Molecular markers in plants for analysis of genetic. Using five soybean cultivars, a complete issr and rapd based molecular id system is described in this study, which can be easily used and expanded with much more information. Issr genotyping of endangered plants using an optimized work. Ten issr markers were profiled for fifty three individual phenotypes with two replications in the seedling seed orchard sso population. The objective of this study is an attempt to develop a plant molecular identity system in soybean. Pdf intersimple sequence repeats issrs are regions in the genome flanked by. Use of efficacious molecular markers to obtain the required knowledge of the genetic diversity within the local and regional germplasm collections can enhance the overall effectiveness of cowpea improvement programs. A total of 73 bands produced were found polymorphic from these 4 markers using capillary electrophoresis. Comparison of rapd, issr, and aflp molecular markers to. Data was scored as 1 for being present and 0 for the absence of dna band in each micropropagated and mother plant. Molecular characterization of thymus species using issr markers article pdf available in journal of animal and plant sciences 254 august 2015 with 888 reads how we measure reads. Intersimple sequence repeat issr pcr is a fast and inexpensive genotyping technique. The shortcomings of biochemically derived markers such as isozymes, drove the development of markers based on dna polymorphism kan and dozy, 1978. Assessment of genetic diversity of vigna unguiculata l.
Evaluation of phylogenetic relationship among ficus palmata. Apr 12, 2016 issr markers comprise a few highly informative multiallelic loci. Inter simple sequence repeat issr markers were used to. Few molecular studies have been performed on this species. Issr plant genotyping issr genotyping of endangered plants using an optimized workflow applied biosystems 3500xl genetic analyzer genemapper v4. Comparative assessment of rapd and issr markers to study. Nine of these fourteen primers were individually able to. Inter simple sequence repeat issrpcr is a technique, which involvesthe use of microsatellite sequences as primers in a polymerase chainreaction to generate multilocus markers. It is important to conserve haloxylon community in kuwait as it can tolerate extreme adverse conditions of drought and salinity to be potentially used in the desert and urban revegetation and greenery national programs.
The rapd and issr markers have been used to study the genetic variability. Rapd and issr markers have been used successfully in diversity analysis of a number of crops aliyev et al. Gupta pk and varshney rk 2000 the development and use of microsatellite markers for genetic analysis and plant breeding with emphasis. However, both ssr primers couldnt authenticate two plants and further work is needed to develop issr dna profiles for the authentication. Walp cowpea accessions using informative molecular markers is imperative for their genetic improvement and conservation. Of these three techniques, while aflp is the most labour intensive and timeconsuming, it is also the most reliable. Issr has been proved to be an effective tool for the determination of phylogenetic relationship among closely related species. Determination of local tobacco cultivars using issr molecular. 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. Genetic homogeneity revealed using scot, issr and rapd. Issr marker based population genetic study of melocanna.
In addition, issr11 and microsatellite markers have been used to assess the genetic diversity and relationships of date palm varieties in sudan12, morocco, oman14, nigeria15 and qatar16. Microsatellites or simple sequence repeats ssrs are ubiquitous in eukaryotic genomes. Diversity index of issr markers issr marker di issr marker di issr marker di ubc8560 0. Pdf application of issr marker in pharmacognosy researchgate. A comparative phylogenetic analysis of medicinal plant.
Development of rapd, damd and issr markers for authentication. Pdf issr markers as a tool for the assessment of genetic. Lycopersicon esculentum, lycopersicon pennellii, lycopersicon cheesmanii, lycopersicon humboldtii, lycopersicon hirsutum and two lycopersicon esculentum substitution lines il 63 and wsl 6 were analyzed. Inter simple sequence repeat issr markers for assessment of.
