Doctor of Philosophy (Ph.D.)
Virginia Institute of Marine Science
An analytical technique was developed to measure adenine nucleotide levels (ATP, ADP, AMP) and adenylate energy charge (EC) in Zostera marina (eelgrass), a submerged marine angiosperm. A tissue comparison and seasonal survey provide baseline information on natural adenylate variability. The methodology developed can be suitably adapted to other macrophyte species as well. Plants were frozen, lyophilized, scraped free of epiphytes, and homogenized. Adenylates were extracted with boiling 1 mM EDTA + 5% (w/v) PVPP (pH 7.6), and assayed by enzymic conversion of AMP and ADP to ATP, followed by quantitative analysis of ATP via the firefly bioluminescent reaction. ATP, ADP, total adenylates (AT), and EC were highest in leaf tissue (photophosphorylating source), while all adenylates were lowest in root plus rhizome. Monthly time series with aboveground tissue show ATP concentration highest in August and lowest in April, corresponding to periods of senescence (decreased ATP utilization) and growth (increased ATP utilization), respectively. Response of adenine nucleotides and EC in Z. marina to nutrient enrichment, light reduction, and herbicide (atrazine) exposure was evaluated as a monitor of metabolic state. Nutrient enrichment over 2 weeks increased epiphyte colonization, which in turn, appeared to negatively impact Z. marina adenylate content, net productivity, and growth. Z. marina ATP, AT, and EC were weakly and positively correlated with nutrients and light, but decreased over time. Short-term (6 hr) atrazine stress reduced ATP and AT at both 10 and 100 ppb, but EC remained constant. Net productivity decreased at 100, but not at 10 ppb atrazine over 6 hrs. Long-term (21 day) atrazine stress was evident from growth inhibition and 50% mortality near 100 ppb. EC was reduced at 0.1, 1.0, and 10 ppb atrazine, but ATP and EC increased with physiological adaptation to severe stress (100 ppb) after 21 days. Apparently, ATP and AT decrease over the short-term but rebound over the long-term with severe atrazine stress, increasing beyond control levels before plant death results. Supplementing adenine nucleotide and EC results with more conventional quantitative analyses would afford greater knowledge of physiological response to environmental variation.
© The Author
Delistraty, Damon A., "Adenine nucleotide levels and adenylate energy charge in Zostera marina (eelgrass): determination and application" (1982). Dissertations, Theses, and Masters Projects. William & Mary. Paper 1539616629.