EST-PCR, EST-SSR and ISSR markers to identify a set of wild cranberries and evaluate their relationships

2015 ◽  
Vol 95 (6) ◽  
pp. 1155-1165 ◽  
Author(s):  
Dong An ◽  
Natalia V. Bykova ◽  
Samir C. Debnath

An, D., Bykova, N. V. and Debnath, S. C. 2015. EST-PCR, EST-SSR and ISSR markers to identify a set of wild cranberries and evaluate their relationships. Can. J. Plant Sci. 95: 1155–1165. The cranberry (Vaccinium marcrocarpon Ait.) is a woody, evergreen, perennial vine with great potential for economic and health benefits. Selection and use of genetically diverse genotypes are key factors in any crop breeding program to develop cultivars with a broad genetic base. Molecular markers play a major role in selecting diverse genotypes. One hundred and two wild cranberry clones collected from four Canadian provinces and five cultivars were screened with inter simple sequence repeat (ISSR), expressed sequence tag–simple sequence repeat (EST-SSR) and EST–polymerase chain reaction (PCR) markers to validate the genetic diversity and relationships among them. EST-PCRs (0.54) and EST-SSRs (0.35) generated higher frequency of major alleles than ISSRs (0.08), but ISSRs presented a higher level of polymorphism and greater polymorphic information content and expected heterozygosity than EST-SSRs and EST-PCRs. Combined cluster analysis by the unweighted pair-group method with arithmetic averages (UPGMA) separated the wild clones and cultivars into four main clusters, which was in agreement with the principal coordinate (PCo) analysis. Analysis of molecular variation detected sufficient variations among genotypes within communities and among communities within provinces with ISSR (66 and 36%, respectively), EST-PCR (72 and 34%, respectively) and EST-SSR (72 and 34%, respectively) markers. These values were 71 and 35%, respectively, for combined analysis. Combined use of three types of molecular markers, for the first time in Vaccinium species, detected a sufficient degree of variation among cranberry genotypes, allowing for differentiation and rendering these technologies valuable for genotype identification in a diverse cranberry germplasm and for more efficient parental choice in the current cranberry breeding program.

2015 ◽  
Vol 22 (2) ◽  
pp. 67-75 ◽  
Author(s):  
Leila Samiei ◽  
Mahnaz Kiani ◽  
Homa Zarghami ◽  
Farshid Memariani ◽  
Mohammad Reza Joharchi

In this study genetic diversity and interspecific relationships of 11 Allium L. species from Khorassan province of Iran including 32 accessions were investigated by inter simple sequence repeat (ISSR) markers. Nine ISSR primers produced a total of 80 polymorphic markers and revealed high polymorphism among the studied species. The average gene diversity, effective number of alleles and Shannon’s information index were 0.2, 1.28 and 0.3, respectively. Allium kuhsorkhense exhibited the greatest level of variation (He: 0.18), whereas A. stipitatum demonstrated the lowest level of variability (He: 0.05). UPGMA (Unweighted Pair Group Method with Arithmetic mean) analysis showed that Allium accessions have a similarity range of 0.60 to 0.95. Allium scapriscapum composed the most distant group in the dendrogram. The clustered groups of Allium species clearly reflect the recent taxonomic concept of the genus at the subgenus and section levels. The present study showed that the ISSR technique is an effective molecular approach for analyzing genetic diversity and relationship in Allium species.Bangladesh J. Plant Taxon. 22(2): 67-75, 2015 (December)


2008 ◽  
Vol 88 (2) ◽  
pp. 313-322 ◽  
Author(s):  
S. C. Debnath ◽  
S. Khanizadeh ◽  
A. R. Jamieson ◽  
C. Kempler

The goal of this study was to determine the level of genetic diversity and relatedness among 16 strawberry (Fragaria H ananassa Duch.) cultivars and 11 breeding lines developed in Canada, using Inter Simple Sequence Repeat (ISSR) markers. Seventeen primers generated 225 polymorphic ISSR-PCR bands. Cluster analysis by the unweighted pair-group method with arithmetic averages (UPGMA) revealed a substantial degree of genetic similarity among the genotypes ranging from 63 to 77% that were in agreement with the principal coordinate (PCO) analysis. Geographical distribution for the place of breeding program explained only 1.4% of total variation as revealed by analysis of molecular variance (AMOVA). The ISSR markers detected a sufficient degree of polymorphism to differentiate among strawberry genotypes, making this technology valuable for cultivar identification and for the more efficient choice of parents in current strawberry breeding programs. Key words: Fragaria × ananassa, DNA fingerprinting, multivariate analysis, breeding, genetic similarity


2006 ◽  
Vol 86 (1) ◽  
pp. 251-257 ◽  
Author(s):  
Zhao Weiguo ◽  
Zhou Zhihua ◽  
Miao Xuexia ◽  
Wang Sibao ◽  
Zhang Lin ◽  
...  