Issr techniques for evolutionary biology sciencedirect. To construct a population level genetic profile for investigation and exploitation of genetic diversity of ficus palmata, 25 ecotypes of. A comparative phylogenetic analysis of medicinal plant tribulus terrestris in northwest india revealed by rapd and issr markers. The level of polymorphism in tomato was studied using issr pcr. Apart, in several plants too, issr markers have been successfully used for the assessment of genetic diversity in corn 8 and for linkage to a specific gene 9, for cultivar identification in oil seed rape and potatoes 10,11, for mapping of plant chromosomes 12. High correlation between the rapd and issr marker systems was shown using a mantel test r 0. Assessment of genetic diversity in vigna unguiculata l. Issr markers for genetic relationships in caricaceae and sex. An external file that holds a picture, illustration, etc. Inter ssr issr fingerprinting was developed such that no sequence knowledge was required.
They provide highly discriminating information with good reproducibility, and are relatively abundant 21, 22. Several dna marker systems and associated techniques are available today for fingerprinting of plant varieties. Inter simple sequence repeats issr description issrs are dna fragments of about 1003000 bp located between adjacent, oppositely oriented microsatellite regions. Application of inter simple sequence repeat issr markers. The issr marker is based on a pcramplification of 1003,000 basepair bp regions between. High genetic diversity indices for both marker types 0. In addition, issrs are considered useful in gene mapping studies. By rapd and issr markers maryam talebi haghighi, ehsan shahdoust dos in biotechnology science, university of mysore, manasagangotri, mysore, india abstract india has immense variety of plants with endophytic organisms within them without any risk of life. Pdf evaluation of genetic fidelity of in vitro raised. Only 4 issr markers were found polymorphic which were used to score 180 f 2 plants. Analysis on genetic diversity of radix astragali by issr. They are therefore intensely used in phylogenetic studies and sex differentiation of some economically interesting plant species. Use of issr markers for cultivar identification in durum wheat.
Because of the multilocus fingerprinting profiles obtained, issr analysis can be applied in studies involving genetic identity, parentage, clone and strain identification, and taxonomic studies of closely related species. Rapd and issr markers of black and green colour of blackcurrant ribes nigrum fruits sylwia keller przyb ylkowicz, malgorzata korbin and jozef gwozdecki research institute of pomology and floriculture pomologiczna 18, 96100 skierniewice, poland email. Genetic diversity and genetic relationships among curcuma. Pdf issr markers in the genetic diversity of chrysanthemum. Secondly if the issr markers used in study are specifically amplifying more in female plants and resulting more number of amplicons then it can also be a probable reason. Issr marker analysis of acacia auriculiformis in first. The couplingphase issr markers detected in that study could be used to select for some levels of quantitative traits associated with fin and p genes in common bean. Primers based on a repeat sequence, such as can, can be made. The aim of this study was to evaluate the efficiency of rapd and issr markers in assessing genetic diversity of date. Efficiency of rapd and issr markers for the detection of. Bunge ex boiss rimth is one of the native plants of kuwait, extensively depleting through the anthropogenic activities. A total of 30 issr markers were used to screen both parents.
The application of issr markers in genetic variance detection among durian durio zibethinus murr. Investigations have to continue for discovering more closely related issr markers with fin and p genes. The genetic marker chosen for this study to solve the problem of tobacco cultivar classification is the issr technique. Issr markers show differentiation among italian populations. Department of plant breeding and production, faculty of engineering and. Determination of suitable gammaray dose for mutation. Rootstocks using issr molecular markers hellenicplants.
Molecular markers have been used for genetic study and identification of cultivars for a long time. Assessment of genetic diversity in costus pictus accessions based. Pdf molecular characterization of thymus species using issr. Genetic diversity and population structure of haloxylon.
Nonanchored inter simple sequence repeat issr markers. Issrs are amplified by pcr using microsatellite core sequences as primers with a few selective nucleotides as anchors into the nonrepeat adjacent regions 1618 bp. Application of inter simple sequence repeat issr markers to. Issr genotyping of endangered plants using an optimized workflow. Pdf the application of issr markers in genetic variance. Issr molecular markers are widely used for population genetic analysis of different plants generating more reliable and reproducible bands than rapd 30, 31. Safari 3 1 department of agronomy and plant breeding, campus of agriculture and natural resources, razi university, kermanshah, iran.
