• Title/Summary/Keyword: inter-simple sequence repeat markers

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Analysis of the Genetic Relationship among Mulberry (Morus spp.) Cultivars Using Inter-Simple Sequence Repeat (ISSR) Markers

  • Park, Eun-Ju;Kang, Min-Uk;Choi, Myoung-Seob;Sung, Gyoo-Byung;Nho, Si-Kab
    • International Journal of Industrial Entomology and Biomaterials
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    • v.41 no.2
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    • pp.56-62
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    • 2020
  • Mulberry (Morus spp. family: Moraceae) has prime importance in the sericulture industry, and its foliage is the only natural feed of the silkworm Bombyx mori L. Traditional classification methods using morphological traits were largely unsuccessful in assessing the diversity and relationships among different mulberry species because of environmental influences on the traits of interest. For these reasons, it is difficult to differentiate between the varieties and cultivars of Morus spp. In the present study, inter-simple sequence repeat (ISSR) markers were used to investigate the genetic diversity of 48 mulberry samples genotyped using nine ISSR primers. The ISSR markers exhibited polymorphisms (53.2%) among mulberry genotypes. Furthermore, similarity coefficient estimated for these ISSR markers was found to vary between 0.67 and 0.99 for the combined pooled data. The phenogram drawn using the UPGMA cluster method based on combined pooled data of the ISSR markers divided the 48 mulberry genotypes into seven major groups. No genetic association was found in the collection area, and there was a mixed pattern between the mulberry lines. The hybridization between different mulberry species is highly likely to be homogenized due to natural hybridization.

Evaluation of Nonanchored Inter Simple Sequence Repeat (ISSR) Marker to Detect DNA Damage in Common Bean (Phaseolus vulgaris L.) Exposed to Acrylamide

  • Enan, Mohamed R.
    • Journal of Forest and Environmental Science
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    • v.24 no.2
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    • pp.61-68
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    • 2008
  • Acrylamide is present as a contaminant in heated food products, predominantly from the precursor asparagine. Nonanchored inter simple sequence repeats (ISSRs) are arbitrary multiloci markers produced by PCR amplification with a microsatellite primer. In order to assess the feasibility of microsatellite primers as markers for DNA damage, the study was conducted on common bean (Phaseolus vulgaris L.) exposed to different concentrations of acrylamide. Polymorphisms were abundant among plant samples treated with acrylamide in comparison to control (untreated one) tested with 4- tri-nucleotide, 2 tetra-nucleotide, and 3- dinucelotide primers. The primer (CCG)4 was the best tested primer to generate polymorphism between the DNA of plants treated or not by acrylamide. Polymorphisms became evident as the presence and absence of DNA fragments in treated samples compared with the untreated one. The highest number of DNA variation on ISSR patterns was observed at the micromollar concentrations of acrylamide. Acrylamide was able to induce DNA damage in non concentration-dependent manner with effectiveness at micromollar concentrations. This study demonstrated that ISSR markers can be highly reliable for identification of DNA damage induced by acrylamide.

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Analysis of Inter-simple sequence repeat (ISSR) markers in cultivars and collected strains of button mushroom (Agaricus bisporus) (양송이 품종과 수집 균주간의 Inter-simple sequence repeat (ISSR) 마커 분석)

  • Nam, Youn-keol;Kong, Won-Sik;Jang, Kab-Yeul;Shin, Pyung-Gyun;Oh, MinJi;Im, Ji-Hoon;Koo, Chang-Duck;Oh, Youn-Lee
    • Journal of Mushroom
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    • v.15 no.3
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    • pp.139-144
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    • 2017
  • A. bisporus is the fifth most cultivated mushroom in Korea, and approximately 10,757 tons were cultivated in 2015. The genetic diversity of collected strains in Korea and commercial cultivars was analyzed using inter-simple sequence repeat (ISSR) markers. ISSR markers known to be comparable among A. bisporus spp. were selected from various markers. Totally, 16 markers, namely the ISSR markers 807, 808, 810, 811, 834, 835, 836, 841, 842, P3, P8, P17, P22, P30, P38, and P39, were evaluated to discriminate between ASI 1110, 1114, 1115, 1238, 1246, 1365, 1366, and 1369 for selecting suitable markers in 16 markers. The ISSR markers P31, P38 and P39 exhibited various fingerprints that could help classify the strains in species. Using the three markers, genetic relationships among 39 strains, including commercial cultivars, such as SaeA and SaeYeon, were analyzed using the UPGMA method. The results of the analysis of the genetic relationships between commercial cultivars and collected strains in Korea confirmed that the commercial cultivars were different from the collected strains in Korea. These results suggested that the ISSR markers P31, P38, and P30 could be used for selecting the commercial cultivars of A. bisporus.

Inter Simple Sequence Repeat (ISSR) Polymorphism and Its Application in Mulberry Genome Analysis

  • Vijayan Kunjupillai
    • 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.

