Article

Application of inter simple sequence (ISSR) markers to plant genetics

Department of Agriculture, University of Queensland, Brisbane, Australia.
Electrophoresis (Impact Factor: 3.16). 01/1997; 18(9):1524-8. DOI: 10.1002/elps.1150180906
Source: PubMed

ABSTRACT Microsatellites or simple sequence repeats (SSRs) are ubiquitous in eukaryotic genomes. Single-locus 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 5'-anchors for polymerase chain reaction (PCR) primers. Inter SSR (ISSR) fingerprinting was developed such that no sequence knowledge was required. Primers based on a repeat sequence, such as (CA)n, can be made with a degenerate 3'-anchor, such as (CA)8RG or (AGC)6TY. The resultant PCR reaction amplifies the sequence between two SSRs, yielding a multilocus marker system useful for fingerprinting, diversity analysis and genome mapping. PCR products are radiolabelled with 32P or 33P via end-labelling or PCR incorporation, and separated on a polyacrylamide sequencing gel prior to autoradiographic visualisation. A typical reaction yields 20-100 bands per lane depending on the species and primer. We have used ISSR fingerprinting in a number of plant species, and report here some results on two important tropical species, sorghum and banana. Previous investigators have demonstrated that ISSR analysis usually detects a higher level of polymorphism than that detected with restriction fragment length polymorphism (RFLP) or random amplified polymorphic DNA (RAPD) analyses. Our data indicate that this is not a result of greater polymorphism genetically, but rather technical reasons related to the detection methodology used for ISSR analysis.

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    • "[2] Polymorphic markers based on short DNA sequences reveal genome variations among expressed and non-expressed regions and are quick, reliable and reproducible.[3] Among the different molecular markers, inter simple sequence repeat (ISSR) markers are rapid, cost effective and do not require any sequence information of the genome under study [4] or any radioactive labelling based assay.[5] ISSR-polymerase chain reaction (PCR) analysis involves gene amplification of a region between two inversely oriented microsatellites placed at an amplifiable distance.[2] ISSR markers have been used to resolve polymorphisms among plant accessions by generating a large number of markers that target multiple microsatellite loci distributed across the genome [6] and also among highly related species.[7] "
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    • "Inter-simple sequence repeat (ISSR) is a PCR-based technique that involves the amplification of DNA sequences between simple sequence repeats (SSR) by means of anchored or non-anchored SSR homologous primers (Zietkiewicz et al. 1994). ISSR does not require information about genome sequence in advance and can detect a greater number of polymorphisms than RFLP or RAPD (Godwin et al. 1997). ISSR is therefore an efficient tool for analysing genetic diversity within closely related species (Yu et al. 2008), and for studying genetic populations of fungi (Menzies et al. 2003; Chadha and Gopalakrishna 2007). "
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