Plant & Animal Genomes XIII Conference

January 15-19, 2005
Town & Country Convention Center
San Diego, CA

Construction Of A Quinoa BAC Library And Its Use In Identifying The 11S Seed Storage Protein Gene

Mikel R. Stevens¹ , Craig E. Coleman¹ , Susan E. Parkinson² , Hong-Bin Zhang³ , Marie B. Balzotti¹ , David Kooyman⁴ , K. Arumuganathan⁵ , Alejandro Bonifacio⁶ , Daniel J. Fairbanks¹ , Eric N. Jellen¹ , Peter J. Maughan¹ , JoLynn Stevens¹

¹  Department of Plant and Animal Sciences Brigham Young University Provo, UT 84602-5157

²  Department of Plant and Microbial Biology 111 Koshland Hall Berkeley, CA 94720

³  Department of Soil and Crop Sciences and Institute for Plant Genomics and Biotechnology 2123 TAMU, Texas A&M University College Station, TX 77843-2123

⁴  Department of Physiology & Developmental Biology Brigham Young University Provo, UT 84602

⁵  Flow and Image Cytometry Core Laboratories Benaroya Research Inst. at Virginia Mason 1201 Ninth Ave. Seattle, WA 98101

⁶  The Foundation for the Promotion and Investigation of Andean Products (PROINPA) La Paz, Bolivia

Quinoa (Chenopodium quinoa Willd.) is a pseudograin that provides nutritional value for high Andean plateau farmers and commercial value for many South American countries. Present interest in quinoa is due to its tolerance of harsh environments and its well balanced nutritional quality. A BAC library, containing 74,496 clones with approximately 10X coverage of the quinoa genome, was constructed by partial digestion of the genome with BamHI and EcoRI. Individual clones were arrayed in 384-well plates for long term storage at Brigham Young University and the Arizona Genomics Institute where they are available for public distribution. Double-spotted, high-density arrays were prepared and probed to characterize the quality of the library. A probe containing a mixture of chloroplast DNA fragments identified 2035 positive clones or 2.76% of the total library. Hybridization using 12 probes representing single copy sequences identified from a quinoa EST collection resulted in an average of 12.4 positive hits per probe which is in close agreement to the 10X estimated coverage of the genome. Five BACs representing two genetic loci containing the 11S seed storage protein gene were identified and characterized following screening of the library with a probe prepared from a quinoa 11S gene cDNA. Restriction fragments containing the 11S gene from these BACs were compared to fragments of genomic DNA on a Southern blot, revealing a potential polymorphism in the 11S gene between highland and coastal varieties of quinoa. This may have important implication for assessing the protein quality of quinoa grain.