Markham 2 Effects of the loss of the Apical Meristem on Growth and Tissue Composition in Macrocystis pyrifera Abstract Changes to the physiology or chemistry of basal producers can have direct consequences for consumers and the transfer of energy between trophic levels. As a foundation species, giant kelp Macrocystis pyrifera sustains high growth rates and primary productivity under dense shading canopies through an efficient system of translocation that provides energy and nutrients to growing tissues. This study investigated what impact removing the growing point or apical meristem from M. pyrifera, as occurs in storms, commercial harvest and historically in grazing by the Stellar's sea cow, has on the physiology of the remaining immature sub-apical blades. Blades from fronds experienced increased growth and erosion rates within the first week after removal of the apical meristem as compared with controls. Äfter two weeks growth and erosion rates were no longer significantly different between manipulated blades and control but substantial differences in concentrations of the sugar mannitol as well as nitrogen and carbon content persisted. The study revealed physiological effects of apical meristem loss that merit further investigation as to their causal mechanisms and suggest the potential for meristem removal to alter interactions with primary consumers. Markham 3 Introduction In many different ecosystems changes to the physiology or chemistry of basal producers can have direct consequences for consumers and the transfer of energy between trophic levels (Moe et al. 2005). Changes in tissue chemistry for autotrophs particularly have the potential to alter palatability to herbivores through shifts in concentrations of secondary metabolites, nutritional content, and structural stability (Hemmi & Jormalainen, 2002; Poore & Steinberg, 1999). Understanding changes in basal producers that may take place as a result of environmental shifts or anthropogenic impacts can yield important insights into the flow of energy through and nutrient cycling between trophic levels (Graham 2004). The giant kelp Macrocystis pyrifera functions as a foundation species forming dense forests upon which diverse communities of organisms rely for food and shelter in temperate coastal waters around the globe(Arkema, Reed, & Stephen C Schroeter, 2009; Dayton, 1985; Graham, 2004; Mann, 2000; Steneck et al., 2003). Its astonishingly high rates of growth and primary productivity form the basis for numerous natural food webs and support expansive canopies that sustain commercial harvests for various human uses (Buschmann & Hernandez- Gonzalez, 2005; Doty, 1987; Gutierrez et al., 2006; Mann, 1973). Kelp growth is sustained by an efficient system of translocation that moves energy and nutrients from mature portions of the kelp to the growing points at the end of the fronds—the apical meristems (Lobban, 1978b; North, 1971). Detailed studies with "C tracers and fluorescein dye demonstrated that photosynthates move at rates of up to 78 cm/hr both up and down the fronds of Macrocystis (Parker, 1963; 1965). The translocated substances— the sugar mannitol, amino acids, and inorganic ions¬ follow a “source-sink" dynamic: fully developed blades export these materials while immature, Markham 4 growing blades import them to aid in growth (Schmitz & Lobban, 1976; Schmitz & Srivastava, 1979). Apical meristems serve as an important sink drawing metabolites up the fronds toward the rapidly growing blades at the apex (Lobban 1978a; Schmitz & Srivastava 1979). Early work suggested that fronds without apical meristems no longer translocate materials up the frond but instead send them down toward the base (Lobban, 1978a). However, little work has been conducted explicitly to understand how the loss of the apical meristem affects blades remaining on Macrocystis fronds. The apical meristems of Macrocystis are commonly lost to storm damage (Seymour, M J Tegner, Dayton, & Parnell, 1989) and many more are removed during canopy cutting for commercial harvest (Doty, 1987). Moreover, historically, canopy grazing by the immense Stellar’s sea cow (once abundant from California to Japan, now extinct) would have frequently removed Macrocystis apical meristems (Burger, 1995). Reduced translocation could potentially result in physiological changes to the remaining blades with rippling ecological effects with direct and indirect consequences for organisms that interact with Macrocystis blades such as microbial biofilms (Bengtsson, Sjøtun, Storesund, & Ovreås, 2011; Duggins, Simenstad, & J. Estes, 1989; Norderhaug, Fredriksen, & Nygaard, 2003), epiphytic sessile invertebrates (Dixon. S.C. Schroeter, & Kastendiek, 1981; Hepburn & Hurd, 2005), and invertebrate mesograzers (Duggins, Eckman, Siddon, & Klinger, 2001; Foster & Schiel, 1985; McMillan, 2010; Norderhaug et al., 2003; Watanabe, 1984) In this study we sought to address what effect the removal of the apical meristem has on the growth rate and chemical composition of the remaining sub-apical blades. We hypothesized that the loss of the apical meristem would halt translocation of mannitol to the remaining sub¬ Markham 5 apical blades, thereby slowing growth rates and reducing the C:N ratio as a consequence of the nitrogen built up in the tissue relative to faster growing portions. We further postulated that a reduced C:N ratio could make blades more palatable to microbes and grazers, thus resulting in greater rates of erosion. Methods The experiment was conducted in the shallow kelp forests of the Hopkins Marine Life Refuge in Monterey, CA (36° 36’ N, 121° 54' W). Pairs of kelp fronds of similar length were selected from twenty Macrocystis pyrifera sporophytes. The apical meristems were removed from one frond of each pair by severing the stipe 25cm below the first dividing pneumatocyst on the apical scimitar (Figl). The paired un-cut frond was used as a control. Growth and deterioration rates were determined by the commonly used hole-punch technique (Rothäusler et al., 2009) in paired blades 30 cm below the apical meristem (Fig.1) and compared in situ after seven days and in the laboratory after sixteen days. Tissue was collected from each blade from three holes approximately 17.5mm in diameter punched consecutively proceeding from the growth measuring hole toward the pneumatocyst. The algal tissue was dried in an oven at 120°C for 48hours and then ground with a ball mill (will check name) for thirty seconds per sample to a fine powder for use in chemical analyses. A colorimetric method (sensu White et al. 2010) was employed in order to compare concentrations of mannitol in fronds with and without apical meristems. Mannitol was extracted by heating 30mg of powdered kelp in 1.5 ml of water at 80 °C, centrifuging at 14000rpm for 30min, and removing the supernatant. 100 uL of this mannitol extract was added to 500 uL formate buffer (0.5 mol L", pH 2.85). After mixing, 300 uL of Reagent 1 (5 mmol L* sodium periodate) was added followed by 300 uL of Reagent 2 (consisting of 0.1 mol L * acetylacetone, Markham 6 2 mol L“ ammonium acetate and 20 mmol L-1 sodium thiosulfate) was added. Reagent 2 was always added 15 s after Reagent 1. The tube was then closed and heated in boiling water for 2 min, cooled under running tap water and measured with a spectrophotometer at 412 nm. Each sample was diluted with water at: 1:10, 1:15, 1:20, and 1:50 vol/vol. Mannitol concentrations were determined for each sample by cross-referencing with standards of varied concentrations from which a linear relationship between absorbance and mannitol concentrations were obtained. Due to the relatively low concentration of mannitol in M. pyrifera as compared with those reported for other kelps by White et al. (2010) the full strength solution was used to calculate percent mannitol. In order to quantify Carbon and Nitrogen content, dried kelp powder in duplicate samples was weighed on a Sartorius supermicro balance, and assayed for total N and C with a Carlo Erba Na-1500 elemental analyser calibrated with acetanilide. All data were analyzed using