Marker-Assisted Verification of Hybrids in Pearl Millet-Napiergrass (Pennisetum glaucum [L.] R. Br. x Pennisetum purpureum Schumach.). (December 2011) Charlie D. Dowling, III, B.S., College of Agriculture and Life Sciences Chair of Advisory Committee: Dr. Russell W. Jessup A high-biomass perennial grass that is directly seeded using existing farm equipment can reduce both planting and overall input costs. Three cytoplasmic male-sterile cms A-lines and four fertile genotypes of pearl millet (Pennisetum glaucum [L.] R. Br.) and one novel pearl millet selection from the Perennial Grass Breeding Program at Texas A&M University were selected to cross with napiergrass (Pennisetum purpureum Schumach.). The pearl millet parents were chosen based on characteristics such as basal tillering, plant height, and days to anthesis. Three napiergrass accessions from the Perennial Grass Breeding Program and the cultivar Merkeron were used as pollinators for these crosses. The cms and fertile pearl millet accessions produced full heads of seed when pollinated with napiergrass. There were a large range of seed sizes and weights for each hybrid family, and the seed were separated into four size classes. The weight differences from the largest to smallest class of seed varied by more than 30%. All of the seed classes germinated, and seed size, in this case, was completely unrelated to the ability to germinate. 100% germination was observed in five seed size classes for both PMN iv hybrids, and 90% germination was observed in three of the eight classes. Essentially all of the hybrid seed recovered from the original pearl millet x napiergrass crosses germinated, but all of the F 1 hybrids were sterile in that none of them produced viable seed. Flow cytometry could not be used to identify the hybrids because the DNA content of pearl millet and napiergrass were essentially the same even though distinct 2C and 4C peaks were seen from the diploid pearl millet. From the 58 EST-SSRs surveyed in the bulked segregate analysis, several were heterozygous dominant and many were homozygous dominant and hemizygous at its particular loci. Seven hemizygous EST-SSRs were identified for Merkeron, seven for PEPU09FL01, eight for PEPU09FL02, and six for PEPU09FL03. These markers are extremely valuable to any pearl millet x napiergrass hybridization program because they provide a means whereby the hybrids can be easily identified. Identification of hemizygous pearl millet markers will also assist in future DNA sequencing and also in a marker-assisted breeding program.

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