Rene Toolson INTRODUCTION Competition among benthic marine invertegrates revolves importantly around finding a place to settle and, maintaining that space and acquiring new space for growth during the life of the organism (Connell,1961; Dayton, 1971; Jackson,1977; Paine, 1974). Buss and Jackson's examination of associations and interactions of coral reef invertebrates (1979). Connell's account of interspecific competition among barnacles, as well as Dayton's study of community organization (1971) introduced exper¬ imental approaches to a previously descriptive field. Their research has suggested that competition for space in the intertidal is complex, involving definite adaptive strategies. Colonial animals from such groups as sponges, coelenterates, bryozoa, and tunicates are often found as the dominant animal cover on benthic substrates. Jackson (1977) makes the case for the strategy of the indeterminate growth and acquisition of space by asexual reproduction of these colonial forms being one of the most effective methods of space competition. The colony can survive damage and continue to spread (Buss, 1979). Thus the abundant and diversified colonial ascidians are an interesting group to examine for competitive interactions for living space. In the rocky intertidal regions of the mid-California coast, full utilization of all available space by competing algae and invertebrates, both solitary and colonial, shows the importance of space in the community. That so many competitors here are colonial gives reason to believe they are successful (Jackson 1977). The lower intertidal is marked by the presence of colonial ascidians who occupy a significant portion of the rocky substrate (Morris, Abbott and Haderlie, 1980). p.2 Rene Toolson The purpose of this study was to examine the competitive associations between colonial ascidians at Cabrillo Point and evaluate competitive strategies. Classification of species followed the categories of growth forms in order to determine a relationship between structure and competitive strategy. Finally, growth forms were evaluated according to their competitive abilities. METHODS The study was conductedin the intertidal at Hopkins Marine Station, Cabrillo Pt., Pacific Grove, California in April and May of 1981. The study area was located northwest of Bird Rock and sites were chosen for high tunicate density and variation in physical conditions of exposure and tidal height. Thirteen undisturbed areas were photographed weekly over a four week period of time to note any changes that occur in the field over time. Other areas were observed for ongoing competitive interactions and selected borders between ascidians were sectioned and examined under a dissecting scope to observe the results of previous competitive interactions. Two follow-up experiments were performed to lead to an understanding of growth strategies of individual species. 1. Entire colonies of different species were brought from the field and tied down onto glass plates,alone and in species pairs. Responses were observed, and each plate was photographed over time. 2. In the field, eight species were isolated by removing all veget¬ ation and invertebrates that were within a two-inch radius around the colonies, and these were photographed over time. In both experiments, colony sizes were traced from photos and compared over time. p.3 Rene Toolson RESULTS The results of the field study detailing competitive interactions and the observations from sectioning interactive borders are summarized in Table 1. The interactions are listed by pairs of species of ascidians, They are classified by growth strategies (as introduced in the discussion), and the relative proportion of times that a given response was observed out of all contacts analyzed is listed. The results of the growth responses observed from the laboratory experiment done on glass plates are presented in Table 2 for comparison. The following is a description of the common interactions, categorized by growth forms. Encrusting Sheets Meeting Encrusting Sheets Colonies of Aplidium californicum met frequently since the species is abundant and occurs in a variety of habitats and tidal heights. It was never observed to overgrow another colony of the same species in the field,and the colonies would meet and maintain their borders, frequently accumulating sand in the space between contiguous colonies. However, when two colonies were paired on glass plates, overgrowth without fusing, was observed. In all observed contacts, A. Californicum grew over Archidistoma psammion. When overgrown, A. psammion often had a thick test, encrusted with sand and empty of zooids in the area of overgrowth, but it might also have been unaffected. A. californicum colonies moved 8 mm and overgrew colonies of A. psammion in seven days on glass plates. Due to its large range, A. psammion met often in the field with colonies of the same species, one-third of the time growing over it. Whether they met or overlapped, they had thick, zooidless tests in the area of contact that were often encrusted with sand. On glass plates when two A. psammion colonies were paired, the colonies spread out but p. Rene Toolson grew too slowly to note any response to neighboring colonies. Encrusting Sheets Meet Encrusting Mounds Interactive borders of A. psammion and Archidistoma molle were often found. Both leaned and overgrew each other with the same frequency while A. psammion zooids were occasionally found at the base of the tunic of A. molle. The test of A. psammion, when underneath an A. molle colony was thick and empty of zooids. When colonies were paired on glass plates, A. molle projected its tunic over A. psammion and attached to it. When colonies of Aplidium solidum and A. psammion grew together in the field, the outcome was quite variable with displacement, leaning and mutual overgrowth all occurring. Both colonies responded to the contact with thicker, sand-encrusted tests, and sand was found in between the two colonies. In the field, when colonies of A. californicum overgrew both mound