two-tailed, paired T-tests between blades from fronds with and without apical meristems from each sporophyte. Results Äfter one week of growth, blades from fronds where the apical meristem was removed grew significantly more quickly than intact controls (t=2.332, n-20, p=.0308) (Fig. 2). However, after between days 7 and 14 the difference in growth rates was no longer significant (t=1.1136, n=17, p=.281). The measure of erosion revealed that elongation occurred between the punched hole and the blade tip. Äfter one week, blades from fronds with the apical meristems removed had an average increase of distance from the punched hole to the blade tip of 3.Scm which was significantly less than the average 6.6 cm of controls (t-2.124, n-20, p=.0467) (fig. 3). After 14 days the difference was no longer significant (t=.7804, n-17, p=.4445). Markham 7 Analysis of Mannitol concentrations revealed that blades from fronds without apical meristems had a significantly lower percent mannitol by dry weight as compared with controls =4.2079, n=10, p=.0018) (Fig.4). However, the average percent carbon of blades from manipulated fronds was significantly higher than that of controls (t-3.9597, n=17, p=.0011)(Fig.5). No significant difference in percent Nitrogen was found between blades from manipulated fronds (t=1.1807, n=17, p=.2549). Accordingly, the C:N ratio was significantly higher for blades from manipulated fronds than those of controls (t-2.1648, n-17, p-.0448) (Fig.6). Discussion High growth rates (Fig. 1) and an increased C:N ratio (Fig. 6)in blades from fronds without apical meristems as compared with controls contradicted our initial hypotheses. While reduced mannitol (Fig. 4) and increased erosion in blades from manipulated fronds (Fig.3) were consistent with initial predictions, the causes of these changes are likely different than those that were previously anticipated. The increased growth rates of blades from fronds without apical meristems may still be consistent with results from Parker (1965) and Lobban (1978) where fronds without their apex no longer had any acropetal translocation (solute movement up the frond) but only basipetal translocation (toward the holdfast). Translocation in Macrocystis is driven by osmotic gradient whereby mature, "source" blades with a relative excess of mannitol and amino acids uptake water which then flows with the solutes to immature, "sink" blades which excrete water to encourage the flow (Schmitz 1982). If translocation upwards were halted with the loss of the apical meristems, it would follow that the higher growth rates in blades from manipulated fronds resulted from the fact that the manipulated blade would receive no imports but also would no Markham 8 longer have blades above them with to export mannitol and amino acids to. By contrast as control stipes continued to add blades, the focal blades became more and more distant from the apical meristem, and may have shifted from primarily importers to partial exporters. This bidirectional translocation, both importing and exporting, for blades between.75-1.1m from the apical meristem as the control blades rapidly grew to in distance would be consistent with previous work (Lobban 1978a). By exporting a portion of their photo-assimilates and amino acids to the faster-growing blades above them on the fronds, control blades may have experienced reduced growth rates. Under these conditions whatever mannitol the blades from manipulated fronds produced would be used in situ for growth or to a lesser extent be drawn weakly toward the base and the distant sink of new fronds. This hypothesis of halted acropetal translocation is consistent with the lower concentrations of mannitol found in blades from manipulated fronds as compared with controls (fig. 4). In controls the powerful "sink" represented by the apical meristem and immature blades above the blades of interest would draw a flow of mannitol acropetally from both the mature blades below and the mid-maturity blades we measured (Lobban 1978b). This flow could keep levels of