紧密连锁的分子标记.结果表明,黄瓜黄色子叶性状是由1对核基因控制的稳定遗传的隐性性状,绿色子叶相对于黄色为完全显性.通过BSA法和AFLP分子标记,应用1024对引物组合EcoR I-NN/Mse I-NNN对黄色、绿色子叶亲本进行筛选,筛选到了1个与黄瓜黄色子叶性状相关的共显性标记E-AG/M-AAG.测序结果表明,片段长度分别为258bp和257bp,为发生插入/缺失或突变的同源序列;在黄色子叶个体中只

  17. Highly Informative Single-Copy Nuclear Microsatellite DNA Markers Developed Using an AFLP-SSR Approach in Black Spruce (Picea mariana) and Red Spruce (P. rubens)

    Yong-Zhong Shi; Natascha Forneris; Rajora, Om P


    Background Microsatellites or simple sequence repeats (SSRs) are highly informative molecular markers for various biological studies in plants. In spruce (Picea) and other conifers, the development of single-copy polymorphic genomic microsatellite markers is quite difficult, owing primarily to the large genome size and predominance of repetitive DNA sequences throughout the genome. We have developed highly informative single-locus genomic microsatellite markers in black spruce (Picea mariana)...

  18. Identification and characterization of some aromatic rice mutants using amplified fragment length polymorphism (AFLP) technique

    Accurate identifying of the genotypes is considered one of the most important mechanisms used in the recording or the protection of plant varieties. The investigation was conducted at the experimental form belonging to the egyptian Atomic Energy Authority, Inshas. The aim was to evaluate grain quality characteristics and molecular genetic variation using Amplified Fragment Length Polymorphism (AFLP) technique among six rice genotypes, Egyptian Jasmine aromatic rice cultivar and five aromatic rice mutants in (M3 mutagenic generation). Two mutation (Egy22 and Egy24) were selected from irradiated Sakha 102 population with 200 and 400Gy of gamma rays in the M2 generation, respectively, and three mutations ( Egy32, Egy33, and Egy34) were selected from irradiated Sakha 103 population with 200, 300, 400Gy of gamma rays in the M2 generation, respectively. The obtained results showed that the strong aroma was obtained for mutant Egy22 as compared with Egyptian Jasmine rice cultivar (moderate aroma). Seven primer combinations were used through six rice genotypes on the molecular level using AFLP marker. The size of AFLP Fragments Were Ranged from 51- 494bp. The total number of amplified bands was 997 band among them 919 polymorphic bans representing 92.2%. The highest similarity index (89%) was observed between Egyptian Jasmine and Egy32 followed by (82%) observed between Egyptian Jasmine and Egy34. On the other hand, the lowest similarity index was (48%) between Egyptian Jasmine and Egy24. In six rice genotypes, Egy24 produced the highest number of the AFLP makers giving 49 unique markers (23 positive and 26 negative), then Egy22 showed 23 unique markers (27 positive and 6 negative) while Egy33 was characterized by 17 unique markers (12 positive and 5 negative). At last Egyptian Jasmine was discriminated by the lowest number of markets, 10 (6 positive and 4 negative). The study further confirmed that AFLP technique was able to differentiate rice genotypes by a higher number

  19. Mutant germplasm characterization using molecular markers. A manual

    Plant biotechnology applications must not only respond to the challenge of improving food security and fostering socio-economic development, but in doing so, promote the conservation, diversification and sustainable use of plant genetic resources for food and agriculture. Nowadays the biotechnology toolbox available to plant breeders offers several new possibilities for increasing productivity, crop diversification and production, while developing a more sustainable agriculture. This training course focuses on one of the most promising set of techniques used in modern crop improvement programmes, i.e. on molecular markers. These are rapidly being adopted by plant breeders and molecular biologists as effective and appropriate tools for basic and applied studies addressing biological components in agricultural production systems. Their use in applied breeding programmes can range from facilitating the appropriate choice of parents for crosses, to mapping/tagging of gene blocks associated with economically important traits (often termed 'quantitative trait loci' (QTLs)). Gene tagging and QTL mapping in turn permit marker-assisted selection (MAS) in backcross, pedigree, and population improvement programmes, thereby facilitating more efficient incremental improvement of specific individual target traits. And through comparative genomics, molecular markers can be used in ways that allow us to more effectively discover and efficiently exploit biodiversity and the evolutionary relationships between organisms. Substantial progress has been made in recent years in mapping, tagging and isolating many agriculturally important genes using molecular markers due in large part to improvements in the techniques that have been developed to help find markers of interest. Among the techniques that are particularly promising are Restriction Fragment Length Polymorphism (RFLPs), Amplified Fragment Length Polymorphism (AFLPs), Random Amplified Polymorphic DNA (RAPDs), Microsatellites

  20. AFLP studies on downy-mildew-resistant and downy-mildew-susceptible genotypes of opium poppy.

    Dubey, Mukesh K; Shasany, Ajit K; Dhawan, Om P; Shukla, Ashutosh K; Khanuja, Suman P S


    Downy mildew (DM) caused by Peronospora arborescens, is a serious disease in opium poppy (Papaver somniferum), which has a world-wide spread. The establishment of DM-resistant cultivars appears to be a sustainable way to control the In this paper, we present the results of a study aimed at the identification of amplified fragment length polymorphism (AFLP) markers for DM-resistance in opium poppy. Three opium poppy genotypes (inbred over about 10 years): Pps-1 (DM-resistant), Jawahar-16 (DM-susceptible) and H-9 (DM-susceptible) were crossed in a diallel manner and the F(1) progeny along with the parents were subjected to AFLP analysis of chloroplast (cp) and nuclear DNA with seven and nine EcoRI / MseI primer combinations, respectively. cpDNA AFLP analysis identified 24 Pps-1 (DM-resistant)-specific unique fragments that were found to be maternally inherited in both the crosses, Pps-1 x Jawahar-16 and Pps-1 x H-9. In the case of nuclear DNA AFLP analysis, it was found that 17 fragments inherited from Pps-1 were common to the reciprocal crosses of both (i) Pps-1 and Jawahar-16 as well as (ii) Pps-1 and H-9. This is the first molecular investigation on the identification of polymorphism between DM-resistant and DM-susceptible opium poppy genotypes and development of DM-resistant opium poppy genotypespecific AFLP markers. These AFLP markers could be used in future genetic studies for analysis of linkage to the downy mildew resistance trait. PMID:20505242


    I. S. Kunin


    Full Text Available The search of molecular markers of metastasing and prognosis in prostate cancer remains an urgent task. In this study, we investigated the relationship of gene expression heparanase-1 (HPSE1 and D-glucuronil C5-epimerase (GLCE with early disease relapse and metastasis of a 2,5−3 years after diagnosis. It was shown that the ratio of the expression levels of genes HPSE1/GLCE > 1 may serve as a prognostic relapse marker and trends of the tumour to metastasis. The data obtained suggest to use this option as a molecular marker for the diagnostics of metastatic process and the disease prognosis.

  2. AFLP markers for the R-gene in the flea beetle, Phyllotreta nemorum, conferring resistance to defenses in Barbarea vulgaris

    Breuker, C.J.; Victoir, K.; Jong, de P.W.; Meijden, van der E.; Brakefield, P.M.; Vrieling, K.


    A so-called R-gene renders the yellow-striped flea beetle Phyllotreta nemorum L. (Coleoptera: Chrysomelidae: Alticinae) resistant to the defenses of the yellow rocket Barbarea vulgaris R.Br. (Brassicacea) and enables it to use it as a host plant in Denmark. In this study, genetic markers for an auto

  3. Molecular markers of tumor invasiveness in ameloblastoma: An update

    Zhong, Yi; Guo, Wei; Wang, Li; CHEN, XINMING


    The aim of the present article was to review the current new knowledge on the molecular markers of tumor invasion in ameloblastoma. In this review, tumor molecular markers were identified and allocated to the following six groups according to their functions: (I) Markers involved in extracellular matrix degradation, (II) Molecular markers involved in cell adhesion lost, (III) Molecular markers involved in bone remodeling, (IV) Cytokines involved in angiogenesis, (V) Molecular markers related ...

  4. Some AFLP amplicons are highly conserved DNA sequences mapping to the same linkage groups in two F2 populations of carrot

    Santos Carlos A.F.


    Full Text Available Amplified fragment length polymorphism (AFLP is a fast and reliable tool to generate a large number of DNA markers. In two unrelated F2 populations of carrot (Daucus carota L., Brasilia x HCM and B493 x QAL (wild carrot, it was hypothesized that DNA 1 digested with the same restriction endonuclease enzymes and amplified with the same primer combination and 2 sharing the same position in polyacrylamide gels should be conserved sequences. To test this hypothesis AFLP fragments from polyacrylamide gels were eluted, reamplified, separated in agarose gels, purified, cloned and sequenced. Among thirty-one paired fragments from each F2 population, twenty-six had identity greater than 91% and five presented identity of 24% to 44%. Among the twenty-six conserved AFLPs only one mapped to different linkage groups in the two populations while four of the five less-conserved bands mapped to different linkage groups. Of eight SCAR (sequence characterized amplified regions primers tested, one conserved AFLP resulted in co-dominant markers in both populations. Screening among 14 carrot inbreds or cultivars with three AFLP-SCAR primers revealed clear and polymorphic PCR products, with similar molecular sizes on agarose gels. The development of co-dominant markers based on conserved AFLP fragments will be useful to detect seed mixtures among hybrids, to improve and to merge linkage maps and to study diversity and phylogenetic relationships.

  5. AFLP genome scan in the black rat (Rattus rattus) from Madagascar: detecting genetic markers undergoing plague-mediated selection.

    Tollenaere, C.; Duplantier, J.-M.; Rahalison, L.; Ranjalahy, M.; Brouat, C.


    The black rat (Rattus rattus) is the main reservoir of plague (Yersinia pestis infection) in Madagascar's rural zones. Black rats are highly resistant to plague within the plague focus (central highland), whereas they are susceptible where the disease is absent (low altitude zone). To better understand plague wildlife circulation and host evolution in response to a highly virulent pathogen, we attempted to determine genetic markers associated with plague resistance in this species. To this pu...

  6. Large-scale Gene Ontology analysis of plant transcriptome-derived sequences retrieved by AFLP technology

    Ramina Angelo


    Full Text Available Abstract Background After 10-year-use of AFLP (Amplified Fragment Length Polymorphism technology for DNA fingerprinting and mRNA profiling, large repertories of genome- and transcriptome-derived sequences are available in public databases for model, crop and tree species. AFLP marker systems have been and are being extensively exploited for genome scanning and gene mapping, as well as cDNA-AFLP for transcriptome profiling and differentially expressed gene cloning. The evaluation, annotation and classification of genomic markers and expressed transcripts would be of great utility for both functional genomics and systems biology research in plants. This may be achieved by means of the Gene Ontology (GO, consisting in three structured vocabularies (i.e. ontologies describing genes, transcripts and proteins of any organism in terms of their associated cellular component, biological process and molecular function in a species-independent manner. In this paper, the functional annotation of about 8,000 AFLP-derived ESTs retrieved in the NCBI databases was carried out by using GO terminology. Results Descriptive statistics on the type, size and nature of gene sequences obtained by means of AFLP technology were calculated. The gene products associated with mRNA transcripts were then classified according to the three main GO vocabularies. A comparison of the functional content of cDNA-AFLP records was also performed by splitting the sequence dataset into monocots and dicots and by comparing them to all annotated ESTs of Arabidopsis and rice, respectively. On the whole, the statistical parameters adopted for the in silico AFLP-derived transcriptome-anchored sequence analysis proved to be critical for obtaining reliable GO results. Such an exhaustive annotation may offer a suitable platform for functional genomics, particularly useful in non-model species. Conclusion Reliable GO annotations of AFLP-derived sequences can be gathered through the optimization

  7. Variation in Some Lycopersicon esculentum and Capsicum annuum Cultivars Revealed by RAPD and AFLP Markers%应用RAPD和AFLP标记的方法对甜(辣)椒和番茄品种的多态性分析

    李丽; 郑晓鹰; E.Klocke


    Twenty-five accessions of pepper (Capsicum annuum) and twenty-two accessions of tomato (Lycopersicon esculentum) cultivated in primary province where the main agricultural breeding insititutes breed the majority improved varieties of pepper and tomato in China were analysed. RAPD and AFLP markers were performed and compared for their effectiveness to discriminate near relatives. In pepper varieties, AFLP markers generated by two primer combinations distinguished every pepper accessions. The percentage of polymorphic markers for AFLP was lower than that for RAPD(9% and 35 % respectively). However, the average numbers of polymorphic fragments were 2.2 per primer for RAPD and 5.1 per AFLP primer combination,respectively. Therefore AFLP primers were two times more efficient than RAPD primers in their ability to generate polymorphic markers in pepper varietes. In tomato varieties,the percentages of polymorphic markers were 5.5% for AFLP and 61% for RAPD single primer and 58% for RAPD two-primer,the percentage of polymorphic markers of AFLP was lower than that for RAPD. The average numbers of informative bands per primer were 4.2 for RAPD single primer and 4.4 for RAPD two-primer and 2.6 for AFLP primer combinations,respectively. The study demonstrated that RAPD markers were more useful than AFLP markers among nearly related tomato varieties. RAPD markers generated by one single primer and one two-primer reactions can discriminate 21 our of 22 tomato varieties. The effectiveness of various marker techniques for the two species C. annuum and L. esculentum is discussed.%应用RAPD和AFLP的DNA指纹图谱方法分析25个甜(辣)椒和22个番茄品种的真实性,并进一步比较RAPD和AFLP的DNA指纹图谱在鉴定亲缘关系较近的品种之间的真实性时的有效性.对于甜(辣)椒品种,AFLP方法中,2个引物组合扩增反应的多态性片段即能将25个品种完全分开.虽然,每个样品的AFLP的扩增产物中多态性片段的百分率为9%,

  8. Use of molecular marker techniques in seed testing by Brazilian seed companies

    Della Vecchia P.T.


    Full Text Available Seed market is becoming global and globalization is growing very fast. To compete favourably in this new global seed world, quality and cost are and will be certanly the key issues. High seed quality can only be obtained by a thorough control of the entire seed production process, step by step from planning to final delivery. That requires science, technology, expertise, experience, good management and certanly, the most important, an absolute and unconditional commitment with quality. Seed testing for quality assurance is one important step in the process of production of high quality seed. In the late years a considerable amount of research has been published, particularly on the use of some Polymerase Chain Reaction DNA based new technologies (RAPD, microsatelites, AFLP for genetic purity determinations in seed testing. As far as we know, no Brazilian seed company is using, on regular basis, RAPD or other molecular marker techniques in the determination of genetic purity in seed testing. Most of these are using morphological or physiological traits expressed by seed, seedling or mature plant and/or electrophoresis of seed or seedling proteins/isoenzymes for that purpose. Main reasons for that are: DNA molecular marker techniques are relatively new; lack of specialized personnel to run DNA molecular marker assays on routine basis; higher cost/sample when compared to proteins/isoenzymes electrophoresis.

  9. Similaridade genética entre cultivares de cebola de diferentes tipos e origens, baseada em marcadores AFLP Genetic similarity among onion cultivars of different types and origins, based on AFLP markers

    CAF Santos


    Full Text Available Foi estimada a similaridade genética entre cultivares de cebola de diferentes tipos e regiões geográficas, de forma a orientar programas de recursos genéticos e melhoramento da espécie no Nordeste brasileiro. Foram analisadas 41 cultivares, adotando-se para a visualização da similaridade genética o fenograma UPGMA gerado da matriz de distâncias genéticas estimadas pelo coeficiente de Jaccard e baseadas em 146 bandas polimórficas de Pst1 e Mse1 de AFLP. A correlação cofenética foi de 0,91, indicando boa confiabilidade da representação gráfica para a interpretação dos resultados. Foram observados dois grupos principais no fenograma, no ponto de corte de 0,55 de similaridade: 1 grupo formado por cultivares predominantemente brasileiras, com algumas inclusões de cultivares estrangeiras; e 2 grupo formado por três cultivares estrangeiras (Mercedes, Perfect e TEG 502 PRR. Rijnsburger Jumbo e IPA 8 apresentaram a maior similaridade (85%, enquanto Madrugada foi a mais divergente em relação às demais cultivares. As cultivares da série IPA se dividiram em subgrupos no grupo das cultivares brasileiras (IPA 8, IPA 10 e IPA 11; IPA 12, IPA 7, IPA 2 e IPA 6; IPA 3, IPA 4 e IPA 9, indicando haver variabilidade genética a ser explorada entre aquelas situadas em subgrupos distintos. Bola Precoce e BRS Cascata apresentaram a maior similaridade entre as cultivares de origem brasileira. Foi observada similaridade superior a 39%, refletindo a alta variabilidade genética da coleção de cebola estudada. A introdução de novos acessos deve considerar procedências outras que não norte americanas, para aumentar a variabilidade de germoplasma de cebola disponível no Nordeste do Brasil.The genetic similarity among onion cultivars of different origins was evaluated, in order to carry out genetic resources and breeding programs for this species on the Brazilian Northeast. Forty-one onion cultivars were analyzed for 146 polymorphic Pst1/Mse1

  10. Functional molecular markers for crop improvement.

    Kage, Udaykumar; Kumar, Arun; Dhokane, Dhananjay; Karre, Shailesh; Kushalappa, Ajjamada C


    A tremendous decline in cultivable land and resources and a huge increase in food demand calls for immediate attention to crop improvement. Though molecular plant breeding serves as a viable solution and is considered as "foundation for twenty-first century crop improvement", a major stumbling block for crop improvement is the availability of a limited functional gene pool for cereal crops. Advancement in the next generation sequencing (NGS) technologies integrated with tools like metabolomics, proteomics and association mapping studies have facilitated the identification of candidate genes, their allelic variants and opened new avenues to accelerate crop improvement through development and use of functional molecular markers (FMMs). The FMMs are developed from the sequence polymorphisms present within functional gene(s) which are associated with phenotypic trait variations. Since FMMs obviate the problems associated with random DNA markers, these are considered as "the holy grail" of plant breeders who employ targeted marker assisted selections (MAS) for crop improvement. This review article attempts to consider the current resources and novel methods such as metabolomics, proteomics and association studies for the identification of candidate genes and their validation through virus-induced gene silencing (VIGS) for the development of FMMs. A number of examples where the FMMs have been developed and used for the improvement of cereal crops for agronomic, food quality, disease resistance and abiotic stress tolerance traits have been considered. PMID:26171816

  11. Fecal Molecular Markers for Colorectal Cancer Screening

    Rani Kanthan


    Full Text Available Despite multiple screening techniques, including colonoscopy, flexible sigmoidoscopy, radiological imaging, and fecal occult blood testing, colorectal cancer remains a leading cause of death. As these techniques improve, their sensitivity to detect malignant lesions is increasing; however, detection of precursor lesions remains problematic and has generated a lack of general acceptance for their widespread usage. Early detection by an accurate, noninvasive, cost-effective, simple-to-use screening technique is central to decreasing the incidence and mortality of this disease. Recent advances in the development of molecular markers in faecal specimens are encouraging for its use as a screening tool. Genetic mutations and epigenetic alterations that result from the carcinogenetic process can be detected by coprocytobiology in the colonocytes exfoliated from the lesion into the fecal matter. These markers have shown promising sensitivity and specificity in the detection of both malignant and premalignant lesions and are gaining popularity as a noninvasive technique that is representative of the entire colon. In this paper, we summarize the genetic and epigenetic fecal molecular markers that have been identified as potential targets in the screening of colorectal cancer.

