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United States Department of Agriculture

Agricultural Research Service

Research Project: INTEGRATING FORAGE SYSTEMS FOR FOOD AND ENERGY PRODUCTION IN THE SOUTHERN GREAT PLAINS

Location: Forage and Livestock Production Unit

Title: Genotypic evaluation of tall fescue dihaploids by capillary electrophoresis

Author
item Kindiger, Bryan

Submitted to: Plant and Animal Genome Conference
Publication Type: Proceedings
Publication Acceptance Date: January 4, 2013
Publication Date: December 26, 2012
Citation: Kindiger, B.K. 2013. Genotypic evaluation of tall fescue dihaploids by capillary electrophoresis. Plant and Animal Genome Conference. January 12-16 2013, San Diego, CA. P0099.

Interpretive Summary: Methods are necessary to discriminate between various products generated in a novel breeding approach that produces inbred lines of tall fescue. When compared to field grow-outs, molecular markers are an efficient and rapid application for discriminating between inbred and hybrid or open-pollinated individuals. A capillary electrophoresis apparatus was employed to determine the frequency of inbred vs open-pollinated individuals generated by the novel breeding process and determine the frequency of such products. Thirty lines were submitted for the analysis which included two well characterized open-pollinated checks. Of these, twenty individuals were identified as dihaploid and eight were identified as outcrosses hybrids. The use of a capillary electrophoresis system represents an effective, rapid and inexpensive method for discriminating between inbred lines and open-pollinated materials. The approach is also effective in determining the quality of inbred line frequency during the breeding process.

Technical Abstract: Recent innovations in tall fescue breeding and selection allow for the generation of dihaploid tall fescue lines. During the dihaploid generation process, two possible products can be generated. These being tall fescue hybrids generated from outcrossing and homozygous dihaploid tall fescue lines. As a means to rapidly differentiate between these two types of products, an automated Advanced Analytical CE capillary electrophoresis system was employed to evaluate various EST-SSR marker profiles for selected individuals. Thirty lines were submitted for the analysis which included two well characterized open-pollinated checks. Of these, twenty individuals were identified as dihaploid and eight were identified as outcrosses hybrids. The 2-5 bp fragment separation resolution for smaller EST-SSR fragments (100-250bp) was found to provide an acceptable level for discriminating between dihaploid and outcross lines. The use of a an automated capillary electrophoresis device can be efficiently utilized to enhance selection of tall fescue dihaploid lines. The analysis also provided valuable information with regard to the viability and receptivity of embryos to fertilization by cross or open-pollination and the necessity for plant isolation during the dihaploid generation process.

Last Modified: 4/23/2014
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