mannitol high as was found in our mannitol assays. Alternatively it is possible that the findings of Parker (1965) and Lobban (1978a) that blades without apical meristems have no acropetal translocation were not applicable to this experiment because of spatial and temporal differences in the experimental setups. Parker removed not only the apical meristem but the preceding 30 blades as well in order to demonstrate that fronds without apical meristems had only basipetal translocation. In doing so, he likely remoyed all immature blades and therefore the osmotic gradient driving translocation. The blades measured in this experiment would have been amongst the immature blades Parker Markham 9 removed in his experiment. Likewise, Lobban simply conducted his experiments bathing blades of Macrocystis in the field with radio-labeled C-14, on fronds as he encountered them in the field with or without apical meristems. Considering the growth rates of Macrocystis blades, it seems quite likely that would not have conducted his experiments in the short period between fronds losing their apical meristem and the remaining subapical blades becoming mature. It remains a distinct possibility that the removal of the apical meristems may have caused accelerated translocation of mannitol and amino acids into the immature sub-apical blades until the osmotic gradient was dissipated. As the last blade on a severed stipe, the blades from manipulated fronds in our experiment would be free from competing "sinks" higher on the stipe, therefore having a considerably weaker draw to export mannitol than controls. These factors may explain the accelerated growth of blades from manipulated fronds in the first week. Parker’s work (1965) would have missed this pattern because he removed the immature portion of the frond entirely. Likewise Lobban, could have failed to observe such translocation by missing the narrow window in which accelerated acropetal translocation followed meristem loss can be measured, before it halts as the blades mature and the osmotic gradient dissipates. Future experimentation could easily resolve these conflicting hypotheses by testing concentrations of mannitol and amino acids during the period of rapid growth in the week following apical meristem removal. Regardless of the cause of the accelerated growth rates in blades from fronds without apical meristems, this change in growth rate was associated with further shifts in chemical composition (fig. 5 & 6). The higher percent carbon (fig. 5) coupled with lower concentrations of mannitol (fig. 4) in blades from fronds without apical meristems, suggest that another form of carbon different from mannitol is more prevalent as compared with controls. Whereas, mannitol Markham 10 is known to be the ephemeral photoassimilate used in translocation, laminarin is known to be a long-term storage carbohydrate (Lee, 1989). Lee notes that "Laminarin may be formed from mannitol and during active growth mannitol can be formed faster than its rate of conversion into laminarin so that both substances increase in amount. When growth slows down or stops...laminarin increases with loss of mannitol". As the blades from both treatment groups slowed in their growth over the second week the blades from manipulated fronds may have converted what mannitol they had to laminarin. This increased conversion may have been due to longer retention times of mannitol in blades from manipulated fronds without a flow of mannitol or due to the comparatively greater maturity of these blades compared with those of controls. In either case, the accretion of the more stable storage carbohydrate laminarin would explain the higher percent carbon in blades from manipulated fronds. Moreover, a halting of translocation would mean that the standing stock of mannitol would not be as actively replenished in blades from manipulated fronds consistent with the reduced concentrations of mannitol. A simple assay of laminarin concentration should be able to resolve whether the difference in percent carbon can be attributed