Using inter simple sequence repeat multiloci markers for. Amplification was performed in a thermal cycler eppendorf, germany. Bisht1 and shashi bhalla1 icarindian agricultural research institute, new delhi110 012, india. Genetic diversity among wild resources of the genus. Issrs are amplified by pcr using microsatellite core sequences as primers with a few selective nucleotides as anchors into the non repeat adjacent regions 1618 bp. Morphological and inter simple sequence repeat issr markers analyses of corynespora cassiicola isolates from rubber plantations in malaysia.
Identification of closely related citrus cultivars with inter. Inter simple sequence repeat issr pcr is a technique, which involvesthe use of microsatellite sequences as primers in a polymerase chainreaction to generate multilocus markers. Issr inter simple sequence repeat analysis which target the flanking region of. Development of simple sequence repeat ssr markers in. The best markers for genome mapping, marker assisted selection, phylogenic studies, and crop conservation have low cost and labour requirements and high reliability. Inter simple sequence repeat issr analysis of somaclonal variation in artemisia amygdalina decne. About 1060 fragments from multiple loci are generated simultaneously, separated by gel electrophoresis and scored as the presence or absence of fragments of particular size. Microsatellites or simple sequence repeat ssr marker amplified tandemely di, tri and tetra nucleotide repeats that are widely distributed on genome. Issr inter simple sequence repeats as molecular markers to study genetic diversity in tarantulas. In this study, the ability inter simple sequence repeat markers issr were used to identify molecular relationships among wild, local and commercial genotypes of strawberry. Issr inter simple sequence repeats as genetic markers in.
Issr for inter simple sequence repeat is a general term for a genome region between microsatellite loci. It is a simple and quick methodthat combines most of the advantages of microsatellites ssrs andamplified fragment length polymorphism aflp to the universality ofrandom amplified polymorphic dna rapd. Inter simple sequence repeat an overview sciencedirect topics. Genetic relationships were evaluated using 27 of 86 issr markers, yielding 307 polymorphic bands with polymorphism contents ranging from 0.
The number of alleles per locus ranged from 1 to 5 with an average of 3. Inter simple sequence repeat issr markers were employed to investigate the genetic diversity and differentiation of 24 okra genotypes. Issr and rapd profiling were performed with 19 and 12 primers, respectively. Molecular characterization of cultivated bromeliad accessions. Issr markers as a tool for the assessment of genetic diversity in. Assessment of genetic diversity and fingerprinting of. Issr has a few advantages because issr primers anneal directly to simple sequence repeats and, thus, unlike ssr markers, no prior knowledge of a target sequence is required wang et al. Pdf inter simple sequence repeat issr polymorphism and its. Inter simple sequence repeat issr polymorphism and its. Pdf development of simple sequence repeat ssr markers in. The complementary sequences to two neighboring microsatellites are used as pcr primers. Pdf inter simple sequence repeat issr markers for genetic. Pdf inter simple sequence repeat issr polymorphism and.
Genetic diversity analysis of okra abelmoschus esculentus. Each issr band corresponds to a dna sequence delimited by two inverted microsatellites. Random molecular markers such as random amplified polymorphic dna 27, inter simple sequence repeat 29 and amplified fragment length polymorphism 26 are widely used in genetic diversity evaluation of medicinal plants. Genotyping and molecular identification of date palm. Applicability of start codon targeted scot and inter. Inter simple sequence repeat markers to assess genetic. Issr markers are highly polymorphic and are useful in studies on genetic diversity, phylogeny, gene tagging, genome mapping and evolutionary biology. Apr 06, 2021 start codon targeted polymorphism scot and inter simple sequence repeats issr markers were used to assess the diversity within and among all the populations. Issr inter simple sequence repeat analysis which target the flanking region of identical ssr in inverse direction.
Intersimple sequence repeat issr dna fingerprinting was used to assess the genetic stability of synthetic seeds of c. Molecular characterization of cultivated bromeliad. Identification of closely related citrus cultivars with. Gaur, in animal biotechnology, 2014 applications of issr.
The dna polymorphism among 92 selected tef genotypes belonging to eight origin groups was assessed using eight inter simple sequence repeat issr primers. Genotyping and molecular identification of date palm cultivars. In this study, inter simple sequence repeat issr amplification was employed to assess the genetic diversity in tef germplasm. In this study, the pcr products were separated by electrophoresis on 8% nondenaturing polyacrylamide gel and visualized by silver staining. Corn rust fungi are considered as major pathogens in corn production of thailand. Issr markers as a tool for the assessment of genetic. Inter simple sequence repeat polymerase chain reaction issr pcr zietkiewicz et al.