  12. The prognostic molecular markers in hepatocellular carcinoma

    Lun-Xiu Qin; Zhao-You Tang


    The prognosis of hepatocellular carcinoma (HCC) stillremains dismal, although many advances in its clinicalstudy have been made. It is important for tumor control toidentity the factors that predispose patients to death. Withnew discoveries in cancer biology, the pathological andbiological prognostic factors of HCC have been studied quiteextensively. Analyzing molecular markers (biomarkers) withprognostic significance is a complementary method. A largenumber of molecular factors have been shown to associatewith the invasiveness of HCC, and have potential prognosticsignificance. One important aspect is the analysis ofmolecular markers for the cellular malignancy phenotypeThese include alterations in DNA ploidy, cellularproliferation markers (PCNA, Ki-67, Mcm2, MIB1, MIA, andCSE1L/CAS protein), nuclear morphology, the p53 geneand its related molecule MDM2, other cell cycle regulators(cyclin A, cyclin D, cyclin E, cdc2, p27, p73), oncogenesand their receptors (such as ras, c-myc, c-fms, HGF, c-met, and erb-B receptor family members ), apoptosisrelated factors (Fas and FasL), as well as telomeraseactivity. Another important aspect is the analysis ofmolecular markers involved in the process of cancerinvasion and metastasis. Adhesion molecules (E-cadherin,catenins, serum intercellular adhesion molecule-1, CD44variants), proteinases involved in the clegradation ofextracellular matrix (MMP-2, MMP-9, uPA, uPAR, PAl), aswell as other molecules have been regarded as biomarkersfor the malignant phenotype of HCC, and are related toprognosis and therapeutic outcomes. Tumor angiogenesisis critical to both the growth and metastasis of cancersincluding HCC, and has drawn much attention in recentyears. Many angiogenesis-related markers, such as vascularendothelial growth factor (VEGF), basic fibroblast growthfactor (bFGF), platelet-derived endothelial cell growth factor( PD-ECGF ), thrombospondin ( TSP ), angiogenin,pleiotrophin, and endostatin (ES) levels, as well asinratumor

  13. Identificação de marcas moleculares associadas à ausência de sementes em videira Identification of molecular markers associated to the absence of seeds in grapevine

    Ana Veruska Cruz da Silva


    Full Text Available A ausência de sementes tem sido uma característica bastante exigida pelos consumidores de uvas de mesa. O objetivo deste trabalho foi identificar marcas moleculares associadas à ausência de sementes, utilizando as técnicas RAPD e fAFLP. Foram utilizadas folhas jovens de 19 cultivares. Na análise RAPD 30, iniciadores possibilitaram amplificação de todas as amostras, produzindo 392 bandas polimórficas. Foi possível encontrar uma marca específica para a ausência de sementes, utilizando o iniciador UBC 443, que poderá futuramente ser utilizado para o desenvolvimento de marcadores SCAR, possibilitando a criação de um teste de identificação rápida e precoce de apirenia em videira. A análise fAFLP proporcionou a visualização de um dendrograma com grupos específicos de cultivares com sementes, sem sementes e porta enxertos.Seedless has been an important characteristic of table grapes required by consumers. The objective was to identify molecular markers associated to seedless, by RAPD and fAFLP techniques with young leaves samples of 19 cultivars. Thirty primers were used for RAPD analysis, producing a total of 337 polymorphic bands. It was also to find a specific mark for seedless, using UBC 443 primer. This mark would be transformed in a scar marker, making possible the early identification of seedless grape possible. The fAFLP analysis provided a visualization of a dendrogram with specific groups, separated in three different cultivars: with seeds, without seeds and the rootstocks.

  14. Molecular markers for detection, surveillance and prognostication of bladder cancer.

    Vrooman, O.P.; Witjes, J.A.


    Many markers for the detection of bladder cancers have been tested and almost all urinary markers reported are better than cytology with regard to sensitivity, but they score lower in specificity. Currently molecular and genetic changes play an important role in the discovery of new molecular marker

  15. Identification of DNA sequence variation in Campylobacter jejuni strains associated with the Guillain-Barré syndrome by high-throughput AFLP analysis

    Endtz Hubert P


    Full Text Available Abstract Background Campylobacter jejuni is the predominant cause of antecedent infection in post-infectious neuropathies such as the Guillain-Barré (GBS and Miller Fisher syndromes (MFS. GBS and MFS are probably induced by molecular mimicry between human gangliosides and bacterial lipo-oligosaccharides (LOS. This study describes a new C. jejuni-specific high-throughput AFLP (htAFLP approach for detection and identification of DNA polymorphism, in general, and of putative GBS/MFS-markers, in particular. Results We compared 6 different isolates of the "genome strain" NCTC 11168 obtained from different laboratories. HtAFLP analysis generated approximately 3000 markers per stain, 19 of which were polymorphic. The DNA polymorphisms could not be confirmed by PCR-RFLP analysis, suggesting a baseline level of 0.6% AFLP artefacts. Comparison of NCTC 11168 with 4 GBS-associated strains revealed 23 potentially GBS-specific markers, 17 of which were identified by DNA sequencing. A collection of 27 GBS/MFS-associated and 17 enteritis control strains was analyzed with PCR-RFLP tests based on 11 of these markers. We identified 3 markers, located in the LOS biosynthesis genes cj1136, cj1138 and cj1139c, that were significantly associated with GBS (P = 0.024, P = 0.047 and P Conclusion This study shows that bacterial GBS markers are limited in number and located in the LOS biosynthesis genes, which corroborates the current consensus that LOS mimicry may be the prime etiologic determinant of GBS. Furthermore, our results demonstrate that htAFLP, with its high reproducibility and resolution, is an effective technique for the detection and subsequent identification of putative bacterial disease markers.

  16. The role of Molecular Markers in Improvement of Fruit Crops

    Zahoor Ahmad BHAT


    Full Text Available Markers have been used over the years for the classification of plants. Markers are any trait of an organism that can be identified with confidence and relative easy, and can be followed in a mapping population on another hand markers be defined as heritable entities associated with the economically important trait under the control of polygenes. Morphological markers can be detected with naked eye (naked eye polymorphism or as difference in physical or chemical properties of the macromolecules. In other words, there are two types of genetic markers viz. morphological markers or naked eye polymorphism and non-morphological markers or molecular markers. Morphological markers include traits such as plant height, disease response, photoperiod, sensitivity, shape or colour of flowers, fruits or seeds etc. Molecular markers include biochemical constituents. Morphological markers have many limitations for being used as markers particularly in fruit crops because of long generation time and large size of fruit trees besides being influenced by environment. Consequently, molecular markers could be appropriate choice to study and preserve the diversity in any germplasm. Molecular markers have diverse applications in fruit crop improvement, particularly in the areas of genetic diversity and varietal identification studies, gene tagging, disease diagnostics, pedigree analysis, hybrid detection, sex differentiation and marker assisted selection.

  17. Biological (molecular and cellular) markers of toxicity

    Shugart, L.R.; D' Surney, S.J.; Gettys-Hull, C.; Greeley, M.S. Jr.


    Several molecular and cellular markers of genotoxicity were adapted for measurement in the Medaka (Oryzias latipes), and were used to describe the effects of treatment of the organism with diethylnitrosamine (DEN). NO{sup 6}-ethyl guanine adducts were detected, and a slight statistically significant, increase in DNA strand breaks was observed. These results are consistent with the hypothesis that prolonged exposure to high levels of DEN induced alkyltransferase activity which enzymatically removes any O{sup 6}-ethyl guanine adducts but does not result in strand breaks or hypomethylation of the DNA such as might be expected from excision repair of chemically modified DNA. Following a five week continuous DEN exposure with 100 percent renewal of DEN-water every third day, the F values (DNA double strandedness) increased considerably and to similar extent in fish exposed to 25, 50, and 100 ppM DEN. This has been observed also in medaka exposed to BaP.

  18. Biological (molecular and cellular) markers of toxicity

    Several molecular and cellular markers of genotoxicity were adapted for measurement in the Medaka (Oryzias latipes), and were used to describe the effects of treatment of the organism with diethylnitrosamine (DEN). NO6-ethyl guanine adducts were detected, and a slight statistically significant, increase in DNA strand breaks was observed. These results are consistent with the hypothesis that prolonged exposure to high levels of DEN induced alkyltransferase activity which enzymatically removes any O6-ethyl guanine adducts but does not result in strand breaks or hypomethylation of the DNA such as might be expected from excision repair of chemically modified DNA. Following a five week continuous DEN exposure with 100 percent renewal of DEN-water every third day, the F values (DNA double strandedness) increased considerably and to similar extent in fish exposed to 25, 50, and 100 ppM DEN. This has been observed also in medaka exposed to BaP

  19. Análise da diversidade genética por AFLP e identificação de marcadores associados à resistência a doenças em videira Genetic diversity and identification of AFLP markers associated with diseases resistance in grapevine

    Paulo Ricardo Dias de Oliveira


    Full Text Available Com objetivo de estudar diversidade genética e identificar marcadores associados à resistência ao míldio (Plasmopara viticola e ao oídio (Uncinula necator, foram analisadas as cultivares de videira A 1976, CG 87746, CNPUV 154-27, Crimson Seedless, Gota de Ouro, Itália, Seyve Villard 12327 e Seyve Villard 12375. As análises de polimorfismo de comprimento de fragmento amplificado (AFLP seguiram as etapas de digestão, ligação, pré-amplificação e amplificação. Efetuou-se a separação dos fragmentos amplificados em gel de 7% de poliacrilamida desnaturante (uréia 7M e coloração com nitrato de prata. A similaridade foi estimada com base no coeficiente de Jaccard, e a agregação, por UPGMA. Foi gerada uma matriz de similaridade com bom ajustamento da agregação (r = 0,84. Os agrupamentos obtidos corresponderam à origem e classificação botânica das cultivares. Foram identificadas 15 marcas dissimilares associadas à resistência, sendo oito para míldio e sete para oídio.The grapevine cultivars A 1976, CG 87746, CNPUV 154-27, Crimson Seedless, Gota de Ouro, Itália, Seyve Villard 12327 and Seyve Villard 12375 were analyzed to establish the genetic relationship and to identify markers associated with resistance to downy mildew (Plasmopara viticola and powdery mildew (Uncinula necator. The amplified fragment lenght polymorphism (AFLP analysis comprise the steps of digestion, ligation, preamplification and amplification. The amplified fragments were separeted on a 7% denaturing polyacrylamide gel (7M urea and silver stained. The similarity was estimated using Jaccard's coefficient and the clustering was estimated by UPGMA. A similarity matrix was generated with a good fit for aggregation (r = 0,84. The clusters corresponded to origin and botany classification of the cultivars. Fifteen markers were associated with resistance, eight for downy mildew and seven for powdery mildew.

  20. Application of molecular markers in livestock improvement

    Teneva A.; Petrović M.P.


    With recent developments in DNA technologies, a large number of genetic polymorphisms at DNA sequence level has been introduced over the last decades as named DNA-based markers. The discovery of new class of DNA profiling markers has facilitated the development of marker-based gene tags, mapbased cloning of livestock important genes, variability studies, phylogenetic analysis, synteny mapping, marker-assisted selection of favourable genotypes, etc. The most commonly used DNA-based markers hav...

  1. The role of molecular markers and marker assisted selection in breeding for organic agriculture

    Lammerts van Bueren, E.T.; Backes, G.; de Vriend, H.;


    Plant geneticists consider molecular marker assisted selection a useful additional tool in plant breeding programs to make selection more efficient. Standards for organic agriculture do not exclude the use of molecular markers as such, however for the organic sector the appropriateness of molecular...... markers is not self-evident and is often debated. Organic and low-input farming conditions require breeding for robust and flexible varieties, which may be hampered by too much focus on the molecular level. Pros and contras for application of molecular markers in breeding for organic agriculture was the...... topic of a recent European plant breeding workshop. The participants evaluated strengths, weaknesses, opportunities, and threats of the use of molecular markers and we formalized their inputs into breeder’s perspectives and perspectives seen from the organic sector’s standpoint. Clear strengths were...

  2. Molecular marker-based prediction of hybrid performance in maize using unbalanced data from multiple experiments with factorial crosses.

    Schrag, Tobias A; Möhring, Jens; Maurer, Hans Peter; Dhillon, Baldev S; Melchinger, Albrecht E; Piepho, Hans-Peter; Sørensen, Anker P; Frisch, Matthias


    In hybrid breeding, the prediction of hybrid performance (HP) is extremely important as it is difficult to evaluate inbred lines in numerous cross combinations. Recent developments such as doubled haploid production and molecular marker technologies have enhanced the prospects of marker-based HP prediction to accelerate the breeding process. Our objectives were to (1) predict HP using a combined analysis of hybrids and parental lines from a breeding program, (2) evaluate the use of molecular markers in addition to phenotypic and pedigree data, (3) evaluate the combination of line per se data with marker-based estimates, (4) study the effect of the number of tested parents, and (5) assess the advantage of haplotype blocks. An unbalanced dataset of 400 hybrids from 9 factorial crosses tested in different experiments and data of 79 inbred parents were subjected to combined analyses with a mixed linear model. Marker data of the inbreds were obtained with 20 AFLP primer-enzyme combinations. Cross-validation was used to assess the performance prediction of hybrids of which no or only one parental line was testcross evaluated. For HP prediction, the highest proportion of explained variance (R (2)), 46% for grain yield (GY) and 70% for grain dry matter content (GDMC), was obtained from line per se best linear unbiased prediction (BLUP) estimates plus marker effects associated with mid-parent heterosis (TEAM-LM). Our study demonstrated that HP was efficiently predicted using molecular markers even for GY when testcross data of both parents are not available. This can help in improving greatly the efficiency of commercial hybrid breeding programs. PMID:19048224


    The historicity of marker development, map construction, and the utility of marker-assisted selection is presented. Experimental results indicate that the identification of marker-trait associations will continue to be extremely expensive and time consuming, but will likely pay large dividends for ...

  4. Estimating genetic diversity and sampling strategy for a wild soybean (Glycine soja) population based on different molecular markers

    CHEN Zhong; ZHAO Ru; GU Senchang; YAN Wen; CHENG Zhou; CHEN Muhong; LU Weifeng; WANG Shuhong; LU Baorong; LU Jun; ZHANG Fan; XIANG Rong; XIAO Shangbin; YAN Pin


    Genetic diversity is the basic and most important component of biodiversity. It is essential for the effective conservation and utilization of genetic resources to accurately estimate genetic diversity of the targeted species and populations. This paper reports analyses of genetic diversity of a wild soybean population using three molecular marker technologies (AFLP, ISSR and SSR), and computer simulation studies of randomly selected subsets with different sample size (5-90 individuals) drawn 50 times from a total of 100 wild soybean individuals. The variation patterns of genetic diversity indices, including expected heterozygosity (He), Shannon diversity index (/), and percentage of polymorphic loci (P), were analyzed to evaluate changes of genetic diversity associated with the increase of individuals in each subset. The results demonstrated that (1) values of genetic diversity indices of the same wild soybean population were considerably different when estimated by different molecular marker techniques; (2) genetic diversity indices obtained from subsets with different sample sizes also diverged considerably; (3) P values were relatively more reliable for comparing genetic diversity detected by different molecular marker techniques; and (4) different diversity indices reached 90% of the total genetic diversity of the soybean population quite differently in terms of the sample size (number of individuals) analyzed.When using the P value as a determinator, 30-40individuals could capture over 90% of the total genetic diversity of the wild soybean population. Results from this study provide a strong scientific basis for estimating genetic diversity and for strategic conservation of plant species.

  5. AFLP marker analysis revealing genetic structure of the tree Parapiptadenia rigida (Benth. Brenan (Leguminosae-Mimosoideae in the southern Brazilian Tropical Rainforest

    Laís Bérgamo de Souza


    Full Text Available Parapiptadenia rigida is a tropical early secondary succession tree characteristic of the Tropical Atlantic Rainforest. This species is of great ecological importance in the recovery of degraded areas. In this study we investigated the variability and population genetic structure of eight populations of P. rigida. Five AFLP primer combinations were used in a sample of 159 individuals representing these eight populations, rendering a total of 126 polymorphic fragments. The averages of percentage of polymorphic loci, gene diversity, and Shannon index were 60.45%, 0.217, and 0.322, respectively. A significant correlation between the population genetic variability and the population sizes was observed. The genetic variability within populations (72.20% was higher than between these (22.80%. No perfect correlation was observed between geographic and genetic distances, which might be explained by differences in deforestation intensities that occurred in these areas. A dendrogram constructed by the UPGMA method revealed the formation of two clusters, these also confirmed by Bayesian analysis for the number of K cluster. These results show that it is necessary to develop urgent management strategies for the conservation of certain populations of P. rigida, while other populations still preserve reasonably high levels of genetic variability.

  6. Molecular markers for use in plant molecular breeding and germplasm evaluation

    A number of molecular marker technologies exist, each with different advantages and disadvantages. When available, genome sequence allows for the development of greater numbers and higher quality molecular markers. When genome sequence is limited in the organism of interest, related species may serve as sources of molecular markers. Some molecular marker technologies combine the discovery and assay of DNA sequence variations, and therefore can be used in species without the need for prior sequence information and up-front investment in marker development. As a prerequisite for marker-assisted selection (MAS), there must be a known association between genetic markers and genes affecting the phenotype to be modified. Comparative databases can facilitate the transfer of knowledge of genetic marker-phenotype association across species so that discoveries in one species may be applied to many others. Further genomics research and reductions in the costs associated with molecular markers will continue to provide new opportunities to employ MAS. (author)

  7. Molecular genetic studies in flax (Linum usitatissimum L.)

    Vromans, J


    In this thesis five molecular genetic studies on flax ( Linum usitatissimum L.) are described, of which two chapters aim to characterize the genetic structure and the amount of genetic diversity in the primary and secondary gene pool of the crop species. Three chapters describe the development of AFLP markers, linkage map construction and QTL analysis of resistance and quality traits.Genetic diversity in the primary gene pool was studied by AFLP fingerprinting 110 varieties representing linse...

  8. Comparison of levels of genetic diversity detected with AFLP and microsatellite markers within and among mixed Q. petraea (MATT.) LIEBL. and Q. robur L. stands

    Mariette, S.; Cottrell, J.; Csaikl, U.M.; Goikoechea, P.; Konig, A.; Lowe, A.J.; Dam, van B.C.; Barreneche, T.; Bodenes, C.; Streiff, R.; Burg, K.; Groppe, K.; Munro, R.C.; Tabbener, H.; Kremer, A.


    The aim of this study was to compare genetic diversity within and among Quercus spp. populations based on two contrasting types of nuclear markers. Seven mixed stands of Quercus petraea and Quercus robur were analysed using six highly polymorphic and codominantly inherited microsatellite markers as

  9. Genetic structure in cultivated and wild carrots (¤Daucus carota¤ L.) revealed by AFLP analysis

    Shim, S.I.; Bagger Jørgensen, Rikke


    Genetic variation within and among five Danish populations of wild carrot and five cultivated varieties was investigated using amplified fragment length polymorphism (AFLP). Ten AFLP primer combinations produced 116 polymorphic bands. Based on the marker data an UPGMA-cluster analysis and principal...... markers specific to the cultivated carrot makes it possible to detect introgression from cultivated to wild types....

  10. Diversidad genética intra e inter-específica de ñame (Dioscorea spp. de la región Caribe de Colombia mediante marcadores AFLP Genetic diversity intra and inter-specific yam (Dioscorea spp. from the colombian caribbean region by AFLP markers

    Hernando Javier Rivera-Jiménez


    Full Text Available Conocer la variabilidad genética del ñame, Dioscorea spp., permite apoyar estrategias de mejoramiento y conservación de este recurso fitogenético. El objetivo de este estudio fue la caracterización molecular de 20 accesiones de Dioscorea spp. mediante la técnica molecular de AFLP para determinar cómo se distribuye la variabilidad genética de manera intra e inter-específica. Los datos fueron analizados mediante los métodos de agrupación de correspondencia múltiple y análisis de similaridad de Dice, estableciendo los niveles de confiabilidad de los grupos genéticos mediante remuestreos. En términos de diversidad interespecífica, los valores promedios de similitud variaron entre 41.81% entre D. alata L. y D. rotundata Poir., y 33.51% entre D. trifida L.f. y D. esculenta (Lour. Burkill, lo que sugiere alta diversidad genética entre las accesiones estudiadas, que formaron cuatro grupos genéticos: D. alata, D. rotundata, D. esculenta y D. trifida, confirmando correspondencia entre la caracterización morfológica, clasificación botánica y la caracterización molecular. En términos de diversidad intraespecífica para la especie D. alata, el análisis también reveló una composición heterogénea en la región Caribe colombiana. Estos estudios ayudarán a definir una estrategia adecuada para fines de conservación y apoyar los esfuerzos futuros en los programas de mejoramiento genético.Knowing the genetic variability of yams, Dioscorea spp., is a good tool to support development and conservation strategies of this plant as genetic resource. The aim of this study was to carried out the molecular characterization of 20 accessions of Dioscorea spp. using the AFLP molecular technique to determine how genetic variation is distributed intra-and inter-specifically. Using multiple correspondence analysis and level of reliability of the genetic groups by resampling, the results showed high genetic variability among the accessions studied

  11. Construction of AFLP Molecular Marking System in Mangifera indica L.%芒果AFLP分子标记体系的建立

    王园; 金志强; 陈业渊; 雷新涛


    物组合多态性最大,达100%,可用于芒果品种指纹图谱构建.[结论]利用AFLP可以高效检测芒果种质资源分子标记多样性.%[Objective]The study aimed to construct the AFLP molecular marking system in Mangifera indica. [Method] Four varieties of Mangifera indica were used to explore new ways for high-quality DNA, and AFLP analysis of 31 varieties of Mangifera indica was carried out to detect the varietal genetic diversity. [Result] 14 pairs of primers with stronger polymorphism, better banding patterns and higher resolution were screened out from 64 pairs of selective amplification primers. Then they were used to analyse the fingerprint of 31 varieties of Mangifera indica, the results showed that the ratio of polymorphic bands amplificated by the 14 pairs of primers reached 97% in 31 varieties of Mangifera. [Conclusion] It was suggested that AFLP was suitable for detecting the polymorphism of Mangifera indica resources.

