DNA changes in naturally DNA changes in naturally and artificially aged and artificially aged longleaf pine ( longleaf pine ( Pinus Pinus palustris palustris Mill) seeds Mill) seeds E.L. Tolentino, Jr E.L. Tolentino, Jr 1 ., W.W. Elam ., W.W. Elam 2 and F.T. Bonner and F.T. Bonner 3 1 University of the Philippines Los University of the Philippines Los Baños, Baños, 2 Mississippi State Mississippi State University, University, 3 US Forest Service US Forest Service
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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 - PowerPoint PPT Presentation
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DNA changes in naturally and DNA changes in naturally and artificially aged longleaf pine artificially aged longleaf pine ((Pinus palustrisPinus palustris Mill) seeds Mill) seeds
E.L. Tolentino, JrE.L. Tolentino, Jr11., W.W. Elam., W.W. Elam22 and F.T. and F.T. BonnerBonner33
11University of the Philippines Los Baños, University of the Philippines Los Baños, 22Mississippi State University, Mississippi State University, 33US Forest US Forest
ServiceService
Outline of PresentationOutline of Presentation
ObjectivesObjectives Germination changesGermination changes DNA changesDNA changes Natural vs artificial agingNatural vs artificial aging Summary and ConclusionsSummary and Conclusions
ObjectivesObjectives
Determine if AAT conditions simulate Determine if AAT conditions simulate natural seed aging processnatural seed aging process
Compare DNA changes in naturally Compare DNA changes in naturally and artificially-aged longleaf pine and artificially-aged longleaf pine seedsseeds
Germination of artificially aged Germination of artificially aged seedsseeds
DNA content of naturally-aged DNA content of naturally-aged seedsseeds
LOT STORAGECONDITION
DNA content (g g-1 seed)
0 days 90 days 180 days 270 days 360 days
LA Forestconditions
a567.67b a675.00a nd nd nd
4oC a567.67a b341.00b b421.25b b525.75a a521.67a
30oC a567.67a a725.83a a696.67a a679.17a a583.17a
MS Forestconditions
a498.83a b340.25b nd nd nd
4oC a498.83a a618.33a b570.25a a494.00a b502.00a
30oC a498.83a a707.17a a694.67a a542.00a a715.25a
Natural vs Artificial AgingNatural vs Artificial Aging
In AAT-treated seeds:In AAT-treated seeds: Very low and insignificant correlations between Very low and insignificant correlations between
DNA and % GDNA and % G In Naturally-aged seedsIn Naturally-aged seeds
No clear pattern of relationshipNo clear pattern of relationship Naturally aged and artificially aged seeds Naturally aged and artificially aged seeds
similar % G when projected DNA value is similar % G when projected DNA value is 0.0.
Rate of change between aging regimes not Rate of change between aging regimes not the samethe same
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).
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).
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).
Summary and ConclusionsSummary and Conclusions
For AAT-treated seedsFor AAT-treated seeds Low-high-low pattern of DNA change Low-high-low pattern of DNA change
coinciding with viable-non-viable cycle.coinciding with viable-non-viable cycle. After 192 hr, abrupt increase in DNA After 192 hr, abrupt increase in DNA Sudden drop in DNA after 240 and 288 hrSudden drop in DNA after 240 and 288 hr Fragmented DNA after 144 hrFragmented DNA after 144 hr High MW fragments in seeds after 240 High MW fragments in seeds after 240
and 288 hr (crosslink between DNA and and 288 hr (crosslink between DNA and lipid peroxidation products)lipid peroxidation products)
Summary and ConclusionsSummary and Conclusions
Storage under forest conditionsStorage under forest conditions Increase in DNA after 90 days (dead seeds)Increase in DNA after 90 days (dead seeds) Fragmented DNAFragmented DNA
Storage in 4Storage in 4ooC and 30C and 30ooCC No distinct changes in DNANo distinct changes in DNA Fragmentation began at 270 days of Fragmentation began at 270 days of
storage at 30storage at 30ooCC No fragmentation for seeds stored at 4No fragmentation for seeds stored at 4ooCC
Summary and ConclusionsSummary and Conclusions
AAT may not be useful for examine AAT may not be useful for examine aging process in seedsaging process in seeds
AAT still valid as vigor test for AAT still valid as vigor test for longleaf pine and even other species.longleaf pine and even other species.