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Flow cytometric analysis of nuclear DNA content,mitotic chromosome number and protein separation by SDS-PAGE in three accessions of African locust bean (Parkia biglobosa Benth.)
Authors:Edak Aniedi Uyoh  Ikootobong Sunday Urua  Valentine Otang Ntui  Elza Cletus Okpako
Affiliation:1.Department of Genetics and Biotechnology,University of Calabar,Calabar,Nigeria;2.Laboratory of Plant Cell Technology, Graduate School of Horticulture,Chiba University,Chiba,Japan
Abstract:Nuclear DNA of three accessions of Parkia biglobosa collected from three locations in northern Cross River State was investigated using a Patec PA II flow cytometer equipped with an argon ion laser (488 nm), and pictures of mitotic chromosomes were taken using a digital micro-camera (Canon) placed on the eye piece of a binocular microscope at 100X oil immersion. Metaphase chromosome counts of 2n = 22 for accessions A and C and 2n = 24 for accession B, were obtained and through flow cytometry, the three accessions were confirmed to be diploids. The nuclear DNA content and genome size for the accessions were 1.5085, 1.489, and 1.5266 pg (737.7054, 728.121, and 746.5074 Mbp) for accessions A, B, and C, respectively. In another experiment, leaf samples from greenhouse-germinated seeds were analyzed for variation in the banding pattern of the protein by SDS-PAGE in the three accessions. Protein was resolved into three banding groups according to their electrophoretic mobility: slow, medium, and fast, clustering between 100–200, 40–70, and 10–25 kDa, respectively. There was 76% similarity in the banding pattern between the accessions.
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