m -y KNOX LIBRARY ' -Mtr, CALIF. 93940 NAVAL POSTGRADUATE SCHOOL Monterey, California THESIS ESTIMATING THE DISTRIBUTION AND PRODUCTION OF MICROPLANKTON IN A COASTAL UPWELLING FRONT FROM THE CELLULAR CONTENT OF GUANOSINE- 5' -TRIPHOSPHATE AND ADENOSINE-5 ' - TRIPHOSPHATE by Carol Diane Jori September 1981 Thesis Advisor: Dr. E. D. Traganza Approved for public release: distribution unlimited l20h:2A UNCLASSIFIED SeCUHITY CLASSiriCATlON Or This ^*CE (Whrnm 0« Triphosphate Carol Diane Jori 1. ^tmwomuma ono«NiZATiON name ano aoomcss Naval Postgraduate School Monterey, California 93940 READ INSTRUCTTONS BEFORE COMPLETINO FORM 1 ^eCl^lCMT'S CATALOG NUM8EI 5 TYPE OF HEPOBT ft PERIOO COV6RCO Master's Thesis September 1981 •. pe«ro«MiNG o«G. mmomr NuMsen • . CONTRACT 0« OWAHT Nt^MaCNCti )0. •WOOHAM ELEMENT. PROJECT TASK AMCA * WOHK UNIT NUMtERS 1 1 coNTnoLLiNC orricE name ano aoohess Naval Postgraduate School Monterey, California 9 3940 12. REPORT DATE I) NUMBER OF PAGES 123 14. MONITOMINC AGENCY NAME * AOONESS^I dtlldtmmt trom CarnmUnf OHIem) IS. SECURITY CLASS, (ol tHI» rri»orf> Unclassified I5«. OECLASSIFIC ATI on/ DOWN GRADING SCHEDULE l«. DISTRIBUTION STATEMENT (•! IMIt Kftt) Approved for public release: distribution unlimited 17. DISTRIBUTION STATEMENT (ot thm mhatrm«t mtfnd In Btdck 30, II dlllmrmtl fron /l«par«> IS SUPPLEMENTARY NOTES IS. KEY WORDS (Canilnum an rmw»r»» aid* II n«c««a«rr an4 idrnntlly *r Mac* nua*«0 guano sine triphosphate, GTP adenosine triphosphate, ATP upwelling productivity biomass 20. ABSTRACT (Canilnud on r*v«ra« midd II nveaaaarr —* Iddniltf *r Woe* m^^dr) This thesis examines the distribution and production of micro-organisms within a coastal upwelling front located off ?t. Sur , California. Underway measurements of adenosine-5 '- triphosphate (ATP) and pigment fluorescence (principally chlorophyll a) were used to estimate the amount of living biomass present at 2.5 m. Specific and absolute productivity were measured by the nucleotide ratio of guanosine-5 ' - DD FORM I JAN 73 1473 EDITION OW t NOV SS IS OBSOLETE S/N 0 10 J-0 1«- 6601 Unclassified SECURITY CLASSiriCATlOM OF THIS PAOE (9t>mn Data Knlfdd) Approved for public release; distribution unlimited Estimating the Distribution and Production of Microplankton in a Coastal Opwelling Front from the Cellular Content of Guanosine-5 '-Triphosphate and Adenosine-5* -Triphosphate by Carol Diane >Jori Lieutenant, United States Navy B.S., University of Maryland, 1973 Submitted in partial fulfillment of the requirements for the degree of MASTER OF SCIENCE IN METEOROLOGY AND OCEANOGRAPHY from the NAVAL POSTGRADUATE SCHOOL September 1981 -\ teufTo ec At>i»»c*iTtow o» twit wa«r«*i«ii DUDLEY KNOX LIBRARY n«fa (■••««4 triphosphate (GTP) to ATP and GTP , respectively. This inves- tigation was conducted to determine the s ignif icance and applicability of these productivity indices in studying the relationship of production and distribution of microplankton (principally algae) to frontal features in the coastal upwelling zone. The highest concentration of biomass associated with the highest rate of absolute productivity was preferentially located in the strong thermo-nutrient gradient in the warmer stratified water at the equatorward edge of the feature. The measurement of specific productivity using the GTP to ATP ratio was significantly correlated with assimilation numbers (productivity index), lending support to the hypothesis that this ratio is a good indicator of specific community productivity in microplankton (principally phytoplankton) . No correlation existed between the GTP concentration which was enzymatically determined, and the concentration of GTP which was inferred from the calculated values of A ATP. This finding indicates that A ATP is not a good estimator of GTP in community assemblages of microplankton. DD Forrn 1473 1 Jan 73 Unclassified ABSTRACT This thesis examines the distribution and production of micro-organisms within a coastal upwelling front located off Pt. Sur, California. Underway measurements of adenosine- s' -triphosphate (ATP) and pigment fluorescence (principally chlorophyll a) were used to estimate the amount of living biomass present at 2.5 m. Specific and absolute productivity were measured by the nucleotide ratio of guanosine-5 ' - triphosphate (GTP) to ATP and GTP , respectively. This investigation was conducted to determine the significance and applicability of these productivity indices in studying the relationship of production and distribution of micro- plankton (principally algae) to frontal features in the coastal upwelling zone. The highest concentration of biomass associated with the highest rate of absolute productivity was preferentially located in the strong thermo-nutrient gradient in the warmer stratified water at the equatorward edge of the feature. The measurement of specific productivity using the GTP to ATP ratio was significantly correlated with assimilation numbers (productivity index), lending support to the hypothesis that this ratio is a good indicator of specific community productivity in microplankton (principally phytoplankton) . No correlation existed between the GTP concentration which was enzymatically determined, and the concentration of GTP which was inferred from the calculated values of A ATP. This finding indicates that a ATP is not a good estimator of GTP in community assemblages of microplankton. TABLE OF CONTENTS I. INTRODUCTION 1^ A. SPATIAL DISTRIBUTION OF BIOMASS AND PRODUCTIVITY ^^ B. GUANOSINE TRIPHOSPHATE IS 1. The Role of GTP in Protein Synthesis ^^ 2. GTP as a i"^ieasure of Productivity 20 II. METHODS 23 A. CRUISE STRATEGY 23 3. MEASUREMENT TECHNIQUES 28 1. Temperature 28 2. Plant Pigments (Principally Chlorophyll a.) 31 3. Nutrients -^2 4. ATP and GTP 32 C. LABORATORY ANALYSES ^^ 1. Theory of ATP Analysis -^^ 2. Theory of GTP Analysis ^^ 3. Procedures for GTP Enzymatic Conversion 36 4. Procedure for Photometric Determination of ATP or GTP ^0 a. Enzyme Preparation ^0 b. ATP and GTP Analysis 41 D. CARBON-14 UPTAKE EXPERIMENT 46 E. TIGRIOPUS CALIFORNICUS PRODUCTIVITY EXPERIMENT 47 F. DATA REDUCTION 50 III. RESULTS 53 A. OCTOBER 1980 CRUISE FINDINGS 53 B. RESULTS OF THE CARBON-14 UPTAKE EXPERIMENT 7 5 C. RESULTS OF THE TIGRIOPUS CALIFORNICUS PRODUCTIVITY EXPERIMENT 76 IV. DISCUSSION 78 A. OCTOBER 1980 CRUISE 78 B. CARB0N-i4 UPTAKE EXPERIMENT 93 C. TIGRIOPUS CALIFORNICUS PRODUCTIVITY EXPERIMENT - 94 V. CONCLUSIONS 96 APPENDIX A Listing of Surface Data: Time, Latitude, Longitude, Elapsed Distance, Nitrate, Phosphate, GTP , ATP, Chlorophyll a_, Temperature 99 LIST OF REFERENCES 108 INITIAL DISTRIBUT?ION LIST 121 LIST OF TABLES I. TRACK POSITIONS FOR 27-28 OCTOBER 1980 29 II. TRACK POSITIONS FOR 29 OCTOBER 1980 30 III. BIOMASS AND PRODUCTIVITY STATISTICS 54 IV. CORRELATION OF NUTRIENTS, TEMPERATURE, AND BIOMASS INDICATORS 70 V. C-14 UPTAKE RESULTS 75 VI . CORRELATION OF PRODUCTIVITY INDICATORS 76 VII. RELATIVE RATES OF PRODUCTIVITY FOR TIGRIOPUS CALIFORNICUS 77 VIII. GTP/ATP RATIOS IN ENVIRONMENTAL SAMPLES 82 8 LIST OF FIGURES 1. CRUISE TRACK FOR 27-28 OCTOBER 1980 26 2. CRUISE TRACK FOR 29 OCTOBER 1980 27 3. ATP LUMINESCENT REACTION 34 4. KINETICS OF ATP AND GTP REACTIVITY FOR THE HEXO- KINASE/GLUCOSE-6-PHOSPHATE DEHYDROGENASE COUPLED REACTION 39 5. KINETICS OF ATP, GTP, UTP, AND CTP-DEPENDENT LIGHT EMISSION USING CRUDE LUCIFERASE PREPARATIONS ^"^ 6. GTP-DEPENDENT LIGHT EMISSION STANDARD CURVES 45 7. GTP REACTION KINETICS FOR SAMPLES RANGING FROM 0 TO 20 NG/ML OF GTP 45 3. SURFACE NITRATE DISTRIBUTION FOR 29 OCTOBER 1980 CRUISE DATA 56 9. SURFACE PHOSPHATE DISTRIBUTION FOR 29 OCTOBER 1980 CRUISE DATA 57 10. SEA SURFACE TEMPERATURE DISTRIBUTION FOR 29 OCTOBER 1980 CRUISE DATA 58 11. SURFACE DISTRIBUTION OF ATP FOR 29 OCTOBER 1980 CRUISE DATA 59 12. SURFACE DISTRIBUTION OF CHLOROPHYLL A FOR 29 OCTOBER 1980 CRUISE DATA ^^ 13. SURFACE DISTRIBUTION OF GTP FOR 29 OCTOBER 1980 CRUISE DATA 61 14. SLT^ACE DISTRIBUTION OF GTP/ATP FOR 29 OCTOBER 1960 CRUISE DATA 62 15. NITRATE, PHOSPHATE, AND TEMPERATURE VERSUS ELAPSED DISTANCE ALONG THE 29 OCTOBER 1980 CRUISE TRACK 63 16. CHLOROPHYLL A/ ATP, AND, TEMPERATURE VERSUS ELAPSED DISTANCE ALONG THE 29 OCTOBER 1980 CRUISE TRACK 64 17. ATP. NITRATE, AND PHOSPHATE VERSUS ELAPSED DISTANCE ALONG THE 29 OCTOBER 1980 CRUISE TRACK — 65 18. GTP/ATP, NITRATE, AND PHOSPHATE VERSUS ELAPSED DISTANCE ALONG THE 29 OCTOBER 1980 CRUISE TRACK 66 19. GTP/ATP AND TEMPERATURE VERSUS ELAPSED DISTANCE ALONG THE 29 OCTOBER 1980 CRUISE TRACK 67 20. GTP/ATP AND ATP VERSUS ELAPSED DISTANCE ALONG THE 29 OCTOBER 1980 CRUISE TRACK 68 21. NITRATE VERSUS PHOSPHATE FOR 29 OCTOBER 1980 CRUISE DATA 71 22. NITRATE VERSUS TEMPERATURE FOR 29 OCTOBER 1980 CRUISE DATA "72 23. PHOSPHATE VERSUS TEMPERATURE FOR 29 OCTOBER 1980 CRUISE DATA 7 3 10 LIST OF PHOTOGRAPHIC PLATES PLATE 1. TIROS SATELLITE IR IMAGE OF THE CALIFORNIA COAST, 29 OCTOBER 1980 25 11 ACKNO WLEDGMENTS Research for this thesis was conducted as part of the ongoing chemical mesoscale research project headed by Dr. Eugene Traganza under the cognizant authority of the Office of Naval Research, NSTL , Bay St. Louis, Mississippi. I wish to take this opportunity to thank my advisor. Dr. Eugene Traganza, Department of Oceanography, for his advice, assistance, and support during every phase of research for and preparation of this thesis. Special thanks to Mr. Dana M. Austin, Department of Oceanography, whose laboratory expertise, unflagging assistance, and patience proved invaluable to successful completion of this thesis. I would also like to acknowledge Ms. Andrea McDonald, Physics and Chemistry Department; Ms. Bonita Hunter, Oceanography Department: Captain W.W. Reynolds and the crew of the R/V ACANIA: and Mr. Larry Breaker of the National Environmental Satellite Service, at Redwood City, California, for their assistance and support. Lastly, I would like to thank Dist. Prof. Eugene C. Haderlie, Department of Oceanography, for his constructive evaluation of the final manuscript. 12 I. INTRODUCTION A. SPATIAL DISTRIBUTION OF BIOMASS AND PRODUCTIVITY This thesis examines the distribution and production of micro-organisms within a coastal upweliing front located off Pt. Sur , California. Underway measurements of adenosine- s' -triphosphate (ATP) and pigment fluorescence (principally chlorophyll a) were used to estimate the amount of living biomass present at 2.5 m. Specific (mg C mg Chla h ) and -3 -i absolute ( mg C m h ) productivity were measured by a new method using the nucleotide ratio of guanosine-5 ' - triphosphate (GTP) to AT? and GTP , respectively. Currently there is no measure of productivity which can easily be applied to large numbers of samples in the field. It was the aim of this thesis to evaluate this new technique to determine if the GTP/ATP ratio would prove to be a viable alternative to -^ C uptake measurements in the field. There- fore, investigations were conducted to determine the signi- ficance and applicability of the productivity indice, GTP/ATP, to a study of the relationship of production and distribu-cion of microplankton (principally algae) to frontal features in the coastal upweliing zone. The patchy distribution of plankton arises from the irregular structure of many ocean variables. Thus distri- bution is directly affected by factors such as the intensity of mixing and stratification of waters, advection 13 and turbulent diffusion (which alter the physical and chemical boundary conditions by varying light, temperature, and salinity gradients) , rate of phytoplankton growth, and zooplankton predation patterns /_ Ref . 