Penstemon cardwelliiHyobanche rubraPenstemon debilis

Penstemon clevelandiiHyobanche rubra Penstemon debilis

Why the pictures of these three taxa? The diploid hybrid species status of P. clevelandii was documented with ISSR markers (Wolfe et al. 1998a, b) and we've used ISSR markers successfully to examine relationships within the holoparasitic plant genus Hyobanche and the conservation genetics of P. debilis.

Welcome to the ISSR Resource Website.

Designed and maintained by Andrea D. Wolfe

Department of Evolution, Ecology and Organismal Biology
The Ohio State University
1735 Neil Avenue
Columbus OH 43210

Index to ISSR Resource Website:

Bibliography (ISSR citations, other references of interest, some notes on selected citations)
Data Gathering and Scoring (general considerations, types of data needed for selected studies)
Introduction (overview of this website, history of method)
Protocols: DNA Miniprep extraction
Protocols: Elu-Quik clean-up of DNA extract
Protocols: ISSR primers and PCR(NEW primers listed as of 1/17/00)
Protocols: ISSR primer optimization and troubleshooting (NEW!!! All photos labeled as of 1/17/00)
Protocols: Sephaglas clean-up of DNA extract
Resources (links and addresses for useful sites, including software)
Software and Algorithms (so many choices, so little memory)
What's New (a chronology of ISSR Resource Website development)

What's New?

19 July 2000

Updated the bibliography - new citations include ISSR references for insect, mammal, algae, and fungi publications.

17 January 2000

New primers are listed and the opitimization page now includes the labeled gel pics.

29 October 1999

I've added a page on optimization and trouble shooting ISSR gels. This page includes pictures and explanations of each step.

17 November 1998

I've added a Sephaglas protocol, a page on resources, and a page on software and algorithms. The latter page includes examples of input and output for Vera S. Ford's programs, which we have cited in several manuscripts. I've included instructions for obtaining these programs through anonymous FTP.

Except for adding examples from case studies and a FAQ page, this website includes the relevant information I included in my 1998 workshop at AIBS. I'll continue to update the bibliography on a regular basis, and I will start developing the add-ons as I have time (oh sure, like I'm really going to have time).

19 September 1998

I've added the Elu-Quik protocol we use.

24 August 1998

There's a section for data gathering/scoring.

20 August 1998

I've added miniprep DNA extraction and ISSR protocols.

Send me feedback! via this link or by leaving a message in the guestbook (click on the Guest book button of the banner below).

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The first studies employing ISSR markers were published in 1994 (Zietkiewicz et al. 1994; Gupta et al. 1994). The initial studies focussed on cultivated species, and demonstrated the hypervariable nature of ISSR markers (reviewed in Wolfe and Liston 1998). To test the utility of the method in natural populations, Wolfe et al. (1998a, b) reexamined a known hybrid complex of four species of Penstemon for which three other molecular data sets were available. Their results clearly demonstrated the utility of ISSR markers for addressing questions of hybridization and diploid hybrid speciation. In addition, several other studies from A. Liston's lab at Oregon State University, A. Wolfe's and D. Crawford's labs at Ohio State University and H. Ballard's lab at Ohio University ranging from conservation biology to molecular ecology and systematics have demonstrated the utility of the technique in a wide range of applications and plant families (Asteraceae, Brassicaceae, Hippocastanaceae, Orchidaceae, Poaceae, Scrophulariaceae, Violaceae).

ISSR techniques are nearly identical to RAPD techniques except that ISSR primer sequences are designed from microsatellite regions and the annealing temperatures used are higher than those used for RAPD markers. Based on the published, unpublished and in-progress studies that have been conducted using ISSR markers, it is clear that ISSR markers have great potential for studies of natural populations (reviewed in Wolfe et al. 1998b).

Gupta M, Chyi Y-S, Romero-Severson J and Owen JL (1994) Amplification of DNA markers from evolutionarily diverse genomes using single primers of simple-sequence repeats. Theoretical and Applied Genetics 89, 998-1006.
Wolfe AD and Liston A (1998) Contributions of PCR-based methods to plant systematics and evolutionary biology. In: Plant Molecular Systematics II eds. D. E. Soltis, P. S. Soltis and J. J. Doyle. pp. 43-86. Kluwer.
Wolfe AD, Xiang Q-Y and Kephart SR (1998a) Diploid hybrid speciation in Penstemon (Scrophulariaceae). Proceedings of the National Academy of Sciences, USA 95, 5112-5115.
Wolfe AD, Xiang Q-Y and Kephart SR (1998b) Assessing hybridization in natural populations of Penstemon (Scrophulariaceae) using hypervariable inter simple sequence repeat markers. Molecular Ecology 7, 1107-1125.
Zietkiewicz E, Rafalski A and Labuda D (1994) Genome fingerprinting by simple sequence repeat (SSR)-anchored polymerase chain reaction amplification. Genomics 20, 176-183.

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Go to:

Wolfe Homepage

Wolfe Lab Homepage

Penstemon Website

Please send your suggestions, comments, corrections to
Last updated January 17, 2000.