to laminarin production (White, Coveny, Robertson, & Clements, 2010). Despite being confounded by elongation, the measurements of changes in distance between the punched hole and the blade tip seem to suggest higher rates of erosion in blades without apical meristems as compared with controls. It seems unlikely that the growth trends observed in the proximal portion of the blades would be reversed in the distal portions of the blade since Macrocystis blades lack structural subdivisions and most differences in the tissue occur in a gradient (Lee 1989). Therefore, the smaller change in average length between the punched hole and blade tip observed in manipulated blades as compared with controls suggests that greater erosion occurred in blades from fronds without apical meristems. Additionally, the Markham 11 higher growth rates of blades measured at the base from manipulated fronds as compared with controls (fig. 2) suggests that the differences observed in erosion may be fairly conservative. However, contrary to our initial hypotheses it is unlikely that this increased erosion is caused by greater palatability for microbes associated with a reduced C:N ratio since manipulated blades, in fact, had an elevated C:N ratio and we would expect blades with high microbial abundance to have a lower C:N ratio (Duggins et al. 1989). Future work should address whether microbes are in fact responsible for this accelerated breakdown and whether it is perhaps associated with lower concentrations of polyphenols or higher concentrations of laminarin. Alternatively increased erosion may be associated with structural difference associated with more rapid growth rates or higher concentrations of laminarin. Investigating the cause of this accelerated erosion and whether it occurred in blades that were already mature when the meristem was severed could reveal interesting insights into the ecological ramifications of apical meristem loss and whether such loss leads to an accelerated degeneration of cropped fronds. In focusing on the consequences of changes in translocation for Macrocystis pyrifera associated with apical meristem loss, this study took a substantially different approach than previous work focusing on patterns of translocation under typical growing conditions (Parker 1963, 1965; Schmitz & Lobban 1976; Lobban 1978 etc.). The study reveals surprising ramifications of the removal of apical meristems that merit further investigation into both their underlying physiological mechanisms and ecological consequences. Our results lead to fürther questions of whether the changes in erosion rates and chemical composition occur throughout larger portions of the frond and whether such changes affect interactions with organisms associated with the kelp, such as microbial biofilms and invertebrate consumers. Answering these questions may in turn reveal important insights into the mechanisms by which kelp carbon Markham 12 is made available to consumers, the impacts of harvesting, and the historic influence of canopy grazing by Sellar’s sea cow (Fig.7). Acknowledgements I would like to thank Drs. Sarah Lee and Vanessa Michelou for their tireless efforts and support, as well as Dr. Steve Palumbi for his invaluable insight. Special thanks also to Walker Clayton and Nicole Sarto for being excellent snorkel buddies and lifeguards. Thanks also to the faculty and students of Hopkins Marine Station who contributed to my project along the way. Markham 13 Arkema, K., Reed, D. C., & Schroeter, Stephen C. (2009). Direct and indirect effects of giant kelp determine benthic community structure and dynamics. Ecology, 90(11), 3126-37. Bengtsson, M., Sjøtun, K., Storesund, J., & Øvreås, J. (2011). Utilization of kelp-derived carbon sources by kelp surface-associated bacteria. Aquatic Microbial Ecology, 62(2), 191-199 Burger, R. (1995). Competition, Predation, and the Evolution and Extinction of Stellar’s Sea Cow, Hydrdamalis Gigas. Marine Mammal Science, 11(3), 391-394. Buschmann, A., & Hernandez-Gonzalez, M. C. (2005). Seaweed cultivation, product