1_7- Growth or productivity is primarily affected by the rate of entry of nutrients into the photo synthetic layer, available insolation, and variations in the physical environment pro- duced by fluctuations in the winds and currents / Ref. 2_/ . Various studies have been initiated to try to under- stand the patchy distribution of biomass and the dynamics which are involved in producing this heterogeneous regime. It is logical to assume that the dispersal of phytoplankton would be controlled to some extent by the physical dynamics of their fluid environment. Some studies of oceanic water which have examined the spatial heterogeneity of biomass have found little evidence of any identifiable structural relationship / Ref. 3, 4, 5_/ . The lack of information about the vertical structure generally limits inferences which can be made about the observed horizontal variability. It may very well be the inhomogeneities in the environ- ment which produce the patchy character of the phytoplankton and in turn the patchy zooplankton distribution, which ultimately ensure the stability of the system. One laboratory study which supports this view investigated limiting nutrient patchiness in an ammonia limited con- tinuous culture and found that organisms were best able to 14 utilize their environment, determined by maximal uptake rate, in a patchy nutrient regime /""Ref . 6_7. Areas of oceanic fronts, however, appear to exhibit some consistent structural relationships between the location of phytoplankton biomass and physical and chemical variables. Although different mechanisms are involved in the generation and maintenance of frontal boundaries (which are produced by upwelling, eddies, and the lateral juxtaposition of adjacent water masses), the generation of strong thermal and chemical gradients indicates that these are likely to be regions of high productivity where a large standing crop will be concentrated. The following studies, both observational and theore- tical, provide supporting evidence for the preferential occurrence of biomass in or adjacent to frontal boundaries. The region southwest of Pt. Sur , California, has been shown to be an active area of frequent upwelling advancing the theories of earlier investigators who proposed that the phenomenon of upwelling is intensified equatorward of capes and points along west coasts in both hemispheres /~Ref. 7_7. Previous research investigating features occurring in this region suggests that biomass is pre- ferentially located adjacent to frontal boundaries defined by strong gradients of nutrients and temperature / Ref . 8, 9/ It is not presently known whether this apparent relationship is simply the result of dynamic processes physically 15 accumulating biomass through the horizontal and vertical movement of water parcels at this location, or if the conditions produced by the upwelling feature itself pro- vide optimal conditions for growth through formation of a "natural chemostat" /""Ref. 10, 11, 12_7. Plankton have been found to be concentrated at the frontal boundary of a warm-core eddy in the southwest Tasman Sea / Ref. 13_7« Seasonal thermal fronts occur in the western Irish and Celtic Seas. The western Irish Sea front is a shallow front which is a zone of transition between mixed and stratified waters. An investigation of the microvariations occurring at the front noted increases in biomass at the front, measured by chlorophyll a, peaking on the western edge in the warmer stratified waters. Microbial activity, measured through urea utilization was found to be highest at the front itself /""Ref. 14_7. The relationship between biomass location, measured by concentrations of chlorophyll a, and primary productivity, measured by the carbon-14 uptake method, was studied in the Celtic Sea / Ref. 15_/. Increases in both biomass and productivity were found at the thermal discontinuity. A persistant salinity front occurs in Liverpool Bay. It has been shown that a marked increase in biomass, measured by chlorophyll a, and growth, by increases in primary productivity and Assimilation Index, occurs at the front /~Ref . 16 7- The frontal boundaries which occur in the 16 southwest approach to the English Channel have not only been found to be sites of high phytoplankton production but under the right conditions red tides may develop. The highest values of chlorophyll a were f ound to extend into the stratified side of the frontal boundary /~Ref . 17_7. Further evidence supporting these in situ observations has been advanced through satellite imagery. SKYLAB obser- vations of the spectral properties of upwelled waters off the northwest coast of Africa found a strong correlation between ocean color ratio gradients, which are indicative of productivity, and sea surface temperature gradients. Ground truth data supported these findings / Ref. 18_/ . Similar structure has been observed in features occurring off Pt. Sur , California. Satellite information from the NIMBUS7 CZCS (Coastal Zone Color Scanner) which has been corrected to show the visible spectral properties associated with the pigment chlorophyll a have confirmed the presence of biomass on the equatorward edge of a cyclonic feature in June 1980 / Ref. 19_7. These satellite images con- firmed previous interpretation of surface contours which were constructed using in situ data / Ref. 20_/ which had suggested the presence of such structure. One simulation study examined phytoplankton patchiness using a spatial model in which an initial phytoplankton patch was subjected to the stresses existing in the natural oceanic environment (turbulent diffusion, nutrient limita- tion, diurnal variations, and nocturnal or continuous 17 grazing) . This model was numerically integrated for several oceanic states. One case of interest examined an oceanic region where abundant nutrients were homogeneously distributed. The immediate consequence was the appearance of a phytoplankton bloom which proliferated as it diffused into the nutrient rich water. An interesting result of this case was the appearance of sharp gradients in the nutrient field near the edge of the phytoplankton patch / Ref . 21_7« The three-dimensional spatial structure of the bio- logical community is extremely complex. Through increased appreciation and understanding of the three-dimensional interactions and identification of the controlling mechanisms which operate in areas of oceanic fronts such as in the relatively contained region where upwelling is occurring, it may be possible to extend these insights to diverse oceanic areas. B. GUANCSINE TRIPHOSPHATE 1, The Role of GTP in Protein Synthesis Because of its crucial role in protein biosynthesis, the intracellular concentration of GTP fluctuates in direct proportion to the increases or decreases in ribosome synthesis and thus growth /~Ref . 22_7- Therefore, the ratio of GTP to ATP has been proposed as an index of potential growth representing the instantaneous productivity per unit biomass / Ref. 23, 24 ~J . 18 Guanosine triphosphate (GTP) is an essential fac- tor required for protein biosynthesis. Its presence is needed to initiate synthesis, for aminoacyl transfer ribonucleic acid (tRNA) binding to ribosomes, in trans- location and elongation processes, and in the termination of the polypeptide chain. During each of these processes GTP is hydrolyzed. No evidence exists for its use in the formation of any covalent bond, so it does not act like an ATP energy donor. In both bacteria and eukaryotes, the binding and release of initiation factors from ribosomes occurs only when all of the components of the initiation complex are present, one of which is GTP. Since initiation is just the first event which begins protein synthesis, GTP is only significantly hydrolyzed during aminoacyl- tRNA binding and translocation processes. After the first aminoacyl- tRNA bond has formed, the acceptor site is blocked by the tRNA. Until this tRNA is translocated and a new codon on the ribosome is exposed, the site cannot accept an additional aminoacyl -tRNA and peptide synthesis is halted. GTP is one factor which is involved in trans- ferring the peptidyl tRNA to a donor site, freeing the acceptor site to receive another aminoacyl -tRNA complex /""Ref. 25_7. GTP thus plays an important role in the noncovalent binding of the translocational factors to the ribosomal surface. Splitting of the high energy phosphate bond is necessary for the movement. Based on 19 experimental evidence, one GTP molecule is hydrolyzed for every translocation act. During the elongation process of protein biosynthesis in prokaryotic systems, a ternary complex is formed, consisting of an elongation factor, GTP, and aminoacyl-tRNA. This complex binds to the ribosome, GTP hydrolysis occurs, and a new peptide bond is formed / Ref . 26_7. During the elongation of a poly- peptide chain, two molecules of GTP must be hydrolyzed for every one molecule of amino acid which is incorporated / Ref. 27_/. For polypeptide chain termination the presence of a GTP-bound release factor is necessary l_ Ref. 28_7» Protein inhibitors suppress nucleotide synthesis, reducing the availability of newly synthesized GTP and GTP { cytosine triphosphate), and thus the sub- sequent incorporation of GTP into RNA and of d-GTP ( deoxy-guanosine triphosphate) into DNA / Ref. 29_/. 2, GTP as a Measure of Productivity The combined supply of nutrients available for the synthesis of new organic matter and the availability of energy for photosynthesis in the form of insolation deter- mines the upper limit of productivity for a particular marine ecosystem. Growth, defined as an increase in phytoplankton substance, measured by carbon-i4 uptake 14 -1 -1 (g Cm d ) has been well documented. The assumption is that carbon^4 uptake measures the net increase in new particulate cell carbon /~Ref . 30, 31, 32_/. This method. 20 although well established, is time-consuming and difficult to apply in the field. Studies which have measured both growth rate and nucleotide concentrations of micro-organisms tend to support the hypothesis that the ratio of GTP to ATP can be equated to a measure of productivity or growth rate such as doublings per hour. Most confirm that the relative amounts of ATP present per unit biomass remain constant; since this level is maintained independent of growth, while the levels of GTP per unit biomass increase. A study of the growth rate of the cyanobacterium Anacystis nidulans grown at different light intensities exhibited an exponential decrease in the GTP concentration per unit biomass with decreasing growth rate while the ATP levels per unit biomass remained relatively constant / Ref . 33_7« A similar decline in GTP levels per unit biomass was reported by Sokawa / Ref. 34_7 in his studies of Escherichia coli . Inversely , Franzen and Binkley have shown GTP levels per unit biomass to increase with growth while ATP levels per unit biomass remain constant in this species /_ Ref. 35_/. In another study which measured GTP in micromoles per gram of dry weight as a function of growth rate in the bacteria Salmonella typhimurium , GTP per unit biomass increased with increasing growth rate ( although a simple relation- ship could not be deduced) while ATP levels per unit bio- mass remained constant for all media "except in the broth where it doubled" /~Ref . 36_7. 21 A field study of the vertical distribution of biomass and productivity in the waters of the Black Sea found simultaneous increases in biomass (ATP), metabolic activity (measured by the energy charge) , and growth (measured by the ratio of particulate nucleic acids to ATP) in the anoxic region of this water body / Ref . 37_7. Results reported in a later study showed a similar increase in the productivity ratio, GTP/ATP , at similar depths /~Ref. 38_7. 