  12. Separation of the genera in the subtribe Cassiinae (Leguminosae: Caesalpinioidae using molecular markers Separação dos gêneros na subtribo Cassiinae (Leguminosae: Caesalpinioidae utilizando marcadores moleculares

    Laxmikanta Acharya


    Full Text Available Random amplified polymorphic DNA (RAPD, Inter simple sequence repeat (ISSR and Amplified fragment length polymorphism (AFLP markers were used to verify the segregation of the genus Cassia L. senso lato into three distinct genera namely Chamaecrista Moench., Senna P. Mill. and Cassia L. sensostricto Eighteen representatives of the three taxa were characterized using the molecular markers. 25 RAPD, six ISSR primers and six AFLP primer combinations resulted in the amplification of 612, 115 and 622 bands (loci respectively. Most of the loci are found to be polymorphic, showing high degrees of genetic diversity among the different taxa studied. The dendrogram constructed on the basis of the RAPD, ISSR and AFLP data using SHAN clustering, divided Cassia L. senso lato. into three different clusters as Chamaecrista Moench. Senna P. Mill. and Cassia L. senso stricto High bootstrap value revealed that all the clusters were stable and robust. It was observed from the present investigation that these genera have their identity at molecular level, which supports the elevation of the genus Cassia L. senso lato to the level of subtribe Cassiinae and segregation into three distinct genera instead of intrageneric categories.Técnicas de Random amplified polymorphic DNA (RAPD, Inter simple sequence repeat (ISSR e Amplified Fragment Length Polymorphism markers (AFLP foram utilizadas para verificar a segregação do gênero Cassia L. senso lato em três diferentes gêneros, Chamaecrista Moench., Senna P. Mill. e Cassia L. senso stricto Dezoito representantes dos três táxons foram caracterizados com o uso de marcadores moleculares: 25 RAPD, seis iniciadores ("primers" ISSR e seis AFLP combinações de iniciadores, resultando na amplificação de 612, 115 e 622 bandas (loci, respectivamente. A maioria dos loci apresentou-se como polimórfico, mostrando um alto grau de diversidade genética entre os táxons estudados. O dendrograma construído com base nos dados de

  13. Modified techniques used for microsatellite and AFLP for the population study of divers species at the Sinu river fish, Colombia

    Lamprea Natalia


    Full Text Available Few genetic conservation studies carried out in Colombia have applied molecular biology techniques. Protocols and modifications arising from the genetic study of four fish species from the Sinú river basin are presented as guidelines for future work in this field (and with other species as laboratory standardisation processes are complex. The original commercial kit's DNA extraction protocols were modified as were those for PCR reactions for obtaining micro-satellite markers and AFLPs, as well as allele genotypification. Particular variations were made f or each of the species studied. Key words: DNA extraction; molecular biology; genotypification; ichthyology.

  14. Promise and pitfalls of molecular markers of thyroid nodules

    S Jadhav


    Full Text Available Thyroid nodules are common in the general population with a prevalence of 5-7% The initial evaluation of thyroid nodules commonly involves thyroid function tests, an ultrasound (USG and fine needle aspiration biopsy (FNAB. The optimal management of patients with thyroid nodules with indeterminate cytology is plagued by the lack of highly sensitive and specific diagnostic modalities In this article we attempt to review the available literature on the molecular markers which are increasingly being studied for their diagnostic utility in assessing thyroid nodules. The various molecular markers consist of gene mutations, gene re arrangements, RNA based assays and immunohistochemical markers. The molecular markers definitely would help to optimise the management of such patients.

  15. New molecular marker technologies for pearl millet improvement

    MD Gale


    Full Text Available The first molecular marker-based genetic linkage map of pearl millet (Pennisetum glaucum was built with restriction fragment length polymorphisms (RFLP, the marker system of choice in the early 1990s. This map has served as the basis for subsequent pearl millet marker-based studies at the John Innes Centre. The RFLP framework in the consensus map is presented and is based on 173 (out of 500 available mapped PstI genomic clones from inbred line Tift 23DB, which has now become the base genotype for pearl millet molecular genetics. Molecular marker techniques have now moved on particularly with the development of polymerase chain reaction (PCR and a programme at ICRISAT has developed the first microsatellite markers (simple sequence repeats; SSR. Some 100 markers, of which 60 are mapped, are available either as DNA primers or as DNA sequences of the flanking regions of the SSR. The integration of the pearl millet map in the grass consensus map is briefly described along with the establishment of the plant genome database MilletGenes. MilletGenes collates as genome related data (maps, markers, DNA sequences and images on pearl millet, finger millet (Eleusine coracana, foxtail millet (Setaria italica and tef (Eragrostis tef.

  16. Productivity improvement in cotton using molecular and nuclear strategies

    This is the first research performed in Peru with the objective of knowing the genetic variability among 27 cotton accessions belonging to the Canete Germplasm Bank using molecular markers (AFLP). Two of out nine AFLP combinations (E1 M1 and E2 M2) were selected as candidates of a panel of molecular markers for the molecular characterization of Peruvian cotton. Moreover, 36,000 cotton Tanguis seeds were irradiated with γ-rays by using three doses: 150 Gy, 250 Gy, and 350 Gy. Mutant plants belonging to the first generation (M1) showed morphological variations however the fiber of these plants kept their high quality. M2 plants with signs of early maturity and high yield were selected and harvested. During the writing of this article, M3 seeds were collected for their future sew and also their respective molecular characterization by using SSR and AFLP. (author)

  17. Sequence-related amplified polymorphism (SRAP) markers: A potential resource for studies in plant molecular biology1

    Robarts, Daniel W. H.; Wolfe, Andrea D.


    In the past few decades, many investigations in the field of plant biology have employed selectively neutral, multilocus, dominant markers such as inter-simple sequence repeat (ISSR), random-amplified polymorphic DNA (RAPD), and amplified fragment length polymorphism (AFLP) to address hypotheses at lower taxonomic levels. More recently, sequence-related amplified polymorphism (SRAP) markers have been developed, which are used to amplify coding regions of DNA with primers targeting open readin...

  18. Optimization of AFLP fingerprinting of organisms with a large-sized genome: a study on Alstroemeria spp

    Han, T.H.; Eck, van H.J.; Jeu, de M.J.; Jacobsen, E.


    The recently introduced PCR-based DNA fingerprinting technique AFLP (amplified fragment length polymorphism) allows the selective amplification of subsets of genomic restriction fragments. AFLP has been used for multiple purposes such as the construction of linkage maps, marker saturation at specifi

  19. Molecular markers of phase transition in locusts



    The changes accompanying the transition from the gregarious to the solitary phase state in locusts are so drastic that for a long time these phases were considered as distinct species. It was Boris Uvarov who introduced the concept of polyphenism. Decades of research revealed that phase transition implies changes in morphometry, the color of the cuticle, behavior and several aspects of physiology. In particular, in the recent decade, quite a number of molecular studies have been undertaken to uncover phase-related differences.They resulted in novel insights into the role of corazonin, neuroparsins, some protease inhibitors, phenylacetonitrile and so on. The advent of EST-databases of locusts (e.g. Kang et al., 2004) is a most encouraging novel development in physiological and behavioral locust research. Yet, the answer to the most intriguing question, namely whether or not there is a primordial molecular inducer of phase transition, is probably not within reach in the very near future.

  20. Application of molecular markers in germplasm enhancement of Cassava (Manihot esculenta L. Crantz) and Yams (Dioscorea spp.) at IITA

    The genetic variation among 28 varieties of cassava (Manihot esculenta L. Crantz), collected from different parts of the Republic of Benin was determined using random amplified polymorphic DNA (RAPD) markers. A set of ten primers out of the one hundred that were screened, detected polymorphisms. Thirty-five cassava landraces from three countries of West Africa, along with five improved varieties and one genetic stock (58308), were analysed using both micro satellite markers and nine selected random primers which generated fifty-four polymorphic markers. Based on the unweighted pair group method with arithmetic averages (UPGMA) and Principal Component Analysis (PCA), six major groups of clusters were identified among the forty one genotypes. Clone 58308, the original source of resistance to African Cassava Mosaic Disease (ACMD) in IITA's cassava breeding program, and TMS 30572, an improved cultivar derived from clone 58308, were found in the same cluster group. All 34 of the landraces that are known to be resistant to ACMD were genetically distant from 58308 and TMS 30572. A diallel mating programme has been initiated to elucidate the genetics of these new sources of resistance to ACMD and determine their complementarity as well as allellism for resistance. A set of eight random primers for RAPD and two combinations of enzymes and specific primers for AFLP were used to generate DNA fingerprinting of twenty varietal groups among the 32 described for cultivated yams in the region. The results obtained confirm that a given varietal group is a mixture of different genotypes. The molecular taxonomy of 30 accessions of cultivated yams, D. rotundata and D. cayenensis, and 35 accessions of wild yams from Nigeria was established using RAPD and micro satellite markers. The cultivated yams separated into two distinct groups corresponding to the two species. D. rotundata genotypes showed relationship to the wild species D. abyssinica and D. praehensilis, whereas D. cayenensis

  1. Pilocytic astrocytoma: pathology, molecular mechanisms and markers.

    Collins, V Peter; Jones, David T W; Giannini, Caterina


    Pilocytic astrocytomas (PAs) were recognized as a discrete clinical entity over 70 years ago. They are relatively benign (WHO grade I) and have, as a group, a 10-year survival of over 90%. Many require merely surgical removal and only very infrequently do they progress to more malignant gliomas. While most show classical morphology, they may present a spectrum of morphological patterns, and there are difficult cases that show similarities to other gliomas, some of which are malignant and require aggressive treatment. Until recently, almost nothing was known about the molecular mechanisms involved in their development. The use of high-throughput sequencing techniques interrogating the whole genome has shown that single abnormalities of the mitogen-activating protein kinase (MAPK) pathway are exclusively found in almost all cases, indicating that PA represents a one-pathway disease. The most common mechanism is a tandem duplication of a ≈2 Mb-fragment of #7q, giving rise to a fusion between two genes, resulting in a transforming fusion protein, consisting of the N-terminus of KIAA1549 and the kinase domain of BRAF. Additional infrequent fusion partners have been identified, along with other abnormalities of the MAP-K pathway, affecting tyrosine kinase growth factor receptors at the cell surface (e.g., FGFR1) as well as BRAF V600E, KRAS, and NF1 mutations among others. However, while the KIAA1549-BRAF fusion occurs in all areas, the incidence of the various other mutations identified differs in PAs that develop in different regions of the brain. Unfortunately, from a diagnostic standpoint, almost all mutations found have been reported in other brain tumor types, although some retain considerable utility. These molecular abnormalities will be reviewed, and the difficulties in their potential use in supporting a diagnosis of PA, when the histopathological findings are equivocal or in the choice of individualized therapy, will be discussed. PMID:25792358

  2. Toxigenic profile and AFLP variability of Alternaria alternata and Alternaria infectoria occurring on wheat

    María Silvina Oviedo


    Full Text Available The objectives of this study were to evaluate the ability to produce alternariol (AOH, alternariol monomethyl ether (AME and tenuazonic acid (TA by A. alternata and A. infectoria strains recovered from wheat kernels obtained from one of the main production area in Argentina; to confirm using AFLPs molecular markers the identify of the isolates up to species level, and to evaluate the intra and inter-specific genetic diversity of these two Alternaria species. Among all the Alternaria strains tested (254, 84% of them were able to produce mycotoxins. The most frequent profile of toxin production found was the co-production of AOH and AME in both species tested. TA was only produced by strains of A. alternata. Amplified fragment polymorphism (AFLPs analysis was applied to a set of 89 isolates of Alternaria spp (40 were A. infectoria and 49 were A. alternata in order to confirm the morphological identification. The results showed that AFLPs are powerful diagnostic tool for differentiating between A. alternata and A. infectoria. Indeed, in the current study the outgroup strains, A. tenuissima was consistently classified. Characteristic polymorphic bands separated these two species regardless of the primer combination used. Related to intraspecific variability, A. alternata and A. infectoria isolates evaluated seemed to form and homogeneous group with a high degree of similarity among the isolates within each species. However, there was more scoreable polymorphism within A. alternata than within A. infectoria isolates. There was a concordance between morphological identification and separation up to species level using molecular markers. Clear polymorphism both within and between species showed that AFLP can be used to asses genetic variation in A. alternata and A. infectoria. The most important finding of the present study was the report on AOH and AME production by A. infectoria strains isolated from wheat kernels in Argentina on a semisynthetic

  3. Role of emerging molecular markers in pancreatic cyst fluid.

    Al-Haddad, Mohammad


    Cystic lesions of the pancreas (CLPs) are increasingly diagnosed due to the growing utilization of cross-sectional imaging modalities. The differentiation between true cysts (epithelial tumors) and nonepithelial lesions (such as pseudocysts) relies on clinical and imaging characteristics, but more reliably obtained by endoscopic ultrasound (EUS) fine-needle aspiration. Due to their malignant potential, some of the true pancreatic cysts require further assessment and periodic follow-up. Therefore, it is important to establish a solid diagnosis at the time of detection of the various types of pancreatic cysts. Due to the limitations of cytology and biochemical markers in accurately classifying cyst pathology, the search for specific molecular markers associated with each type of cyst is ongoing. In this chapter, we will review some of the emerging molecular markers in pancreatic cystic fluid and their potential impact on endosonography and pancreatic cyst management. PMID:26643693

  4. The phylogenetic relationship of the family Lutjanidae based on analyses of AFLP and mitochondrial 12S rRNA sequences

    ZHANG Junbin; LIU Xin


    Fishes of the family Lutjanidae are commercially important in South China Sea. However,the phylogeny of Lutjanids is still unclear and there are many controversies over it. Herein, studies about the phylogeny of Lutjanids were performed based on Amplified Fragment Length Polymorphism (AFLP) analysis of genome DNA and sequence analysis of mitochondrial 12S rRNA gene, and 10 Lutjanidae species and 1 Lethrinidae species were employed.The topologies of minimum evolution (ME) trees based on the two analyses respectively were congruent except for positions of genera Pristipomoides and Caesio. The optimal substitution model TrN + G for DNA sequences of 12S rRNA genes in Lutjanids was obtained using MODELTEST 3.6 software and maximum likelihood (ML) analysis supports the topology displayed by the ME tree. The test of log-likelihood suggests that the use of molecular clock calibrations to estimate species divergence time appeared valid. Phylogenetic analyses using AFLP data and DNA sequences of mitochondrial 12S rRNA genes indicated the monophyly of Lutjanus genra. However, further studies are required to reveal the phylogenetic relationship among other genera. In addition, the results demonstrated that AFLP genetic marker was suitable for the phylogenetic analysis of Lutjanids.

  5. Molecular pathology in adult gliomas: diagnostic, prognostic, and predictive markers.

    Jansen, Michael


    Over the past 10 years, there has been an increasing use of molecular markers in the assessment and management of adult malignant gliomas. Some molecular signatures are used diagnostically to help pathologists classify tumours, whereas others are used to estimate prognosis for patients. Most crucial, however, are those markers that are used to predict response to certain therapies, thereby directing clinicians to a particular treatment while avoiding other potentially deleterious therapies. Recently, large-scale genome-wide surveys have been used to identify new biomarkers that have been rapidly developed as diagnostic and prognostic tools. Given these developments, the pace of discovery of new molecular assays will quicken to facilitate personalised medicine in the setting of malignant glioma.

  6. AFLP fingerprinting of elite varieties (clones) from the genus Populus

    GAO Jianming; ZHANG Shougong; QI Liwang; ZHANG Yong; WANG Chunguo; CHEN Ruiyang; SONG Wenqin


    Accurate identification of varieties (clones) and knowledge of their genetic relationships are essential for poplar breeding and variety management.In this study,21 elite poplar varieties of Tacamahaca and Aigeiros in China were fingerprinted using amplified fragment length polymorphism (AFLP) markers.Four AFLP primer pairs developed generated totally 181 AFLP polymorphic fragments,and in particular,each primer pair generated fingerprint profiles specific to each of the tested varieties.The genetic relationships among the varieties were evaluated by dendrograms and multidimensional scaling (MDS).The results showed that the tested poplar can be classified into five groups,and indicated the clear separation of varieties of different sections of poplar and the primary distinction between native and exotic poplar varieties.This study indicated that tested poplar varieties could be identified by their fingerprint profiles and that genetic relationships deduced from the study were consistent with their genealogy.In addition,our results demonstrated that AFLP could be used to construct DNA fingerprints of poplar clones at a large-scale level and to determine genetic relationships of poplar varieties.

  7. Clinical Applications of Molecular Markers in Bone Tumors.

    Hameed, Meera


    Pathologic diagnosis of primary bone neoplasms can be challenging primarily due to rarity of the disease entities, overlapping imaging and histologic findings, and lack of tumor-specific immunohistochemical stains. Although slow to evolve, in recent years there has been a rapid advance in the discovery of new and novel molecular markers in primary bone neoplasms, which has enhanced diagnostic accuracy and has shed light into their pathogenesis. Modern technological approaches such as next-generation sequencing including RNA sequencing are serving as "rapid discovery platforms" for new and novel mutations and translocations with diagnostic, prognostic, and possible therapeutic applicability. As next-generation sequencing technologies are finding their place in clinical laboratories, one could envision routine testing for mutations spanning across a gene or translocations with multiple breakpoints and partner genes. This review will focus on the clinical relevance and applicability of molecular markers in primary bone neoplasms with examples. PMID:26452209

  8. Promise and pitfalls of molecular markers of thyroid nodules

    Jadhav, S.; Anurag Lila; Tushar Bandgar; Nalini Shah


    Thyroid nodules are common in the general population with a prevalence of 5-7% The initial evaluation of thyroid nodules commonly involves thyroid function tests, an ultrasound (USG) and fine needle aspiration biopsy (FNAB). The optimal management of patients with thyroid nodules with indeterminate cytology is plagued by the lack of highly sensitive and specific diagnostic modalities In this article we attempt to review the available literature on the molecular markers which are increasingly ...

  9. Beyond an AFLP genome scan towards the identification of immune genes involved in plague resistance in Rattus rattus from Madagascar

    Tollenaere, C.; Jacquet, S.; Ivanova, S.; Loiseau, A; Duplantier, Jean-Marc; Streiff, R; Brouat, Carine


    Genome scans using amplified fragment length polymorphism (AFLP) markers became popular in nonmodel species within the last 10 years, but few studies have tried to characterize the anonymous outliers identified. This study follows on from an AFLP genome scan in the black rat (Rattus rattus), the reservoir of plague (Yersinia pestis infection) in Madagascar. We successfully sequenced 17 of the 22 markers previously shown to be potentially affected by plague-mediated selection and associated wi...

  10. The usefulness of molecular techniques to assess the presence of Aeromonas spp. harboring virulence markers in foods.