development and integrated aquaculture studies in Chile. WORLD AQUACULTURE-, 36(3), 51-53. Dayton, P. K. (1985). Ecology of Kelp Communities. Annual Review of Ecology and Systematics, 16, 215-245. Dixon, J., Schroeter, S.C., & Kastendiek, J. (1981). Effects of the encrusting bryozoan, Membranipora membranacea, on the loss of blades and fronds by the giant kelp, Macrocystis. Journal of Phycology, 17(4), 341-345. Wiley Online Library. Doty, M. S. (1987). Case studies of seven commercial seaweed resources (281-282 ed., p. 311). Food & Agriculture Org. Duggins, D., Eckman, J., Siddon, C., & Klinger, T. (2001). Interactive roles of mesograzers and current flow in survival of kelps. Marine Ecology Progress Series, 223, 143-155. Duggins, D., Simenstad, C., & Estes, J. (1989). Magnification of Secondary Production by Kelp Detritus in Coastal Marine Ecosystems. Science, 245(4914), 170. American Association for the Advancement of Science. Foster, M. S., & Schiel, D. R. (1985). Ecology of giant kelp forests in California: a community profile. Graham, M. H. (2004). Effects of Local Deforestation on the Diversity and Structure of Southern California Giant Kelp Forest Food Webs. Ecosystems, 7(4), 341-357. Gutierrez, A., Correa, T., Munoz, V., Santibanez, A., Marcos, R., Cáceres, C., et al. (2006). Farming of the Giant Kelp Macrocystis Pyrifera in Southern Chile for Development of Novel Food Products. Journal of Applied Phycology, 18(3-5), 259-267. Markham 14 Hemmi, A., & Jormalainen, V. (2002). Nutrient Enhancement Increases Performance of a Marine Herbivore via Quality of Its Food Alga. Ecology, 83(4), 1052. doi: 10.2307/307191 Hepburn, C., & Hurd, C. (2005). Conditional mutualism between the giant kelp Macrocystis pyrifera and colonial epifauna. Marine Ecology Progress Series, 302, 37-48. Lee, R. E. (1989). Phycology. Phycology (p. 146). New York: Cambridge University Press. Lobban, C. S. (1978a). Translocation of C in Macrocystis pyrifera (Giant Kelp). Plant physiology, 61(4), 585-9. Lobban, C. S. (1978b). The growth and death of the Macrocystis sporophyte (Phaeophyceae, Laminariales). Phycologia, 17(2), 196-212. The International Phycological Society Phycologia Business Office, Allen Press, 810 East 10th Street, P.O. Box 1897, Lawrence, KS 66044-889 Mann, K. (1973). Seaweeds: Their Productivity and Strategy for Growth. SCIENCE, VOL 182, NO 4116, P 975-981, DECEMBER 7, 1973. Mann, K. (2000). Ecology of coastal waters: with implications for management (2nd ed.). Malden, Mass.: Blackwell Science,. McMillan, S. (2010). Trophic interactions among Chlorostoma brunnea, Macrocystis pyrifera, and fungi. Society. Norderhaug, K., Fredriksen, S., & Nygaard, K. (2003). Trophic importance of Laminaria hyperborea to kelp forest consumers and the importance of bacterial degradation to food quality. Marine Ecology Progress Series, 255, 135-144. North, W. J. (1971). The biology of giant kelp beds (Macrocystis) in California,. Lehre,: J. Cramer, Parker, B. C. (1963). Translocation in the Giant Kelp Macrocystis. Science, 140(3569), 891. American Association for the Advancement of Science. Parker, B. C. (1965). Translocation in the Giant Kelp Macrocystis I. Rates, direction, quantity of C14-lablled products and fluorescein. Journal of Phycology, 1(2), 41-46. Wiley Online Library Poore, A. G. B., & Steinberg, P. D. (1999). Preference-Performance Relationships and Effects of Host Plant Choice in an Herbivorous Marine Amphipod. Ecological Monographs, 69(4), 443-464. Eco Soc America. Markham 15 Rothäusler, E., Gómez, I., Hinojosa, I. a, Karsten, U., Tala, F., & Thiel, M. (2009). Effect of Temperature and Grazing on Growth and Reproduction of Floating Macrocystis Spp. (Phaeophyceae) Along a Latitudinal Gradient. Journal of Phycology, 45(3), 547-559. Schmitz, K., & Lobban, C. S. (1976). A survey of translocation in laminariales (Phaeophyceae). Marine Biology, 36(3), 207-216. Schmitz, K., & Srivastava, L. (1979). Long Distance Transport in Macrocystis integrifolia: I. Translocation of C-labeled Assimilates. Plant Physiology, 63(6), 