22 II. METHODS A. CRUISE STRATEGY One purpose of this thesis was to study the distri- bution and productivity of microplankton within areas of strong thermal gradients. To accomplish this goal, data were acquired from the Naval Postgraduate School's research vessel R/V ACANIA during 27-29 October 1980 in an area located southwest of Pt. Sur , California, which is known to be a region of recurrent upwelling events. Measurements of adenosine-5 '-triphosphate (ATP) and chlorophyll a were used to estimate the amount of living biomass present. Productivity was determined using a new technique, viz., the enzymatic determination of the nucleotide ratio of guanosine-5 '-triphosphate (GTP) to ATP. Another purpose was to gain some insight into the biological significance of this ratio, and to investigate its applicability as a measure of microplanktonic pro- ductivity. To study these questions two experiments were performed. The first, a field experiment, compared carbon-14 (■^"^C ) uptake and assimilation numbers (mg-'-'*C mg chla hr ) to GTP and the GTP/ATP productivity ratio, respectively. The second experiment related rates of protein synthesis inferred from the relative RNA ( ribonucleic acid) to DNA ( deoxy-ribonucleic acid) ratio for different population 23 groups of Tigriopus californicus to the GTP/ATP ratio of similar groupings in order to determine if this productivity index is relevant to zooplankton. Prior to and during the October 1980 cruise, satellite infrared imagery of the area of interest was relayed to the scientific party by Mr. Larry Breaker, the staff oceanographer for the National Environmental Satellite Service (NESS) office located in Redwood City, California. The images provided the location, orientation, and dimen- sions of the feature of interest as well as the apparent strength of the associated thermal gradients which defined the upwelling area. This information proved invaluable for initial location and for designing a cruise track for optimal underway sampling strategy. Most of the sampling effort was concentrated near the equatorward boundary of the feature extending into the stratified waters of the surrounding region. This strategy was principally designed to select prime transects through areas of strong thermal gradients to investigate the levels of biomass and productivity within and adjacent to these areas. Subsequent to the completion of the cruise, enhanced satellite imagery coincident with the cruise dates was fowarded for project documentation. The proposed cruise track was zoom-transferred onto an image (Plate 1) of the California coast, taken 29 October 1980. This view proved useful in interpreting ground truth data. Outer track positions are indicated for orientation (refer to Fig. 1, 2 ). 24 Plate 1. TIROS Satellite IR image of the California coast, 29 October 1980. 25 I 'Oi^ni, I 123 W 122W Figure 1. Cruise Track for 27-28 October 1980. Scale 1:216.116 26 122W Figure 2. Cruise Track for 29 October 1980 Scale 1:216,116 27 Once in the area, the cruise track on 27-28 October was an expanding square (Fig. 1). Table I gives naviga- tional positions for this portion of the cruise track. A continuous record of temperature and fluorescence pin- pointed the areas of sharp gradients and high biological activity validating the satellite imagery. The R/V ACAivIlA used Loran C to fix its position every half hour, while maintaining a speed varying between six to nine knots during the transit of each leg. Once the feature and areas of prime interest were defined, the cruise track on October 29th (Fig. 2) followed prime transects which allowed a more detailed view of the feature's major chemical and biological structure to be obtained. Table II gives navigational positions for this portion of the cruise track. During this phase of operations, the dissolved nutrients- nitrate and phosphate, fluorescence, and sample collections for chlorophyll, ATP, and GTP were taken, and the Carbon-14 experiment was begun. B. iMEASUREMENT TECHNIQUES 1 . Temperature Temperature was measured continuously in situ by a thermistor immediately adjacent to the pump intake located in a sea chest at a depth of 2.5 meters. A strip chart recording was later calibrated through the use of bucket thermometer readings which were taken every half hour. 28 TABLE I Track Positions for 27 -28 October 1980 i 1 Date Position Identification GMT Latitude Longitude i 1 10/27/80 1 0520 36^03. 8'N 122°06.0'W 1 2 0646 36°00.0'N 122^13. I'W 1 110/28/80 3 0743 36°06.5'N 122°17.9'W 1 4 0919 36013.7'N 122°03.6'W 1 5 1050 36*^02.1 'N 121^54.9 'W 1 6 1315 35^51.0 'N 122°15.2'W 1 7 1546 36°08.0'N 1 22° 29.5 'W ] 8 1847 36^22. I'N 122O01.0'W i 9 2157 36^00.1 'N 121*^42.9 'W 1 10 0156 35^40.9 "N 122<=^18.3'W 1 11 0519 36*^05.2 'N 122^^36.4"^ ', 29 TABLE II Track Positions for 29 October 1980 1 Date Position Identification GxMT Latitude Longitude { •10/29/90 9 1646 35^59.9 'N 121^42.9 'wl 8 1946 36°22.1'N 122^01. 0'W| 6 2335 35050.7 'N 122°15.4'wi 7 0200 36*^07.9 'N 122^29.5 'Wi 8 0500 36^21.0 'N 122^02. a 'W{ I 30 2. Plant Pigments (Principally Chlorophyll a) Seawater was pumped from the keel intake to the dry lab where a debubbler removed air bubbles from the water in the line before it was shunted to the fluorescence measuring device. A Turner III Fluorometer provided a continuous record of fluorescence , which was later calibrated following the procedures outlined by Lorenzen /~Ref . 39_7/ using concentrations of discrete chlorophyll a samples. The basic assumption of in vivo fluorometry is that there is a constant ratio between the fluorescence of an in vivo sample and the extractable chlorophyll a pigment. This value is the calibration ratio of the instrument, which is subsequently used to approximate the chlorophyll a concentration. This assumption does not always provide accurate information. Fluorescence can vary by a factor of two to five for a fixed chlorophyll a concentration when phytoplankton are exposed to varying conditions / Ref . 40_7» Seawater samples of 275 ml each were taken in triplicate hourly from the debubbler outflow. Each sample was vacuum fil- tered through a Whatman G/FC glass fiber filter with a pore size of approximately 0.45^m. These filters are able to separate phytoplankton and other larger organisms. The filters were folded, placed in polyethelene bags labeled with the time and ship's position, and stored frozen at -30°G until analyzed according to the procedures outlined by Strickland and Parsons / Ref. 41_/. 31 3. Nutrients A Technicon Autoanalyzer AA-II (Technicon Corporation, Tarrytown, New York) was used to colorometrically determine the reactive inorganic phosphorus and nitrogen in the surface water (2.5m) at a sampling rate of once every two minutes in accordance with the procedures outlined in /~Ref. 42; Ref . 43, Ref. 44_7. Cadmium columns were packed and conditioned for nitrate analysis in accordance with the procedures detailed in Ref. 45. The autoanalyzer reduces the nitrate to nitrite before measurements are made. Therefore traces of nitrite in the upper portion of the water column could erroneously elevate the levels of nitrate recorded. According to Paulson / Ref. 46_7 this error would be unlikely, since his studies of the nutrient variations in this area have indicated that there is almost no interference from surface nitrite. 4. ATP and GTP Vacuum filtration is the most practical and effec- tive method of cell concentration but, it also has a detrimental effect on the concentration of ATP / Ref. 47_7. Therefore, a minimal sample extraction volume of 50 ml was used for concentration and subsequent extraction of the nucleotides in order to reduce the effects of the filtra- tion procedure. To determine ATP, ^ ATP , and GTP at a later date these 50 ml sample volumes which had been pref iltered through a 200/«jn nylon screen were vacuum filtered 32 through a microfine glass fiber filter (Reeve Angel 984-H with a pore size of approximately 0.45 jam). This filter is capable of separating algae, microzooplankton, and marine bacterial cells. Immediately after the final amount of liquid had passed through the filter, the vacuum was released and the filter quickly immersed in 10 ml of boiling Trizma (Tris ( hydroxymethyl ) aminomethane and hydrochloride) buffer (pH 7.7) in order to extract the nucleotides present in the water sample. The test tubes containing the extracts were labeled and stored frozen at -30^C for later analysis. C. LABORATORY ANALYSES 1. Theory of ATP Analysis The ATP concentration in microplankton (algae, microzooplankton, and bacteria) is a reasonably good measure of the total living organic cellular carbon present /~Ref. 48_7. The amount of ATP present within a given sample is determined by the amount of light emitted during a bioluminescent reaction catalyzed by lucif erase. Fig. 3. In the first reaction, lucif erin reacts with ATP in the presence of magnesium producing the bound luciferyl adenylate which then reacts with oxygen to form oxylucif erin , carbon dioxide, adenosine monophosphate and light / Ref . 50_7 According to the kinetics of this reaction the quantum yield of visible light as measured by Seliger and McElroy 33 N N COOH Mr** ATP ^=i E LH^ AMP + PPi 2 HO 0 LUCIFERIN CO N N__OH + AMP + hv HO OXYLUCIFERIN Figure 3: ATP luminescent reaction (from DeLuca / Ref . 49_y) where E-LH2-AMP is the bound luciferyl adenylate; PPi , inorganic pyrophosphate: 0 2/ oxygen: CO 2* carbon dioxide: AMP, adenosine monophosphate: hv, light /~Ref. 51_7 is of the order of 0.88 emitted for each ATP molecule which is hydrolyzed / Ref. 52_7 , Concentrations of ATP can be determined by comparing the light emitted (counts per minute) by the standards with that of the unknown samples. 2. Theory of GTP Analysis GTP was determined by the method outlined by Karl /""Ref. 53_7. GTP is measured using an NDPK (nucleoside diphosphate kinase-f iref ly lucif erase) coupled bioluminescent reaction. In this method GTP is transphosphorylated to an 34 equivalent amount of ATP. The concentration of ATP is then determined by the amount of light produced when it is reacted with a crude substrate-enzyme mixture of firefly luciferase-luciferin as described above. The pertinent reactions are: HK 1. D-glucose + ATP P=^ D- glucose 6-P + ADP + h"*" 2. D-glucose 6-P NADP"^ G6P-DH g^^^onate 6-P+ NADPH + H^ UDPG-PP 3. ^ -D-glucose 1-P + UTP 7 UDP-glucose + pp. NDPK 4. ADP + GTP 1 GDP + ATP L 5. ATP + luciferin y product + PP. + light where HK is hexokinase: G6P-DH, glucose- 6 phosphate dehydrogenase: UDPG-PP, UDP-glucose pyrophos- phorylase: UTP, uridine triphosphate: PP., inorganic pyrophosphate: NDPK, nucleoside- 5 '-diphosphate kinase: and L, firefly lucif erase. Reactions (1) and (2) eliminate all of the ATP present within the sample extracts , preventing any inter- ference from this nucleotide in the quantitative assay of GTP. Reaction (1) removes a phosphate from an ATP molecule and transfers it to D-glucose producing D-glucose-6-phos- phate and ADP. Reaction (.2) oxidizes D-glucose-6-phosphate to gluconate-6-phosphate and simultaneously reduces NADP to NADPH. Reaction (2) drives reaction (1} to completion 35 ensuring that all the ATP in a given sample extract will be converted to ADP. In reaction (3), a phosphate is removed from UTP forming UDP which is combined with a glucose molecule from the D-glucose-1-phosphate to form UDP-glucose, thus effectively preventing interference from UTP. In reaction (4), a phosphate is transferred from GTP to ADP in the presence of excess ADP producing one molecule of ATP for each GTP molecule v^iich is trans- phosphorylated. The assay for GTP is based on the linear luminescent response of crude firefly lantern extracts to the addition of ATP (produced in reaction (4)). In reaction (5) ATP in the presence of the luciferin/lucif erase enzyme preparation gives off light which is then measured by a photometer. This method is sensitive to picomolar (pM) concentrations of GTP /~Ref . 54_7. 3. Procedures for GTP Enzymatic Conversion For the GTP determination, 0.8 ml of each sample extract was pipetted into a series of disposable glass cuvettes (12x75mm) and placed in a test tube rack in an incubating water bath at 30^C. Each tube was allowed to come to temperature and then 0.2 ml of an enzyme solution containing 75mM potassium phosphate buffer (p H 7.4), 15mM MgCl2.0.5 mM .sIADP , 0 . 5mM d-glucose, 0.5 mM K-D-glucose -1-phosphate, HK/G6P-DH (2 units/ml), and UDPG-PP (5 units/ ml) was pipetted into each vial. To obtain the proper proportions, four separate solutions were prepared as follows: 36 1. A 60 mM solution of MgCl2 was prepared adding 5 ml of distilled H^O to .13g MgCl^ 2. A 300 mM solution of K-PO. was prepared by adding 5 ml of distilled H2O to .261g K^PO. 3. A 2mM solution of D-glucose and << -D-glucose-1- phosphate was prepared by adding .01802g of D-glucose and .03363g of •< -D-glucose-1-phosphate to 50 ml H^O ( "nanopure" ) 4. A 2mM solution of NADP was prepared by adding 3.1 ml distilled H2O to a preweighed vial containing 5 mg NADP Then 2.5 ml of each of the above four solutions were com- bined to produce a total solution of lOml to which .236mg HK/G6P-DH and 12.5mg UDPG-PP was added. This produces a final enzyme solution with the desired colarities and required activity levels. Each vial was swirled to mix the reactants and then allowed to react at 36° C for 15 minutes. The enzymes were then deactivated by placing the culture tubes in a boiling water bath (100 C) for three minutes to ensure that the GTP would not in time be hydrolyzed enzymatically . The rack was then removed from the boiling water bath and the cuvettes were allowed to come to room temperature (25*^0) at which time they were sealed with parafilm and stored frozen at -30 C until analyzed. An experiment was conducted similar to one per- formed by Karl /~Ref . 