    Bin Kingombe, César I; Huys, Geert; Howald, Denise; Luthi, Elisabeth; Swings, Jean; Jemmi, Thomas


    A total of 78 raw and 123 processed and ready-to-eat retail food samples were used to assess the presence of motile Aeromonas spp. harboring virulence genes (cytotoxic enterotoxin and hemolysin genes) using a recently described PCR method in comparison with the conventional cultivation method based on the use of Ampicillin-Dextrin Agar (ADA) medium. With the ADA-based method, 65/201 (32.3%) samples showed presumptive Aeromonas spp. colonies whereas the PCR method revealed the presence of Aeromonas spp. harboring the targeted virulence genes in 51/201 (25.4%) of the tested samples. The rate of contaminated samples and the presence of pathogenic Aeromonas were significantly lower with both methods for processed than in case of raw samples. A polyphasic identification approach including biochemical and molecular techniques was applied to a selection of 34 PCR-positive presumptive Aeromonas isolates. Following fatty acid methyl ester (FAME) analysis and amplified fragment length polymorphism (AFLP) fingerprinting, a total of 33 isolates (97%) could be identified to the DNA hybridization group (HG) level. The majority of these isolates belonged to the species Aeromonas hydrophila HG3 (50%) and Aeromonas veronii biovar sobria (HG8/10) (38%). Molecular characterization of PCR amplicons obtained from these strains by PCR-Restriction Fragment Length Polymorphism (PCR-RFLP) fingerprinting and PCR-Amplicon Sequence Analysis (PCR-ASA) allowed classification of all strains in a known PCR-RFLP and PCR-ASA type. In conclusion, the current findings demonstrate that the combined use of PCR-based virulence marker detection, PCR-RFLP and PCR-ASA offers a rapid, sensitive, and specific system to assess the presence and prevalence of Aeromonas spp. harboring virulence markers in food samples. PMID:15193799

  11. Biochemical and genetic variation of some Syrian wheat varieties using NIR, RAPD and AFLPs techniques

    This study was performed to assess chemical components and genetic variability of five Syrian wheat varieties using NIR, RAPD and AFLP techniques. NIR technique showed that Cham6 was the best variety in term of wheat grain quality due to their lowest protein (%), hardness, water uptake and baking volume and the highest starch (%) compared to the other tested varieties. PCR amplifications with 21 RAPD primers and 13 AFLP PCs primer combinations gave 104 and 466 discernible loci of which 24 (18.823%) and 199 (45.527%) were polymorphic for the both techniques respectively. Our data indicated that the three techniques gave similar results regarding the degree of relatedness among the tested varieties. In the present investigation, AFLP fingerprinting was more efficient than the RAPD assay. Where the letter exhibited lower Marker Index (MI) average (0.219) compared to AFLP one (3.203). The pattern generated by RAPD, AFLPs markers or by NIR separated the five wheat varieties into two groups. The first group consists of two subclusters. The first subcluster involved Cham8 and Bohous6, while the second one includes Cham6 that is very closed to precedent varieties. The second group consists of Bohous9 and Cham7 that were also closely related. Based on this study, the use of NIR, RAPD and AFLP techniques could be a powerful tool to detect the effectiveness relationships of these technologies. (author)

  12. Genetic diversity of Chilean and Brazilian alstroemeria species assessed by AFLP analysis.

    Han, T H; de Jeu, M; van Eck, H; Jacobsen, E


    One to three accessions of 22 Alstroemeria species, an interspecific hybrid (A. aurea x A. inodora), and single accessions of Bomarea salsilla and Leontochir ovallei were evaluated using the AFLP-marker technique to estimate the genetic diversity within the genus Alstroemeria. Three primer combinations generated 716 markers and discriminated all Alstroemeria species. The dendrogram inferred from the AFLP fingerprints supported the conjecture of the generic separation of the Chilean and Brazilian Alstroemeria species. The principal co-ordinate plot showed the separate allocation of the A. ligtu group and the allocation of A. aurea, which has a wide range of geographical distribution and genetic variation, in the middle of other Alstroemeria species. The genetic distances, based on AFLP markers, determined the genomic contribution of the parents to the interspecific hybrid. PMID:10849081

  13. Microsatellite markers- A new practice of DNA based markers in molecular genetics

    Neha Mittal


    Full Text Available Microsatellites are randomly interspersed within eukaryotic genomes. These are also known as SSRs (Simple sequence repeats, STRs (Short tandem repeats, STMs (Sequence tagged microsatellites, and VNTRs (Variable number of tandem repeats, are short 1-8 bp long monomer sequences, which randomly repeated in the DNA sequences. Conservation of the flanking sequence of each microsatellite locus allows the design of primers for PCR amplification. Then amplified products are separated by electrophoresis (either on high resolution agarose or acrylamide gels to detect the polymorphism in repeat length. These markers don′t require the radioactivity for detection and these are extremely robust as well as easily exchanged between the laboratories. Multiplex reactions of SSRs can easily be run to speed up the assay, when the products have non-overlapping size ranges. Microsatellites have emerged as the marker of choice for plant genetic resources and molecular genetic applications due to their abundant and uniform distribution throughout the genome, highly variable nature with regard to repeat number, show co-dominant inheritance, ease of transferability and reproducibility, and found highly efficient in the DNA fingerprinting analysis, as well as can also be utilized in the pedigree analysis of different plant species. This marker system has been proven beneficial for crop improvement and for breeding applications in many species. These are also found the useful tools to detect the polymorphisms in low level of intraspecific diversity. Consequently, in the present review we have described the various characteristic features and properties of these highly polymorphic microsatellite markers, which found beneficial in several molecular genetic and breeding applications.

  14. Identification of early molecular markers for breast cancer

    Schlag Peter M; Schoenegg Winfried; Siedentopf Friederike; Sterner-Kock Anja; Kretschmer Céline; Kemmner Wolfgang


    Abstract Background The ductal carcinoma in situ (DCIS) of the mammary gland represents an early, pre-invasive stage in the development of invasive breast carcinoma. Since DCIS is a curable disease, it would be highly desirable to identify molecular markers that allow early detection. Mice transgenic for the WAP-SV40 early genome region were used as a model for DCIS development. Gene expression profiling was carried out on DCIS-bearing mice and control animals. Additionally, a set of human DC...

  15. Estimation of genetic relationship in rice using molecular markers

    correlation of the marker estimates to fij indicate their validity as measures of genetic relatedness, while their immunity to assumptions of the pedigree method and their appropriateness to classification imply their superiority. If data from molecular markers are to be useful for varietal identification and protection, they have to be translated to some meaningful measurement, especially when the number of both marker variants and genotypes under comparison are increased. A measure to directly quantify the degree of similarity, thus, becomes important and indispensable. This study has demonstrated that marker based estimates of coefficient of coancestry can address this need in rice

  16. Determination of molecular markers associated with anthesis-silking interval in maize

    Maize lines contrasting in anthesis-silking, interval (ASI), a trait strongly linked to drought tolerance, have been analyzed under different water stress conditions in the field and with molecular markers. Correlation of marker and field data has revealed molecular markers strongly associated with flowering and yield traits. (author)

  17. A molecular marker of artemisinin-resistant Plasmodium falciparum malaria

    Ariey, Frédéric; Witkowski, Benoit; Amaratunga, Chanaki; Beghain, Johann; Langlois, Anne-Claire; Khim, Nimol; Kim, Saorin; Duru, Valentine; Bouchier, Christiane; Ma, Laurence; Lim, Pharath; Leang, Rithea; Duong, Socheat; Sreng, Sokunthea; Suon, Seila; Chuor, Char Meng; Bout, Denis Mey; Ménard, Sandie; Rogers, William O.; Genton, Blaise; Fandeur, Thierry; Miotto, Olivo; Ringwald, Pascal; Le Bras, Jacques; Berry, Antoine; Barale, Jean-Christophe; Fairhurst, Rick M.; Benoit-Vical, Françoise; Mercereau-Puijalon, Odile; Ménard, Didier


    Plasmodium falciparum resistance to artemisinin derivatives in southeast Asia threatens malaria control and elimination activities worldwide. To monitor the spread of artemisinin resistance, a molecular marker is urgently needed. Here, using whole-genome sequencing of an artemisinin-resistant parasite line from Africa and clinical parasite isolates from Cambodia, we associate mutations in the PF3D7_1343700 kelch propeller domain (`K13-propeller') with artemisinin resistance in vitro and in vivo. Mutant K13-propeller alleles cluster in Cambodian provinces where resistance is prevalent, and the increasing frequency of a dominant mutant K13-propeller allele correlates with the recent spread of resistance in western Cambodia. Strong correlations between the presence of a mutant allele, in vitro parasite survival rates and in vivo parasite clearance rates indicate that K13-propeller mutations are important determinants of artemisinin resistance. K13-propeller polymorphism constitutes a useful molecular marker for large-scale surveillance efforts to contain artemisinin resistance in the Greater Mekong Subregion and prevent its global spread.

  18. MicroRNAs as molecular markers in lung cancer

    Javier Silva


    Full Text Available Lung cancer is the most common cause of cancer death in the western world for both men and women. Lung cancer appears to be a perfect candidate for a screening program, since it is the number one cancer killer, it has a long preclinical phase, curative treatment for the minority of patients who are diagnosed early and a target population at risk (smokers and it is also a major economic burden. The earliest approaches to identifying cancer markers were based on preliminary clinical or pathological observations, although molecular biology is a strong candidate for occupying a place among the set of methods. In search of markers, several alterations, such as mutations, loss of heterozygosity, microsatellite instability, DNA methylation, mitochondrial DNA mutations, viral DNA, modified expression of mRNA, miRNA and proteins, and structurally altered proteins have all been analysed. MicroRNAs (miRNA are small RNA molecules, about 19-25 nucleotides long and encoded in genomes of plants, animals, fungi and viruses. It has been reported that miRNAs may have multiple functions in lung development and that aberrant expression of miRNAs could induce lung tumorigenesis. We review here the role of miRNAs in lung tumorigenesis and also as a novel type of biomarker.-----------------------------------Cite this article as:Silva J, Garcia V, Lopez-Gonzalez A, Provencio M. MicroRNAs as molecular markers in lung cancer. Int J Cancer Ther Oncol 2013;1(1:010111. DOI:

  19. Identification of early molecular markers for breast cancer

    Schlag Peter M


    Full Text Available Abstract Background The ductal carcinoma in situ (DCIS of the mammary gland represents an early, pre-invasive stage in the development of invasive breast carcinoma. Since DCIS is a curable disease, it would be highly desirable to identify molecular markers that allow early detection. Mice transgenic for the WAP-SV40 early genome region were used as a model for DCIS development. Gene expression profiling was carried out on DCIS-bearing mice and control animals. Additionally, a set of human DCIS and invasive mammary tumors were analyzed in a similar fashion. Enhanced expression of these marker genes in human and murine samples was validated by quantitative RT-PCR. Besides, marker gene expression was also validated by immunohistochemistry of human samples. Furthermore in silico analyses using an online microarray database were performed. Results In DCIS-mice seven genes were identified that were significantly up-regulated in DCIS: DEPDC1, NUSAP1, EXO1, RRM2, FOXM1, MUC1 and SPP1. A similar up-regulation of homologues of the murine genes was observed in human DCIS samples. Enhanced expression of these genes in DCIS and IDC (invasive ductal carcinoma was validated by quantitative RT-PCR and immunohistochemistry. Conclusions By comparing murine markers for the ductal carcinoma in situ (DCIS of the mammary gland with genes up-regulated in human DCIS-samples we were able to identify a set of genes which might allow early detection of DCIS and invasive carcinomas in the future. The similarities between gene expression in DCIS and invasive carcinomas in our data suggest that the early detection and treatment of DCIS is of utmost relevance for the survival of patients who are at high risk of developing breast carcinomas.

  20. Intelligent DNA-based molecular diagnostics using linked genetic markers

    Pathak, D.K.; Perlin, M.W.; Hoffman, E.P.


    This paper describes a knowledge-based system for molecular diagnostics, and its application to fully automated diagnosis of X-linked genetic disorders. Molecular diagnostic information is used in clinical practice for determining genetic risks, such as carrier determination and prenatal diagnosis. Initially, blood samples are obtained from related individuals, and PCR amplification is performed. Linkage-based molecular diagnosis then entails three data analysis steps. First, for every individual, the alleles (i.e., DNA composition) are determined at specified chromosomal locations. Second, the flow of genetic material among the individuals is established. Third, the probability that a given individual is either a carrier of the disease or affected by the disease is determined. The current practice is to perform each of these three steps manually, which is costly, time consuming, labor-intensive, and error-prone. As such, the knowledge-intensive data analysis and interpretation supersede the actual experimentation effort as the major bottleneck in molecular diagnostics. By examining the human problem solving for the task, we have designed and implemented a prototype knowledge-based system capable of fully automating linkage-based molecular diagnostics in X-linked genetic disorders, including Duchenne Muscular Dystrophy (DMD). Our system uses knowledge-based interpretation of gel electrophoresis images to determine individual DNA marker labels, a constraint satisfaction search for consistent genetic flow among individuals, and a blackboard-style problem solver for risk assessment. We describe the system`s successful diagnosis of DMD carrier and affected individuals from raw clinical data.

  1. Diagnostically Relevant Molecular Markers in Head and Neck Neoplasms

    Soma Susan Varghese; Philips Mathew; Jithin Jose


    Tumor markers are grouped into diagnostic and prognostic markers. Specific diagnostic markers appear extensively in cells of a particular neoplasm and not in other tumors. These markers can be used to assess the cellular lineage and histogenic origin of various neoplasms. Thus, diagnostic markers can be used for the confirmatory diagnosis of various tumors. This paper reviews the literature on various diagnostic markers and aims to group them based on the cellular lineage of neoplasms.

  2. Molecular Pathogenesis and Diagnostic, Prognostic and Predictive Molecular Markers in Sarcoma.

    Mariño-Enríquez, Adrián; Bovée, Judith V M G


    Sarcomas are infrequent mesenchymal neoplasms characterized by notable morphological and molecular heterogeneity. Molecular studies in sarcoma provide refinements to morphologic classification, and contribute diagnostic information (frequently), prognostic stratification (rarely) and predict therapeutic response (occasionally). Herein, we summarize the major molecular mechanisms underlying sarcoma pathogenesis and present clinically useful diagnostic, prognostic and predictive molecular markers for sarcoma. Five major molecular alterations are discussed, illustrated with representative sarcoma types, including 1. the presence of chimeric transcription factors, in vascular tumors; 2. abnormal kinase signaling, in gastrointestinal stromal tumor; 3. epigenetic deregulation, in chondrosarcoma, chondroblastoma, and other tumors; 4. deregulated cell survival and proliferation, due to focal copy number alterations, in dedifferentiated liposarcoma; 5. extreme genomic instability, in conventional osteosarcoma as a representative example of sarcomas with highly complex karyotype. PMID:27523972

  3. Evaluación de la variación somaclonal en vitroplantas de caña de azúcar mediante marcadores moleculares Evaluation of somaclonal variation in in vitro produced sugarcane plants through molecular markers

    María F. Perera

    Full Text Available El cultivo in vitro de tejidos vegetales puede producir variación somaclonal, fenómeno que consiste en modificaciones genéticas en las células y tejidos cultivados. Esto puede limitar la aplicación de dicha técnica para la micropopagación masiva, especialmente si la variación provoca un cambio fenotípico de importancia agronómica. En este trabajo se optimizó una metodología basada en la comparación de perfiles de marcadores moleculares AFLP (del inglés "Amplified Fragment Length Polymorphism", para la detección de la variación somaclonal en vitroplantas de caña de azúcar. Para la optimización de la técnica de AFLP en caña de azúcar, se utilizaron plantas de seis genotipos propagados convencionalmente y dos tipos de muestras: hojas tiernas y meristemas. La variación somaclonal fue evaluada en líneas de vitroplantas de los mismos genotipos al final del cultivo in vitro, luego de seis meses de micropropagación. Con las 19 combinaciones de cebadores utilizadas, se diferenciaron los perfiles moleculares de los seis genotipos. En los plantines micropropagados se detectaron perfiles diferenciales en las variedades LCP85-384 y TUCCP77-42 con 3 de las 19 combinaciones de cebadores utilizadas. Este resultado muestra la validez de la técnica para detectar variantes somaclonales, y deja en evidencia la diferencia de susceptibilidad de los genotipos al cultivo in vitro. Esto permitió ajustar la metodología de micropropagación para cada genotipo multiplicado y asegurar la pureza genética de cada vitroplanta.In vitro culture of plant tissue can produce somaclonal variation, which consists of genetic modifications in cultured cells and tissues. This may constrain the use of this technique in massive micropopagation, especially if such change causes an agronomically relevant phenotypical modification. In this work, a methodology based on the comparison of AFLP (Amplified Fragment Length Polymorphism molecular marker profiles was

  4. Genetic diversity of Chilean and Brazilian Alstroemeria species assessed by AFLP analysis

    Han, T.H.; Jeu, de M.J.; Eck, van H.J.; Jacobsen, E.


    One to three accessions of 22 Alstroemeria species, an interspecific hybrid (A. aurea ́ A. inodora), and single accessions of Bomarea salsilla and Leontochir ovallei were evaluated using the AFLP-marker technique to estimate the genetic diversity within the genus Alstroemeria. Three primer combinati

  5. Bootsie: estimation of coefficient of variation of AFLP data by bootstrap analysis

    Bootsie is an English-native replacement for ASG Coelho’s “DBOOT” utility for estimating coefficient of variation of a population of AFLP marker data using bootstrapping. Bootsie improves on DBOOT by supporting batch processing, time-to-completion estimation, built-in graphs, and a suite of export t...

  6. Molecular characterization of Cryptococcus gattii genotype AFLP6/VGII isolated from woody debris of divi-divi (Caesalpinia coriaria), Bonaire, Dutch Caribbean

    Hagen, F.; Chowdhary, A.; Prakash, A.; Yntema, J.B.; Meis, J.F.G.M.


    BACKGROUND: The basidiomycetous yeast Cryptococcus gattii is an emerging and primary pathogen. There is a lack of information about its environmental spread outside outbreak regions in Mediterranean Europe, North and South America. Environmental sampling for C. gattii and molecular characterization

  7. Does Primula intricata Gren. et Godr. Merit Species Rank? A Taxonomic Revision Based on nrDNA, cpDNA and AFLP Data

    Dana ŞUTEU


    Full Text Available Primula intricata Gren. et Godr. has an unclear taxonomic status: it was originally described as a distinct species but subsequently was considered a subspecies (Primula elatior subsp. intricata or even a variety (P. elatior var. intricata of Primula elatior (L. Hill. No prior genetic studies were performed on this group of Primulaceae, therefore we considered useful to investigate taxonomies boundaries within the P. elatior-intricata group. We explored genetic differences between Primula intricata and Primula elatior group by applying three different types of molecular markers: nuclear ribosomal DNA (ITS1, chloroplast DNA (spacer trnH-psbA and intron trnL and Amplified Fragment Length Polymorphisms (AFLP. We found a solid differentiation between P. intricata and P. elatior group, differentiation that was confirmed by all the employed molecular markers. This finding enabled us to propose a valid species rank for Primula intricata, as a separate taxon from the P. elatior group.

  8. The role of Molecular Markers in Improvement of Fruit Crops

    Zahoor Ahmad BHAT; Wasakha Singh DHILLON; Rashid, Rizwan; Javid Ahmad BHAT; Waseem Ali DAR; Mohammad Yousf GANAIE


    Markers have been used over the years for the classification of plants. Markers are any trait of an organism that can be identified with confidence and relative easy, and can be followed in a mapping population on another hand markers be defined as heritable entities associated with the economically important trait under the control of polygenes. Morphological markers can be detected with naked eye (naked eye polymorphism) or as difference in physical or chemical properties of the macromolecu...

  9. Molecular markers for identifying municipal, domestic and agricultural sources of organic matter in natural waters.

    Harwood, John J


    Molecular markers can be used to determine the sources of organic pollution in water. This review summarizes progress made during the last two decades in identifying reliable molecular markers to distinguish pollution from sewage, animal production, and other sources. Two artificial sweeteners, sucralose and acesulfame-K, are sufficiently stable to be molecular markers and easily associated with domestic wastewater. Waste from different animal species may be distinguished by profiling fecal sterols and bile acids. Other markers which have been evaluated, including caffeine, detergent components, and compounds commonly leached from landfills are discussed. PMID:24200048

  10. Recent advances in high-throughput molecular marker identification for superficial and invasive bladder cancers

    Andersen, Lars Dyrskjøt; Zieger, Karsten; Ørntoft, Torben Falck


    individually contributed to the management of the disease. However, the development of high-throughput techniques for simultaneous assessment of a large number of markers has allowed classification of tumors into clinically relevant molecular subgroups beyond those possible by pathological classification. Here......, we review the recent advances in high-throughput molecular marker identification for superficial and invasive bladder cancers....

  11. Molecular markers for urothelial bladder cancer prognosis: Toward implementation in clinical practice

    Rhijn, B.W. van; Catto, J.W.; Goebell, P.J.; Knuchel, R.; Shariat, S.F.; Poel, H.G. van der; Sanchez-Carbayo, M.; Thalmann, G.N.; Schmitz-Drager, B.J.; Kiemeney, L.A.L.M.