Genetic diversity assessment of yams dioscorea spp. Genetic diversity analysis and molecular characterization of. Optimization of issr markers for molecular dna fingerprinting in. Application of inter simple sequence repeat issr markers. Nonanchored inter simple sequence repeat issr markers. Characterization of plants using molecular markers is an ideal approach for improvement and conservation of plant genetic resources. Sixteen of the issr genotypes were shared by more that one individual in a population, representing potential clones. Assessment of genetic diversity in contrasting sugarcane varieties using inter simple 427 sequence repeat issr markers in agarose gel 1.
The use of inter simple sequence repeat issr pcr markers is a molecular technique that overcomes most of these limitations pradeep reddyet al. Preliminary study on genetic diversity in moehringia. Inter simple sequence repeat markers issr, anchored. Morphological and inter simple sequence repeat issr markers. Molecular characterisation and similarity relationships. Apr 14, 2005 inter ssr issr fingerprinting was developed such that no sequence knowledge was required. Issr analysis involves amplification of regions between adjacent and. These generated 271 polymorphic fragments with an average of 18. Inter simple sequence repeat issr marker system was first describes by zietkiewics et al. In the present study, a selection of cultivated bromeliads was characterized via inter simple sequence repeat issr markers with an emphasis on genetic diversity and population structure. Reproductive biology and genetic diversity of a cryptoviviparous mangrove aegiceras corniculatum myrsinaceae using allozyme and intersimple sequence repeat issr analysis. Ten of the 16 shared genotypes were not limited to clusters of plants groups of plants growing from the same point.
Puccinia polysora in thailand based on inter simple sequence repeat issr markers analysis. Application of inter simple sequence repeat issr markers to plant genetics godwin, i. Pdf molecular characterization of cultivated bromeliad. Issr and ssr markers for determining genetic relationships. Preliminary study on genetic diversity in moehringia janka. A nonlethal technique was used to collect samples from six populations in the yucatan peninsula, and seven issr primers were tested. Primers based on a repeat sequence, such as ca n, can be made with a degenerate 3anchor, such as ca 8 rg or agc 6 ty. Inter simple sequence repeat issr was employed to investigate genetic diversity and differentiation. The aim of this study was to evaluate the diversity of kermani sheep using issr markers. Molecular markers are widespread tools in the study of genetic diversity. Tanya et al 2011 employed inter simple sequence repeats issrs to assess genetic variation among 30 accessions of jatropha.
Singlelocus ssr markers have been developed for a number of species, although there is a major bottleneck in developing ssr markers whereby flanking sequences must be known to design 5anchors for polymerase ch. Intersimple sequence repeat issr analysis of somaclonal. Issr markers generated a total number of 156 bands with 92. Since 1994, a new molecular marker technique called inter simple sequence repeat issr. The resultant pcr reaction amplifies the sequence between two ssrs, yielding a multilocus marker system useful for. Inter simple sequence repeat an overview sciencedirect.
Issr markers are easy to use, lowcost, and methodologically less demanding compared. Eighteen issr primers were used to evaluate 45 accessions. Among dna markers, inter simple sequence repeat issr was preferred in genetic diversity, phylogeny, genomics and evolutionary studies due to their easiness to use, cost effectiveness, and their multiple amplification products reddy et al. The objectives were to examine the possibility of using issr markers for unravelling genetic diversity in tef, and to assess the extent and pattern of genetic diversity in the test germplasm with respect to origin groups. The maximum number of alleles was observed at the a7, 818, 825 and 849 loci, and their size ranged from 300 to 2500 bp. Nghia na1, kadir j, sunderasan e, puad abdullah m, malik a, napis s.
Pdf characterization of genetic diversity in jatropha. Genetic diversity analysis of okra abelmoschus esculentus l. Phylogenetic relationship of four ficus species using random. Start codon targeted polymorphism scot and inter simple sequence repeats issr markers were used to assess the diversity within and among all the populations. Inter simple sequence repeats issr or microsattelite dna this lecture explains the issr which is a type of microsattelite dna in human genome. Inter simple sequence repeat issr and random amplified polymorphic dna rapd markers by sarah alamri master of science msc in biology a thesis submitted in partial fulfillment of the requirement for the degree of masters of science msc in biology the school of graduate studies laurentian university sudbury, ontario, canada. Morphological and inter simple sequence repeat issr. Weber and may 1989, and inter simple single repeat issr. Inter simple sequence repeats issr or microsattelite dna.