species, A. solidum and Archidistoma diaphanes, the mounds showed empty and deteriorating tunics. A.solidum was also observed to have a thick test, sometimes encrusted with sand. Colonies of A. californicum and A. molle were never observed to meet in the field but when paired on glass plates, colonies of A. californicum were seen to spread out zooid systems and overgrow the adjacent colony of A. molle. The same results were seen when A. californicum was paired with A. solidum on glass plates. Lobe-shaped Species Meet Encrusting Sheets A. californicum was observedto overgrow the sandy-lobed species, Synoicum parfustis, Aplidium propinquum, and Aplidium arenatum. p.5 Rene Toolson A. psammion would displace, but rarely overgrow, Synoicum and A. propin- quum lobes. Both of these sandy lobes, and colonies of Polyclimum planum, were commonly observed to use A. psammion as substrate while A. arenatum did not. Most often, A. arenatum was surrounded by A. psammion, or the sandy lobe was observed leaning over the encrusting sheet - in which case the test of Archidistoma psammion was thick and sand encrusted. Lobe-shaped Species with Encrusting Mounds In general, the sandy lobes most commonly were found being leaned upon and overgrown by the mound species, A. solidum,A. molle, or A. diaphanes. The lobes were usually displaced,and the mound species frequently had a thick layer of test at these borders. The mounds were never used as substrate by the lobes. Social Colonials, In General A. californicum was the only species that was observed to overgrow any of the social colonials, and this was Clavelina huntsmani. Since its growth only extended around the base of the Clavelina colony, no damage was observed. Frequently, both Clavelina and Euherdmania claviformes attached stolons to colonies of A. psammion, using it as substrate, but these social colonial species were more often displaced when located at the borders of this encrusting sheet. When in contact with species of encrusting mounds, both Clavelina and Euherdmania either showed no response or were displaced. When social colonials were in contact with lobes, they were generally tol¬ erant, and showed little competitive interaction. In the field, when Euherdmania and Clavelina come in contact, they showed no visible p.6 Rene Toolson response to each other, but their stolons were all intermingled at the base of the colonies. Perophora annectens, (with the exception of A. molle) extended its stolons out over the tunic of the neighbor colonies it encountered, and the neighbor colony showed no response to the interaction. All of the social colonial species exhibited budding in contact situations and did no damage to the colony they bordered. Of the colonies isolated in the field or placed singly on glass plates, all increased their colony sizes within ten days, except Synoicum parfustis, Archidistoma diaphanes, and Clavelina huntsmani. Results are summarized in Table 3 with sample growth plates in the Appendix. A. californicum showed the fastest increase in size. A. californicum, A. molle, and A. solidum showed the fasted repair to damage done by the string used to tie down the colonies with. Repair consisted of separating the tunic and zooids at the point of contact with the string and moving the test up and around the string until it was completely covered. The colony appeared normal after covering the string. Rearrangement of zooid systems was observed in both A. californicum and A. solidum. In A.californicum, systems would spread out, and more test was observed in between systems. The systems of A. solidum broke apart into lobes. Clavelina, when isolated on glass plates, was observed to have grown buds that were two to seven millameters long within two weeks. p.7 Rene Toolson All colonies flattened out onto the plates within one week, and fusion to the glass plates occurred within two weeks. When groups of colonies of A. molle or A. psammion were isolated in the field, the colonies would move closer together when different competing species were removed from between them. DISCUSSION In observing the variety of interactions involved in space competit¬ ion between the intertidal colonial ascidians, it became apparent that there was a relationship between the growth formsof the ascidians and the strategies empled. This led to classifying the colonies into four categories: 1)encrusting sheets, 2) encrusting mounds, 3) lobe-shaped colonies, and 4)social colonials. Encrusting sheets are compound colonials that are often large, cortinuous colonies that sit low on the surface of their substrate. Encrusting mounds are compound species that are attached to the substrate and slightly "mushroom' out beyond their attachment, usually growing taller than the sheets. Lobes are attached to the substrate at one small point at their bases, and the remainder of the colony is projected outwards. Social colonials are groups of individuals that are attached at the base by stolons which often include parts of the colony's circulatory system, in contrast to compound ascidians that are composed of a group of individuals bound together in a common test. Each of these groups has a distinctive set of competitive strategies though individual members of the set may be shared with other growth forms. In addition, each strategy may be developed to different degrees by species of the same or different groups. Table 4 summarizes p.8 Rene Toolson these observations for the intertidal ascidians examined for this study. The strategies are discussed below in relation to their effectiveness, and the summary of their success is presented in Table 5 in the form of a dominance matrix. Sheets Among the growth forms of ascidians, encrusting sheets show relatively good competitive abilities (Buss, 1979), as evidenced by the large anomt of space they occupy in the intertidal (Pulliam, 1981). Encrusting sheets use the most direct techniques for obtaining space: by creating new substrate out of living ascidians through overgrowth. In the case of A. californicum, growth rate, on top of