Mendelian Inheritance of Inter-Simple Sequence Repeats Markers in Abies Koreans Wilson (구상나무에 있어서 Inter-Simple Sequence Repeats Marker의 유전양식(遺傳樣式))

  • Hong, Yong-Pyo;Cho, Kyung-Jin;Kim, Yong-Yul;Shin, Eun-Kyeong
    • Journal of Korean Society of Forest Science
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    • v.87 no.3
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    • pp.422-428
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    • 1998
  • Polymerase chain reaction(PCR)-based inter-simple sequence repeats(I-SSR) markers were analyzed in 48 megagametophytes of a single tree of Abies koreana $W_{ILS}$. Nineteen of the 35 primers, screened with 6 megagametophyte DNA and produced the clearest amplification products in the preliminary experiment, were used for PCR with 48 megagametophyte DNAs sampled from a single tree. On the basis of the chi-square test, a total of 51 amplicons, amplified by the 19 primers, were revealed to be segregated according to the Mendelian ratio(i.e., 1 : 1 segregation ratio) in the 48 megagametophytes at 5% significance level. Based on the linkage analysis, the observed 51 Mendelian loci turned out to be unlinked each other, which suggested that they are evenly distributed in the genome. However, majority of RAPD markers are known to belong to the independent linkage blocks, which frequently results in the amplification of RAPD markers from the restricted regions of the genome. Owing to the nature of even distribution of the 51 loci observed in this study, the I-SSR markers could give better resolution of estimating genetic diversity from the whole genome than RAPD markers. And I-SSR markers are also more suitable than RAPD markers for reconstructing phylogenetic relationship by a cladistic method which requires to fulfil the assumption of independent evolution of the different characters.

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DNA Polymorphism and Assessments of Genetic Relationships in genus Zoysia Based on Simple Sequence Repeat Markers (ISSR에 의한 잔디속 식물의 DNA 다형성과 유전적 관계 평가)

  • Huh, Man Kyu
    • 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 (Pp) 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 (HT) 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 (HS) was low (0.170). On a per-locus basis, the proportion of total genetic variation due to differences among species (GST) 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 GST, was very low among species of the genus Zoysia (Nm = 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.

Genetic Diversity Studies and Identification of Molecular and Biochemical Markers Associated with Fusarium Wilt Resistance in Cultivated Faba Bean (Vicia faba)

  • Mahmoud, Amer F.;Abd El-Fatah, Bahaa E.S.
    • The Plant Pathology Journal
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    • v.36 no.1
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    • pp.11-28
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    • 2020
  • Faba bean (Vicia faba L.) is one of the most important legume crops in Egypt. However, production of faba bean is affected by several diseases including fungal diseases. Fusarium wilt incited by Fusarium oxysporum Schlecht. was shown to be the most common wilt disease of faba bean in Assiut Governorate. Evaluation of 16 faba bean genotypes for the resistance to Fusarium wilt was carried out under greenhouse conditions. Three molecular marker systems (inter-simple sequence repeat [ISSR], sequence related amplified polymorphism [SRAP], and simple sequence repeat [SSR]) and a biochemical marker (protein profiles) were used to study the genetic diversity and detect molecular and biochemical markers associated with Fusarium wilt resistance in the tested genotypes. The results showed that certain genotypes of faba bean were resistant to Fusarium wilt, while most of the genotypes were highly susceptible. The percentage of disease severity ranged from 32.83% in Assiut-215 to 64.17% in Misr-3. The genotypes Assiut-215, Roomy-3, Marut-2, and Giza2 were the most resistant, and the genotypes Misr-3, Misr-1, Assiut-143, Giza-40, and Roomy-80 performed as highly susceptible. The genotypes Assiut-215 and Roomy-3 were considered as promising sources of the resistance to Fusarium wilt. SRAP markers showed higher polymorphism (82.53%) compared with SSR (76.85%), ISSR markers (62.24%), and protein profile (31.82%). Specific molecular and biochemical markers associated with Fusarium wilt resistance were identified. The dendrogram based on combined data of molecular and biochemical markers grouped the 16 faba bean genotypes into three clusters. Cluster I included resistant genotypes, cluster II comprised all moderate genotypes and cluster III contained highly susceptible genotypes.

Polymorphism of inter simple sequence repeat markers in Hypsizygus marmoreus (Inter Simple Sequence Repeat(ISSR) 마커를 활용한 느티만가닥버섯(Hypsizigus marmoreus) 종내 다형성 분석)

  • Oh, Youn-Lee;Nam, Yun-Geul;Jang, Kab-Yeul;Kong, Won-Sik;Oh, Min ji;Im, Ji-Hoon;Choi, In-Geol
    • 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.

Improved characterization of Clematis based on new chloroplast microsatellite markers and nuclear ITS sequences

  • Liu, Zhigao;Korpelainen, Helena
    • Horticulture, Environment, and Biotechnology : HEB
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    • v.59 no.6
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    • pp.889-897
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    • 2018
  • Currently, there is a lack of genetic markers capable of effectively detecting polymorphisms in Clematis. Therefore, we developed new markers to investigate inter- and intraspecific diversity in Clematis. Based on the complete chloroplast genome of Clematis terniflora, simple sequence repeats were explored and primer pairs were designed for all ten adequate repeat regions (cpSSRs), which were tested in 43 individuals of 11 Clematis species. In addition, the nuclear ITS region was sequenced in 11 Clematis species. Seven cpSSR loci were found to be polymorphic in the genus and serve as markers that can distinguish different species and be used in different genetic analyses, including cultivar identification to assist the breeding of new ornamental cultivars.