995-1002. Seymour, R.J., Tegner, MJ, Dayton, P. K., & Parnell, P. E. (1989). Storm wave induced mortality of giant kelp, Macrocystis pyrifera, in Southern California. Estuarine, Coastal and Shelf Science, 28,277-292. Steneck, R. S., Graham, M. H., Bourque, B. J., Corbett, D., Erlandson, J. M., Estes, J. a, et al. (2003). Kelp forest ecosystems: biodiversity, stability, resilience and future. Environmental Conservation, 29(04), 436-459. Watanabe, J. (1984). Food preference, food quality and diets of three herbivorous gastropods (Trochidae: Tegula) in a temperate kelp forest habitat. Oecologia, 62(1), 47-52. Springer. White, W. L., Coveny, a H., Robertson, J., & Clements, K. D. (2010). Utilisation of mannitol by temperate marine herbivorous fishes. Journal of Experimental Marine Biology and Ecology, 391(1-2), 50-56. Elsevier B.V. Arkema, K. K., Reed, D. C., & Schroeter, Stephen C. (2009). Direct and indirect effects of giant kelp determine benthic community structure and dynamics. Ecology, 90(11),3126-37. Bengtsson, M., Sjøtun, K., Storesund, J., & Øvreås, J. (201 1). Utilization of kelp-derived carbon sources by kelp surface-associated bacteria. Aquatic Microbial Ecology, 62(2), 191-199 doi: 10.3354/ame01477 Burger, R. (1995). Competition, Predation, and the Evolution and Extinction of Stellar’s Sea Cow, Hydrdamalis Gigas. Marine Mammal Science, II(3), 391-394. Buschmann, A., & Hernandez-Gonzalez, M. C. (2005). Seaweed cultivation, product development and integrated aquaculture studies in Chile. WORLD AQUACULTURE¬, 36(3), 51-53. Dayton, P. K. (1985). Ecology of Kelp Communities. Annual Review of Ecology and Systematics, 16,215-245. Markham 16 Dixon, J., Schroeter, S.C., & Kastendiek, J. (1981). Effects of the encrusting bryozoan, Membranipora membranacea, on the loss of blades and fronds by the giant kelp, Macrocystis. Journal of Phycology, 17(4), 341-345. Wiley Online Library. Doty, M. S. (1987). Case studies of seven commercial seaweed resources (281-282 ed., p.311). Food & Agriculture Org. Duggins, D., Eckman, J., Siddon, C., & Klinger, T. (2001). Interactive roles of mesograzers and current flow in survival of kelps. Marine Ecology Progress Series, 223, 143-155. doi: 10.3354/meps223143. Duggins, D., Simenstad, C., & Estes, J. (1989). Magnification of Secondary Production by Kelp Detritus in Coastal Marine Ecosystems. Science, 245(4914), 170. American Association for the Advancement of Science. Foster, M. S., & Schiel, D. R. (1985). Ecology of giant kelp forests in California: a community profile. Graham, M. H. (2004). Effects of Local Deforestation on the Diversity and Structure of Southern California Giant Kelp Forest Food Webs. Ecosystems, 7(4), 341-357. doi: 10.1007/810021- 003-0245-6. Gutierrez, A., Correa, T., Munnoz, V., Santibannez, A., Marcos, R., Cáceres, C., et al. (2006) Farming of the Giant Kelp Macrocystis Pyrifera in Southern Chile for Development of Novel Food Products. Journal of Applied Phycology, 18(3-5), 259-267. doi: 10.1007/8 1081 1-006-9025-y Hemmi, A., and V. Jormalainen. (2002). Nutrient Enhancement Increases Performance of a Marine Herbivore via Quality of Its Food Alga. Ecology, 83(4), 1052. Hepburn, C., & Hurd, C. (2005). Conditional mutualism between the giant kelp Macrocystis pyrifera and colonial epifauna. Marine Ecology Progress Series, 302,37-48. doi: 10.3354/meps302037. Lee, R. E. (1989). Phycology. Phycology (p. 146). New York: Cambridge University Press. Lobban, C. S. (1978a). Translocation of C in Macrocystis pyrifera (Giant Kelp). Plant physiology, 61(4), 585-9. Lobban, C. S. (1978b). The growth and death of the Macrocystis sporophyte (Phaeophyceae, Laminariales). Phycologia, 17(2), 196-212. The International Phycological Society Phycologia Business Öffice, Allen Press, 810 East 10th Street, P.O. Box 1897, Lawrence, KS 66044-8897. doi: 10.2216/10031-8884-17-2-196.1. Mann, K. (1973). Seaweeds: Their Productivity and Strategy for Growth. SCIENCE, VOL 182, NO 4116, P 975-981, DECEMBER 7, 1973.5 FIG, 2 TAB, 57 REF., 182(4116). Markham 17 Mann, K. (2000). Ecology of