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ARS Home » Southeast Area » Gainesville, Florida » Center for Medical, Agricultural and Veterinary Entomology » Chemistry Research » Research » Publications at this Location » Publication #340003
Title: Laboratory techniques in plant molecular biology taught with UniformMu insertion alleles of maize

Author
item CATHEY, REBECCA - University Of Florida
item COSKAN, SEVGI - University Of Florida
item MAXWELL, DUNCAN - University Of Florida
item ELLIOTT, KIONA - University Of Florida
item JOSEPH, RYAN - University Of Florida
item LIM, ADRIENNE - University Of Florida
item MCMANUS, THOMAS - University Of Florida
item RODGERS, JOHN - University Of Florida
item SHIMANSKY, ANDREW - University Of Florida
item VEREEN, CHRISTINA - University Of Florida
item XIE, YUCONG - University Of Florida
item Hunter, Charles
item SUZUKI, MASAHARU - University Of Florida
item KOCH, KAREN - University Of Florida
item MCCARTY, DONALD - University Of Florida
item CHAMUSCO, KAREN - University Of Florida
item CHASE, CHRISTINE - University Of Florida

Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: 3/12/2017
Publication Date: N/A
Citation: N/A

Interpretive Summary:

Technical Abstract: An undergraduate course - Laboratory Techniques in Plant Molecular Biology - was organized around our research application of UniformMu insertion alleles to investigate mitochondrial functions in plant reproduction. The course objectives were to develop students’ laboratory, record keeping, bioinformatics, and critical thinking skills, enabling them to work comfortably and safely in a molecular biology laboratory environment and use current web-based tools for the analysis of nucleic acid and protein sequences. S-type cytoplasmic male sterility (CMS-S) in maize is characterized by a mitochondria-encoded pollen collapse phenotype that can be reversed by nuclear genetic mutations. These include numerous restorer-of-fertility-lethal (rfl) alleles that additionally condition homozygous-lethal seed phenotypes. Students investigated UniformMu insertion alleles of 10 nuclear genes having predicted mitochondrial functions as candidate rfl alleles. Expression patterns of non-mutant alleles were investigated in pollen and seed development through end-point, reverse-transcriptase PCR (RT-PCR). Mitochondrial targeting leaders were predicted for the non-mutant protein products through amino acid sequence comparisons with orthologous proteins. PCR assays were developed for the insertion alleles, and plants heterozygous for insertion alleles were self or sib pollinated to investigate seed phenotypes. Insertion positive plants were also used to pollinate CMS-S plants for analysis of pollen transmission frequencies in normal cytoplasm and pollen fertility restoration in S-cytoplasm. Funding acknowledgement: National Science Foundation (NSF), United States Department of Agriculture (USDA)