Pre-germination genotypic screening using PCR amplification of half-seeds (original) (raw)

Abstract

A simple and rapid PCR-based method has been developed for determining the genotype of seeds before germination. Single half-seeds of rice (Oryza sativa L.) and wheat (Triticum aestivum L. em. Thell.) were preincubated, without grinding, in an aqueous extraction buffer. The resulting supernatants were then used in polymerase chain reaction (PCR) with oligonucleotide primers corresponding to rice single-copy sequences or a wheat microsatellite repeat. PCR products of identical size were amplified using either the half-seed extract or DNA isolated from leaf tissue. The remnant half-seeds can be maintained in ordered arrays using microtiter plates allowing the recovery of selected genotypes. Pre-germination genotypic screening of seed populations as described in this report should be useful for a variety of applications in plant breeding and genetics studies.

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Author notes

1. G. B. Martin
   Present address: Department of Agronomy, Purdue University, 47907-1150, West Lafayette, IN, USA

Authors and Affiliations

1. Department of Plant Breeding and Biometry, Cornell University, 14853-1902, Ithaca, NY, USA
   J. Chunwongse, G. B. Martin & S. D. Tanksley

Authors

1. J. Chunwongse
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2. G. B. Martin
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3. S. D. Tanksley
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Communicated by G. E. Hart

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Chunwongse, J., Martin, G.B. & Tanksley, S.D. Pre-germination genotypic screening using PCR amplification of half-seeds.Theoret. Appl. Genetics 86, 694–698 (1993). https://doi.org/10.1007/BF00222658

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• Received: 13 August 1992
• Accepted: 04 January 1993
• Issue Date: July 1993
• DOI: https://doi.org/10.1007/BF00222658

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