55_7 to confirm that these procedures would produce the desired enzymatic reactions. The enzymatic procedures outlined in the preceding paragraph 37 were followed using 0.8ml of 2x10 M solutions of GTP and ATP. The 30*^0 reaction times were allowed to vary. These were zero, 2 min. , 4 min. , 8 min. , 10 min. , 20 min. , and 30 min. Percent light emission for both ATP and GTP were plotted as a function of reaction time (Fig. 4). The con- centration of solutions composed of 0.8 ml of ATP and GTP, respectively, to which 0.2 ml of Trizma had been added were used as the maximum to which all other values were normalized when percent light emission was calculated. The ATP levels dropped to zero after four minutes indicating that all interference from the ATP had been eliminated from the sample as anticipated. The GTP levels remained relatively constant as expected. For the zero reaction value the percent light enission is only about 46% vice 100% because it took a small amount of time before the mixture could come to deactivation temperature in the boiling water bath and halt the reaction. This delay was enough to significantly reduce the levels of ATP present within the sample. Had the enzyme preparation been injected into a sample which had already come to deactivation temperature the reaction would not have been allowed to proceed and the percent light emission would have been more nearly 100%. The experiment confirmed that the enzymatic pro- cedures do in fact eliminate all interference from ATP which may be present in a sample extract. 38 100. 10. 2 O — (0. X 2 10. UJ ' ' ' i ' 1 ' ' ' I i IS. TIME (MINUTES) Figure 4. Kinetics of ATP and GTP reactivity for the Hexokinase/glucose-6-phosphate dehydrogenase coupled reaction. 39 4. Procedure for Photometric Determination of ATP or GTP a. Enzyme Preparation Each vial of lyophilized firefly lantern extract, Sigma FLE-50 (Sigma Chemical Co., St. Louis, Kissouri), was reconstituted with 5 ml of distilled water, 10 ml of J.iM arsenate buffer ( pH 7.4), and 10 ml of D4 A r-igSC^. This mixture was allowed to sit in the dark at room temperature for approximately 16 hours in order to allow the level of background light emission to subside. To remove the insoluble residue, the enzyme was centrifuged for 15 minutes. It was then filtered through a Whatman #2 paper filter. Immediately before use, 500 /uLg of luciferin and 25 ml of Trizma ( .02 M) were added to the bottle. Spiking the enzyme preparation with exogenous luciferin increases the net light emission per unit ATP in solution / Ref . 56_/. This increased reactivity allows subpicogram levels of ATP to be detected, which in practice allows a 50 ml sam- ple of seawater to be used. A larger volume would take too long to filter because stress produced on the organisms would cause them to alter their ATP levels (therefore, their GTP/ATP ratio would be suspect). When the luciferase enzyme was being prepared for a GTP assay, the enzyme preparation was saturated with ADP using 10.0 /^.g. The enzyme preparation was mixed and stored in a 250ml plastic bottle to lessen the decay rate which was found to be significant in glass containers 40 / Ref . 57_7 due to shearing of the enzyme molecules which occurs at the glass-liquid interface. The assay was carried out under reduced lighting conditions to minimize the des- tructive effects of normal laboratory lighting on the enzyme preparation. b. ATP and GTP Analysis Both ATP and GT? concentrations were analyzed using the light dependent reaction of ATP in the presence of the luciferin/lucif erase enzyme preparation. The amount of light emitted in this reaction is proportional to the concentration of ATP in the unknown sample. The total measuring system was designed and engineered by Biospherical Instruments, Inc., San Diego, California. The primary components include an Industrial Micro Systems 5000 Microcomputer: a SOROC IQ-120 CRT terminal; an Anadex DP9500 Dot Matrix Printer with extended buffer: an ATP integrating photometer, 3AI model 3000, with an automatic pipette: software programs on five inch diskettes which control individual runs and perform the required calculations. A strip chart recorder was inter- faced with the photometer to monitor the reaction kinetics. From observation of the kinetics it was possible to deter- mine if the enzyme was stable from one sample to the next, if the pipette tip dripped and a drop of the enzyme solution initiated the reaction prematurely, and in the GTP deter- minations, if there was any residual interference from ATP which would appear as an initial peak. 41 During each run 0.2 ml of an unknown or standard solution was automatically injected into a polystryene cuvette (10x50 mm) containing 0.5 ml of the enzyme prepara- tion. A collar which was locally engineered to fit inside the photometer held each vial exactly centered ensuring consistant optical conditions from one sample measurement to the next. The reliability of the peak height values depends upon the quick and total mixing of the reactants. This is ensured through the use of an automatic pipette which provides consistant mixing of all samples throughout a run. Actual injection of samples is controlled by the computer. A microswitch in the photometer senses when the shutter is closed, automatically measuring the dark count. when the shutter is opened the computer then measures the enzyme background for five seconds. The sample is then injected. The peak value is recorded in counts s based upon the largest 0.33 second count recorded during the first five seconds after injection. Integral light levels are deteirmined from a series of one-second measurements that are scanned by the computer after the desired delay. In accordance with the assay procedures developed for ATP /~Ref. 58, Ref. 59, Ref . 60_7/ a 15 second delay and 50 second assay period were used. The analysis section of the ATP calculation software developed by Biospherical Instruments Inc., San Diego, Ca. uses the measurement of enzyme background level as a method of tracking and 42 adjusting for the decline in enzyme activity. The rela- tionship between raw counts (uncorrected for endogenous background emission or blank) and standard values is not linear. With increasing time there is a smaller change in counts at lower concentrations than at higher concentrations of ATP. This relationship can be approximated by a second degree polynomial which is used to calculate a completely new standard curve for each sample according to its enzyme background emission / Ref . 61_7. This multiple curve fitting is used to compute the integrated and peak values. Counts per minute are then converted into concentration "1 values in units of ng ml . When samples were analyzed, duplicates of each were run. If these values, in counts per minute, did not agree within 10%, a third sample was analyzed. A complete set of nucleotide standards were prepared using the sodium salt of the nucleotide and .02M Trizma. Standards prepared in the following concentrations: .3, l.,3.,5., 9., 15., and 30. ng ml were run at least every two hours. The final values of both ATP and GTP concentrations were the averaged values of the replicates. The ATP values were determined from the peak height measurements in order to reduce the interference of GTP and other nucleotides which may be present within the sample. If integrated light readings are used, the kinetics of the other nucleotides would interfere with the reaction resulting in falsely elevated levels of ATP /~?ig. 5_7. 43 lOO r- 1 z 1 g 80 - 1 UJCO 1 >VJ 60 — \ cri 1 o -1 40 20 - - V r? 7*^ ---^-jr:::::::^:;;^ •^ ATP 1 — L'TO 1 *~- GTP J rTP 0 - iC ^^ TRIS 1 1 1 1 1 1 1 1 1 1 , BUFFER c ) 20 40 60 eo 100 I2C TIME AFTER INJECTION (sec) 1 Figure 5: Kinetics of ATP, GTP , UTP , 1 and CTP-dependent | light emission using crude luciferase pre- | 1 par tie ■at le ion. was 3. The 4 X 10^ concentration of each nucleo- i M (from Karl / Ref. 62_/) ', Figure 6 shows the reaction kinetics for varying concentrations of GTP through standard light emission curves. Integral values of light emitted were used to deter- mine the concentrations of GTP present within the samples. Figure 7 shows that either the initial rise, peak, or integrated values can be used to calculate GTP concentra- tions. Although graphical analyses of integrated light values have been used in the past to calculate ATP concen- trations, greater precision is achieved through the use of sophisticated computer software as described above /~Ref. 65_7. 44 100 I h 80 I- 20 ng ml' 10 nq ml"' 6 nq ml*' 4 nq ml"' 2 nq ml"' TRIS BUFFER 20 40 60 60 lOO TIME AFTER INJECTION (tec) 120 Figure 6: GTP-Dependent Light Emission Standard Curves (from Karl / Ref . 63 7) 4 — too - oo '" • - L o INTEG«AT€D «t y so 80 — y/ 'o > L / n PEAK 2 a. u o Ul < O 2 1 >60 - sc < a . 40 20 S - .60 £ - <40 z 20 / y jO WtTlAL 0 — A _ O ^1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 I 1 ,1 J ( J 5 10 15 20 CTP ,nq ml" Figure 7 : GTP reaction kinetics for samples rangi ng 1 from f Rel 0 to :. 64 20 ng/ml of GTP i from Karl -7) 45 D. CARBON-14 UPTAKE EXPERIMENT Five water samples were collected at different locations along the October cruise track using a two liter PVC Van Dorn bottle at 1136, 1150, 1339. 1610, and 1933 hours local time. These locations were chosen from observation of the fluorescence record so that areas with differing amounts of biomass could be studied. From these samples, subsamples 14 were taken to determine C uptake, ATP, GTP , and chlorophyll a. Measurement of carbon-14 uptake is the most direct approach used to measure primary productivity. The uptake 14 of CO^ is considered to be a reasonable indicator of primary production or growth. In this experiment a known 14 amount of radioactive carbon, G, was added to three 125ml 14 water subsamples. The C was added to the phytoplankton 14 culture in the form of sodium carbonate, Na CO^) , which contained approximately five microcuries of radioactivity per ml per subsample. Two of these subsamples were incubated at surface temperature for approximately four hours on board the R/V ACANIA using a fluorescent light incubator which provided about .06 langley per minute of illumination. The third bottle was kept in the dark and used as a control . To eliminate the contribution of the non-photo- synthetic fixation of carbon from -the experiment, the dark bottle counts were subtracted from those of the light bottles. After incubation was completed 100ml of each 46 subsample was filtered and stored in accordance with the procedures outlined by Rowney /~Ref . 66_7 to await shore- based calibration. (Dana M. Austin performed these analyses using a liquid scintillation counter as outlined by Jitts and Scott /~Ref. 67_7) . Primary production was determined using the following formula from Strickland and Parsons /~Ref . 68_7: 1. mg C m"^ hr"-^ = (Rs-Rb) (1.05w)/(RN) where Rs is the radionucleotide activity in the light bottle (CPM): Rb, the dark bottle count (CPM): R, the absolute activity in one ampule (CPM): W, weight of the carbonate carbon in water which is assumed constant at 24,000 mg C ; N, the number of hours of incubation: 1.05, the isotope discrimination factor. Assimilation 14 -1 -1 numbers ( mg G uptake mg Chla hr ) were determined 14 using the C uptake information and the results of the discrete chlorophyll a. determinations. 