• International Journal of Industrial Entomology and Biomaterials
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• v.10 no.2
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• pp.79-86
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• 2005

Molecular markers have increasingly been used in plant genetic analysis, due to their obvious advantages over conventional phenotypic markers, as they are highly polymorphic, more in number, stable across different developmental stages, neutral to selection and least influenced by environmental factors. Among the PCR based marker techniques, ISSR is one of the simplest and widely used techniques, which involves amplification of DNA segment present at an amplifiable distance in between two identical microsatellite repeat regions oriented in opposite direction. Though ISSR markers are dominant like RAPD, they are more stable and reproducible. Because of these properties ISSR markers have recently been found using extensively for finger printing, pohylogenetic analysis, population structure analysis, varietal/line identification, genetic mapping, marker-assisted selection, etc. In mulberry (Morus spp.), ISSR markers were used for analyzing phylogenetic relationship among cultivated varieties, between tropical and temperate mulberry, for solving the vexed problem of identifying taxonomic positions of genotypes, for identifying markers associated with leaf yield attributing characters. As ISSR markers are one of the cheapest and easiest marker systems with high efficiency in generating polymorphism among closely related varieties, they would play a major role in mulberry genome analysis in the future.

• Korean Journal of Plant Resources
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• v.19 no.4
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• pp.509-514
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• 2006

For the variant identification of Korean dogwood, Cornus kousa Buerg., we investigated useful genetic marker using the ISSR primer. The overall amplicons was 58 in six primers and the mean number of amplicons per primer was 9.67. The UPGMA dendrogram based on the genetic distance showed two major clusters composed of pink-bracted and white-bracted groups respectively. And white-bracted group was divided to two groups, island and inland groups. This result showed that ISSR analysis provides useful markers for the variant identification even in the seedling or young tree of Cornus kousa, especially pink-bracted individuals of Isl. Oenaro.

• Journal of Microbiology and Biotechnology
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• v.20 no.4
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• pp.683-692
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• 2010

Morphologically, nine different slow-growing protoclones were screened from regenerated protoplasts of heterokaryotic Agaricus bisporus. As such, the present study is the first report on differentiating homo- and heterokaryotic protoclones using random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR) markers. Among 80 primers tested, the seven ISSR and seven RAPD primers selected for the analysis generated a total of 94 ISSR and 52 RAPD fragments, respectively. The ISSR fingerprinting also detected more polymorphic loci (38.29%) than the RAPD fingerprinting (34.61%). A principal coordinate analysis (PCA) was employed to evaluate the resolving power of the markers as regards differentiating protoclones. As a result, the mean polymorphism information content (PIC) for each marker system (i.e., 0.787 for RAPD and 0.916 for ISSR) suggested that ISSR is more effective for determining polymorphisms. The dendrograms constructed using RAPD, ISSR, and an integrated RAPD and ISSR marker system were highly correlated with one another as revealed by a high Mantel correlation (r= 0.98). The pairwise similarity index values also ranged from 0.64 to 0.95 (RAPD), 0.67 to 0.98 (ISSR), and 0.67 to 0.98 (RAPD and ISSR), whereas the mean similarity index values of 0.82, 0.81, and 0.84 were obtained for the RAPD, ISSR, and combined data, respectively. As there was a good correspondence between the RAPD and ISSR similarity matrices, ISSR would appear to be an effective alternative to RAPD in the genetic diversity assessment and accurate differentiation of homo- and heterokaryotic protoclones of A. bisporus.

• Journal of Life Science
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• v.16 no.2 s.75
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• pp.219-224
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• 2006

Inter-simple sequence repeat (ISSR) analysis were used to assess genetic diversity among 18 genotypes of watermelon (Citrullus lanatus Thunb.) including breeding lines and commercial varieties. The 21 ISSR primers selected from 100 primers were showed the amplification of 105 reproducible fragments ranging from about 200 bp to 5000 bp. A total of 58 DNA fragments were polymorphic with an average 2.7 polymorphic bands per primer. The polymorphic primers were divided into 18 anchored primers and 3 non anchored primers. All of the anchored primers were di-nucleotide repeat motif, and was more polymorphic than non anchored primers. Eighteen watermelon genotypes were classified into two large groups. Clustering was in some accordance with the division of fruit shape into 18 watermelon. Therefore, ISSR markers may be suitable for variety discrimination and for constructing a linkage map of watermelon.

