Title: Can high quality DNA be extracted and utilized from Arachis seeds in long term storage with zero percent germination? Authors
Submitted to: American Peanut Research and Education Society Abstracts
Publication Type: Abstract Only
Publication Acceptance Date: May 18, 2011
Publication Date: July 15, 2011
Citation: Barkley, N.L., Wang, M.L., Pittman, R.N. 2011. Can high quality DNA be extracted and utilized from Arachis seeds in long term storage with zero percent germination?. American Peanut Research and Education Society Abstracts. SanAntonio, TX 7/15/2011. Technical Abstract: Plant germplasm collections are useful resources for both researchers and breeders. These collections provide a source of new gene combinations that can be used in breeding and molecular studies to thwart disease, introduce novel traits, and enhance nutritional benefits of a crop. The USDA maintains a germplasm collection of Arachis species which contains over 9,000 accessions of cultivated and wild peanuts. Many of the accessions in this repository stored in cold storage are old and some have no germination data collected. However, due to the high oil content in these seeds, the germination rate is known to drastically decline after 15 years or more of cold storage; therefore, shelf life for peanut seeds is time limited. The goal of this work was to acquire 0% germination seeds from cultivated and wild species and test their efficacy with molecular markers in comparison to 10 accessions with normal germination. Six seeds from five different accessions (4 cultivated and 1 wild) with 0% germination were obtained and DNA was extracted. DNA was quantified and subsequently diluted for PCR. Primers for SSR, SNP markers and several large fragments (0.9-1.5kb) from Ara h 1 and Arah 5 genes were tested on the DNA extracts from seeds with a 0% germination rate and seeds with a normal germination rate. Overall, this work demonstrated that high quality DNA can be obtained from 0% germination seeds in wild and cultivated peanuts. Further, the extracted DNA can be successfully used as a PCR template for large gene targets and to reveal polymorphic fragments from both SSR and SNP markers.