14 Measurements of productivity based on C uptake and assimilation number were then correlated with GTP and the GTP/ATP productivity ratio y respectively. E. TIGRIOPUS GALIFORNICUS PRODUCTIVITY EXPERIMENT Tigriopus calif ornius is a marine copepod found in the splash zone above the mean high water mark along Monterey 3ay. Tigriopus calif ornicus hatches from eggs and grows through six naupliar and six copepodid stages, 47 each of which can be microscopically characterized by distinctive developmental features /~Ref . 69_7. Research by Baugh / Ref . 70_7 related the growth stages of this copepod to the RNA/DNA ratios at various phases in its life cycle. Since RNA is a necessary ingredient required for pro- tein synthesis, rapidly growing cells contain relatively large amounts of this nucleic acid. In contrast, cells which are growing at a slower rate contain little RNA. A direct correlation between RNA synthesis and protein synthesis has been demonstrated in populations of exponentially grow- ing cells / Ref. 71_7. Sutcliffe / Ref. 7 2_/ showed that this relationship existed in 24 diverse species. Pease / Ref. 73_/ was only able to verify these results in the exponential phase of growth. Work by Leick / Ref. 74_7 studying bacteria, yeast, and protozoa showed that for a given micro-organism the ratios of RisIA to protein and RNA to DNA are linear functions of the growth rate. Tiqriopus californicus were chosen as an experimental organism since data was available on the RNA to DNA ratios for various population subgroups of this species. An experiment was performed to determine if similar trends would be observed in the d^ta when the GTP/ATP ratios were compared to the RNA/DNA ratios for different popula- tion subgroups. 48 Tigriopus californicus were collected with a number 10 plankton bucket from splash pools above the mean high water mark near the Great Tide Pool south of Monterey Bay. Once collected these copepods were kept in plastic con- tainers at 23 C. Individuals were then separated into three experimental groups, similar to those described by Baugh / Ref . '75_/. An acryllic separation column with nylon screens was used to separate a mixed population of individuals between 400-500 /a m in size. A Pasteur pipette was used to separate an all-gravid female population, the all-but-gravid population were those individuals which were left. Once separated into groups the populations were allowed to rest for two days to recover from the stress of separation in order that their ATP levels would return to normal [_ Ref. 76_/. Immediately prior to extraction the copepods were concentrated by using a 100 ^ m nylon screen in the separation column. The organisms were then extracted by immersion in 40ml of boiling Trizma and by simultaneous homogenization with a pestle homogenizer for five minutes. The test tubes were removed and allowed to cool before the extraction fluid was filtered and the filtrate stored frozen at -30°C to await nucleotide analysis. GTP and ATP analyses were performed in accordance with the procedures outlined above. The trend of the productivity ratio, GTP/ATP , was then compared to the trend of the RNA/DNA ratio for similar population groups. 49 F. DATA REDUCTION To prepare the data for statistical analysis, the strip chart recordings of fluorescence, temperature, nitrate, and phosphate were hand digitized at a constant interval of two minutes. This sampling interval represents approxi- mately 0.6 km along the cruise track when at full speed. To generate organic carbon equivalents for comparison of biomass concentrations (determined through ATP and chlorophyll a analyses) , the conversion factors originally proposed by Holm-Hansen / Ref . 77_/ were used. Determination of these factors was based on exhaustive laboratory obser- vations. These values are nevertheless just averages which can vary depending on species composition and changing environmental conditions /_ Ref. 78_/. Chlorophyll a. concen- trations were converted to carbon units by using the average conversion factor of 100. ATP values were converted to organic carbon using the average carbon to AT? conversion factor of 250 which is believed to be representative of community microbial biomass (i.e., representing the com- bined contributions from bacteria, algae, and microzoo- plankton) . In Ref. 79 Holm-Hansen reviews various studies which relate ATP to total cellular organic carbon in a wide variety of fresh and marine organisms giving credence to this average conversion factor. Another study which verifies this conversion factor used direct microscopy to estimate the microbial biomass comparing these values 50 to those obtained from ATP determinations /~Ref . 80_7. These investigators found an average conversion factor of 245 for community microbial populations not only in euphotic areas of upwelling but also in the euphotic zone in areas of equatorial divergence, and in aphotic layers. Studies of ATP levels in nutrient-deficient phy topi ank tonic organisms have shown some effect on ATP levels, but not enough to significantly affect biomass determinations based on this assay procedure /""Ref . 81_7. A ATP values were computed by subtracting the peak ATP values from the integrated ATP values /~Ref . 82_7. The A ATP and the GTP values were normalized to the biomass by dividing these values by the amount of ATP present within the sample. Population correlation coefficients were derived using the equation: r = n 2 x^y^ - CS x^ ) i^y^) ( / nSx. ^ - (S X )^_7 / nil y^ (•Sy^)^_7)^ Nitrate, phosphate, temperature, biological and productivity indicators were first correlated for all the data points. Then the values were separated into two groups based upon their nitrate concentration. Correlations were then run en these two distinct groups whose separation of values (Fig. 21) suggest the presence of two distinct water masses 51 during the October 1980 cruise. The implications of the results of these correlations in relation to the biological variables will be discussed in the following chapter. The standards which were used in determining the GTP concentrations for the cruise data were not treated in a manner similar to the samples. To correct for this systematic oversight an experiment was conducted in which one set of standards was prepared and subsampled into two groups. One group was enzymatically treated, the other was not. The relative activity of GTP in the treated standards was an average of 25.6 percent of the untreated standards. The cruise data were adjusted upwards in accordance with this finding. 52 Ill . RESULTS A. OCTOBER 1980 CRUISE FINDINGS On the October 1980 cruise, temperature, nutrient, biomass , and productivity data were collected and analyzed. A listing of these data appears in Appendix A. Results presented in this chapter are associated with a minimum of description. Interpretive discussion is reserved for the following chapter. Ranges for the biomass and pro- ductivity indicators are given in Table III. Surface maps and linear track plots from this cruise were constructed using the IBM 3033 AP computer's VERSATEC plotter (Fig. 8 to 20). To create the surface contour maps a grid was constructed on a planar surface assigning the origin to the head of the Sur submarine canyon. Each data point was then assigned a cartesian coordinate on this grid based on its latitude and longitude / Ref. 83_/. Using this data the C0NI3D library subroutine from the W.R. Church Computer Center was able to construct surface contours of the indicated variable. Because of the low sampling density the hashed contours are at best a first approximation. The contour maps for chlorophyll a, nitrate, phosphate, and temperature extend further north than those which were constructed for ATP, GTP , and ATP/GTP because sampling for these variables continued from station 8 on the return transit to Monterey. 53 ATP -1. TABLE III Biomass and Productivity Statistics (ng 1"^) GHLA (mg m" ) GTP (pM) GTP/ATP ^ ATP/ATP High 3392 56 906 .3 1.6 Low 97 ean Standard Deviation No. of Samples 529 ±520 57 9 i 9 398 102 il35 57 4 .1 + .1 51 4 .5 + .3 57 where * indicates that the values were below the level of detection 54 The two-dimensional surface spatial distribution of nitrate and phosphate relative to the California coastline is depicted in Fig. 8 and Fig. 9, respectively. The contour plots are constructed from data accumulated on 29 October 1980. The higher nutrient values and the sharpest tem- perature gradients were coincident (compare Fig. 8 and Fig. 9 with Fig. 10). Comparison of the surface temperature map (Fig. 10) with satellite IR imagery from the same day (Plate 1) reveals the strong thermal gradients located between the upwelling colder water along the periphery and the warmer oceanic water. The ATP biomass shows several cells (Fig. 11). The highest concentration of biomass is located in the gra- dients at the equatorward edge of the feature. The level of ATP-biomass in this cell is approximately three times higher than that in either of the other two cells. The highest concentration of chlorophyll a biomass was found in the same geographical location (Fig. 12). The highest observed concentrations of both nitrate and phosphate were also located in this region (Fig. 8, 9) , as was the highest level of GTP (Fig. 13). Although a relatively high level of protein synthesis was occurring in this area, the highest value of the productivity indicator, GTP/ATP , did not simultaneously occur at this location (Fig. 14). The large standing crop observed adjacent to the strong chemical/thermal gradients of the upwelling feature sup- ports the "natural chemostat" hypothesis which maintains 55 Figure 8. Surface Nitrate Distribution for 29 October 1980 Cruise data. Contour intervals of 1 yuum. 56 Figure 9. Surface Phosphate Distribution for 29 October 1980 Cruise Data. Contour intervals of .1 /*. M 57 Figure 10. Sea Surface Temperature Distribution for 29 October 1980 Cruise Data. Contour intervals of .5 C. 5S Picpare 11 Surface Distribution of AT? for 29 October 1560 Cruise Data. Contour intervals of 300 ng 1"^. 59 Figure 12. Surface Distribution of Chlorophyll a for 29 October 1980 ^Cruise Data. Contour inte: vals of 4 mg m"-^. 60 Figure 13. Surface Distribution of GTP for 29 October 1980 Cruise Data, Contour intervals of 100 oicomoles I""'-. 61 Figure 14. Surface Distribution of GTP/.-.TP for 29 October 1980 Cruise x^ata. Contour intervals of .04. 62 :**•-"' ^ U :i5"SNC£. -if* , .1 J\' i_-P5EL, 3:?TqNCE. •*" 3 J- *- 5 t- =nce:. -"1 Figure 15. Mitrate, Phosphate, and temperature versus elapsed distance along the 29 October I960 cruise track. 63 ccT . • ATP a • H CHLP ■ •— ^P5EG CIsr-^NCE. ^^ >» **'^,' ..-•. /A- -". ELOPSED 3!STfiNC£. ■<" Figure 16. Chlorophyll a, .-.TP, and temperature versus elapsed distance along the 29 October 1980 cruise track. 64 ELHP5ED aiSTPNCE. KM -f5P5ED DISTANCE. r^M Q. ' ^ *« ELRPSED QISTONCE. KM Figure 17. ATP, Nitrate, and Phosphate versus elapsed distance along the 29 October 1980 cruise track . 65 2 ELPPSED □IS''=lNC£. .^^i ELflPSEO DlSrfiNCE. ^M ELRPSEO QISTHNCE. KM Figure 13. GTr/ATP , Nitrate, and .-hosphate versus elapsed distance along rhe 29 October 1980 cruise track. 66 CTP/RTP^ ^•^/-^ I \< \ ELPPSED OISTRNCE. KM ELAPSED OISTflNCE. KM Figure 19 GTP/ATP and Temperature versus elapsed distance along the 29 October 1980 cruise track. 67 cLflPSEO OlSTPNCE. KM Elapsed oistsnce. -(m Figure 20. GTP/.hTP and rvTP versus elapsed distance along the 29 October 196C cruise track. 68 that conditions for optimal growth are produced in these regions / Ref . 84_7. Correlation coefficients for various parameters were computed and are summaried in Table IV. Since the correla- tions between variables depend not only on the initial con- ditions of the source water, which evolves over time as a result of both conservative and nonconservative processes such as advection, diffusion, mixing, heat and salt transfer, biological uptake, and release of nutrients /^Ref . 