• Plant Biotechnology Reports
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• v.2 no.4
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• pp.239-243
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• 2008

Simmondsia chinensis (Link) Schneider, a multipurpose and monogeneric dioecious shrub from arid zones, has emerged as a cash crop all over the globe. Its seed propagation poses severe problems due to its male-biased population: the male:female ratio is 5:1. Investigations have been carried out to generate a sex-specific Inter-simple sequence repeat (ISSR) marker for the early detection of male and female plants. Of the 42 primers analysed with a bulk sample of pooled male DNA and a bulk sample of pooled female DNA, only one primer, UBC-807, produced a unique ~1,200 base-pair fragment in the male DNA. To validate this observation, this primer was re-tested with individual male and female samples from eight cultivars. A similar unique ~1,200 bp fragment was present in the male individuals of all eight cultivars and completely absent in the female individuals tested. This is the first report of the use of ISSR markers to ascertain sex in physiologically mature S. chinensis plants.

• International Journal of Industrial Entomology and Biomaterials
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• v.9 no.2
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• pp.207-215
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• 2004

The present study was carried out to evaluate the ISSR and RAPD markers for their efficiency as genetic marker systems to establish the relationships between 18 mulberry genotypes. A total of 36 from 56 (64%) RAPD primers and 12 from 48 (25%) ISSR primers produced reproducible amplification patterns. A high proportion of polymorphic bands ranging from 44 to 91% was observed respectively with RAPD and ISSR markers. The average Resolving Power (Rp) of ISSR primers was higher than RAPD primers. The ISSR primers, UBC 825, 868 and 873, and RAPD primers, UBC 712, 720 and 729, possessed the highest Rp values and could in each instance distinguish all the 18 genotypes. Similarity matrix values were estimated based on Jaccards coefficient, considering 109 polymorphic ISSR and 212 polymorphic RAPD bands and two dendrograms were constructed. The dendrograms obtained with ISSR and RAPD markers distinguished the eight exotic genotypes from the ten indigenous (Indian) genotypes. A significant correlation value (r=0.959; p=0.001) for the cophenetic matrix between the RAPD and ISSR matrices was observed. The results indicated that the ISSR and RAPD markers could assist in the differentiation of genotypes and permit the determination of genetic distances that might be exploited by mulberry breeders in improvement programs.

• Journal of Mushroom
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• v.15 no.4
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• pp.273-278
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• 2017

Hypsizygus marmoreus is a mushroom with abundant flavor and medicinal properties. However, its application is limited by problems such as long cultivation period, low biological efficiency, and microbiological contamination; therefore, there is a substantial need for development of new cultivars of this species. In this study, 55 strains of H. marmoreus were subjected to inter simple sequence repeat (ISSR) analysis to identify markers for the selection of mother strains for breeding from the collected germplasm. ISSR 13 and 15 were confirmed as polymorphic markers. The three strains (KMCC03106, KMCC03107, and KMCC03108) with white cap color were found to be genetically closely related upon UPGMA analysis of both ISSR 13 and 15. Based on the PCR analysis results for ISSR 15, the collected germplasm were differentiated into three groups according to the strain collection year. Thus, ISSR 15 could be a marker for determining the phylogeny of cap color and genetic variations according to the strain collection year. These results suggest that ISSR markers can be effective tools for the selection of mother strains for breeding of H. marmoreus.