    OBJECTIVES: To summarize the current status of clinicopathological and molecular markers for the prediction of recurrence or progression or both in non-muscle-invasive and survival in muscle-invasive urothelial bladder cancer, to address the reproducibility of pathology and molecular markers, and to

  12. Molecular Markers to Characterize Genetic Variability in Brazilian Cochliomyia hominivorax

    The screwworm fly Cochliomyia hominivorax is one of the most important agents of traumatic myiasis throughout neotropical regions. In Brazil this pest is devastating, causing great profit losses for cattle breeders (around U$ 180 million annually). In South America there are no preventive methods to control natural populations of screwworm fly. The basic knowledge of the genetic variability and evolution within a species is necessary information to understand the structure and evolution of populations. In the case of screwworm populations we are, in our laboratory, conducting analyses with different types of molecular markers in the mitochondrial and nuclear genomes using RFLP, PCR and sequencing procedures and protein electrophoresis to characterize the genetic polymorphism and population structure of screwworms in Brazil. Based on the fragment patterns for the five marker enzymes, 15 mtDNA composite haplotypes were detected among the individuals of the seven populations of screwworm analysed. The average of nucleotide diversity was 0.92%. The nucleotide divergence estimates between pairs of haplotypes ranged from 0.3% to 2.7%. The analysis of the geographical distribution among the observed haplotypes suggests that the sampled populations probably belong to a single evolutionary lineage interconnected by reduced gene flow. The RAPD-PCR technique was used to detect genetic polymorphism and to select genetic markers to discriminate seven populations, including one from northern Argentina. In general, results of both mitochondrial, RAPD analysis and allozymes are concordant in suggesting divergence among screwworm populations. The Esterase system was the most polymorphic (with ten alleles) and was polymorphic in all the studied populations. The genetic differentiation, Fst value, was Fst=0.214. The estimated rate of gene flow from the total sample of screwworm was low Nm=0.92. Our data show a great amount of genetic variability as revealed by isozymes. In addition

  13. Phylogeography and molecular diversity analysis of Jatropha curcas L. and the dispersal route revealed by RAPD, AFLP and nrDNA-ITS analysis

    Sudheer Pamidimarri, D. V N


    Jatropha curcas L. (Euphorbiaceae) has acquired a great importance as a renewable source of energy with a number of environmental benefits. Very few attempts were made to understand the extent of genetic diversity and its distribution. This study was aimed to study the diversity and deduce the phylogeography of Jatropha curcas L. which is said to be the most primitive species of the genus Jatropha. Here we studied the intraspecific genetic diversity of the species distributed in different parts of the globe. The study also focused to understand the molecular diversity at reported probable center of origin (Mexico), and to reveal the dispersal route to other regions based on random amplified polymorphic DNA, amplified fragment length polymorphism and nrDNA-ITS sequences data. The overall genetic diversity of J. curcas found in the present study was narrow. The highest genetic diversity was observed in the germplasm collected from Mexico and supports the earlier hypothesis based on morphological data and natural distribution, it is the center for origin of the species. Least genetic diversity found in the Indian germplasm and clustering results revealed that the species was introduced simultaneously by two distinct germplasm and subsequently distributed in different parts of India. The present molecular data further revealed that J. curcas might have spread from the center of the origin to Cape Verde, than to Spain, Portuguese to other neighboring countries and simultaneously to Africa. The molecular evidence supports the Burkill et al. (A dictionary of the economic products of the Malay Peninsula, Governments of Malaysia and Singapore by the Ministry of Agriculture and Co-operatives. Kuala Lumpur, Malaysia, 1966) view of Portuguese might have introduced the species to India. The clustering pattern suggests that the distribution was interfered by human activity. © Springer Science+Business Media 2014.

  14. Variabilidade genética em populações de pitangueira oriundas de autopolinização e polinização livre, acessada por AFLP Genetic variability in surinam cherry populations originated from self-pollination and cross pollination, estimated by AFLP

    Rodrigo Cezar Franzon


    Full Text Available Foram utilizados marcadores AFLP para a avaliação de populações de plantas de pitangueira (Eugenia uniflora oriundas de autopolinização e de polinização livre, com o objetivo de verificar a variabilidade existente entre e dentro dessas populações, visando a fornecer mais informações que ajudem no entendimento do modo de reprodução dessa espécie. O material vegetal utilizado foi oriundo de duas seleções de pitangueira ("Pit 15" e "Pit 52", mantidas na Embrapa Clima Temperado. De cada seleção, foram obtidas duas populações F1, por meio de autopolinização e de polinização livre, totalizando quatro populações. Foram analisados 18 indivíduos de cada população e as duas plantas-mãe, totalizando 74 indivíduos. Foram utilizadas três combinações de primers AFLP e calculada a similaridade genética entre plantas pelo coeficiente de Jaccard. Uma estimativa da variabilidade genética entre e dentro das populações foi estimada pela AMOVA. As três combinações de primers AFLP utilizadas amplificaram um total de 178 locos AFLP, dos quais 114 (64,0% foram polimórficos entre todos os indivíduos. Não houve separação clara entre populações descendentes da mesma planta-mãe. Foi observado maior polimorfismo de marcadores AFLP em populações de polinização livre. A proporção da variabilidade genética total entre populações foi significativa, embora tenha sido menor do que aquela observada dentro das populações. A reprodução da pitangueira é decorrente tanto da autofertilização quanto da polinização cruzada, sendo necessário, no entanto, novos estudos para determinar qual a estratégia de reprodução mais eficiente.AFLP molecular markers were used aiming to study the genetic variability within and between Surinam cherry (Eugenia uniflora populations, originated from self-pollination and open pollination of two selections (Pit 15 and Pit 52 of the Embrapa Clima Temperado collection. The objective was to

  15. Genetic Confirmation of Mungbean (Vigna radiata) and Mashbean (Vigna mungo) Interspecific Recombinants using Molecular Markers

    Abbas, Ghulam; Hameed, Amjad; Rizwan, Muhammad; Ahsan, Muhammad; Asghar, Muhammad J.; Iqbal, Nayyer


    Molecular confirmation of interspecific recombinants is essential to overcome the issues like self-pollination, environmental influence, and inadequacy of morphological characteristics during interspecific hybridization. The present study was conducted for genetic confirmation of mungbean (female) and mashbean (male) interspecific crosses using molecular markers. Initially, polymorphic random amplified polymorphic DNA (RAPD), universal rice primers (URP), and simple sequence repeats (SSR) markers differentiating parent genotypes were identified. Recombination in hybrids was confirmed using these polymorphic DNA markers. The NM 2006 × Mash 88 was most successful interspecific cross. Most of true recombinants confirmed by molecular markers were from this cross combination. SSR markers were efficient in detecting genetic variability and recombination with reference to specific chromosomes and particular loci. SSR (RIS) and RAPD identified variability dispersed throughout the genome. In conclusion, DNA based marker assisted selection (MAS) efficiently confirmed the interspecific recombinants. The results provided evidence that MAS can enhance the authenticity of selection in mungbean improvement program. PMID:26697053

  16. Molecular Markers of Lung Cancer in MAYAK Workers

    Steven A. Belinsky, PhD


    The molecular mechanisms that result in the elevated risk for lung cancer associated with exposure to radiation have not been well characterized. Workers from the MAYAK nuclear enterprise are an ideal cohort in which to study the molecular epidemiology of cancer associated with radiation exposure and to identify the genes targeted for inactivation that in turn affect individual risk for radiation-induced lung cancer. Epidemiology studies of the MAYAK cohort indicate a significantly higher frequency for adenocarcinoma and squamous cell carcinoma (SCC) in workers than in a control population and a strong correlation between these tumor types and plutonium exposure. Two hypotheses will be evaluated through the proposed studies. First, radiation exposure targets specific genes for inactivation by promoter methylation. This hypothesis is supported by our recent studies with the MAYAK population that demonstrated the targeting of the p16 gene for inactivation by promoter methylation in adenocarcinomas from workers (1). Second, genes inactivated in tumors can serve as biomarkers for lung cancer risk in a cancer-free population of workers exposed to plutonium. Support for this hypothesis is based on exciting preliminary results of our nested, case-control study of persons from the Colorado cohort. In that study, a panel of methylation markers for predicting lung cancer risk is being evaluated in sputum samples from incident lung cancer cases and controls. The first hypothesis will be tested by determining the prevalence for promoter hypermethylation of a panel of genes shown to play a critical role in the development of either adenocarcinoma and/or SCC associated with tobacco. Our initial studies on adenocarcinoma in MAYAK workers will be extended to evaluate methylation of the PAX5 {alpha}, PAX5 {beta}, H-cadherin, GATA5, and bone morphogenesis 3B (BMP3B) genes in the original sample set described under Preliminary studies. In addition, studies will be initiated in SCC


    I. Hrytsyniak


    Full Text Available Purpose. The application of molecular genetic markers has been widely used in modern experimental fish-farming in recent years. This methodology is currently presented by a differentiated approach with individual mechanisms and clearly defined possibilities. Numerous publications in the scientific literature that are dedicated to molecular genetic markers for the most part offer purely practical data. Thus, the synthesis and analysis of existing information on the general principles of action and the limits of the main methods of using molecular genetic markers is an actual problem. In particular, such a description will make it possible to plan more effectively the experiment and to obtain the desired results with high reliability. Findings. The main types of variable parts of DNA that can be used as molecular genetic markers in determining the level of stock hybridization, conducting genetic inventory of population and solving other problems in modern fish-farming are described in this paper. Also, the article provides an overview of principal modern methods that can be used to identify molecular genetic markers. Originality. This work is a generalization of modern ideas about the mechanisms of experiments with molecular genetic markers in fish-farming. Information is provided in the form of consistent presentation of the principles and purpose of each method, as well as significant advances during their practical application. Practical value. The proposed review of classic and modern literature data on molecular genetic markers can be used for planning, modernization and correction of research activity in modern fish-farming.

  18. Molecular markers for leaf rust resistance genes in wheat.

    Chełkowski, J; Stepień, L


    Over 100 genes of resistance to rust fungi: Puccinia recondita f. sp. tritici, (47 Lr - leaf rust genes), P. striiformis (18 Yr - yellow rust genes) and P. graminis f. sp. tritici (41 Sr - stripe rust genes) have been identified in wheat (Triticum aestivum L.) and its wild relatives according to recent papers. Sixteen Lr resistance genes have been mapped using restriction fragments length polymorphism (RFLP) markers on wheat chromosomes. More than ten Lr genes can be identified in breeding materials by sequence tagged site (STS) specific markers. Gene Lrk 10, closely linked to gene Lr 10, has been cloned and its function recognized. Available markers are presented in this review. The STS, cleaved amplified polymorphic sequence (CAPS) and sequence characterized amplified regions (SCAR) markers found in the literature should be verified using Triticum spp. with different genetic background. Simple sequence repeats (SSR) markers for Lr resistance genes are now also available. PMID:14564046

  19. Molecular evaluations of thirty one clones of poplar based on RAPD and SSR molecular markers

    Singh M.K.


    Full Text Available Poplar is an important tree species valued all over the world for its wood importance. Despite limited knowledge of the levels of genetic diversity and relatedness, their cultivation as a source of plywood is widespread. In order to facilitate reasoned scientific decisions on its management and conservation and prepare for selective breeding programme, genetic analysis of 31 genotypes was performed using RAPD and SSR molecular markers. Twenty six RAPD primers and 14 SSR primers amplified a total of 236 and 85 scoreable bands of which 86.44% and 86.02% were polymorphic. The mean coefficient of gene differentiation (Gst was 0.388 and 0.341 indicating that 61.2% and 65.9% of the genetic variation resided within the populations. Analysis of molecular variance (AMOVA indicated that majority of genetic variation (94.6% using RAPD and 89% using SSR occurred among genotypes, while the variation between the three groups (categorized as tall, medium and small plants height was 5.4% (using RAPD and 11% (using SSR. The dendrogram obtained from NJ and STRUCTURE analysis revealed splitting of genotypes into four clusters with clear distinction between short, medium and tall height genotypes, indicated that genetic differentiations measure with respect to RAPD and SSR. However, both the markers were equally useful in providing some understanding about the genetic relationship of different genotypes of poplar that are important in the conservation and exploitation of poplar genetic resources.

  20. Indel Group in Genomes (IGG) Molecular Genetic Markers.

    Toal, Ted W; Burkart-Waco, Diana; Howell, Tyson; Ron, Mily; Kuppu, Sundaram; Britt, Anne; Chetelat, Roger; Brady, Siobhan M


    Genetic markers are essential when developing or working with genetically variable populations. Indel Group in Genomes (IGG) markers are primer pairs that amplify single-locus sequences that differ in size for two or more alleles. They are attractive for their ease of use for rapid genotyping and their codominant nature. Here, we describe a heuristic algorithm that uses a k-mer-based approach to search two or more genome sequences to locate polymorphic regions suitable for designing candidate IGG marker primers. As input to the IGG pipeline software, the user provides genome sequences and the desired amplicon sizes and size differences. Primer sequences flanking polymorphic insertions/deletions are produced as output. IGG marker files for three sets of genomes, Solanum lycopersicum/Solanum pennellii, Arabidopsis (Arabidopsis thaliana) Columbia-0/Landsberg erecta-0 accessions, and S. lycopersicum/S. pennellii/Solanum tuberosum (three-way polymorphic) are included. PMID:27436831

  1. Molecular Markers for Breast Cancer: Prediction on Tumor Behavior

    Bruna Karina Banin Hirata; Julie Massayo Maeda Oda; Roberta Losi Guembarovski; Carolina Batista Ariza; Carlos Eduardo Coral de Oliveira; Maria Angelica Ehara Watanabe


    Breast cancer is one of the most common cancers with greater than 1,300,000 cases and 450,000 deaths each year worldwide. The development of breast cancer involves a progression through intermediate stages until the invasive carcinoma and finally into metastatic disease. Given the variability in clinical progression, the identification of markers that could predict the tumor behavior is particularly important in breast cancer. The determination of tumor markers is a useful tool for clinical m...

  2. Genetic confirmation of mungbean (Vigna radiata) and mashbean (Vigna mungo) interspecific recombinants using molecular markers

    Ghulam eAbbas; Amjad eHameed; Muhammad eRizwan; Muhammad eAhsan; Muhammad Jawad eAsghar; Nayyer eIqbal


    The present study was conducted with the aim to investigate recombination between mungbean (female) and mashbean (male) interspecific crosses using molecular markers i.e., URP (Universal Rice Primers), RAPD (Random Amplified Polymorphic DNA) and SSR (Simple Sequence Repeats). As a first step parental screening was performed and polymorphic markers differentiating parent genotypes were identified. Recombinations were then confirmed through polymorphic DNA markers in many of the hybrids. The N...

  3. Tracking neuronal marker expression inside living differentiating cells using molecular beacons

    Ilieva, Mirolyuba; Della Vedova, Paolo; Hansen, Ole;


    Monitoring gene expression is an important tool for elucidating mechanisms of cellular function. In order to monitor gene expression during nerve cell development, molecular beacon (MB) probes targeting markers representing different stages of neuronal differentiation were designed and synthesized...

  4. Molecular markers and conservation of plant species in Latin America: the case of Phaedranassa viridflora (Amaryllidaceae)

    Microsatellites are molecular markers with great potential for investigating genetic structure of populations. This information is valuable for generating effective conservation plans. We studied the endemic and endangered Phaedranassa viridiflora (Amaryllidaceae) to show the utility of microsatelli...

  5. Kazusa Marker DataBase: a database for genomics, genetics, and molecular breeding in plants.

    Shirasawa, Kenta; Isobe, Sachiko; Tabata, Satoshi; Hirakawa, Hideki


    In order to provide useful genomic information for agronomical plants, we have established a database, the Kazusa Marker DataBase ( This database includes information on DNA markers, e.g., SSR and SNP markers, genetic linkage maps, and physical maps, that were developed at the Kazusa DNA Research Institute. Keyword searches for the markers, sequence data used for marker development, and experimental conditions are also available through this database. Currently, 10 plant species have been targeted: tomato (Solanum lycopersicum), pepper (Capsicum annuum), strawberry (Fragaria × ananassa), radish (Raphanus sativus), Lotus japonicus, soybean (Glycine max), peanut (Arachis hypogaea), red clover (Trifolium pratense), white clover (Trifolium repens), and eucalyptus (Eucalyptus camaldulensis). In addition, the number of plant species registered in this database will be increased as our research progresses. The Kazusa Marker DataBase will be a useful tool for both basic and applied sciences, such as genomics, genetics, and molecular breeding in crops. PMID:25320561

  6. Studies on the AFLP Molecular Markers of Kava and Piper nigrum%卡瓦胡椒及胡椒的荧光AFLP分子标记研究

    施江; 辛莉; 杨彦; 郑学勤




    I. Hrytsyniak; O. Zaloilo; I. Zaloilo; N. Borysenko


    Purpose. The application of molecular genetic markers has been widely used in modern experimental fish-farming in recent years. This methodology is currently presented by a differentiated approach with individual mechanisms and clearly defined possibilities. Numerous publications in the scientific literature that are dedicated to molecular genetic markers for the most part offer purely practical data. Thus, the synthesis and analysis of existing information on the general principles of action...

  8. World Antimalarial Resistance Network (WARN) III: Molecular Markers for Drug Resistant Malaria

    Sibley Carol H; Shafer Robert W; Price Ric N; Naidoo Inbarani; Mugittu Kefas; Meshnick Steven R; Mbacham Wilfred; Joshi Hema H; Happi Christian T; Barnwell John W; Roper Cally; Plowe Christopher V; Sutherland Colin J; Zimmerman Peter A; Rosenthal Philip J


    Abstract Molecular markers for drug resistant malaria represent public health tools of great but mostly unrealized potential value. A key reason for the failure of molecular resistance markers to live up to their potential is that data on the their prevalence is scattered in disparate databases with no linkage to the clinical, in vitro and pharmacokinetic data that are needed to relate the genetic data to relevant phenotypes. The ongoing replacement of older monotherapies for malaria by new, ...

  9. World antimalarial resistance network (WARN) III: Molecular markers for drug resistant malaria

    plowe, cv; Roper, C.; Barnwell, JW; Happi, CT; Joshi, HH; Mbacham, W.; Meshnick, SR; Mugittu, K; Naidoo, I; Price, RN; Shafer, RW; Sibley, CH; Sutherland, CJ; Zimmerman, PA; Rosenthal, PJ


    Molecular markers for drug resistant malaria represent public health tools of great but mostly unrealized potential value. A key reason for the failure of molecular resistance markers to live up to their potential is that data on the their prevalence is scattered in disparate databases with no linkage to the clinical, in vitro and pharmacokinetic data that are needed to relate the genetic data to relevant phenotypes. The ongoing replacement of older monotherapies for malaria by new, more effe...

  10. Beyond an AFLP genome scan towards the identification of immune genes involved in plague resistance in Rattus rattus from Madagascar.

    Tollenaere, C; Jacquet, S; Ivanova, S; Loiseau, A; Duplantier, J-M; Streiff, R; Brouat, C


    Genome scans using amplified fragment length polymorphism (AFLP) markers became popular in nonmodel species within the last 10 years, but few studies have tried to characterize the anonymous outliers identified. This study follows on from an AFLP genome scan in the black rat (Rattus rattus), the reservoir of plague (Yersinia pestis infection) in Madagascar. We successfully sequenced 17 of the 22 markers previously shown to be potentially affected by plague-mediated selection and associated with a plague resistance phenotype. Searching these sequences in the genome of the closely related species Rattus norvegicus assigned them to 14 genomic regions, revealing a random distribution of outliers in the genome (no clustering). We compared these results with those of an in silico AFLP study of the R. norvegicus genome, which showed that outlier sequences could not have been inferred by this method in R. rattus (only four of the 15 sequences were predicted). However, in silico analysis allowed the prediction of AFLP markers distribution and the estimation of homoplasy rates, confirming its potential utility for designing AFLP studies in nonmodel species. The 14 genomic regions surrounding AFLP outliers (less than 300 kb from the marker) contained 75 genes encoding proteins of known function, including nine involved in immune function and pathogen defence. We identified the two interleukin 1 genes (Il1a and Il1b) that share homology with an antigen of Y. pestis, as the best candidates for genes subject to plague-mediated natural selection. At least six other genes known to be involved in proinflammatory pathways may also be affected by plague-mediated selection. PMID:23237097

  11. Identification of QTLs associated with callogenesis and embryogenesis in oil palm using genetic linkage maps improved with SSR markers.

    Ngoot-Chin Ting

    Full Text Available Clonal reproduction of oil palm by means of tissue culture is a very inefficient process. Tissue culturability is known to be genotype dependent with some genotypes being more amenable to tissue culture than others. In this study, genetic linkage maps enriched with simple sequence repeat (SSR markers were developed for dura (ENL48 and pisifera (ML161, the two fruit forms of oil palm, Elaeis guineensis. The SSR markers were mapped onto earlier reported parental maps based on amplified fragment length polymorphism (AFLP and restriction fragment length polymorphism (RFLP markers. The new linkage map of ENL48 contains 148 markers (33 AFLPs, 38 RFLPs and 77 SSRs in 23 linkage groups (LGs, covering a total map length of 798.0 cM. The ML161 map contains 240 markers (50 AFLPs, 71 RFLPs and 119 SSRs in 24 LGs covering a total of 1,328.1 cM. Using the improved maps, two quantitative trait loci (QTLs associated with tissue culturability were identified each for callusing rate and embryogenesis rate. A QTL for callogenesis was identified in LGD4b of ENL48 and explained 17.5% of the phenotypic variation. For embryogenesis rate, a QTL was detected on LGP16b in ML161 and explained 20.1% of the variation. This study is the first attempt to identify QTL associated with tissue culture amenity in oil palm which is an important step towards understanding the molecular processes underlying clonal regeneration of oil palm.

