Morphological and Biochemical Analysis of Variation in Diploid Glycine Tomentella Hayata (2N = 38, 40)
Hill, Jerry Leon
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https://hdl.handle.net/2142/87679
Description
Title
Morphological and Biochemical Analysis of Variation in Diploid Glycine Tomentella Hayata (2N = 38, 40)
Author(s)
Hill, Jerry Leon
Issue Date
1999
Doctoral Committee Chair(s)
Hymowitz, Theodore
Department of Study
Agronomy
Discipline
Agronomy
Degree Granting Institution
University of Illinois at Urbana-Champaign
Degree Name
Ph.D.
Degree Level
Dissertation
Keyword(s)
Chemistry, Biochemistry
Language
eng
Abstract
"Great strides have been made in the past two decades in understanding and resolving the structure of the genus Glycine. One species, G. tomentella, has remained somewhat problematic because of its great variability. It has been termed a ""species complex"". This research was designed to help sort out that complexity by gaining a greater understanding of the diploid members of the species. A better understanding of the diploids would then be a basis for understanding the complex of tetraploids and for clarifying relationships to other species. A total of 76 diploid accessions plus three outgroup species were studied. Measurements were made at the levels of gross morphology, cytology (somatic chromosome count), and macromolecules---total protein, isozymes, and DNA. Measurements or observations regarding 54 morphological traits were recorded. Twenty-three were deemed sufficiently reliable for statistical use and were analyzed by Principal Components Analysis followed by clustering. Terminal leaflet shape only was analyzed in a very precise manner by use of a machine vision system and singular value decomposition. Seed proteins were extracted under both denaturing (SDS) and native conditions. Denatured proteins were examined by polyacrylamide gel electrophoresis (PAGE). Some native proteins were examined by two methods: (1) General trypsin inhibitors separated by PAGE and visualized with negative staining, and (2) Bowman-Birk inhibitors (BBI) separated by isoelectric focusing (IEF) and visualized by immuno-staining of Western blots using anti-BBI primary antibodies and a phosphatase linked secondary antibody. Genomic DNA was extracted from fresh, young leaves and digested by three restriction enzymes. After separation on agarose gels, the Southern blots were probed with three P32 labeled, genomic DNA probes (Chalcone synthase, soybean seed urease, and soybean lectin). Genomic DNA was also amplified with the Polymerase Chain Reaction (PCR) using primers tailored to one intron of the a' subunit of b -conglycinin These PCR products were digested with restriction enzymes and the fragment polymorphisms analyzed. The groupings defined by all levels of measurements agreed perfectly; three well-defined groups were identified from Queensland and one more variable group from Western Australia and Northern Territory, Australia."
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