• Journal of Life Science
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• v.25 no.3
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• pp.257-262
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• 2015

The genetic variability of four species of the genus Zoysia collected from South Korea was analyzed using an inter-simple sequence repeat (ISSR) marker system. Polymerase chain reactions (PCR) with eight ISSR primers generated 86 amplicons, 76 (87.1%) of which were polymorphisms. The polymorphism information content (PIC) value of the ISSR marker system was 0.848. The percentage of polymorphic loci (P_p) ranged from 41.2% to 44.7%. Nei’s gene diversity (H) ranged from 0.149 to 0.186, with an average overall value of 0.170. The mean of Shannon’s information index (I) value was 0.250. Total genetic diversity values (H_T) varied between 0.356 (ISSR-1) and 0.418 (ISSR-16), for an average overall polymorphic loci of 0.345. Interlocus variation in within-species genetic diversity (H_S) was low (0.170). On a per-locus basis, the proportion of total genetic variation due to differences among species (G_ST) was 0.601. This indicated that about 60.1% of the total variation was among species. Thus, about 39.9 of genetic variation was within species. The estimate of gene flow, based on G_ST, was very low among species of the genus Zoysia (N_m = 0.332). The phylogenic tree showed three distinct groups: Z. macrostachya and Z. tenuifolia clades and other species were formed the separated clusters. In conclusion, the ISSR assay was useful for detecting genetic variation in the genus Zoysia, and its discriminatory power was comparable to that of other genotyping tools.

• International Journal of Industrial Entomology and Biomaterials
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• v.24 no.2
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• pp.57-61
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• 2012

The Indian Oak Tasar silkworm, Antheraea proylei J. is a beneficial insect with great economic importance in India for its silk production. In this study, six populations of Antheraea proylei and A. frithi Moore (as an out group) were subjected to inter simple sequence repeat (ISSR) marker analysis in order to assess its genetic diversity. Fifteen ISSR primers produced 91 markers among different breeds of A. proylei and A. frithi of which 89 are polymorphic, generating 97.8% polymorphism. The dendrogram constructed using the Unweighted Pair Group Method with Arithmetic Mean (UPGMA) and cluster analysis made using Nei's genetic distance resulted in the formation of one major group containing four sub-groups separating the breeds. This result suggests that ISSR amplification is potentially useful for molecular characterization of oak tasar silkworm genotypes.

• Journal of Korean Society of Forest Science
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• v.108 no.3
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• pp.302-310
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• 2019

Precise and fast identification of crop cultivars is essential for efficient breeding and plant breeders' rights. Traditional methods for identification of jujube cultivars are based on the evaluation of morphological characteristics. However, due to time constraints and environmental influences, it is difficult to distinguish cultivars using only morphological traits. In this study, we cloned fragments from improved inter simple sequence repeats (ISSR) analysis, and developed stably diagnostic sequence-characterized amplified region (SCAR) markers. The specific ISSR bands of jujube cultivars from Dalizao and Boeundaechu were purified, cloned, and sequenced. As a result, four clones labeled 827Dalizao550, 827Boeun750, 846Boeun700, and 847Dalizao850 were identified. In order to investigate whether they were specific for the jujube cultivar, four pairs of SCAR primers were then designed and polymerase chain reaction (PCR) amplifications were conducted to analyze 32 samples, including jujube and sour jujube. In the PCR amplification of the 827Dalizao550 SCAR marker, the specific bands with 550 bp were amplified in six samples (Dalizao, Sandonglizao, Dongzao, Yuanlin No. 2, Suanzao 2, Suanzao 4), but unexpected bands (490 bp) were amplified in the others. Moreover, in the PCR amplification of the 847Dalizao850 SCAR marker, the specific bands with 850 bp were found in three samples (Dalizao, Sandonglizao, and Dongzao) and 900 bp unexpected bands were amplified in five samples (Pozao, Suanzao 1, Suanzao 2, Suanzao 3, Suanzao 4). These results showed that newly developed markers could be useful as a fast and reliable tool to identify jujube cultivars. However, further identification of polymorphic information and the development of SCAR markers are required for the identification of more diverse cultivars.