85_7, the instantaneous relationships are complex and the instan- taneous chemical and biological status of a feature is not easily deduced from point observations. Correlations are useful, however, for determining general relationships and trends among variables. The correlations indicated under the column "all data points" contain correlations between all the points of the indicated variables. Values of variables which were too low to be detected were not included in the correlation calculations. An attempt was made to obtain more insight by subdividing the data into two groups based on nitrate concentrations. Figures 21 and 22 are point plots which show the dissolved nitrate concentrations plotted against those of dissolved phosphate concentration and tem- perature, respectively. These figures depict the separa- tion between the two distinct water masses present. Figure 23 is a point plot of dissolved phosphate versus temperature This figure does not similarly show a distinct separation. 69 TABLE IV Temperature , and Biomass Indicators All Data New Water Old Water i Points 1 ATP:CHLA .67 .94 .62 I 1 ATP : GTP .88 .91 .91 1 1 CKLA:GTP .79 .85 .78 1 1 GTP/ATP:ATP -.05 .65 -.16 1 1 GTP /ATP : GTP .32 .86 .18 1 I GTP/ATP:CHLA .24 .65 .17 1 1 ATP:TE>iP -.54 -.93 -.47 I 1 CHLA : TEMP -.82 -.94 -.85 1 1 GTP : TEMP -.52 -.86 - . 54 [ ] GTP/ ATP: TEMP .02 -.66 .05 1 ] N03:TSi\P -.58 -.21 -.78 j 1 PG4 : TEMP -.77 -.70 -.36 j 1 N03:ATP -.15 .77 .25 1 1 N03:GTP -.31 .69 .12 1 1 NC3:CHLA -.54 .40 .06 j 1 NC3:GTP/ATP -.59 .41 -.43 1 1 NC3:?C4 .43 .55 .82 [ 1 ?04:ATP .26 .65 .34 j 1 PG4;GT? .28 .58 .47 1 1 P04:CHLA .58 .62 .80 j 1 P04:GTP/ATP -.18 .36 .02 j 70 30 20, LU V— (X cc 10, 0. s <§> I I I I I I I J I I I I '■'''''■''■' 0.0 0.5 1.0 1.5 2.0 PHOSPHflTE. /xM 2.5 3.0 Figure 21. Nitrate versus JFhosphate for 29 October 1980 cruise data. 71 30, 2Q. CC 10. o 0. P 38 A 10. 12. 14. IS, TEMPERATURE. DEG.C. 18. Figure 22. Nitrate versus Temperature for 29 Cctcber 1980 cruise data. 72 J. r i — cc X o 0. 6 CSD O 10. 12. 14. TEMPERATURE, DEG.C. 16. 18, Figure 23. Phosphate versus Temperature for 29 October 1930 criiise data. 73 Based on this separation an arbitrary value of 2.0 ^w was used as the dividing mark to separate the values. Water which contained dissolved nitrate in excess of 2.0 yu- M was designated as "New .vater." These higher nutrient values were observed along the transect from station 9 to 8 (Fig. 2) during the first 55.0 km of elapsed distance. Values during this part of the cruise track ranged from 5 to 11 fijuh. The second group, designated as "Old Water," consisted of data points whose dissolved nitrate content was less than 2.0 i*-M. Essentially these values occurred over the rest of the cruise track (Fig. 2) beginning halfway between stations 8 and 6, to station 7, until station 8 was approached where the "New Water" was encountered again, From 55.0 to 81.3: 122.4 to 128.6; 156.6 to 160.4 km elapsed distance the autoanalyzer was inoperative so nitrate and phosphate measurements could not be made and the water at these locations could not be classified as to type. Values for these and other variables which were correlated under the subcategories of "New Water" and "Old Water" were not included from these portions of the cruise track. In some places where the correlations do not show internal consistancy, more points were correlated under the "all data" point category than under the "New" and "Old Water" mass combined categories. Data which could 74 not be placed in either subcategory were not included in the "old" and "new" categories even though they were cor- related when all data values were considered. B. RESULTS OF THE CARBON-14 UPTAKE EXPERIMENT 14 Table V contains the results from the C uptake- productivity experiment. Results listed are the average values determined from duplicate subsamples which were analyzed in tandem. The GTP and ATP concentrations used to determine the productivity ratio were analyzed from subsamples of the light bottles. 1 TABLE V G-14 Uptake ; Results •Sample CHLA (mg m ) 14 C Uptake Assimilation / ^ -3 , -Iv Number ( mgC m h ) , (mgC mgChla" h"" GTP (Pm) GTP /ATP ] 1 #1 21.53 31.9 1.48 134.5 .178 1 1 #2 7.75 22.9 2.95 152.5 .178 ] i ^3 43.06 25.9 .60 112.9 .110 1 1 #4 3.44 .58 .17 69.4 .103 1 1 #5 2.36 .59 .25 60.4 .110 Table 71 illustrates the results of correlation calcu- lations between the specific productivity indicators, assimilation numbers and GTP/ATP ratios, and absolute pro- 14 ductivity indicators, C uptake and GTP. Correlation 75 TABLE VI 1 Correlation of Product ivi ty Indicators i Assimilation GTP/ATP #: i^c Uptake : i GTP 1 1 All Data 1 Samples .89 .89 1 j Sample #1 1 Removed .99 .88 1 Sample #2 1 Removed .96 .99 I Sample ^3 Removed .89 .92 I Sample #4 Removed .86 .85 1 Sample =fr5 Removed .87 .82 1 statistics were first determined for all samples and then successively with the values of each sample removed in turn from the data set to determine if deletion of one sample from the data set would significantly alter the correlation statistics. C. RESULTS OF THE TIGRIGPUS GALIFORNICUS PRODUCTIVITY EXPERIMENT Table VII contains the results of the productivity deter- minations for different population groupings of Tigriopus calif ornicus. Specific productivity, GTP/ATP, is com- pared to the RNA/DNA ratio determined by Baugh for similar population subgroups of Tigriopus calif ornicus. 76 I TABLE VII Relative Rates of Productivity for Tigriopus calif ornicus GTP/ATP RNA/DNA (from Baugh /_ Ref. 85_7) Gravid .125 4.62 All-But-Gravid .099 2.16 Mixed .189 3.15 77 IV. DISCUSSION A. OCTOBER 1980 CRUISE A quasi synoptic approach based on point sampling was used to study the spatial heterogeneity and productivity of biomass. An assumption made when obtaining point samples is that they are representative quantitatively of population and community parameters in the body of water sampled. One study which has examined this problem found that precision was not increased merely by increasing the sample volume / Ref . 87_/. Generally the precision is only increased when the patch size increases so that the probability of sampling the "right" number of patches and of capturing the "correct" number and kinds of individual organisms representative of the general area is increased. This is one of the problems associated with discrete sampling methods for .^T? and GTP. Another problem is encountered when point data collected over an extended time period is used to characterize the biology and productivity of an entire area, since these variables are continuously in flux changing spatially and temporally even as the measurements are made. t\ study, which looked at the variability in the spatial and temporal heterogeneity of phytoplankton biomass in relation to the horizontal spatial structure of physical and chemical 78 variables in surface waters / Ref. 88_/ , found that the time component contributed to 63 percent of the total variance in the data , while the contribution of the space component was only 3.5 percent. The time component included the daily changes in available insolation and water column variables. Another study /~Ref . 89_7 similarly found that rapid temporal variations could occur over a time period of a few days which produced variations in chlorophyll concentration ranging 21 to 45 percent and variations in phosphate concentration ranging from 32 to 64 percent. Similar fluctuations could be expected for ATP and GTP determinations . Although at any moment there exists specific relation- ships between the different variables observed in the environment, data from the October cruise suggest that these interactions are continuously and rapidly changing and that the spatial distribution of phy topi ank ton is successively controlled by different factors or by a com- plex state of equilibrium between these variables. One temporal variation which is difficult to take into account is the diel depth variation which occurs. A possi- ble way to circumvent this difficulty in future experiments would be to pump water from the chlorophyll maximum instead of from the depth of the ship's intake using a depth variable fluorometer. 79 Figure 15 graphically illustrates the relationship between some of the physical and chemical variables of this system. The most significant correlations for these variables occur in the "Old Water" where nitrate and phosphate are highly positively correlated and nitrate to temperature and phosphate to temperature are highly negatively correlated. The high correlation between nitrate and phosphate is due to the uniformly low values of both which are observed in the "Old Water." The high inverse correlation between the temperature and the nutrients indicates, in view of the relatively high temperature values and low nutrient concentrations, that this water has been substantially altered from its initial condition through dynamical and biological processes. The correlations for "all data" points and for the "New Water" indicate sub- stantially the same result, although in these categories it is apparent through the lower correlation coefficients. The highest values of nitrate and phosphate, 11.35 jj^M and 1.29 u.M respectively, were observed in the gradients at the southern edge of the upwelling feature where the nitrate to phosphate ratio was 8.8 to 1. Since phyto- plankton uptake nitrate and phosphate in a ratio of 16 to 1 /~Ref. 90_7» the relationship between the nutrients suggests that the water masses observed are or will be nitrate limited. Since the steady state biomass is regulated by the concentration of the limiting nutrient 80 in the aquatic environment, it is important to identify this critical element to understand the dynamic relation- ship between chemical mesoscale and biological patchiness / Ref . 91_7. Studies have provided evidence that the nutrient and light requirements of phytoplankton follow a pattern similar to the Michaelis-Menten hyperbolic rela- tionship for enzyme kinetics / Ref. 92, 93, 94, 95, 96, 97_/ Results of Maclsaac and Dugdale l_ Ref. 9S_7 have shown that nitrate uptake by phytoplankton will generally increase with nitrate concentration only up to nitrate levels of about four julM at which time any excess in the environment is superfluous. It is apparent that phytoplankton in the "New Water" have sufficient nitrate available to provide optimal nutrient conditions necessary for maximal growth. This state contrasts to that in the "Old Water" where the availability of nitrate falls below detectable limits along large stretches of the cruise track. When the data were divided into two groups based upon their dissolved nitrate concentration, some interesting results emerged. Table III contains the range of values for the biomass and productivity parameters over the area surveyed (Fig. 2). There was a great deal of variability in the data due to the juxtaposition of adjacent water types. Table VIII illustrates the average GTP , ATP, and GTF/ATP levels taken from studies of various water masses at varying depths, According to Karl /"Ref. 100_7 typical surface ATP concentrations range from greater than 500 ng 1 ' for 81 TABLE VIII GTP/ATP Ratios in Environmental Samples (adapted from Karl / Ref . 99_/ ) Sample Description Goncn GTPC ATP GTP/ATP Southern California 7O N, 117^30.1' total - ~f r^ c" incirv ^''m^itvi/mniOO?^ c. ••••••« «•»••«••••••••• «•••«•••••••••• ••«•••••••••• 3!'J -r -f irM/>^.r\ j^imfMritoj^u^-rMr«jMr>j>kr\mn j^irvir>.A^i/>i/Mr>uM/Mr'r>vntrif\ .■N-.n.n.n.'Niniri.'Mninj^.r^ LULJ -<-rf.M-4->,^ .^-<^_<^^M^^_4_4^-<^-< _«.rf^-<_.^.«-<-<^-a^_^.d _4^«_M^.M^^.^_<..iM.iM "* .*0'*(M■«>•'^i^J■>^^0'OOcr. 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Stavn, R.H. , "The Horizontal-Vertical Distribution Hypothesis; Langmuir Circulation and Daphnia Dis- tributions," Limnology and Oceanography, Vol. 16, p. 453-466, 1971. 2. Huntsman, 3. A. and Barber, R.T., "Primary Production off Northwest Africa:, the Relationship to wind and Nutrient Conditions," Deep-Sea Research, Vol. 24, p. 25-33, 1977. 3. Lekan, J.F. and Wilson, R.E. , "Spatial Variability of Phytoplankton Biomass in the Surface waters of Long Island," Estuarine and Coastal i\arine Science, /ol. 6, p. 239-251, 1978. 4. Therriault, J.C. and Flatt, T. , "Spatial Heterogeneity of Phytoplankton Biomass and Related Factors in the Near-Surface Waters of an Exposed Coastal Eiribayment" Limnology and Oceanography, Vol. 23, No. 5, p. 888-C599 , 1978. 