Singlelocus ssr markers have been developed for a number of. Genetic stability assessed by rapd and issr markers showed that almond plantlets. The average polymorphism level with the issr markers 97. Determination of genetic uniformity in transgenic cotton plants using. Issr markers are easy to use, lowcost, and methodologically less demanding. Screening a higher number of anonymous loci in sugar beet using these molecular markers. Identification of microsporederived plants in anther culture of flax linum usitatissimum l. In the present paper, the issr technique was employed to study genetic diversity in italian a. After surface sterilization, shoot tips were used as primary explants. Molecular markers in plants for analysis of genetic diversity.
Materials and methods plant material and in vitro culture conditions healthy and diseased free a. Alishah3 1department of biology, school of basic sciences, science and research branch, islamic azad university, tehran, iran. International journal of environment agriculture and biotechnologyissn. Sugar beet, genetic diversity, molecular markers, rapd, issr received. Molecular identity using intersimple sequence repeat. Methodology genetic diversity analysis in opal cotton hybrids.
Application of issr markers in the genus lycopersicon. Sample files are imported into genemapper software. It is known for its fine flavour and could represent an important resource for cultivation programs in desert areas. 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. Using rapd and issr markers for genetic diversity evaluation among several radiation doses the total genomic dna was isolated from young leaves of normal garlic germinated plants from treated and untreated cloves using the ctab method murray and thompson, 1980. Issr plant genotyping issr genotyping of endangered plants using an optimized work. Comparison of the genome size, endoreduplication, and issr. Article pdf available september 2018 with 176 reads. The fragments were scored as present 1 or absent 0, and those readings were entered in a computer file as a binary matrix one for each marker. Turk j biol 430 prior knowledge about the sequence under investigation is required.
Issrpcr fingerprinting of plant pathogen strains using the. These results clearly discriminated the medicinal plant c. Genetic diversity, molecular markers, rapd, ssr, issr, ests, sts molecular markers molecular marker is a sequence of dna, which are located with. The issr markers were found to correlate with the following alsike clover traits. Methodology genetic diversity analysis in opal cotton hybrids based on ssr, issr, and rapd markers z.
During the processes of improvement of plant using genetic transformation or the mass production using micropropagation, the most crucial aspect is to retain genetic integrity with respect to the mother plants. Efficiency of using rapd and issr markers in evaluation of. The issr primers amplified 91 bands, and 54 showed polymorphism 59. Assessment of genetic variations among highly endangered medicinal plant l. Nodal explants of three months old shoots were used in murashige and skoog medium supplemented with n6benzyladenine ba for shoot proliferation and with naphthaleneacetic acid naa for rooting. One qtl was found linked to photosynthetic rate on chromosome 3a, one to. Ten primers of each rapd and issr table 1 obtained from bio basic canada inc. 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 chain reaction pcr primers.
Issr markers in the genetic diversity of chrysanthemum plants derived via somatic embryogenesis. They are technically simpler as compared to rflp, ssr, and aflp markers as no previous sequence information is required for generating dna amplification products 32 34. About 1060 fragments from multiple loci are generated. Finally, the aflp showed 100 bands, of which 92 were polymorphic 92%. Development of rapd, damd and issr markers for authentication of medicinal plant cassia auriculata and its adulterant cassia surattensis. In this study srap and issr markers were used to evaluate genetic variation and relationships among 25 accessions in china. Molecular analysis of genetic fidelity in micropropagated. Issr markers as a tool for the assessment of genetic diversity in passiflora article pdf available in biochemical genetics 4978.
Both marker systems obtained high polymorphism for the high resolution of capillary electrophoresis. Inter simple sequence repeat issr polymorphism and its. Muhammad idrees, muhammad irshadmolecular markers in plants for analysis of genetic diversity. Dna quality and quantity were established by spectrophotometric measurements and agarose gel electrophoresis. Issr markers were introduced in 1994 gupta 1994, zietkiewicz 1994 for studies of cultivated plants but have been used for studies of hybridization and hybrid speciation archibald 2004, wolfe 1998a, wolfe 1998b, population and conservation genetics culley 2001, esselman 1999, and systematic investigations in natural populations crawford.
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