For the first time in spiders, the inter simple sequence repeat issr technique was used to study the genetic variability of mexican populations of brachypelma vagans. Molecular characterisation and similarity relationships among. The issr is a molecular marker method which does not need the genome sequence information and leads to multiloci and highly polymorphic patterns 7. Inter simple sequence repeat issr analysis of genetic. Kamili 2 and masood akbar 1 1department of biochemistry, university of kashmir, srinagar190006. Genome fingerprinting by simple sequence repeat ssranchored polymerase chain reaction amplification zietkiewicz, e. A total of 11 inter simple sequence repeat issr primers were utilized to estimate molecular variability of genotypes under study table 2. Fifteen inter simple sequence repeat issr markers were used. The number of polymorphic bands per primer varied from 4 to 22, with 12. The issr analysis involves the pcr amplification of dna regions between adjacent, inversely oriented microsatellites using a single simple sequence repeat ssrcontaining primers. Determining the genetic stability of micropropagated. Random amplified polymorphic dna rapd and inter simple sequence repeat issr molecular fingerprinting markers were employed as genetic markers for the assay. Genetic diversity among wild resources of the genus boehmeria.
Issr markers were developed from the need to explore microsatellite repeats without the use of dna sequencing lagercrantz et al. Assessment of genetic diversity in contrasting sugarcane. The ultimate goal of the breeders is to improve yield. Interrelationships of quantitative traits and genetic. Application of aflp, rapd and issr markers to genetic mapping of european and japanese larch. Pcr amplification of these regions using a single primer yields multiple amplification products that can be used as a dominant multilocus marker system for the study of genetic variation in various organisms. Intersimple sequence repeat issr markers are generated by pcr amplification of dna using a single primer composed of a microsatellite sequence that is usually anchored at the 3 or 5 end by two to four arbitrary, often degenerate, nucleotides. Genetic diversity of puccinia polysora in thailand based on. In the present investigation, the genetic relationships of 29 j. Inter simple sequence repeat issr markers generated by 22 primers were tested for their ability to distinguish among samples from 94 trees of 68 citrus cultivars. According to grivet and arruda 2002, molecular markers are important tools that can.
Application of inter simple sequence repeat issr markers to plant genetics electrophoresis. Applicability of start codon targeted scot and inter simple. Inter simple sequence repeats isrr requires no prior genomic information as anchor template in producing multiloci markers of tandem repeats for. Twelve issr primers produced 342 bands, of which 287 84% were polymorphic, with polymorphic bands per primer ranging from 17 to 34. Issr markers are highlypolymorphic and are useful in studies on genetic diversity, phylogeny, genetagging, genome mapping and evolutionary biology. Genetic diversity of puccinia polysora in thailand based. Dendrobium, microsatellite, orchid, simple sequence repeat ssr received 5 january 2006. Evaluation of genetic diversity in wheat cultivars and. Inter simple sequence repeat issr is the genome region between microsatellite loci. Research article genetic profiling of sugarcane genotypes. Thirtyeight alleles were generated at 12 issr loci.
Genetic diversity analysis and molecular characterization. Among molecular markers, issr inter simple sequence repeat markers are considered as informative in genetic diversity studies bornet and blanchard, 2001. Issr markers involve pcr amplification of dna using a. Thirteen scot primers amplified 73 fragments with 97. Within each of the six cultivar groups studied, most of these cultivars are so closely related that they are difficult to distinguish by other molecular marker techniques. Application of inter simple sequence repeat issr markers to.
Rapd markers have been used for the identification of cultivars and for assessing the genetic diversity among cultivars of several crops like bean 15, cowpea 16, pea 17, mungbean 4 18, and vigna angularis 19. It is rapidly being adopted by the research community in various fields of plant improvement. Neis genetic distance between accessions ranged from 0. Two corn rust species, puccinia polysora southern rust and p. Inter simple sequence repeats issrs are regions in the genome flanked by microsatellite sequences. Comparative analysis of genetic diversity among cultivated. Phylogenetic relationship of four ficus species using. Inter ssr issr fingerprinting was developed such that no sequence knowledge was required.
Since 1994, a new molecular marker technique called inter simple sequence repeat issr has been available zietkiewicz et al. Twelve out of the 25 tested primers produced clear and reproducible fragments with high value of polymorphism 96. Issr marker is based on primers having microsatellite sequences, and detects variation in the size of inter. Development of simple sequence repeat ssr markers in eucalyptus from amplified inter simple sequence repeats issr. Pdf development of simple sequence repeat ssr markers. Inter simple sequence repeat issr markers rapidly reveal high polymorphic fingerprints and have been used frequently to determine the genetic diversity among date palm cultivars. Intersimple sequence repeat issr markers rapidly reveal high polymorphic fingerprints and more. Microsatellites or simple sequence repeats ssrs are ubiquitous in eukaryotic genomes. Detection of genetic variations in orobanche crenata using. Issr inter simple sequence repeats as molecular markers to. In this scenario, inter simple sequence repeat issr markers stand out as they can efficiently access variability at the dna level, which contributes to increased heterosis during hybrid development toppa and jadoski, 20. Issr are single primer designed from ssr and amplified region between two ssrs. Issr markers as a tool for the assessment of genetic passi. Molecular markerstypes and applications online biology.
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