5. Denman, K.L., "Convariability of Chlorophyll and Tem- perature in the Sea," Deep-Sea Research, Vol. 23, p. 539-550, 1976. 6. Turpin , D.H. and Harrison, P.J., "Limiting Nutrient Patchiness and its Role in Phytoplankton Ecology," Journal of Experimental harine Biological Ecology , Vol . 39, p. 151-166, 1979. 7. Reid, J.L., Roden , G.I., and Wyllie, J.G. , Studies of the California Current System, Contribution from the Scripps Institution of Oceanography, Progress report- California Cooperative Oceanic Fisheries Investigation, July 1, 19 56- January 1, 19 58. 8. Traganza , Z.D. , Conrad, J.W., and Breaker, L.C. , "Satellite Observations of a 'Cyclonic Upwelling System' and 'Giant Plume' in the California Current," In: Coastal Uuvelling, American Geophysical Union, Washington, D.C. , 1981. 9. Bronsink, 3.H. , ^.icroplanktcnic .-^TP-Biomass and GTP- Productivity Associated with Upwelling off Point Sur , California . >iaster ' s Thesis, Naval Postgraduate School, tMonterey, California, 1980. 10. Traganza, Z.D. , Conrad, J.W. , and Breaker, L.C, "Satellite Observations of a 'Cyclonic Upwelling 108 System' and 'Giant Plume' in the California Current," In: Coastal Upwelling, American Geophysical Union, Washington, D.C. , 1981. 11. Eppley, R.W. and others, "A Study of Plankton Dynamics and Nutrient Cycling in the Central Gyre of the North Pacific Ocean," Limnology and Oceanography , Vol. 18, No. 4, 1973. 12. Goldman, J.C. , McCarthy, J. J. , and Peavey, D.G. , "Growth Rate Influence on the Chemical Composition of Phytoplankton in Oceanic Waters," Nature, Vol. 279, 1979. 13. Tranter, D.J. , Parker, R.R. , and Cresswell , G.R. , "Are Warm-Core Eddies Unproductive?," Nature , Vol. 284, No. 5756, p. 540-542, 1980. 14. Beardall , J. and others, "Phitoplankton Distributions in the Western Irish Sea and Liverpool Bay and their Relation to Hydrological Factors: a progress report," Biologia Contemporanca , Vol. 5, No. 4, p. 163-175, 1978. 15. Savidge , G. and Foster, P., "Phytoplankton Biology of a Thermal Front in the Celtic Sea," Nature, Vol. 271, p. 155-157, 1978. 16. Floodgate, G.D. and others, "Microbiological and Zoo- plankton Activity at a Front in Liverpool Bay , " Nature , Vol. 290, p. 133-136, 1981. 17. Pingree, R.D. and others, "Summer Phytoplankton Blooms and Red Tides Along Tidal Fronts in the Approaches to the English Channel," Nature, Vol. 258, p. 672-677, 1975 18. Szekielda, K.H., and Suszkowski , D.J., and Tabor, P.S. , "Skylab Investigation of the Upwelling off the North- west Coast of Africa," Journal Cons. Int. Explor. Mer. , Vol. 37, No. 3, p. 205-213, 1977. 19. Traganza , E.D. and Austin, D.M. , "Nutrient Mapping and the Biological Structure of Upwelling Systems From Infrared and Ocean Color Imagery," Science (in process), 1981. 20. Bronsink, S.H. , Microplanktonic ATP-Biomass and GTP- Productivity Associated with Upwelling off Pt. Sur , California , Master's Thesis, Naval Postgraduate School, Monterey, California, 1980. 21. Wroblewski , J.S. and O'Brien J. J. , "A Spatial Model of Phytoplankton Patchiness," Marine Biology, Vol. 35, o. 161-175, 1976. 109 22. Pace, N.R. , 'Structure and Synthesis of the .^.ibosomal Ribonucleic .-.cid of Prokaryotes , " Bacterio logical Reviews , p. 562-6C3, December 1973. 23. Karl, D.i'i. , "Occurrence and Ecological Significance of GTP in the Ccean and in I-.icrobial Jells, ' Applied and Environmental t-.icrobioloav , p. 349-355, 1978. 24. .'.arl , Z>..:, , 'Adenosine Triphosphate and Juanosine Triphosphate I^eterminations in ^.ntertidal Sediments," In :-.ethodology for Siomass Determinations and >.icrobial Activities in Sediments, .-.STK STP 573, CD. ^.ittlefield and ?.L. Seyfried, Dds., .-unerican Society for Testing and Materials, p. 5-20, 1979. 25. Lucas-Lenard , J. and Lipmann, ?. , 'Protein Sicsynthesis , " in Z.D. Snell (ed)., .Annual Review of Biochemistry, Annual Reviews Inc., Palo Alto, California, /ol. 40, 409-448, 19 71. 26. Eccleston, J.F, , "Single Turnover Kinetic Studies of Guancsine Triphosphate Hydrolysis and Peptide Formation in the Dlongation Factor Tu-dependent Binding of AuTiincacyl-tR>7A to Bschericnia Coli Ribc somes," The Journal of Biolocical Chemistry, Vol. 255, -\o . 23, p. 11CS6-11090, 193C. 27. Chinali , G. and Parmeggiani , .-. , "The Coupling with Polypeptide Synthesis of the GTPase activity Dependent on Elongation Factor G," The Journal of Biolocical Chemistry , Vol. 255, No. 15, p. 7455-7459, 1980. 28. Watson, J.D. , ;-.olecular Biolocp:/ of zhe Gene, W.A. Benjamin, Inc., p. 331-338, 1977, 29. Vclkin, D. , and others, "Suppression of the Biosynthesis of Guanosine Triphosphate by Protein Synthesis Inhibitors," The Journal of Biological Chem.istry, Vol. 255, No. 19, p. 9105-1909, 1980. 30. Ryther , J.H. , "The Measurement of Primary Production," limnology and Cceancgraohy , vol. 1, p. 72-34, 1956. : d 31. Rytner , J.H., and Menzel , u..s., "Comparison or tne C Technique wirh Direct Measurement of Phctosynthemc Carbon Fixation," Limnology and Cceanograohy , /ol . 10, p. 490-492, 1965, 32. Zppley, R.'.v., "An Incubation Method for estimating "che Darbcn Content of Phytoplankron in Natural Samples, ' limno-ogy and Cceanograohy, "/cl. 13, p. 574-58 2, 1968. 33. Sm.ith, R.J. , "Increasing Guanosine 3 ' -Diphosphate Concentrarion with Decreasing Growth Raiie m Anacys~is 110 nidulans ■ " Journal of General Microbiology, Vol, 113, p. 403-405, 1979. 34. Sokawa , Y. , Sokawa , J., and Kaziro, Y. , "Regulation of Stable RNA Synthesis and ppGpp Levels in Growing Cells of Escherichia coli , " Cell, Vol. 5, p. 69-74, 1975. 35. Franzen, J.S. and Binkley, S.B. , "Comparison of the Acid-soluble Nucleotides in Escherichia coli at Different Growth Rates , " The Journal of Biological Chemistry, Vol. 236, No. 2, p. 515-519, 1961. 36. Smith, R.C. and Maaloe, O. , "Effect of Growth Rate on the Acid-Soluble Nucleotide Composition of Salmonella Typhimurium , " Biochimica et Biophysica Acta, Vol. 86, p. 229-234, 1964. 37. Karl, D.M., "Distribution, Abundance, and Metabolic States of Microorganisms in the Water Column and Sediments of the Black Sea," Limnology and Oceanography, Vol. 23, No. 5, p. 936-949, 1978. 38. Karl, D.M., "Occurrence and Ecological Significance of GTP in the Ocean and in Microbial Cells," Applied and Environmental Microbiology, p. 349-355, 1978. 39. Lorenzen, C. J. , "A Method for the Continuous Measure- ment of In Vivo Chlorophyll Concentration," Deep Sea Research, Vol. 13, p. 223-227, 1966. 40. Slovacek , R.E. and Hannan , P.J. , "In Vivo Fluorescence Determinations of Phytoplankton Clorophyll a" Limnology and Oceanography, Vol. 22 (5), p. 919-925, 1977. 41. Strickland, J.D. and Parsons, T.R. , A Practical Handbook of Seawater Analysis, J.C. Stevenson (ed). Queen's Printer and Comptroller of Stationary, p. 49-62 and 71-76, 1968. 42. Technicon Industrial Systems Industrial Method No. 177-72-WM, Ortho Phosphate in Water and Seawater, January, 1978. 43. Technicon Industrial Systems Technical Publication No. TA 1-0170-20 , Operation Manual for the Technicon auto Analyzer II System, 1972. 44. Technicon Industrial Systems Industrial >iethod No. 100-70-W/3, Nitrate and Nitrite in Water and Waste Water, January 1978. Ill 45. Technicon .-^utoanalyzer Industrial i-iethod No. 175-72- wT-i, Nitrate and Nitrite in >7ater and Seavater, January 197 3. 46. Paulson, G.?., A Study of Nutrient Variations in the Surface and t'lixed i^ayer of i-.onterey Bay using .--automatic P:.nalvsis Techniques, i-iaster ' s Thesis, Naval Post- graduate School, Monterey, California, 1972. 47. Holm-Hansen, O. and Karl, D.M. , "Biomass and .-adenylate Energy Charge Determination in inicrobial Cell Extracts and Environmental Samples," I'lethods in Enzymology , Vol. 57, p. 73-85. 1978. 48. Ibid 49. DeLuca , M. , "Enzyme Reaction and Historical Background," In G.A. Borun (ed.), ATP tnethodology Seminar , Sr^I Technology Co., San Diego, California, p. 1-21, 1975. 50. Ibid, p. 1-3. 51. Ibid, p. 2. 52. Holm-Hansen, C. and Karl, D.M. , "Biomass and .-adenylate Energy Charge Determination in Microbial Geall Extracts and Environmental Samples," I\ethods in Enzymology, 7ol. 57, p. 73-85, 1978. 53. Karl, D.M. , "Determination of GTP , GDP, and GMP in Cell and Tissue Extracts^' In M.A. DeLuca ied.), Methods in Enzymology , Volume LVII, Academic Press, New York, p. 85-94, 1978. 54. Karl, D.M. , "A Rapid and Sensitive Method fer the Measurement of Guanine Ribonucleotides in Bacterial and Environmental Extracts," Analytical Biochemistry, Vol. 89, p. 581-595, 1978. 55. Ibid, p. 587-588. 56. Karl, D.M. and Holm-Hansen, O., "Effects of Luciferin Concentration on Quantitative .-^ssay of ATP Using Crude Lucif erase Preparations," Analytical Biochemistry, Vol. 75, p. 110-112', 1976. 57. Luminescent Review Bulletin 202, ATF Measurements: Theory and Practice- " p. 11, February 1981. 53. Holm-Hansen, O. and Booth, C.R. , "The Measurement of ATP in the Ocean and Its Ecological Significance," Limnology and oceanography, Vol. 11, p. 510-519, 1966. 112 59. Holm-Hansen, O. and Karl, D.3, , "3iomass and Adenylate Energy Charge Determination in Microbial Cell Extracts and Environmental Samples," Methods- in Snzymology, Vol. 57, p. 73-85, 1978. 60. Karl, D.:-.. and Holm-Hansen, O. , "Effects of Luciferin Concentration on Quantitative ^ssay of r.TF Using Crude Lucif erase Preparations," analytical Biochemistry, Vol. 75, p. 110-112', 1976. 61. Booth, C. , '-'Instrument Development and Data Processing for the a.T.?. Photometer," in G.A. Borun (ed.), ATP Methodology Seminar 3AI Technology Co., San Diego, California, p. 104-130, 1975. 62. Karl, D.H. , "A Rapid and Sensitive Method for the Measurement of Guanine Ribonucleotides in Bacterial and Environmental Extracts," analytical Biochemistry, Vol. 89, p. 581-595, 1978. 63. Ibid. p. 590. 64. Ibid, p. 590. 65. Booth, C. , "Instrument Development and Data Processing for the A.T.P. Photometer," In G.,\. Borun (ed.), .^TP Methodology Seminar , SaI Technology Co., San Diego, California, p. 104-130, 1975. 66. Rowney , J./., Gradient Analysis of Fhytoplankton Productivitv and Chemical ir^arameters in Polluted and Other Nearshore Habitats, I'la^ster ' s Thesis, Naval Postgraduate School, Monterey, California, 1973. .14 67. Jitts, H.R. and Scott, B.D. , "The Determination of Zero-Thickness Activity in Geiger Counting of C Solutions Used in Marine Productivity Studies," Limnolooy and Oceanoqraohy , Vol. 6(2), p. 116-123, 1961. 68. Strickland, J.D. and Parsons, T.R. , A Practical Handbook of Seavater Analysis, J.C. Stevenson t,ed) , Queen's Printer and Comptroller of Stationary, 1968. 69. Eglof f , D.A., Ecological Aspects of Sex Ratios and Reproduction in Experimental and Field Populations of the Marine Cooeood Tigriopus calif ornicus . Ph.D. Dissertation, Stanford University, California, 1966. 70. Baugh, D.E. , RNA/'DNA Ratios in the Estimation cf Growth Stages of Oceanic Zcoplankton Population s_, ^--as^cer's Thesis, Naval .-ostgraduate School, :-:onterey, California, 1974. 113 71. Brachet , J., The Biological Role of Ribonucleic i^cids , Elsevier, .slew York, p. 1-49, 1960. 72. Sutcliffe, //.H. , Jr., "The Relationship Between Growth Rate and Ribonucleic Acid Concentration in Bome Inver- tebrates," Journal of Fisheries Research Board of Canada, Vol. 27, p. 606-609, 1969. 73. Pease, A.K. , The Use of Estimations of Ribonucleic .-\cid to Predict the Growth Plates of Zooplanktonic 'organisms, I-aster's Thesis, University of British Columbia, 196c. 74. Leick, V., "Ratios Between Content of DrJA, RiTA, and Protein in Different I-iicro-organisms as a Function of iMaximal Growth Rate," Nature , Vol. 217, o. 1153-1155, 1968. 75. Baugh, D.S. , RNA/PNA Ratios in the Estimation of Growth Stages of Oceanic Zooplankton Populations, i-;aster's Thesis, Naval Postgraduate School, I'lonterey, California, 1974. 76. Skjoldal, H.R. and U. Bamstedt , "Ecochemical Studies on the Deepwater Pelagic Community of Korsf jorden , Western Norway, .-adenine Nucleotides in Zooplankton." Karine Biology, Vol. 42, p. 197-211, 1977. 77. nolm-Hansen, O. , "Determination of r-iicrobial Biomass in Ccean Profiles," Limnology and Oceanography Vol. 14, 740-747, 1969. 