The genetic diversity of 27 mulberry (Morus spp.) genotypes mainly from China was investigated using inter-simple sequence repeat (ISSR) markers to assist in addressing breeding objectives and conserving existing genetic resources. Of the 22 primers screened, 15 produced highly reproducible ISSR bands. Using these 15 primers, 138 discernible DNA fragments were generated with 126 (91.3%) being polymorphic, indicating considerable genetic variation among the mulberry genotypes studied. Genetic similarity ranged from 0.6014 between Yu 2 and Yu 711 to 0.9493 between Cuizhisang and Dejiang 10. The phenetic dendrogram based on ISSR data generated by the unweighed pair group method with arithmetical averages (UPGMA) method grouped the 27 accessions into two major clusters: cluster I, cultivated mulberry species (M. multicaulis Perr., M. alba Linn., M. atropurpurea oxb., M. bombycis Kiodz., M. australis Poir., M. rotundiloba Kiodz., M. alba var. pendula Dipp., M. alba var. macrophylla Loud., and M. alba var. venose Delile.); and cluster II, wild mulberry species (M. cathayana Hemsl., M. laevigata Wall., M. wittiorum Hand-Mazz., M. nigra Linn., and M. mongolica Schneid.). Our molecular analyses agree with the existing morphological classification of Morus and clarify the genetic relationships among mulberry species. Key words: Morus L., genetic diversity, inter-simple sequence repeat, relatedness


Author(s):  
Thanuja Dilrukshi Dharmarathna ◽  
H.M.D.A.K. Herath ◽  
P.A. Weerasinghe ◽  
H.M.V.G. Herath

The genus Dendrobium is one of the largest genera in the family Orchidaceae having more than thousand species over the world with diverse morphological characters. Dendrobium is a popular ornamental plant with complex genetic background which emphasize on the species identification at molecular level. The present study was aimed to identify Inter-Simple Sequence Repeat (ISSR) markers capable of detecting genetic polymorphism to characterize 18 hybrid, commercially available Dendrobium cultivars. Genomic DNA of each cultivar was extracted using CTAB method. A total of 17 different ISSR primers were evaluated. Only the reproducible bands were scored and number of different alleles (Na), number of effective alleles (Ne), Shannon’s Information Index (I), Expected heterozygosity (He), Unbiased expected heterozygosity (UHe), polymorphic percentage and polymorphic information content (PIC) of each primer were calculated. The highest Shannon’s Information Index (0.537±0.08) was recorded by the primer UBC 826 while the highest polymorphic information content (PIC) was generated by primer UBC 807. The PIC values of the primers were ranged from 0.0068 to 0.451, indicating that primers are moderately informative. In total, 631 bands representing 120 loci were amplified showing 85.71% - 100% polymorphism. The genetic similarities between individuals were compiled in the Nei’s genetic identity matrix in order to construct the UPGMA dendrogram. Principle component analysis (PCA) and clustering analysis were done to divide different cultivars into groups. The analysis revealed the presence of four major clusters and two minor clusters among the cultivars. The study suggested that the ISSR markers originated from eight primers 12, 155, UBC 807, UBC 812, UBC 826, UBC 835, UBC 841 and UBC 842 can be used in the detection of molecular variation among cultivars in the genus Dendrobium.


HortScience ◽  
2009 ◽  
Vol 44 (2) ◽  
pp. 293-297 ◽  
Author(s):  
Kadir Uğurtan Yılmaz ◽  
Sezai Ercişli ◽  
Bayram Murat Asma ◽  
Yıldız Doğan ◽  
Salih Kafkas

Inter-simple sequence repeat (ISSR) markers were used to study the genetic diversity and phylogenetic relationships among 16 genotypes from subgenus Prunus (six genotypes from section Prunophora, seven genotypes from section Armeniaca and two plumcot genotypes, and one genotype from subgenus Cerasus) in Prunus genus. From the polymerase chain reaction amplifications with 20 ISSR primers showing polymorphism among subgenera and sections, 180 polymorphic ISSR bands were detected and polymorphism ratio ranged from 57% to 100%. Based on the unweighted pair group method with arithmetic mean (UPGMA) analysis and principal coordinate analysis (PCoA) using the Jaccard coefficient, a dendrogram and three-dimensional plot were constructed including genotypes in Prunus genus. Two main groups formed in the dendrogram; one of them (Cluster I) included Cerasus, whereas Cluster II included Prunus. Cluster II also divided into three subgroups, including sections Prunophora, Armeniaca, and plumcot. Both UPGMA and the PCoA demonstrated that Armeniaca genotypes had lower genetic variation and plumcot genotypes are closer to the plums than the apricots. The ISSR-based phylogeny was generally consistent with Prunus taxonomy based on molecular evidence, suggesting the applicability of ISSR analysis for genotypic and phylogenetic studies in Prunus genus.