coastal waters : with implications for management (2nd ed.). Malden, Mass.: Blackwell Science,. McMillan, S. (2010). Trophic interactions among Chlorostoma brunnea, Macrocystis pyrifera, and fungi. Society. Norderhaug, K., Fredriksen, S., & Nygaard, K. (2003). Trophic importance of Laminaria hyperborea to kelp forest consumers and the importance of bacterial degradation to food quality. Marine Ecology Progress Series, 255, 135-144. doi: 10.3354/meps255135. North, W. J. (1971). The biology of giant kelp beds (Macrocystis) in California,. Lehre,: J Cramer, Parker, B. C. (1963). Translocation in the Giant Kelp Macrocystis. Science, 140(3569), 891. American Association for the Advancement of Science. Parker, B. C. (1965). Translocation in the Giant Kelp Macrocystis I . Rates, direction, quantity of C14-lablled products and fluorescein. Journal of Phycology, 1(2), 41—46. Wiley Online Library. Poore, A. G. B., and P.D. Steinberg. (1999). Preference-Performance Relationships and Effects of Host Plant Choice in an Herbivorous Marine Amphipod. Ecological Monographs, 69(4), 443-464. Eco Soc America. Rothäusler, E., Gómez, I., Hinojosa, I. a, Karsten, U., Tala, F., & Thiel, M. (2009). Effect of Temperature and Grazing on Growth and Reproduction of Floating Macrocystis Spp. (Phaeophyceae) Along a Latitudinal Gradient. Journal of Phycology, 45(3), 547-559. doi: 10.1111/j.1529-8817.2009.00676.x. Schmitz, K., & Lobban, C. S. (1976). A survey of translocation in laminariales (Phaeophyceae). Marine Biology, 36(3), 207-216. Schmitz, K., & Srivastava, L. (1979). Long Distance Transport in Macrocystis integrifolia: 1. Translocation of C-labeled Assimilates. Plant Physiology, 63(6), 995-1002. Seymour, R. J., Tegner, M J, Dayton, P. K., & Parnell, P. E. (1989). Storm wave induced mortality of giant kelp, Macrocystis pyrifera, in Southern California. Estuarine, Coastal and Shelf Science, 28, 277-292. Steneck, R. S., Graham, M. H., Bourque, B.J., Corbett, D., Erlandson, J. M., and J. Estes.(2003). Kelp forest ecosystems: biodiversity, stability, resilience and future. Environmental Conservation, 29(04), 436-459. Watanabe, J. (1984). Food preference , food quality and diets of three herbivorous gastropods (Trochidae: Tegula) in a temperate kelp forest habitat. Oecologia, 62(1), 47-52. Springer. Markham 18 White, W. L., Coveny, H., Robertson, J., and K. Clements. (2010). Utilisation of mannitol by temperate marine herbivorous fishes. Journal of Experimental Marine Biology and Ecology, 391(1-2), 50-56. Elsevier B.V. Markham 19 Fig. 1: Experimental setup in which fronds were cut 30cm from the first division on the apical blade. Growth and erosion were measured on the nearest blade 30cm from the first divison. Figure modified from Hepburn et al. (2007). Apical meristem pe ade 30cm lenenenag - 8 Tag 2 Markham 20 Fig. 2: Average growth rates (cm/day) of kelp blades in the first and second week after removal of the apical meristem. Error bars are standard errors bars of the mean. n=20 3.5 3 2.5 2 Control 515 Apex - Removed 0.5 Week Two Week One Markham 21 Fig. 3: Average change in distance (cm) between the punched hole and the kelp blade tip in the first week after removal of apical meristem. Error bars are standard errors bars of the mean. n=16 10 Apex Removed Control Markham 22 Fig. 4: Average percent dry weight mannitol for blades from fronds with and without apical meristem two weeks after cutting. Error bars are standard errors bars of the mean. n=10 -6 23 8 Control Apex Removed Markham 23 Fig. 5: Percent carbon of dry weight for blades with and without apical meristems two weeks after cutting. Error bars are standard errors bars of the mean. n=16 30.5 30 29.5 29 § 285 28 27.5 27 26.5 26 AM Removed Control Markham 24 Fig. 6: Carbon:Nitrogen ratio of tissue from blades with and without apical meristems two weeks after cutting. Error bars are standard errors bars of the mean. n=16 10.5 10 — 9.5 8.5 AM Removed Control Markham 25 Fig. 7: Stellar's sea cow (Hydrodamalis gigas)grazing on Macrocystis canopies. (Encyclopedia Britannica)