  12. Molecular biology of breast cancer metastasis Molecular expression of vascular markers by aggressive breast cancer cells

    During embryogenesis, the formation of primary vascular networks occurs via the processes of vasculogenesis and angiogenesis. In uveal melanoma, vasculogenic mimicry describes the 'embryonic-like' ability of aggressive, but not nonaggressive, tumor cells to form networks surrounding spheroids of tumor cells in three-dimensional culture; these recapitulate the patterned networks seen in patients' aggressive tumors and correlates with poor prognosis. The molecular profile of these aggressive tumor cells suggests that they have a deregulated genotype, capable of expressing vascular phenotypes. Similarly, the embryonic-like phenotype expressed by the aggressive human breast cancer cells is associated with their ability to express a variety of vascular markers. These studies may offer new insights for consideration in breast cancer diagnosis and therapeutic intervention strategies

  13. Mapping Quantitative Trait Loci Controlling Endosperm Traits with Molecular Marker

    XU Chen-wu; LI Tao; SUN Chang-sen; GU Shi-liang


    Based on the genetic models for triploid endosperm traits and on the methods for mapping diploid quantitative traits loci (QTLs), the genetic constitutions, components of means and genetic variances of QTL controlling endosperm traits under flanking marker genotypes of different generations were presented. From these results, a multiple linear regression method for mapping QTL underlying endosperm traits in cereals was proposed, which used the means of endosperm traits under flanking marker genotypes as a dependent variable, the coefficient of additive effect ( d ) and dominance effect ( h 1 and/or h2 ) of a putative QTL in a given interval as independent variables. This method can work at any position in a genome covered by markers and increase the estimation precision of QTL location and their effects by eliminating the interference of other relative QTLs. This method can also be easily used in other uneven data such as markers and quantitative traits detected or measured in plants and tissues different either in generations or at chromosomal ploidy levels, and in endosperm traits controlled by complicated genetic models considering the effects produced by genotypes of both maternal plants and seeds on them.

  14. Molecular markers for granulovacuolar degeneration are present in rimmed vacuoles.

    Masahiro Nakamori

    Full Text Available BACKGROUND: Rimmed vacuoles (RVs are round-oval cytoplasmic inclusions, detected in muscle cells of patients with myopathies, such as inclusion body myositis (IBM and distal myopathy with RVs (DMRV. Granulovacuolar degeneration (GVD bodies are spherical vacuoles containing argentophilic and hematoxyphilic granules, and are one of the pathological hallmarks commonly found in hippocampal pyramidal neurons of patients with aging-related neurodegenerative diseases, such as Alzheimer's disease and Parkinson's disease. These diseases are common in the elderly and share some pathological features. Therefore, we hypothesized that mechanisms of vacuolar formation in RVs and GVD bodies are common despite their role in two differing pathologies. We explored the components of RVs by immunohistochemistry, using antibodies for GVD markers. METHODS: Subjects included one AD case, eight cases of sporadic IBM, and three cases of DMRV. We compared immunoreactivity and staining patterns for GVD markers. These markers included: (1 tau-modifying proteins (caspase 3, cyclin-dependent kinase 5 [CDK5], casein kinase 1δ [CK1δ], and c-jun N-terminal kinase [JNK], (2 lipid raft-associated materials (annexin 2, leucine-rich repeat kinase 2 [LRRK2], and flotillin-1, and (3 other markers (charged multi-vesicular body protein 2B [CHMP2B] and phosphorylated transactive response DNA binding protein-43 [pTDP43] in both GVD bodies and RVs. Furthermore, we performed double staining of each GVD marker with pTDP43 to verify the co-localization. RESULTS: GVD markers, including lipid raft-associated proteins and tau kinases, were detected in RVs. CHMP2B, pTDP43, caspase 3, LRRK2, annexin 2 and flotillin-1 were detected on the rim and were diffusely distributed in the cytoplasm of RV-positive fibers. CDK5, CK1δ and JNK were detected only on the rim. In double staining experiments, all GVD markers colocalized with pTDP43 in RVs. CONCLUSIONS: These results suggest that RVs of muscle

  15. Characterization of a resistance locus (Pfs-1) to the spinach downy mildew pathogen (Peronospora farinosa f. sp. spinaciae) and development of a molecular marker linked to Pfs-1.

    Irish, B M; Correll, J C; Feng, C; Bentley, T; de Los Reyes, B G


    Downy mildew is a destructive disease of spinach worldwide. There have been 10 races described since 1824, six of which have been identified in the past 10 years. Race identification is based on qualitative disease reactions on a set of diverse host differentials which include open-pollinated cultivars, contemporary hybrid cultivars, and older hybrid cultivars that are no longer produced. The development of a set of near-isogenic open-pollinated spinach lines (NILs), having different resistance loci in a susceptible and otherwise common genetic background, would facilitate identification of races of the downy mildew pathogen, provide a tool to better understand the genetics of resistance, and expedite the development of molecular markers linked to these disease resistance loci. To achieve this objective, the spinach cv. Viroflay, susceptible to race 6 of Peronospora farinosa f. sp. spinaciae, was used as the recurrent susceptible parent in crosses with the hybrid spinach cv. Lion, resistant to race 6. Resistant F(1) progeny were subsequently backcrossed to Viroflay four times with selection for race 6 resistance each time. Analysis of the segregation data showed that resistance was controlled by a single dominant gene, and the resistance locus was designated Pfs-1. By bulk segregant analysis, an amplified fragment length polymorphism (AFLP) marker (E-ACT/M-CTG) linked to Pfs-1 was identified and used to develop a co-dominant Sequence characterized amplified region (SCAR) marker. This SCAR marker, designated Dm-1, was closely linked ( approximately 1.7 cM) to the Pfs-1 locus and could discriminate among spinach genotypes that were homozygous resistant (Pfs-1Pfs-1), heterozygous resistant (Pfs-1pfs-1), or homozygous susceptible (pfs-1pfs-1) to race 6 within the original mapping population. Evaluation of a wide range of commercial spinach lines outside of the mapping population indicated that Dm-1 could effectively identify Pfs-1 resistant genotypes; the Dm-1 marker

  16. Markers

    Healthy Schools Network, Inc., 2011


    Dry erase whiteboards come with toxic dry erase markers and toxic cleaning products. Dry erase markers labeled "nontoxic" are not free of toxic chemicals and can cause health problems. Children are especially vulnerable to environmental health hazards; moreover, schools commonly have problems with indoor air pollution, as they are more densely…

  17. Application of molecular markers for variety protection of ryegrass (Lolium perenne L.)

    Jensen, Louise Bach; Deneken, Gerhard; Roulund, N; Nielsen, K K; Lübberstedt, T


    legislation complies with the Convention of the International Union for the Protection of New Varieties of Plants (UPOV 1991). The corner stone of the UPOV system is that in order to qualify for protection, a newly bred plant variety has to be shown to be distinct (D), uniform (U) and stable (S). At the...... morphological characterization from the DUS trial. 18 SSR markers were selected based on their genome distribution, reproducibility, level of information and ease of scoring. It was found, that for variety discrimination, reducing the number of SSR markers from 18 SSR markers with 262 alleles to six SSR markers...... with 140 alleles gives the same level of information. Furthermore, number of genotypes per variety can be reduced to 20 compared to the original dataset containing 60 genotypes when using all 18 SSR markers but not when using only six SSR markers. Significant association was found between the molecular...

  18. The Effects of Water Matrix on Decay of Human Fecal Molecular Markers and Campylobacter spp.

    Although molecular source tracking for human fecal contamination is used on a wide range of sample types, little is known about comparative decay of proposed molecular markers under different conditions, or correlation with pathogen decay. Our purpose was to measure correlations ...

  19. Genetic Confirmation of Mungbean (Vigna radiata) and Mashbean (Vigna mungo) Interspecific Recombinants using Molecular Markers

    Abbas, Ghulam; Hameed, Amjad; Rizwan, Muhammad; Ahsan, Muhammad; Asghar, Muhammad J.; Iqbal, Nayyer


    Molecular confirmation of interspecific recombinants is essential to overcome the issues like self-pollination, environmental influence, and inadequacy of morphological characteristics during interspecific hybridization. The present study was conducted for genetic confirmation of mungbean (female) and mashbean (male) interspecific crosses using molecular markers. Initially, polymorphic random amplified polymorphic DNA (RAPD), universal rice primers (URP), and simple sequence repeats (SSR) mar...

  20. Molecular Pathology: Prognostic and Diagnostic Genomic Markers for Myeloid Neoplasms.

    Kuo, Frank C


    Application of next-generation sequencing (NGS) on myeloid neoplasms has expanded our knowledge of genomic alterations in this group of diseases. Genomic alterations in myeloid neoplasms are complex, heterogeneous, and not specific to a disease entity. NGS-based panel testing of myeloid neoplasms can complement existing diagnostic modalities and is gaining acceptance in the clinics and diagnostic laboratories. Prospective, randomized trials to evaluate the prognostic significance of genomic markers in myeloid neoplasms are under way in academic medical centers. PMID:27523973

  1. Estandarización de la técnica molecular de aflp en palma de aceite tipo dura (elaeis guineensis jacq.) y estudio preliminar de caracterización

    Galeano, Carlos Hernando


    La palma de aceite (Elaeis guinnensis Jacq.) desempeña un importante papel en la econo­mía de algunos países tropicales; Colombia ocupa el primer puesto en producción en América Latina y el quinto lugar a nivel mundial. En esta especie el mejo­ramiento genético busca generar materiales con alta producción de aceite y adaptados a las características edafoclimáticas de las diferentes zonas palmeras. El objetivo del presente trabajo fue estandarizar la téc­nica de AFLP (Amplified Fragment-Length...

  2. Molecular Markers in Differential Diagnostics of Follicular Neoplasms of the Thyroid

    E Troshina


    Full Text Available Ultrasound-guided fine-needle aspiration biopsy (FNAB is a basic method of morphological diagnostics at the preoperative examination, although it has some limitations. In 10-30 % of the cases, cytological examination results defined as indefinite or suspicious to malignant nodules, including follicular neoplasm, as according to the results of a cytological examination it does not appear to be possible to make the difference between follicular attendance the molecular markers adenomas and follicular cancer. Molecular medicine progress let us put an additional examination in a cytological, or surgical aspirates with the molecular markers. The most effective molecular markers in the clinical practice are thyroid peroxidases (TPO, telomerase and galectin-3. The application FNAB with the following immunocytochemistry examination in the thyroid tissue let us improve a differential diagnostics between benign and malignant nodules of the thyroid.

  3. AFLP and RAPD Analysis of the Boer and Indigenous Breeds of the Goat in Jiangsu

    CAO Shao-xian; YANG Li-guo; JIANG Xun-ping; LIU Hong-lin; LU Wei-zhong; XIANG Yang-hai


    Blood and tissue samples were collected from 105 goats including 60 Boer goats (30 for eachsex), 30 Xuhuai goats (15 for each sex) and 15 Haimen goats (7 stud and 8 does). DNA was extracted andDNA pools were constructed on the basis of goat breeds. In 36 selective primer combinations, 29 combinationsamplified totally 3 253 markers including 92 polymorphic markers by amplified fragment length polymor-phism (AFLP). On average, 3.17 polymorphic markers were amplified per combination, with a polymorphicfrequency of 2.8%. The primer combinations amplifying more polymorphic markers (showed in brackets)were involved in E00+ACG/M00+CAA (13), E00+ACG/M00+CAG (10), E00+AAC/M00+CAC (8)and E00+AAC/M00+ACT (7). A total of 183 markers including 60 polymorphic markers were amplified byRAPD from the pooled DNA of three breeds using 22 primers with strong polymorphism and high reproduc-ibility selected from 93 RAPD primers. On average, 2.73 polymorphic markers were amplified per primer,with a polymorphic frequency of 32.8%. The results of AFLP and RAPD coincidently suggested that the ge-netic distance is the closest between Xuhuai and Haimen goat, next between Xuhuai and Boer goat, and the far-thest between Haimen and Boer goat. According to the UPGMA method, Haimen and Xuhuai goats can begathered together as a cluster, then Boer goat. Both methods can be used to implicate the genetic difference ofthese three breeds, in particular AFLP has more polymorphic markers.

  4. Molecular Markers with Predictive and Prognostic Relevance in Lung Cancer

    Alphy Rose-James; TT, Sreelekha


    Lung cancer accounts for the majority of cancer-related deaths worldwide of which non-small-cell lung carcinoma alone takes a toll of around 85%. Platinum-based therapy is the stronghold for lung cancer at present. The discovery of various molecular alterations that underlie lung cancer has contributed to the development of specifically targeted therapies employing specific mutation inhibitors. Targeted chemotherapy based on molecular profiling has shown great promise in lung cancer treatment...

  5. Molecular markers to study competition and diversity of Rhizobium.

    Sessitsch, A


    The research described in this thesis was directed to the development of molecular identification and detection techniques for studying the ecology of Rhizobium, a nitrogen- fixing bacterium of agricultural importance. Competition of inoculant strains with indigenous microbes is a serious problem in agricultural practice and was therefore addressed in this work using the developed tools. Furthermore, various molecular techniques have been applied to analyse rhizobial populations nodulating co...

  6. An assessment of the use of molecular markers in developing countries

    Four different sources of information were analysed to assess the current uses of molecular markers in crops, forest trees and livestock in developing countries: the FAO Biotechnology in Developing Countries (FAO-BioDeC) database of biotechnology in developing countries; country reports evaluating the current status of applied plant breeding and related biotechnologies; country reports on animal genetic resources management for preparing the First Report on the State of the World's Animal Genetic Resources (SoW-AnGR); and the results of a questionnaire survey on animal genetic diversity studies. Even if still largely incomplete, the current data show that molecular markers are widely used for plant breeding in the developing world and most probably their use will increase in the future. In the animal sector the use of molecular markers seems less developed and limited or absent in most developing countries. Major differences exist among and within regions regarding the application of molecular marker techniques in plant and animal breeding and genetics. These can be explained by the relatively high investments in infrastructure and human resources necessary to undertake research in these fields. The spectrum of application of molecular markers in crop plants is quite wide, covering many plants relevant to the enhancement of food security, but other important plant species are still neglected. The practical results of marker-assisted selection (MAS) in the field are disappointingly modest, possibly due to: low levels of investment; limited coordination between biotechnologists and practical breeders; instable, non-focused or ill-addressed research projects; and the lack of linkages between research and farmers. Partnerships between developed and developing countries may be a means of better realizing the potential of molecular marker techniques for improving both animal and crop production. (author)

  7. Molecular markers for detection of resistance to chemotherapy

    Objectives: The scope of this thesis was to select new biomarkers for the response to standard chemotherapies and new targeted therapies in ovarian cancer. Furthermore the utility of new platforms for the routine testing of biomarkers on RNA and DNA level was evaluated. Such markers are especially interesting for ovarian cancer as after initial good response to chemotherapy most tumors acquire multiple drug resistance (MDR). Material and Methods: Mutational status of KRAS was determined in fresh frozen and formalin fixed paraffin embedded (FFPE) ovarian tissue samples. The experiments were conducted on two different platforms, Gastoxin, a micro array system, and a reverse hybridisation strip assay. Gene expression of nine ATP-binding cassette (ABC) transporters were analysed in recurrent ovarian cancer samples and benign tissue with real-time Pcr. Transporters exhibiting a significant overexpression in recurrent disease were further evaluated in primary cancer tissue. Furthermore real-time Pcr results were validated with two novel platforms. Results: In 15% of ovarian carcinoma samples KRAS was mutated. Mutation rates in fresh and FFPE tissue were approximately the same which leads to the conclusion that both assays are able to process these types of tissue. Four of the ABC transporters were significantly higher expressed in recurrent cancer tissue. Primary lesions compared to benign tissue showed no mentionable differences in gene expression. Therefore the examined transporters are not feasible as prognostic markers but some seem to play a role in MDR of ovarian cancer. Regarding the two tested platforms, the Quantitating 2.0 Reagent System was found to be an adequate alternative to real-time Pcr. For the Approve-B platform the first optimization experiments were promising, further development is currently ongoing. Conclusion: Mutation of KRAS is no prognostic marker for patients under standard therapy, but in the light of the new anti-EGF R therapies, which are

  8. Molecular genetics of vaccinia virus: demonstration of marker rescue.

    Nakano, E.; Panicali, D; E. Paoletti


    Two genomic variants of vaccinia virus isolated from serially propagated stocks were used to demonstrate marker rescue. The smaller (S variant) virus contains a 6.3 megadalton (MDal) deletion of unique DNA sequences present in the 123-MDal larger (L variant) virus. The deletion was mapped at 6.85 MDal from the left terminus of the genome, just outside of the inverted terminal repetition. Rescue of the unique deleted DNA sequences by infectious S variant virus was obtained in CV-1 cells by usi...

  9. Molecular and cellular markers of toxicity in the Japanese Medaka @

    Shugart, L.R.; McCarthy, J.F.; D' Surney, S.J.; Greeley, M.S. Jr.; Hull, C.G.


    The Japanese Medaka (Oryzias latipes) has been recommended for use as a model organism to detect carcinogenic, teratogenic, cytotoxic, and genotoxic compounds in aquatic systems. Because a long latent period often occurs between initial contact with deleterious chemicals and subsequent expression of the pathology, we are investigating early biologically-relevant responses that can be used as genotoxicity markers of exposure and effect. This project focuses on the development of genotoxic bioassays and experimental protocols for exposing Japanese Medaka to genotoxic compounds. 21 refs., 8 figs, 2 tabs.

  10. Metabolomics and molecular marker analysis to explore pepper (Capsicum sp.) biodiversity.

    Wahyuni, Yuni; Ballester, Ana-Rosa; Tikunov, Yury; de Vos, Ric C H; Pelgrom, Koen T B; Maharijaya, Awang; Sudarmonowati, Enny; Bino, Raoul J; Bovy, Arnaud G


    An overview of the metabolic diversity in ripe fruits of a collection of 32 diverse pepper (Capsicum sp.) accessions was obtained by measuring the composition of both semi-polar and volatile metabolites in fruit pericarp, using untargeted LC-MS and headspace GC-MS platforms, respectively. Accessions represented C. annuum, C. chinense, C. frutescens and C. baccatum species, which were selected based on variation in morphological characters, pungency and geographic origin. Genotypic analysis using AFLP markers confirmed the phylogenetic clustering of accessions according to Capsicum species and separated C. baccatum from the C. annuum-C. chinense-C. frutescens complex. Species-specific clustering was also observed when accessions were grouped based on their semi-polar metabolite profiles. In total 88 semi-polar metabolites could be putatively identified. A large proportion of these metabolites represented conjugates of the main pepper flavonoids (quercetin, apigenin and luteolin) decorated with different sugar groups at different positions along the aglycone. In addition, a large group of acyclic diterpenoid glycosides, called capsianosides, was found to be highly abundant in all C. annuum genotypes. In contrast to the variation in semi-polar metabolites, the variation in volatiles corresponded well to the differences in pungency between the accessions. This was particularly true for branched fatty acid esters present in pungent accessions, which may reflect the activity through the acyl branch of the metabolic pathway leading to capsaicinoids. In addition, large genetic variation was observed for many well-established pepper aroma compounds. These profiling data can be used in breeding programs aimed at improving metabolite-based quality traits such as flavour and health-related metabolites in pepper fruits. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s11306-012-0432-6) contains supplementary material, which is available to

  11. Molecular marker-assisted selection for resistance to pathogens in tomato

    Since the 1980s, the use of molecular markers has been suggested to improve the efficiency of releasing resistant varieties, thus overcoming difficulties met with classical breeding. For tomato, a high-density molecular map is available in which more than 40 resistance genes are localized. Markers linked to these genes can be used to speed up gene transfer and pyramiding. Suitable PCR markers targeting resistance genes were constructed directly on the sequences of resistance genes or on restriction fragment length polymorphisms (RFLPs) tightly linked to them, and used to select resistant genotypes in backcross schemes. In some cases, the BC5 generation was reached, and genotypes that cumulated two homozygous resistant genes were also obtained. These results supported the feasibility of using marker-assisted selection (MAS) in tomato and reinforcing the potential of this approach for other genes, which is today also driven by the development of new techniques and increasing knowledge about the tomato genome. (author)

  12. Use of Molecular Markers for Catfish Production and Product Quality

    Channel catfish are a diploid species native to North America and the primary product of U.S. aquaculture, and molecular genetic research supports selective breeding of catfish for commercial production. To date, short tandem repeat loci have been used to characterize DNA sequence variation within ...