2016 ◽  
Vol 59 (4) ◽  
pp. 477-483 ◽  
Author(s):  
Leila Simaei-Soltani ◽  
Alireza Abdolmohammadi ◽  
Alireza Zebarjadi ◽  
Saheb Foroutanifar

Abstract. The aim of this study was to investigate the genetic diversity and structure in three Iranian native goat breeds (Markhoz, Mahabadi and Lori) and the Beetal imported breed using inter-simple sequence repeat (ISSR) markers and also to investigate ISSR markers' potential in order to genetically separate single (S) and twin-birth (T) subpopulations. Blood samples were collected from 210 animals for this purpose. In total, 16 primers were used, and finally 5 primers were selected based on the number of clear bands and the level of polymorphisms. The result of this study showed that 76 of 86 observed fragments were polymorphic. Genetic diversity for each breed ranged from 0.23 in the Beetal breed to 0.26 in the Markhoz breed; this represents a relatively similar genetic diversity in these breeds. An unweighted pair group method with arithmetic mean (UPGMA) dendrogram based on the Nei's standard genetic distance between the breeds studied showed that three Iranian goat breeds (Mahabadi, Lori and Markhoz) were clustered closer together, while the Beetal breed formed a separate cluster. In the constructed dendrogram of the subpopulations, the S and T subpopulations of each breed were clustered together. The constructed dendrogram of the Beetal breed and the S and T subpopulations of all breeds studied showed a separate cluster for the Beetal breed as an imported breed and another cluster for the S and T subpopulations as Iranian native breeds. The current study showed that the ISSR markers studied had no potential to genetically separate S and T subpopulations. On the other hand, these ISSR markers can be used for the clustering of distinct populations.


2007 ◽  
Vol 87 (2) ◽  
pp. 337-344 ◽  
Author(s):  
Samir C. Debnath

Forty-three wild lingonberry [Vaccinium vitis-idaea ssp. minus (Lodd) Hult.] clones collected from four Canadian provinces were assessed for genetic variability by using inter simple sequence repeat (ISSR). Fifteen primers generated 356 polymorphic ISSR-PCR bands. A substantial degree of genetic diversity was found am ong the wild collections. Cluster analysis by the unweighted pair-group method with arithmetic averages (UPGMA) separated the wild clones into four main clusters, and identified the two remaining clones as outliers. Furthermore, within four clusters, the genotypes tended to form sub-clusters that were in agreement with the principal coordinate (PCO) analysis. Geographical distribution explained 10% of total variation as revealed by analysis of molecular variance (AMOVA). The ISSR markers detected a sufficient degree of polymorphism to differentiate among lingonberry clones, making this technology valuable for germplasm management and the more efficient choice of parents in current lingonberry breeding programs. Key words: Vaccinium vitis-idaea, DNA fingerprinting, molecular marker


2010 ◽  
Vol 39 (1) ◽  
pp. 119-122 ◽  
Author(s):  
Mahmudul Islam Nazrul ◽  
Fan Xiao Lin ◽  
Bian Yin-Bing

Among ten slow-growing protoclones of Agaricus bisporus (J. Lge) Imbach, all appressed colonies showed slower growth rate and spawn run, and inability to produce fruiting bodies in substrate. Seven of 40 inter-simple sequence repeat (ISSR) primers amplified 78 reproducible fragments, 48.93% were polymorphic, each producing 7 to 16 bands ranging from 0.10 to 2.10 kbp, sufficient to differentiate the protoclones from each other. Appressed protoclones were homoallelic at a number of loci that were heteroallelic in the parent, suggesting that they represented rare homokaryons. Thus, using morphological characters along with ISSR, polymorphisms could be useful for quick, easy, and accurate in distinguishing homo- and heterokaryotic isolates. Key words: Agaricus bisporus (J. Lge) Imbach; Homokaryon; ISSR; Protoclone DOI: 10.3329/bjb.v39i1.5537Bangladesh J. Bot. 39(1): 119-122, 2010 (June)


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