78. Ibid, p. 744. 79. Holm-Hansea C. , "Carbon/ATP Ratios in Microbial Cultures and in Natural Populations," In G.A. Borun (ed.), ATP Methodology Seminar , SAI Technology Co., San Diego, California, p. 446-473, 1975. 80. Sorokin , Y.I. and Mikheev, V.N. , "On Characteristics of the Peruvian Upwelling Ecosystem," Hydrobiologia , Vol. 62, No. 2, p. 165-189, 1979. 81. Holm-Hansen, O. , "ATP Levels in Algal Cells as Influenced by Environmental Conditions," Plant Cell Physiology, Vol. 11, p. 669-700, 1970. 32. Karl, D.M. , "Occurrence and Ecological Significance of GTP in the Ocean and in Microbial Cells," Applied and Environmental Microbiology, p. 349-355, 1978. 114 \ 83. Hanson, W.E. , Nutrient Study of Mesoscale Thermal Features off Point 3ur , California, Master's Thesis , Naval Postgraduate School, Monterey, California, 1980. 84. Traganza , E.D. , Conrad, J.W. , and Breaker, L.C., "Satellite Observations of a 'Cyclonic Upwelling System' and 'Giant Plume' in the California Current," In: Coastal Uovelling, American Geophysical Union, Washington, D.C. , 1981. 85. Traganza, E.D. , Nestor, D.A. and incDonald, A.K., "satellite Observations of a Nutrient Upwelling Off the Coast of California," Journal of Geophysical Research, Vol. 85, No. C7, p. 4101-4106, July 1980. 86. Baugh, D.E. , RNA/DNA Ratios in the Estimation of Growth Stages of Oceanic Zooplankton Populations, Piaster's Thesis, Naval Postgraduate School, Monterey, California, 1974. 87. Wiebe, P.H., "A Computer Model Study of Zooplankton Patchiness and its Effect on Sampling Error , " Limnology and Oceanography, Vol. 16, No. 1, p. 29-38, 1971. 88. Therriault, J.C. and Piatt, T. , "Spatial Heterogeneity of Phytoplankton Biomass and Related Factors in the Near-Surface Waters of an Exposed Coastal Embayment , " Limnology and Oceanography, Vol. 23, No. 5, p. 888-899, 19 73. 89. Piatt, T. , Dickie, L.M. and Trites, R.W. , "Spatial Heterogeneity of Phytoplankton in a Near-Shore Environment," Journal of Fisheries Research Board of Canada, Vol. 27, p. 1453-1473, 1970. 90. Redfield, I\.C. , "The Biological Control of Chemical Factors in the Environment," American Science, Vol. 46( 3) , p. 205-221, 1958. 91. Goldman, J.G., McCarthy, J.J., and Peavey , D.G. , "Growth Rate Influence on the Chemical Composition of Phytoplankton in Oceanic Waters," Nature , Vol. 279, 1979. 92. :-:acIsaac, J.J. and Dugdale, R.C. , "The kinetics of I^itrate and Ammonia Uptake by Natural Populations of Marine Phytoplankton," Deep Sea Research, Vol. 16, p. 45-57, 1969. 93. Qasim, S.Z., Bhattathiri , P.:-i., and Devassy, V.?., "Growth Kinetics and Nutirent Requirements of Two Trooical Marine Phytoplankters , " .marine Biology , Vol. 21, 'o. 299-304, 1973. lu.3 94. Gaperon, J., "Population Growth in i\icro-Crganisms Limited by Food Supply," Ecology, ^/ol. 48, o. 715-722, 1967. 95. Dugdale, R.G. , "Nutrient Limitation in the Sea; Dynamics, Identification, and Significance," Limnology and Oceanograohv , Vol. 12, p. 685-69 5, 1967. 96. Thomas, W.H. , "Effect of Ammonium and ^^itrate Concentration on Chlorophyll Increases in Matural Tropical Pacific Phytoplankton Populations," Lxmnolcgy and Cceanography , Vol. 15, p. 386-394, 1970. 97. r-iaclsaac, J.J. and Dugdale, R.C, , "Interactions of Light and Inorganic iNiitrogen in Controlling i.\itrogen Uptake in the Sea," Deep- Sea Rp^search, Vol. 19, p. 209-232, 1972. 98. I'laclsaac, J.J. and Dugdale, RmC. , "The Kinetics of Nitrate and Ammonia Uptake by Natural Populations of Inarine Phytoplankton," Deep Sea Research, /ol. 16, o. 45-57, 1969. 99. Karl, D.M. , "Cellular Nucleotide Measurements and Applications in ^:icrobial Ecology," Microbiological Reviews, p. 731, December 1980. 100. Karl, D.M. , "Cellular Nucleotide Measurements and Applications in Microbial Ecology," Microbiological Reviews , p. 739-796, December 1930. 101. Sronsink, S.ii. , i^:icroplanktonic --^TP-5icmass and GTP- Productivity r-.ssociated with Upwelling off Pt. Sur. California , Master's Thesis, I'Javal Postgraduate School, Monterey, California, 1980. 102. Steele, J.H. and Baird, I.E., "Relations Between Primary Production, Chlorophyll and Particulate Carbon," Limnology and Oceanography, Vol. 6, 1960. 103. Eppley, R.W. , Harrison, W.G. , Chisholm, S.W. , and Stewart, E. , "Particulate Organic Matter in Surface /7aters of Southern California and its Relationship to Phytoolankton , " Journal of i'larine Resources, Vol. 35, p. 671-696, 1977. 104. Perry, :-i.J. , Talbot, M.C. , and .^Iberte, R.S., "Phctoadaption in Marine Phytoplankton: Response of the Photosynthetic Unit," :-iarine Biology (in oress 1931). 116 105. Slovacek, R.S. and Hannan , P.J., "In Vivo Fluorescence Determinations of Phytoplankton Glorophyll a," Limnology and Oceanography, Vol. 22(5), o. 919-925, 1977. 106. Traganza , S.D. , Conrad, J.W. , and Breaker, L.G., "Satellite Observations of a 'Cyclonic Upwelling System' and 'Giant Flume' in the California Current," In: Coastal Upwelling, American Geophysical Union, v/ashington, D.C. , 1981. 107. Goldman, J.C. , McCarthy, J.J. , and Peavey , D.G. , "Growth Rate Influence on the Chemical Composition of Phytoplankton in Oceanic Waters," £ature. Vol. 279, 1979. 103. Parsons, P. and Masayuki , T. , Biological Qceanographic Processes, Pergamon Press, Ltd., New York, 1973. 109. Denman, K., Ckubo , A., and Piatt, T. , "The Chlorophyll Fluctuation Spectrum in the Sea , " Limnology and Oceanography, Vol. 22, No. 5, p. 1033-1038, 1977. 110. Sorokin, Y.I. and Mikheev , V.N. , "un Characteristics of the Peruvian Upwelling Ecosystem," Hydrobiologia , Vol. 62, No. 2, p. 165-189, 1979. 111. Shushkina , S.A. and others, "Characteristics of Func- tioning of Planktonic Communities in the Peruvian Upwelling," Okeanologiy a , Vol. 18, No. 5, p. 886-90 2, 1978. 112. Sorokin, Y. , "Characteristics of Primary Production and Heterotrophic Microplankton in the Peruvian Upwelling," Okeanologlya , Vol. 18, No. 1, p. 9 7-110, 19 78^. 113. Karl, D.m. , "Adenosine Triphosphate and Guanosine Triphosphate Deteinni nations in Intertidal Sediments," In i-.ethodology for Biomass Determinations and .-.icrobial Activities in Sediments, ASTi'i STP 673, CD. ^ittlefield and P.L. Seyfried, rids., .-nmerican Society for Testing and i\aterials, p. 5-20, 1979. 114. Karl, D.M. , "Deep-sea i^rimary Production at the Galaoagos Hydrothermal Vents," Science , Vol. 207, o. 1345-1347,1980. 115. Karl, D.tM. "Occurrence and Ecological Significance cf GTP in the Ocean and in Microbial Cells," .applied and Environmental Microbiology, p. 349-355, 1978. 117 116. Ibid. 117. Fogg, G.E., Algal Cultures and ir-hy to plankton Scoloqy, second ed. , University of Wisconsin Press, 1975. 118. Anita, N.J. and others, "Further I^easurements of Primary Production in Coastal Sea v/ater Using a Large Volume Plastic Sphere," Limnology and Oceanography, Vol. 6, p. 237-258, 1963. 119. TiC'^llister , CD. and others, "Measurements of Primary Production in Coastal Sea Water Using a l^arge Volume Plastic Sphere," Limnology and Oceanography, Vol. 6, p. 237-258, 1961. 120. Maclsaac, J.J. and Dugdale, R.C. , "Interactions of Light and Inorganic Nitrogen in Controlling Nitrogen Uptake in the Sea," Deep-Sea Research, Vol. 19, p. 209-232, 1972. 121. Caperon, J. and i\eyer , J., "Nitrogen-Limited Growth of >.arine Phytoplankton. I. Changes in Population Char- acteristics with Steady-State Growth Rate, ' Leep-Sea Research, Vol. 19, p. 601-618, 1972. 122. Collos, Y. and Slawyk, G. , "Significance of Cellular Nitrate Content in Natural Populations of I-.arine Phytoplankton Growing in Shipboard Cultures," :-.arine Biology , Vol. 34, p. 27-32, 1976. 123. Perry, :..J., Talbot, i\.C. , and Alberte , P..S. , "Photoadapticn in I'larine Phytoplankton: Response of the Phctosynthetic Unit," i-;arine Biology (in cress 1981). 124. Fogg, G.E» , Algal Cultures and Phytoplankton Ecology, second ed. , University of Wisconsin Press, 1975. 125. Perry, M.J. Talbot, M.C. , and /^Iberte , R.S., "Photoadapticn in Marine Phytoplankton: Response of the Phctosynthetic Unit," Marine Biology (. m press 1981). 126. Maclsaac, J.J. and Dugdale, R.C, "Interactions of Light and Inorganic Nitrogen in Controlling Nitrogen Uotake in the Sea," Deep- Sea Research, Vol. 19, p. 209-232, 1972. 127. Dppley, R.W. , Renger , D.H. , and Cullen, J.J., ".-^mmcnium Distribution in Southern California Coastal ..'arers and Its Role in the Growth of Phytoplankton," LimnclC'gy and Oceanography, Vol. 24, No. 3, p. 495-50^, i^'Tr. 118 128. Ibid, p. 495. 129. Gaperon, J. and i.eyer, J., "Nitrogen-Limited Gro'//th of Marine Phytoplankton-I . Changes in Population Characteristics with Steady-State Growth .^ate , " Deep-sea Research, Vol. 19, p. 601-61b , 1972. 130. Goldman, J.C. , i^.cCarthy, J.J. , and Peavey , D.G. , "Growth Rate Influence on the Chemical Composition of Phytoplankton in Oceanic Waters", Nature , Vol. 279, 1979. 131. Ibid 132. Ibid 133. Karl, D.M. , "Occurrence and Ecological Significance of GTP in the Ocean and in Microbial Cells," applied and Environmental Microbiology, p. 349-355, 1978. 134. Bronsink, S.H. , Mi croplank tonic ATP-Biomass and GTF- Productivity Associated with Upwelling off Pt. Sur , California , Master's Thesis, Naval Postgraduate School, Monterey, California, 1980. 135. Karl, D.M. , "Occurrence and Ecological Significance of GTP in the Ocean and in Microbial Cells," Applied and Environmental Microbiology, p. 349-355, 1978. 136. Costantini, R.Z. and Sturani , S. , "Levels of the Ribonucleoside Triphosphates and Rate of PJSIA Synthesis in Neurospora Crassa," Biochimica et Biophysica r^cta. . Vol. 476, ^p. 272-278, 1977. 137. Holm-Hansen, O., "Carbon/ATP Ratios in Microbial Cultures and in Natural Populations," In G.A. Borun (ed.), ATP r-iethodology Seminar , SAI Technology Co., San Diego, California, p. 446-47 3, 1975. 138. Baugh, D.E. , RNA/DNA Ratios in the Estimation of Growth Stages of Oceanic Zooplanton Populations, Master ' s Thesis, Naval Postgraduate School, ;'-cnt9rey , California, 1974. 139. Goldman, J.C, McCarthy, J.J. , and Peavey, D.G. , 'Growth Rate Influence on the Chemical Composition of Phytoolankton in ^ceanic Waters, ' Nature , Vol. 279, 1979.^ 140. Traganza , S.D. , Conrad, J.W. , and Breaker, L.C., 'Satellite Observations of a 'Cyclonic Upwelling System' and 'Giant Plume' in the California Current, ' In: Coastal Upwelling, .-jnerican Geo- physical Union, ./asnmgton, D.C. , 1981. 119 141. Eppley, R.W. and others, "^\ Study of Plankton Dynamics and Nutrient Cycling in the Central Gyre of the ITorth Pacific ocean," Limnology and Oceanography, Vol. 18, No. 4, p. 534-551, 1973. 142. Goldman, J.C. , r^cCarthy, J.J. , and Peavey , D.G. , "Growth Rate Influence on the Chemical Composition of Phytoolankton in Oceanic Waters," Nature , Vol. 279, 1979.' 143. Hanson, W.E. , Nutrient Study of r-iesoscale Thermal Features off Point Sur , California, master 's Thesis, Naval Postgraduate School, Monterey, California, 1930. L 120 INITIAL DISTRIBUTION LIST o. Copies 1. 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Chairman, Oceanography Department U.S. Naval Academy Annapolis, :-iaryiand 2140 2 21. :'ir. 3en Gagie Office of Naval Research Branch Office 1030 East Green Street Pasadena, California 91106 22. Dr. Robert S. Stevenson Scientific Liaison Office, ONR Scripps Institution of Oceanography La Jolla, California 92037 23. i-is. Bonita Hunter, Code 58 Department of Oceanography Naval Postgraduate School Monterey, California 93940 24. LCDR Craig S. Nelson Pacific Environmental Group c/o FlvOC honterey , California 9 3940 25. Dr. Edward Green Office of Naval 102 Research (code 432) Ocean Sciences Si Technology Division Chemical Oceanography Program NSTL Station Bay St. Louis, ..ississippi 39529 123 r Thesis 19 w 68 3 J8255 Jori c.l Estimating the dis- tribution and produc- tion of microplankton in a coastal upwelling front from the cellu lar content of guano- sine-5' triphosphate and adenosine-5 ' tri- phosphate. Thesis J8255 c.l 1 Qrr ^-^ X \> V U v«/ \J Jori Estimating the dis- tribution and produc- tion of microplankton in a coastal upwelling front from the cellu- lar content of guano- sine-5' triphosphate and adenosine-5' tri- phosphate.