  13. Genetic Diversity Analysis of 98 Collections of Sugarcane Germplasm with AFLP Markers%98份甘蔗种质资源遗传多样性的AFLP分析

    昝逢刚; 应雄美; 吴才文; 赵培方; 陈学宽; 马丽; 苏火生; 刘家勇


    BURMA;第IV类群有89份种质,在遗传相似性系数0.79处切割时,可将第Ⅳ类群划分为9个亚群(A、B、C、D、E、F、G、H和I)。基于Jaccard系数用PCA法对98份甘蔗种质AFLP标记结果进行主效应分析,主效应分析显示了不同种质的分类位置,分子主效应分析结果与分子聚类结果一致,集中在一个区域的种质亲缘关系较为紧密,澳大利亚种质位置较为分散,最分散材料为蔗茅属和细茎野生种。【结论】98份种质资源的遗传基础差异较小,亲缘关系较近,其中,澳大利亚种质遗传多样性相对较丰富。90-110-9、KN93-06和粤糖00-236较为特殊,在杂交组合选配中应给予重点关注。%Objective]Cane sugar accounts for 92% of sugar production in China, and hybridization is the most widely used and the most effective way for developing new sugarcane cultivars. Sugarcane germplasm is essential for sugarcane breeding. Selecting parental clones and the cross combinations for hybridization contributes directly to the breeding efficiency. Aimed at providing reference for selecting parental clones and cross combinations, the genetic diversity and similarity among 98 sugarcane germplasm were studied.[Method]Good genomic DNA was extracted from young leaves of 98 sugarcane germplasm collected from 10 countries following the CTAB method, and then were amplified by sequence-related amplified polymorphism molecular markers to analyze genetic diversity and genetic similarity. Separation of the amplified fragments was performed on 5% denaturing polyacrylamide gels, the gels were stained with AgNO3, then “0,1” matrix was obtained according to the electrophoresis result. The number of polymorphic loci, percentage of polymorphic loci, quantity of polymorphic information, effective number of alleles and the indexes of genetic diversity were estimated by POPGENE version 32. The genetic similarity that estimated by NTSYS pc-V. 2.1 was used for UPGMA (unweighted

  14. Prediction of Physicochemical Properties of Indonesian Indica Rice Using Molecular Markers



    Full Text Available Physicochemical properties determine the palatability and cooking quality of rice, which must be determined efficiently in order to satisfy consumer demand. To date, little information exists on the use of molecular markers to predict physicochemical properties of the “indica” rice varieties found in Indonesia. The objective of this study was to investigate physicochemical properties and genetic variation of Indonesian rice varieties, and to formulate regression equations to analyze sets of DNA markers which could predict amylose content (AC, protein content (PC and pasting properties of the varieties. A total of 24 Indonesian indica rice varieties were chosen based on their genetic background and agricultural characteristics. We then measured selected physicochemical properties, and genotyped the varieties using 30 DNA markers. The chosen varieties showed favorable values for PC, AC, and six rapid viscosity analyzer (RVA pasting properties, which was supported by molecular data. As demonstrated by principal component analysis (PCA, markers could provide a complementary method for differentiating rice varieties, as an alternative to measuring physicochemical properties. PCA analysis also allowed us to establish marker sets using multiple regression analysis. We formulated eight model regression equations comprising data regarding 15 to 19 markers with high coefficients (R2=0.98-0.99. The formulas provided results that consistently correlated and therefore predicted the physicochemical properties of indica rice. Further validation of these marker sets may provide rapid and efficient means for predicting the physicochemical properties of Indonesian-bred indica rice in the future.

  15. Molecular Markers of Secondary Organic Aerosol in Mumbai, India.

    Fu, Pingqing; Aggarwal, Shankar G; Chen, Jing; Li, Jie; Sun, Yele; Wang, Zifa; Chen, Huansheng; Liao, Hong; Ding, Aijun; Umarji, G S; Patil, R S; Chen, Qi; Kawamura, Kimitaka


    Biogenic secondary organic aerosols (SOA) are generally considered to be more abundant in summer than in winter. Here, polar organic marker compounds in urban background aerosols from Mumbai were measured using gas chromatography-mass spectrometry. Surprisingly, we found that concentrations of biogenic SOA tracers at Mumbai were several times lower in summer (8-14 June 2006; wet season; n = 14) than in winter (13-18 February 2007; dry season; n = 10). Although samples from less than 10% of the season are extrapolated to the full season, such seasonality may be explained by the predominance of the southwest summer monsoon, which brings clean marine air masses to Mumbai. While heavy rains are an important contributor to aerosol removal during the monsoon season, meteorological data (relative humidity and T) suggest no heavy rains occurred during our sampling period. However, in winter, high levels of SOA and their day/night differences suggest significant contributions of continental aerosols through long-range transport together with local sources. The winter/summer pattern of SOA loadings was further supported by results from chemical transport models (NAQPMS and GEOS-Chem). Furthermore, our study suggests that monoterpene- and sesquiterpene-derived secondary organic carbon (SOC) were more significant than those of isoprene- and toluene-SOC at Mumbai. PMID:27045808

  16. Molecular markers for tumor cell dissemination in female cancers

    In the fight against cancer many advances have been made in early detection and treatment of the disease during the last few decades. Nevertheless, many patients still die of cancer due to metastatic spreading of the disease. Tumor cell dissemination may occur very early and usually is not discovered at the time of initial diagnosis. In these cases, the mere excision of the primary tumor is an insufficient treatment. Microscopic tumor residues will remain in the blood, lymph nodes, or the bone marrow and will cause disease recurrence. To improve the patient's prognosis, a sensitive tool for the detection of single tumor cells supplementing conventional diagnostic procedures is required. As the blood is more easily accessible than the bone marrow or tissue biopsies, we intended to identify gene markers for the detection of circulating tumor cells in the blood of cancer patients. We focused on patients with breast, ovarian, endometrial or cervical cancer. Starting from a genome-wide gene expression analysis of tumor cells and blood cells, we found six genes higher expression levels in cancer patients compared to healthy women. These findings suggest that an increased expression of these genes in the blood indicates the presence of circulating tumor cells inducing future metastases and thus the need for adjuvant therapy assisting the primary treatment. Measuring the expression levels of these six genes in the blood may supplement conventional diagnostic tests and improve the patient's prognosis. (author)

  17. Gene Specific Molecular Markers for Hop (Humulus lupulus L.)

    Patzak, J.; Matoušek, Jaroslav

    Ghent: ISHA Acta Horticulturae, 2009 - (DeKeukeleire, D.; Hummer, K.; Heyerick, A.), s. 73-80 ISBN 978-90-6605-722-7. ISSN 0567-7572. [ISHS International Humulus Symposium /2./. Ghent (BE), 01.09.2009-05.09.2009] R&D Projects: GA ČR GA521/08/0740; GA MZe QH81052 Institutional research plan: CEZ:AV0Z50510513 Keywords : cluster analysis * expressed sequence tag microsatellite repeat (EST-SSR) * single nucleotide polymorphism (SNP) Subject RIV: EB - Genetics ; Molecular Biology

  18. Estimating Out-Crossing Rate of Bg 379-2 Using Morphological Markers and Confirmation by Molecular Markers



    Rice is largely self-fertilized and accordingly a field population of rice is completely composed of near homozygotes.Due to the emergence of off-types,homozygosity will be affected.With the time,this will cause the reduction of genetic purity in some rice varieties.One of the reasons has been suspected to be the high out-crossing frequencies of such varieties.Studies were conducted at the Rice Research and Development Institute,Batalagoda,Sri Lanka to estimate the out-crossing rote of Bg 379-2,a variety having the problem of maintaining genetic purity.Bg 379-2 was allowed to out-cross with Bg 450 and the number of outcrossed plants were counted using dominant morphological markers such as short-round grain and purple culm of pollen donor.A molecular confirmation of out-crossing was also performed using sequence tagged site (STS) molecular marker pTA248.The variety Bg 379-2 showed a potential out-crossing rate of 3.41% and an average out-crossing rate of 1.29% using dominant morphological markers.Polymorphism was clearly detected between parents and out-crossed plants as well as selfed plants of Bg 379-2 using their banding patterns.A similar study can be performed to determine the out-crossing rates of other varieties which show high percentage of off-types in the population for the better understanding of the breeding behavior of the varieties.

  19. Molecular markers and targets for colorectal cancer prevention

    Naveena B JANAKIRAM; Chinthalapally V RAO


    Colorectal cancer is the third most prevalent cancer in the world. If detected at an early stage, treatment often might lead to cure. As prevention is better than cure, epidemiological studies reveal that having a healthy diet often protects from pro-moting/developing cancer. An important consideration in evaluating new drugs and devices is determining whether a product can effectively treat a targeted disease. There are quite a number of biomarkers making their way into clinical trials and few are awaiting the preclinical efficacy and safety results to enter into clinical trials. Researchers are facing challenges in modifying trial design and defining the right control population, validating biomarker assays from the bio-logical and analytical perspective and using biomarker data as a guideline for decision making. In spite of following all guidelines, the results are disappointing from many of the large clinical trials. To avoid these disappointments, selection of biomarkers and its target drug needs to be evaluated in appropriate animal models for its toxicities and efficacies. The focus of this review is on the few of the potential molecular targets and their biomarkers in colorectal cancers. Strengths and limitations of biomarkers/surrogate endpoints are also discussed. Various pathways involved in tumor cells and the specific agents to target the altered molecular biomarkerin biomolecular pathwayare elucidated. Importance of emerging new platforms siRNAs and miRNAs technology for colorectal cancer therapeutics is reviewed.

  20. Highly isotopically depleted isoprenoids: Molecular markers for ancient methane venting

    Thiel, Volker; Peckmann, Jörn; Seifert, Richard; Wehrung, Patrick; Reitner, Joachim; Michaelis, Walter


    We propose that organic compounds found in a Miocene limestone from Marmorito (Northern Italy) are source markers for organic matter present in ancient methane vent systems (cold seeps). The limestone contains high concentrations of the tail-to-tail linked, acyclic C 20 isoprenoid 2,6,11,15-tetramethylhexadecane (crocetane), a C 25 homolog 2,6,10,15,19-pentamethylicosane (PME), and a distinctive glycerol ether lipid containing 3,7,11,15-tetramethylhexadecyl (phytanyl-) moieties. The chemical structures of these biomarkers indicate a common origin from archaea. Their extremely 13C-depleted isotope compositions (δ 13C ≈ -108 to -115.6‰ PDB) suggest that the respective archaea have directly or indirectly introduced isotopically depleted, methane-derived carbon into their biomass. We postulate that a second major cluster of biomarkers showing heavier isotope values (δ 13C ≈ -88‰) is derived from sulfate-reducing bacteria (SRB). The observed biomarkers sustain the idea that methanogenic bacteria, in a syntrophic community with SRB, are responsible for the anaerobic oxidation of methane in marine sediments. Marmorito may thus represent a conceivable ancient scenario for methane consumption performed by a defined, two-membered bacterial consortium: (1) archaea that perform reversed methanogenesis by oxidizing methane and producing CO 2 and H 2; and (2) SRB that consume the resulting H 2. Furthermore, the respective organic molecules are, unlike other compounds, tightly bound to the crystalline carbonate phase. The Marmorito carbonates can thus be regarded as "cold seep microbialites" rather than mere "authigenic" carbonates.

  1. Biomedical wellness monitoring system based upon molecular markers

    Ingram, Whitney


    We wish to assist caretakers with a sensor monitoring systems for tracking the physiological changes of homealone patients. One goal is seeking biomarkers and modern imaging sensors like stochastic optical reconstruction microscopy (STORM), which has achieved visible imaging at the nano-scale range. Imaging techniques like STORM can be combined with a fluorescent functional marker in a system to capture the early transformation signs from wellness to illness. By exploiting both microscopic knowledge of genetic pre-disposition and the macroscopic influence of epigenetic factors we hope to target these changes remotely. We adopt dual spectral infrared imaging for blind source separation (BSS) to detect angiogenesis changes and use laser speckle imaging for hypertension blood flow monitoring. Our design hypothesis for the monitoring system is guided by the user-friendly, veteran-preferred "4-Non" principles (noninvasive, non-contact, non-tethered, non-stop-to-measure) and by the NIH's "4Ps" initiatives (predictive, personalized, preemptive, and participatory). We augment the potential storage system with the recent know-how of video Compressive Sampling (CSp) from surveillance cameras. In CSp only major changes are saved, which reduces the manpower cost of caretakers and medical analysts. This CSp algorithm is based on smart associative memory (AM) matrix storage: change features and detailed scenes are written by the outer-product and read by the inner product without the usual Harsh index for image searching. From this approach, we attempt to design an effective household monitoring approach to save healthcare costs and maintain the quality of life of seniors.

  2. Identification of Chromosomes from Multiple Rice Genomes Using a Universal Molecular Cytogenetic Marker System

    Xiaomin Tang; Weidong Bao; Wenli Zhang; Zhukuan Cheng


    To develop reliable techniques for chromosome identification is critical for cytogenetic research, especially for genomes with a large number and smaller-sized chromosomes. An efficient approach using bacterial artificial chromosome (BAG) clones as molecular cytological markers has been developed for many organisms. Herein, we present a set of chromosomal arm-specific molecular cytological markers derived from the gene-enriched regions of the sequenced rice genome. All these markers are able to generate very strong signals on the pachytene chromosomes of Oryza satlva L. (AA genome) when used as fluorescence in situ hybridization (FISH) probes. We further probed those markers to the pachytene chromosomes of O. punctata (BB genome) and O. officinalis (CC genome) and also got very strong signals on the relevant pachytene chromosomes. The signal position of each marker on the related chromosomes from the three different rice genomes was pretty much stable, which enabled us to identify different chromosomes among various rice genomes. We also constructed the karyotype for both O. punctata and O. officinalis with the BB and CC genomes, respectively, by analysis of 10 pachytene cells anchored by these chromosomal arm-specific markers.

  3. A format for databasing and comparison of AFLP fingerprint profiles

    Chuah Aaron; Hong Yan


    Abstract Background Amplified fragment length polymorphism (AFLP) is a PCR-based technique that involves restriction of genomic DNA followed by ligation of adaptors to the fragments generated and selective PCR amplification of a subset of these fragments. The amplified fragments are separated on a sequencing gel and visualized by autoradiography or fluorescent sequencing equipment. AFLP allows high-resolution genotyping but the lack of a format for databasing and comparison of AFLP fingerprin...

  4. Molecular markers as therapeutic targets in lung cancer

    Hsin-Hui Tseng; Biao He


    Lung cancer is responsible for 29% of cancer deaths in the United States and has very low 5-year survival rates of approximately 11% in men and 15% in women.Although the early diagnosis of lung cancer may increase the survival rate with adequate treatment,advanced lung cancers are often metastasized and receive limited benefit from therapeutic regimens.As conventional treatments for lung cancer reach their limitations,researchers have attempted to discover novel drug therapies aimed at specific targets contributing to the progression of tumorigenesis.Recent advances in systems biology have enabled the molecular biology of lung carcinogenesis to be elucidated.Our understanding of the physiologic processes of tumor development provide a means to design more effective and specific drugs with less toxicity,thereby accelerating the delivery of new drug therapies to the patient's bedside.

  5. Appraisal of progenitor markers in the context of molecular classification of breast cancers

    Haviv, Izhak


    Clinical management of breast cancer relies on case stratification, which increasingly employs molecular markers. The motivation behind delineating breast epithelial differentiation is to better target cancer cases through innate sensitivities bequeathed to the cancer from its normal progenitor state. A combination of histopathological and molecular classification of breast cancer cases suggests a role for progenitors in particular breast cancer cases. Although a remarkable fraction of the re...

  6. Transcriptome Analysis and Development of SSR Molecular Markers in Glycyrrhiza uralensis Fisch.

    Yaling Liu; Pengfei Zhang; Meiling Song; Junling Hou; Mei Qing; Wenquan Wang; Chunsheng Liu


    Licorice is an important traditional Chinese medicine with clinical and industrial applications. Genetic resources of licorice are insufficient for analysis of molecular biology and genetic functions; as such, transcriptome sequencing must be conducted for functional characterization and development of molecular markers. In this study, transcriptome sequencing on the Illumina HiSeq 2500 sequencing platform generated a total of 5.41 Gb clean data. De novo assembly yielded a total of 46,641 uni...

  7. Molecular Characterization of Some Turkish Olive Cultivars Using Random Amplified Polymorphic DNA (RAPD Markers

    Ergün KAYA


    Full Text Available Olive (Olea europea L. is one of the oldest cultivated plants characteristic in the Mediterranean area, where it is the most important oilproducing crop. The cultivated olive (O. europaea L. var. europaea is propagated by cutting or grafting, whereas wild olive (O. europaea L. var. sylvestris is reproduced from seeds. These two olive types are interfertile and have led to a large number of varieties. Morphological descriptions are not entirely reliable, due to numerous synonyms and homonyms in designations, labelling mistakes, the presence of varietal clones, and the uncertain identification methods thus far applied. Molecular markers, as random amplified polymorphic DNA (RAPD markers, are environment-independent and efficient to identify olive varieties and to detect synonymous and homonymous. In this study, fifteen selected RAPD markers are used for determination of relationships among twenty individuals belonging to four important Turkish olive cultivars. Our results showed that RAPD markers can be used to differentiate olive cultivars

  8. Evaluating the effects of non-neutral molecular markers on phylogeny inference.

    Dawn M Roje

    Full Text Available Nucleotide substitution models used in molecular phylogenetics do not account for nucleotide sequences evolving under selection, yet selection is rarely tested for. If non-neutral markers violate these models (i.e. non-independence of sites, it is expected that their reconstructed topologies be incongruent with those inferred from neutral ones and conclusions made from those phylogenies should be reexamined. Using rhodopsin as a phylogenetic marker has recently been called into question for exactly this reason. Rhodopsin is assumed to have evolved under strong positive selection for organisms that inhabit similar aquatic environments, making it unsuitable for the phylogenetics of aquatic organisms, but it is unclear what the effects of non-neutrality on phylogeny estimation are. To evaluate potential incongruence of neutral versus non-neutral markers, and the notion that rhodopsin should not be used in the molecular phylogenetics of fishes, a molecular dataset of 78 acanthomorph taxa and sequences from four nuclear, protein coding loci (including rhodopsin, were examined. Only one marker was found to be neutral while the remaining tests, for all other loci, rejected the null hypothesis of neutrality. To evaluate the possible effect(s of positively versus negatively selected sites, the three non-neutral markers were analyzed to determine the presence of positively and negatively selected codons. To determine congruence in topology among ML trees inferred by individual neutral and non-neutral markers, as well as the combined (concatenated dataset, tree, comparisons of distances among trees and hypothesis (topology testing were carried out. Results of the tree distance metrics and topology testing support the notion that neutrality alone does not determine congruence in topology, and those data that are inferred to have evolved under selection should not necessarily be excluded. In addition, the number of sites inferred to have evolved under

  9. Molecular genetic diversity of Punica granatum L. (pomegranate) as revealed by microsatellite DNA markers

    Pomegranate (Punica granatum L.) is one of the oldest known edible fruits and more and more it arouse interest of scientific community given its numerous biological activities. However, information about its genetic resources and characterization using reliable molecular markers are still scarce. In...


    This work focuses on analysis of organic molecular markers in airborne particulate matter (PM) by Gas Chromatography/Ion Trap Mass Spectrometry (GC/IT MS). The particulate samples used in the method development were collected as PM10 in metropolitan Philadelphia during...


    A. A. Zabolotneva


    Full Text Available Bladder cancer (BC is the second most common urinary tract malignancy. Early diagnosis of BC generally increases the probability of successful treatment in a patient. The paper considers noninvasive diagnosis methods for BC and gives a database of the known molecular markers of this disease.


    A. A. Zabolotneva; N. M. Gaifullin; A. A. Buzdin; B. Ya. Alekseyev; Yu. Yu. Andreyeva; P. V. Shegai; D. G. Sokov; I. G. Rusakov


    Bladder cancer (BC) is the second most common urinary tract malignancy. Early diagnosis of BC generally increases the probability of successful treatment in a patient. The paper considers noninvasive diagnosis methods for BC and gives a database of the known molecular markers of this disease.

  13. Application of Genome-Wide Expression Analysis To Identify Molecular Markers Useful in Monitoring Industrial Fermentations

    Higgins, Vincent J.; Rogers, Peter J.; Dawes, Ian W.


    Genome-wide expression analysis of an industrial strain of Saccharomyces cerevisiae identified the YOR387c and YGL258w homologues as highly inducible in zinc-depleted conditions. Induction was specific for zinc deficiency and was dependent on Zap1p. The results indicate that these sequences may be valuable molecular markers for detecting zinc deficiency in industrial fermentations.

  14. The prognostic value of molecular marker analysis in patients treated with trimodality therapy for esophageal cancer.

