Miniature Inverted-Repeat Transposable Element Identi!cation and Genetic Marker Development in Agrostis

Creeping bentgrass (Agrostis stolonifera L.) is an important species to the turfgrass industry because of its adaptation for use in high quality turf stands such as golf course putting greens, tees, and fairways. Creeping bentgrass is a highly outcrossing allotetraploid, making genetic marker development dif!cult. Genetic markers anchored to miniature inverted-repeat transposable elements (MITEs) were developed in Agrostis that could be used in genetic linkage mapping, quantitative trait loci studies, or diversity analyses. The FindMITE software program identi!ed 495 candidate MITEs from 16,122 Agrostis DNA sequences. There was evidence of transposition in 79 of the candidate MITEs based on MITE insertional polymorphisms. Genetic markers were developed by MITE-display, a modi!ed ampli!ed fragment length polymorphism technique that anchors ampli!ed fragments to MITEs. Four MITE-display primer combinations were tested on a creeping bentgrass experimental mapping population and 139 polymorphic markers were developed with a polymorphic information content of 0.33. Twentyeight of the polymorphic genetic markers segregated normally. MITE-display genetic markers are a new class of genetic markers for studies of the Agrostis genome. These genetic markers target transposable elements and provide an easy method of identifying allelic variation between Agrostis accessions that may be used for diversity studies or genetic linkage map development. K. Amundsen and S. Warnke, USDA-ARS, Floral and Nursery Plants Research Unit, Beltsville, MD 20705; D. Rotter, Univ. of Texas Southwestern Medical Center, Dallas, TX 75390; F. Belanger and H.M. Li, Plant Biology and Pathology, Rutgers Univ., New Brunswick, NJ 08901; J. Messing, Waksman Institute of Microbiology, Rutgers Univ., Piscataway, NJ 08854; G. Jung, Plant, Soil and Insect Sciences, Univ. of Massachusetts, Amherst, MA 01003. Received 19 Apr. 2010. *Corresponding author ([email protected]). Abbreviations: AFLP, ampli(ed fragment length polymorphism; EST, expressed sequence tag; MIP, MITE insertional polymorphism; MITE, miniature inverted-repeat transposable element; PIC, polymorphic information content; RAPD, random ampli(ed polymorphic DNA; RS, random sheared genomic DNA; SSR, simple sequence repeat; TE, transposable element; TIR, terminal inverted repeat; TSD, target site duplication. Published in Crop Sci. 51 (2011). doi: 10.2135/cropsci2010.04.0215 Published online 29 Dec. 2010. © Crop Science Society of America | 5585 Guilford Rd., Madison, WI 53711 USA All rights reserved. No part of this periodical may be reproduced or transmitted in any form or by any means, electronic or mechanical, including photocopying, recording, or any information storage and retrieval system, without permission in writing from the publisher. Permission for printing and for reprinting the material contained herein has been obtained by the publisher.