Genetic and molecular analysis of the organellar genomes of cytoplasmically inherited mutants of soybean (Glycine max (L.) Merr)

Shoemaker, Randy
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A genetic analysis of a maternally inherited yellow foliage mutant (cyt-Y(,3)) was conducted. This mutant was characterized by pigment content determinations and by plastid ultrastructure analyses;A molecular analysis of soybean chloroplast DNA (ctDNA) was also conducted. The coding sequences of the genes for the atrazine receptor protein (PS II) and for the large subunit of Ribulose-1,5-bisphosphate carboxylase (LS) were located on the physical restriction map of the chloroplast genome;Nuclease activity associated with soybean chloroplasts was identified and was inhibited using high pH and various concentrations of spermidine, disodium ethylenediamine tetraacetic acid, and adenosine triphosphate. A ctDNA-enrichment procedure was developed and used to isolate ctDNA from cytoplasmically inherited mutants, cultivars, and Plant Introductions. Mitochondrial DNA (mtDNA) and ctDNA was isolated from yellow cotyledon lines (cyt-Y(,1)) and green cotyledon lines (cyt-G(,1)) and from a yellow foliage line (cyt-Y(,2)) and a normal green foliage line (cyt-G(,2)). Restriction fragment polymorphisms (RFP) were seen between organelle DNAs of yellow and green cotyledon lines but were not seen between yellow and green foliage lines. Five endonucleases were used to screen selections of the soybean germplasm collection for ctDNA diversity. The enzymes Cla I and Eco R I unveiled RFPs. Five plastome types were seen and intraspecific ctDNA variation was identified. Diverse cytoplasms can now be incorporated into soybean breeding programs.