Title: Pollen genotyping in cotton for genetic linkage analysis Authors
Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: April 18, 2013
Publication Date: N/A
Technical Abstract: Cotton is an important fiber and oil crop and thus makes very important contributions to US agricultural security and sustainable agriculture. Two species are vital for American cotton industry, i.e., Upland cotton (Gossypium hirsutum) and Pima cotton (G. barbadense) that are prized for high yields and exceptional fiber fineness, respectively. For combining superior fiber quality and diverse adaptability, breeding lines of Upland cotton have been created by deleting some Upland genetic material and replacing it from Pima cotton. This research used cotton aneuploid (G. hirsutum x G. barbadense) plants with G. barbadense chromosomes17 in G. hirsutum background. Segregation analysis of molecular markers from individual pollen grains can generate genetic data for linkage mapping without the need of performing controlled pollinations. These maps can simplify and enhance breeding efficiencies. The individually isolated pollen grains were collected from the CS-B17 mature flowers and then germinated to release their DNA. The limited amount of DNA found in a pollen grain is not adequate for analysis and needs to be increased by using primer extension pre-amplification (PEP) protocol. In this research, Upland cotton substitution line CS-B17 pollen genome was amplified with modified PEP procedures using an array of continuous random 15-mer PEP primers to increase the sizes and numbers of AFLP markers in subsequent AFLP analyses of these PEP products. Use of the MasterAmp™ Extra-Long PCR kit (EPICENTRE®, Madison, WI) substantially increased the sizes and numbers of restriction digestion products. The parental and pollen samples were thus analyzed using IRD-800 and IRD-700 labeled AFLP primer pairs. The parental AFLP markers were scored for their presence or absences in the pollen samples using Saga™ Generation 2 Version 3.1 (Li-Cor, Lincoln, NE) software.