    Harpole, D H; Moore, M B; Herndon, J E; Aloia, T; D'Amico, T A; Sporn, T; Parr, A; Linoila, I; Allegra, C


    The purpose of this study was to define the prognostic value of a group of molecular tumor markers in a well-staged population of patients treated with trimodality therapy for esophageal cancer. The original pretreatment paraffin-embedded endoscopic esophageal tumor biopsy material was obtained from 118 patients treated with concurrent cisplatin + 5-fluorouracil (5-FU) + 45 Gy radiation followed by resection from 1986 until 1997 at the Duke University Comprehensive Cancer Center. Three markers of possible platinum chemotherapy association [metallothionein (MT), glutathione S-transferase-pi (GST-pi), P-glycoprotein (P-gp or multidrug resistance)] and one marker of possible 5-FU association [thymidylate synthase (TS)] were measured using immunohistochemistry. The median cancer-free survival was 25.0 months, with a significantly improved survival for the 38 patients who had a complete response (P GST-pi, P-gp, and TS were associated with a decreased survival. MT was not significant in this population. Multivariate analysis identified high-level expression in two of the platinum markers (GST-pi and P-gp) and the 5-FU marker TS as independent predictors of early recurrence and death. In conclusion, this investigation measured three possible markers associated with platinum and one possible marker associated with 5-FU in a cohort of esophageal cancer patients. Independent prognostic significance was observed, which suggests that it may be possible to predict which patients may benefit most from trimodality therapy. These data need to be reproduced in a prospective investigation. PMID:11297249

  15. Molecular features and genetic markers of gastrointestinal stromal tumors

    N. N. Mazurenko


    Full Text Available Gastrointestinal stromal tumors (GISTs are the most spread mesenchymal tumors located within the gastrointestinal tract that have particular clinico-morphological, immunohistochemical and molecular characteristics. The distinguishing mark of GISTs is the presence of the cell-surface antigen CD117 (KIT receptor tyrosine kinase, identified by immunohistochemistry. GISTs consist of tumors with various activating mutations in KIT (75–80 % or PDGFRA (5–15 % receptor tyrosine kinases. Numerous KIT and PDGFRA mutations are associated with specific GIST morphology, histologic phenotype, metastasizing and prognosis. 10–15 % of GISTs contain KIT and PDGFRA wild type genes, some of them have driver BRAF, IGF1R or PIK3CA mutations. The other GISTs patients have familial syndromes (neurofibromatosis type 1, Carney–Stratakis syndrome, Carney triad and contain germline mutations of NF1 or the genes coding for the succinate dehydrogenase subunits SDHA, SDHB, SDHC, and SDHD. GISTs are first and the most studied model for development of principles and methods of personalized targeted therapy of solid tumors with tyrosine kinase inhibitors.

  16. Ochratoxin A producing Penicillium verrucosum isolates from cereals reveal large AFLP fingerprinting variability

    Frisvad, J C; Lund, F; Elmholt, S.


    Aims: To examine if molecular amplified fragment length polymorphism (AFLP) fingerprinting of the only ochratoxin A-producing species in European cereals, Penicillium verrucosum, can be used as a method in hazard analysis using critical control points (HACCP). Methods and Results: A total of 321 isolates of P. verrucosum were isolated from ochratoxin A contaminated cereals from Denmark (oats), UK (wheat and barley) and Sweden (wheat). Of these, 236 produced ochratoxin A as determined by t...

  17. AFLP analysis of Xanthomonas axonopodis and X. arboricola strains used in xanthan production studies reveal high levels of polymorphism

    Laurí Mayer


    Full Text Available Amplified fragment length polymorphism (AFLP was used to analyze the genetic diversity of 14 strains of Xanthomonas arboricola pv. pruni and seven strains of X. axonopodis pv. phaseoli, which are used in xanthan production studies. Relationships identified by the AFLP profiles were assessed for xanthan production capacity, geographical location and host plant. Strains were isolated from 10 different geographic regions in South and Southeast States in Brazil. Data were analyzed for genetic similarity using the Dice coefficient and subjected to UPGMA cluster analysis. A total of 128 AFLP fragments were generated from four primer combinations: EcoRI+C/MseI+0, EcoRI+A/MseI+0, EcoRI+G/MseI+T and EcoRI+G/MseI+A. Of these, 96.1% were polymorphic. X. axonopodis pv. phaseoli (S D = 0.27 was shown to be more polymorphic than X. arboricola pv. pruni (S D = 0.58. All 14 pathovar pruni strains were included in a single main group (S D = 0.58, while the pathovar phaseoli strains were divided into three separate groups, with one group containing five strains (S D = 0.38 and two isolated groups (S D = 0.31 and 0.27 composed of only one strain each. Species were distinguished by three and eight specific AFLP markers present in the pathovar phaseoli and the pathovar pruni, respectively. For the unique strain without xanthan production capacity (X. axonopodis pv. phaseoli str. 48, nine specific AFLP bands were found. There was no evidence that geographic area or host plant influenced genetic heterogeneity. Correlations between AFLP patterns and xanthan production capacity were found in some strains, but were not consistent enough to establish a relationship.

  18. Origins of the amphiploid species Brassica napus L. investigated by chloroplast and nuclear molecular markers

    Allender Charlotte J


    Full Text Available Abstract Background The amphiploid species Brassica napus (oilseed rape, Canola is a globally important oil crop yielding food, biofuels and industrial compounds such as lubricants and surfactants. Identification of the likely ancestors of each of the two genomes (designated A and C found in B. napus would facilitate incorporation of novel alleles from the wider Brassica genepool in oilseed rape crop genetic improvement programmes. Knowledge of the closest extant relatives of the genotypes involved in the initial formation of B. napus would also allow further investigation of the genetic factors required for the formation of a stable amphiploid and permit the more efficient creation of fully fertile re-synthesised B. napus. We have used a combination of chloroplast and nuclear genetic markers to investigate the closest extant relatives of the original maternal progenitors of B. napus. This was based on a comprehensive sampling of the relevant genepools, including 83 accessions of A genome B. rapa L. (both wild and cultivated types, 94 accessions of B. napus and 181 accessions of C genome wild and cultivated B. oleracea L. and related species. Results Three chloroplast haplotypes occurred in B. napus. The most prevalent haplotype (found in 79% of accessions was not present within the C genome accessions but was found at low frequencies in B. rapa. Chloroplast haplotypes characteristic of B. napus were found in a small number of wild and weedy B. rapa populations, and also in two accessions of cultivated B. rapa 'brocoletto'. Whilst introgression of the B. napus chloroplast type in the wild and weedy B. rapa populations has been proposed by other studies, the presence of this haplotype within the two brocoletto accessions is unexplained. Conclusions The distribution of chloroplast haplotypes eliminate any of the C genome species as being the maternal ancestor of the majority of the B. napus accessions. The presence of multiple chloroplast

  19. Immunohistochemical molecular markers as predictors of curability of endoscopically resected submucosal colorectal cancer


    AIM: To clarify the usefulness of immunohistochemical molecular markers in predicting lymph node metastasis of submucosal colorectal cancer.METHODS: We examined microvessel density, lymphatic vessel density, the Ki-67 labeling index, expression of MUC1 and Matrix metalloproteinase-7 (MMP-7) in tumor cells, and expression of cathepsin D in stromal cells at the invasive front by immunostaining of samples resected from 214 patients with submucosal colorectal cancer.Pathologic features were assessed on hematoxylin-eosinstained samples. We evaluated the relations between clinicopathologic/immunohistochemical features and lymph node metastasis.RESULTS: Lesions of the superficial type, with an unfavorable histologic grade, budding, lymphatic involvement, high microvessel density (≥ 40), high lymphatic vessel density (≥ 9), high Ki-67 labeling index (≥ 42), and positivity of MUC1, cathepsin D, and MMP-7 showed a significantly high incidence of lymph node metastasis. Multivariate analysis revealed that high microvessel density, unfavorable histologic grade,cathepsin D positivity, high lymphatic vessel density,superficial type, budding, and MUC1 positivity were independent risk factors for lymph node metastasis.A combined examination with four independent immunohistochemical markers (microvessel density,cathepsin D, lymphatic vessel density, and MUC1)revealed that all lesions that were negative for all markers or positive for only one marker were negative for lymph node metastasis.CONCLUSION: Analysis of a combination of immunohistochemical molecular markers in endoscopically resected specimens of submucosal colorectal cancer allows prediction of curability regardless of the pathologic features visible of hematoxylin-eosin-stained sections.

  20. Identification of Molecular Marker Linked to Salt Tolerance Gene in Alfalfa


    The study has established the F2 offspring obtained by crossing salt-tolerant with salt-sensitive alfalfa, and appraised the salt-tolerant F2 offspring seedling was evaluated in pot culture. With the F2 segregated population, the research has obtained a molecular marker linked with salt-tolerant genes of alfalfa using the improved BSA combined with RAPD. The RAPD PCR products were excised from the agarose gel and purified using a kit, then were mixed with pMD-18T vector and sequenced. Sequencing result indicated the RAPD marker was 1 438 bp in length. Similarity researches using blast in Genbank indicated that the nucleotide sequence of the RAPD marker showed 93% and 91% similarity with mth2-6el8 gene fragment (347 bp) and mth2-33122 gene fragment (334 bp) of Medicago truncatula respectively. Medicago truncatula is a close relative of alfalfa and Mth2-6e18 is a molecular marker of the gene coding for a cysteine protease which was salt inducible in some plants. These results indicated the RAPD marker was possibly related to cysteine protease genes in alfalfa.

  1. Optimization of ISSR Markers for Molecular DNA Fingerprinting in Aquilaria sp

    Aquilaria sp. belongs to the Thymelaeaceae family and well distributed to Asia region. The species is a multipurpose use from root to shoot and becoming an economic important crop, which generates wide interest in understanding the genetic diversity of the species. Understanding of the effectiveness in differentiating DNA-based markers is an important step towards plant germplasm characterization and evaluation. It is becoming a prerequisite for more effective application of molecular marker techniques in breeding and mapping programs. Polymerase Chain Reaction (PCR)-based approaches are in demanding as its simplicity and requirement for only small quantities of sample genomic DNA. Inter-simple sequence repeats (ISRR) requires no prior genomic information as anchor template in producing multi-loci markers of tandem repeats for polymorphic patterns by PCR amplification which becoming a key of advantageous of ISSR primers. ISSR markers have shown rapid, simple, reproducible and inexpensive means in molecular taxonomy, conservation breeding and genetic diversity analysis. The ISSR for marker applications are essential to facilitate management, conservation and genetic improvement programs towards improvement of standard resin quality for perfume and or pharmaceutical industries. In this paper, a total of 100 ISSR primers were optimized by using Aquilaria malaccensis. Primers optimization resulted, 38 ISSR primers affirmative for the polymorphism evaluation study, which encountered both from specific and degenerate ISSR primers. Marker derived from ISSR profiling is a powerful method for identification and molecular classification of Aquilaria sp from species to accessions and further will useful in identifying any mutant lines derived from nature and/or mutagenesis activities. (author)

  2. Prediction of industrial tomato hybrids from agronomic traits and ISSR molecular markers.

    Figueiredo, A S T; Resende, J T V; Faria, M V; Da-Silva, P R; Fagundes, B S; Morales, R G F


    Heterosis is a highly relevant phenomenon in plant breeding. This condition is usually established in hybrids derived from crosses of highly divergent parents. The success of a breeder in obtaining heterosis is directly related to the correct identification of genetically contrasting parents. Currently, the diallel cross is the most commonly used methodology to detect contrasting parents; however, it is a time- and cost-consuming procedure. Therefore, new tools capable of performing this task quickly and accurately are required. Thus, the purpose of this study was to estimate the genetic divergence in industrial tomato lines, based on agronomic traits, and to compare with estimates obtained using inter-simple sequence repeat (ISSR) molecular markers. The genetic divergence among 10 industrial tomato lines, based on nine morphological characters and 12 ISSR primers was analyzed. For data analysis, Pearson and Spearman correlation coefficients were calculated between the genetic dissimilarity measures estimated by Mahalanobis distance and Jaccard's coefficient of genetic dissimilarity from the heterosis estimates, combining ability, and means of important traits of industrial tomato. The ISSR markers efficiently detected contrasting parents for hybrid production in tomato. Parent RVTD-08 was indicated as the most divergent, both by molecular and morphological markers, that positively contributed to increased heterosis and by the specific combining ability in the crosses in which it participated. The genetic dissimilarity estimated by ISSR molecular markers aided the identification of the best hybrids of the experiment in terms of total fruit yield, pulp yield, and soluble solids content. PMID:27323023

  3. Intestinal microflora molecular markers of spleen-deficient rats and evaluation of traditional Chinese drugs

    Ying Peng; Zhuo Wang; Yuan Lu; Chun-Fu Wu; Jing-Yu Yang; Xiao-Bo Li


    AIM: To find a rapid and efficient analysis method of gastrointestinal microflora in Pi-deficient (spleendeficient) rats and to evaluate traditional Chinese drugs. METHODS: Enterobacterial repetitive intergenic consensus-PCR (ERIC-PCR) based assay was performed to examine changes of intestinal microflora in two Pi-deficienct animal models and to evaluate the efficacy of four traditional Chinese drugs as well as a probiotic recipe and another therapy in Pi-deficient rats. RESULTS: A molecular marker was identified for Pideficiency in rats. The pharmacodynamic evaluation system, including identified molecular markers (net integral area and abundance of DNA bands), Shannon's index for diversity of intestinal microflora, and Sorenson's pairwise similarity coefficient, was established. The four major clinical recipes of traditional Chinese drugs for Pi-deficiency in rats, especially at their medium dose (equivalence to the clinical dose), produced more pronounced recovery activities in Pi-deficient rats, while higher doses of these recipes did not show a better therapeutic effect but some toxic effects such as perturbation deterioration of intestinal microflora. CONCLUSION: Both fingerprint analysis and identified marker can show Pi-deficiency in rats and its difference after treatment. The identified molecular marker may be applied in screening for the active compounds both in relative traditional Chinese drugs and in pharmacodynamic study of Pi-deficiency in rats.

  4. Molecular marker development and genetic diversity exploration by RNA-seq in Platycodon grandiflorum.

    Kim, Hyun Jung; Jung, Jungsu; Kim, Myung-Shin; Lee, Je Min; Choi, Doil; Yeam, Inhwa


    Platycodon grandiflorum, generally known as the bellflower or balloon flower, is the only species in the genus Platycodon of the family Campanulaceae. Platycodon plants have been traditionally used as a medicinal crop in East Asia for their antiphlogistic, antitussive, and expectorant properties. Despite these practical uses, marker-assisted selection and molecular breeding in platycodons have lagged due to the lack of genetic information on this genus. In this study, we performed RNA-seq analysis of three platycodon accessions to develop molecular markers and explore genetic diversity. First, genic simple sequence repeats (SSRs) were retrieved and compared; dinucleotide motifs were the most abundant repeats (39%-40%) followed by trinucleotide (25%-31%), tetranucleotide (1.5%-1.9%), and pentanucleotide (0.3%-1.0%) repeats. The result of in silico SSR analysis, three SSR markers were detected and showed possibility to distinguish three platycodon accessions. After several filtering procedures, 180 single nucleotide polymorphisms (SNPs) were used to design 40 cleaved amplified polymorphic sequence (CAPS) markers. Twelve of these PCR-based markers were validated as highly polymorphic and utilized to investigate genetic diversity in 21 platycodon accessions collected from various regions of South Korea. Collectively, the 12 markers yielded 35 alleles, with an average of 3 alleles per locus. Polymorphism information content (PIC) values ranged from 0.087 to 0.693, averaging 0.373 per locus. Since platycodon genetics have not been actively studied, the sequence information and the DNA markers generated from our research have the potential to contribute to further genetic improvements, genomic studies, and gene discovery in this genus. PMID:26501479

  5. Investigation of Combining Plant Genotypic Values and Molecular Marker Information for Constructing Core Subsets

    Jian-Cheng Wang; Jin Hu; Ning-Ning Liu; Hai-Ming Xu; Sheng Zhang


    In the present study, a strategy was proposed for constructing plant core subsets by clusters based on the combination of continuous data for genotypic values and discrete data for molecular marker information. A mixed linear model approach was used to predict genotypic values for eliminating the environment effect.The "mixed genetic distance" was designed to solve the difficult problem of combining continuous and discrete data to construct a core subset by cluster. Four commonly used genetic distances for continuous data (Euclidean distance, standardized Euclidean distance, city block distance, and Mahalanobis distance)were used to assess the validity of the continuous data part of the mixed genetic distance; three commonly used genetic distances for discrete data (cosine distance, correlation distance, and Jaccard distance) were used to assess the validity of the discrete data part of the mixed genetic distance. A rice germplasm group with eight quantitative traits and information for 60 molecular markers was used to evaluate the validity of the new strategy. The results suggest that the validity of both parts of the mixed genetic distance are equal to or higher than the common genetic distance. The core subset constructed on the basis of a combination of data for genotypic values and molecular marker information was more representative than that constructed on the basis of data from genotypic values or molecular marker information alone. Moreover, the strategy of using combined data was able to treat dominant marker information and could combine any other continuous data and discrete data together to perform cluster to construct a plant core subset.

  6. Genetic analysis of maize by using the AFLP (R) method

    Vuylsteke, M.J.R.


    Methylation AFLP® is a novel PCR-based method to detect methylation of restriction sites randomly over the genome.In framework of the AFLP®-related technology development, a modification of the AFLP method was developed, called the methylation AFLP® method. Several features of the methylation AFLP a

  7. An AFLP-based genome-wide mapping strategy.

    Peters, J.L.; Cnops, G.; Neyt, P.; Zethof, J.; Cornelis, K.; Lijsebettens, M. van; Gerats, A.G.M.


    To efficiently determine the chromosomal location of phenotypic mutants, we designed a genome-wide mapping strategy that can be used in any crop for which a dense AFLP (Amplified Fragment Length Polymorphism) map is available or can be made. The AFLP technique is particularly suitable to initiate ma

  8. Molecular characterization of ochratoxin A producing strains of the genus Penicillium

    Castella, G.; Larsen, Thomas Ostenfeld; Cabanes, J.; Schmidt, H.; Alboresi, A.; Niessen, L.; Farber, P.; Geisen, R.


    Sixty-six strains classified as P. verrucosum based oil morphological criteria were characterized by molecular methods like RAPD, AFLP and ITS sequencing. Two groups could be identified by RAPD and AFLP analyses. The two RAPD as well as the two AFLP groups were completely coincidental. Strains ill...

  9. Use of molecular markers to compare Fusarium verticillioides pathogenic strains isolated from plants and humans.

    Chang, S C; Macêdo, D P C; Souza-Motta, C M; Oliveira, N T


    Fusarium verticillioides is a pathogen of agriculturally important crops, especially maize. It is considered one of the most important pathogens responsible for fumonisin contamination of food products, which causes severe, chronic, and acute intoxication in humans and animals. Moreover, it is recognized as a cause of localized infections in immunocompetent patients and disseminated infections among severely immunosuppressed patients. Several molecular tools have been used to analyze the intraspecific variability of fungi. The objective of this study was to use molecular markers to compare pathogenic isolates of F. verticillioides and isolates of the same species obtained from clinical samples of patients with Fusarium mycoses. The molecular markers that we used were inter-simple sequence repeat markers (primers GTG5 and GACA4), intron splice site primer (primer EI1), random amplified polymorphic DNA marker (primer OPW-6), and restriction fragment length polymorphism-internal transcribed spacer (ITS) from rDNA. From the data obtained, clusters were generated based on the UPGMA clustering method. The amplification products obtained using primers ITS4 and ITS5 and loci ITS1-5.8-ITS2 of the rDNA yielded fragments of approximately 600 bp for all the isolates. Digestion of the ITS region fragment using restriction enzymes such as EcoRI, DraI, BshI, AluI, HaeIII, HinfI, MspI, and PstI did not permit differentiation among pathogenic and clinical isolates. The inter-simple sequence repeat, intron splice site primer, and random amplified polymorphic DNA markers presented high genetic homogeneity among clinical isolates in contrast to the high variability found among the phytopathogenic isolates of F. verticillioides. PMID:24065642

  10. Determination of the molecular markers associated with the anthesis-silking interval in maize

    Water stress in maize is a serious problem, especially in tropical and sub-tropical areas. A short anthesis-silking interval (ASI) has proved to be a good indicator of drought tolerance. A strategy involving closely related maize lines differing principally in ASI has been undertaken in order to determine the molecular markers associated with this character. Parental and F2 materials were grown under different water stress regimes in order to assess the morphological differences. DNA samples were also obtained from these materials. Significant differences were found for ASI, grain yield and grain number between the different genotypes, suggesting that these materials are adequate for use in molecular marker analysis. Data from a survey of parental lines with restriction fragment length polymorphisms and amplified fragment length polymorphisms show a total of 58 polymorphic bands between the two lines. (author). 15 refs, 3 figs