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| Author(s): | Fergudon, J. M.;
Grabe, D. F. |
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| Author Affiliation: | Oregon State University |
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| Title: | Separation of Annual and Perennial Species of Ryegrass by Gel Electrophoresis of Seed Proteins |
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| Source: | Journal of Seed Technology. Vol. 9, No. 2, 1984, p. 137-149. |
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| Publishing Information: | East Lansing, MI: Association of Official Seed Analysts |
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| Keywords: | TIC Keywords: Lolium multiflorum; Lolium perenne; SDS-PAGE; Seed industry; Seed testing
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| Abstract/Contents: | "Rapid and effective techniques that would complement the seedling fluorescence test are needed to differentiate annual (Lolium multiflorum Lam.) from perennial (L. perenne L.) ryegrass. Electrophoresis procedure have been successful in separating the two species, but differences between them are often based only on band intensity. The purpose of this work was to develop an electrophoretic procedure that would, by presence or absence of distinct bands, differentiate between annual and perennial ryegrass, detect mixtures of the two species, and possibly detect ploidy level of annual ryegrass. Proteins were analyzed using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Seveteen annual, three itnermediate (L. hybridum Hausslen), and 14 perennial cultivars were tested. The annual and intermediate cultivars possessed characteristic protein bands at Rf 0.71 and 0.73 that were not found in any of the perennial cultivars. Moreover, bands were present in the perennial cultivars at Rf 0.80 and 0.88 that were absent or very faintly stained in the annual and intermediate cultivars. The banding patterns for annual and intermediate species were similar. To test for contamination of perennial ryegrass seed lots with small percentages of annual ryegrass seed, mixtures of annual and pereniial ryegrass seed were made in concentrations of 0, 1, 3, 5, 10, 25, 50, 75, and 100% annual seed. Annual bands were visible in the samples containing 25% or more annual seed. Densitometer scans could detect the annual bands in the mixtures containing 10% annual seed, but lower percentages of annual seed could not be detected in the bulk seed mixtures. Protein extracts of indicidual seeds were electrophoresed to determine whether species mixtures can be detected on an individual seed basis. When individual seeds were used, the resulting banding patterns were different than those produced from bulk seed extracts from the same cultivar. Furthermore, no two seeds within a cultivar showed the same banding patterns. However, the characteristic annual bands at Rf 0.71 and 0.73 were still evident in most single seeds from annuals. Likewise, the characteristic perennial bands at Rf 0.80 and 0.88 were normally present in individual seeds of the perennials. Individual seeds could be identified by either or both pairs of these bands or bands in other zones. The SDS-PAGE procedure used in this study were successful in differentiating between annual and perennial ryegrass. Bulk seeds and single seeds of the two species show distinct species-specific banding patters. Perennial ryegrass seed lots contaminated with 10% or more annual ryegrass seed could be detected that would allow determination of ploidy level among annual ryegrass cultivars." |
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| Language: | English |
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| References: | 14 |
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| Note: | Pictures
Tables |
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| ASA/CSSA/SSSA Citation (Crop Science-Like - may be incomplete):
Fergudon, J. M., and D. F. Grabe. 1984. Separation of Annual and Perennial Species of Ryegrass by Gel Electrophoresis of Seed Proteins. Journal of Seed Technology. 9(2):p. 137-149. |
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