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DNA changes in naturally and artificially aged longleaf pine ( Pinus palustris Mill) seeds

DNA changes in naturally and artificially aged longleaf pine ( Pinus palustris Mill) seeds. E.L. Tolentino, Jr 1 ., W.W. Elam 2 and F.T. Bonner 3 1 University of the Philippines Los Baños, 2 Mississippi State University, 3 US Forest Service. Outline of Presentation. Objectives

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DNA changes in naturally and artificially aged longleaf pine ( Pinus palustris Mill) seeds

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  1. DNA changes in naturally and artificially aged longleaf pine (Pinus palustris Mill) seeds E.L. Tolentino, Jr1., W.W. Elam2 and F.T. Bonner3 1University of the Philippines Los Baños, 2Mississippi State University, 3US Forest Service

  2. Outline of Presentation • Objectives • Germination changes • DNA changes • Natural vs artificial aging • Summary and Conclusions

  3. Objectives • Determine if AAT conditions simulate natural seed aging process • Compare DNA changes in naturally and artificially-aged longleaf pine seeds

  4. Germination of artificially aged seeds

  5. Naturally-aged seeds (Alabama Lot)

  6. Naturally-aged seeds (Florida Lot)

  7. Naturally-aged seeds (Louisiana Lot)

  8. Naturally-aged seeds (Mississippi Lot)

  9. DNA content of artificially aged seeds

  10. DNA content of naturally-aged seeds

  11. DNA content of naturally-aged seeds

  12. Natural vs Artificial Aging • In AAT-treated seeds: • Very low and insignificant correlations between DNA and % G • In Naturally-aged seeds • No clear pattern of relationship • Naturally aged and artificially aged seeds similar % G when projected DNA value is 0. • Rate of change between aging regimes not the same

  13. Gel electrophoresis of artificially aged long leaf pine seeds. DNA was treated with 1 l RNAase (Legend: Lane 1 – 1 kb DNA ladder; Lane 2 – control/unaged seeds; Lane 3 – 48 hr AAT; Lane 4 – 96 hr AAT; Lane 5 – 144 hr AAT; Lane 6 – 192 hr AAT; Lane 7 – 240 hr AAT; Lane 8 – 288 hr AAT).

  14. Gel electrophoresis of naturally aged long leaf pine seeds. DNA was treated with 1 l RNAase (Legend: Lane 1 – 4oC, 90 days; Lane 2 – 4oC, 180 days; Lane 3 – 4oC, 270 days; Lane 4 – 4oC, 360 days; Lane 5 –30oC, 90 days; Lane 6 – 30oC, 180 days; Lane 7 – 30oC, 270 days; Lane 8 – 30oC, 360 days).

  15. Gel electrophoresis of naturally aged long leaf pine seeds. DNA was treated with 1 l RNAase (Legend: Lane 1 - 1 kb DNA ladder; Lane 2 – forest condition, 90 days, without RNAase; Land 3 – forest conditions, 90 days with RNAase).

  16. Summary and Conclusions • For AAT-treated seeds • Low-high-low pattern of DNA change coinciding with viable-non-viable cycle. • After 192 hr, abrupt increase in DNA • Sudden drop in DNA after 240 and 288 hr • Fragmented DNA after 144 hr • High MW fragments in seeds after 240 and 288 hr (crosslink between DNA and lipid peroxidation products)

  17. Summary and Conclusions • Storage under forest conditions • Increase in DNA after 90 days (dead seeds) • Fragmented DNA • Storage in 4oC and 30oC • No distinct changes in DNA • Fragmentation began at 270 days of storage at 30oC • No fragmentation for seeds stored at 4oC

  18. Summary and Conclusions • AAT may not be useful for examine aging process in seeds • AAT still valid as vigor test for longleaf pine and even other species.

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