structure, is an important strategy influencing competitive success. Both on glass plates and in the field, this seasonal ascidian showed the fastest increase in colony size of all the species examined. Though it has a low stature on the substrate interms of height which would make it easy for a mound species to lean over it, A. californicum was determined to be the most dominant species because it grows faster than any other species it was observed to come in contact with (Buss, 1979; Buss and Jackson, 1979). No other species was ever observed to overgrow or lean onto A. californicum. On the other hand, A. psammion, the other very common encrusting sheet, showed very slow growth rates in the field and on glass plates. Its thick test is unsuitable for fast growing, so it is often suscept¬ ible to the leaning of the mound species who grow both taller and faster. The growth-limiting tunic, though, possesses protective features which also serve as stra egic advantages. The thick test is hardened by sand encrustation and acts as a barrier to movement by the softer species and as a protective shield for the zooids p.9 Rene Toolson beneath. Munds Encrusting mounds use the leaning strategy most often because it conforms most usefully with their mushroom-like structure. They are able to root down in an area, and then extend their tunic, with zooids, beyond their point of attachment. The soft, jelly-like composition of their tests (also like that of A. californicum) probably facilitiates faster growth rates, so when they come into contact with a dominant encrusting sheet like A. psammion, they are not as likely to be overgrown because they grow taller and faster. The combined use of these strategies also proves to be effective when in contact with the less aggressive lobe and social colonial species, as is indicated by the direction of the arrows on Table 5. Lobes Compound ascidians found in lobe forms are efficient with their use of space due to their small point of attachment which is able to support a large colony in very little space. Their only real aggressive strategy is the ability to crowd out another species with a large colony of lobes, though laboratory observations have not shown this response to be as quick as the growth rate of encrusting sheets or mounds. Lobes may also be successful when they use encrusting sheets as substrate by settling on and embedding into them. When confronted with an aggressive neighbor, the lobes will respond (except A. arenatum) by leaning and growing away from the neighbor, a form of displacement. The hard, sand-encrusted tests of the sandy lobe species, A. propinquum. A. arenatum, and Synoicum, serves the same protective purposes as it does for A. psammion. p.10 Rene Toolson Social Colonials The main strategy of the social colonial ascidian, who is structurally much more fragile than any other growth form, is its ability to repro¬ duce asexually very quickly, as seen on glass plates in the laboratory. This allows them to occupy alot of space in a short amount of time evade overgrowth, and even crowd out another species on occasion. The social colonials are also very succcessful, more so than the lobe species, at using other growth forms (including lobes) for substrate. This is a highly advantageous strategy for such easily damaged ascidians. The overall diversity of the rocky intertidal can be attributed at least in part to the variety of differen tially successful competitive strategies used by the organisms within the community structure (Buss and Jackson, 1979). The species' structure, composition, longevity, and growth rate all contribute to its overall success against other ascidians. By using these varying combinations of factors as well as others not investigated, most species are equipped with the means to survive even in the toughest neighborhoods. Diversity in the pattern of distribution of intertidal tunicates may well be the result of this variability of competitive strategies between species, between their effectiveness in different habitats and the life history strategies of growth rate, longevity and successful recruitment. SUMMARY 1. Strategies for space competion between species of colonial ascidians were studied in the rocky intertidal of Hopkins Marine Station, Pacific Grove, California. Rene Toolson 2. A general hierarchy of successful competitors based upon growth forms can be presented (in order of greatest to least success): Encrusting sheets, encrusting mounds, social colonials, and lobe¬ shaped species. Within these categories, however, there are exceptions, so that growth forms alone cannot be considered as a universal factor for determining competitive success. 3. Rate of asexual reproduction, or colony growth rate, based upon a list of individual species, can also be used to produce a hierarchy of successful competitors; however, this conclusion would be even less accurate if applied to all ascidians, and thus can only be considered as a secondary factor influencing competitive success. 4. Composition of the test, ranging from hardness to softness, because it has an effect bo on otective and growth abilities, has a correlation with competitive ability. 5. Protective as well as aggressive strategies are at work during space competition between ascidians, leading to a complex community structure. 6. The diversity of species present and the community and the complex structure of their distribution is a result of the variety of competitive tactics utilized by the individual ascidian. ACKNOWLEDGEMENTS I would like to thank eyeryone at HMS for the opportunity to do this study, especially Dr. Abbott for his enthusiasm, Dr. Gilly for his tolerance, and mostly Chuck Baxter for endless patience. I would also like to express my appreciation for the indispensable help I received from Keith Kohatsu and collaboration from Bill Pulliam. p.12 Table SHEETS + SHEETS Aplidium californicum Aplidium californicum Aplidium californicum Archidistoma psammion Archidistoma psammion Archidistoma psammion SHEETS + MOUNDS Aplidium californicum Archidistoma molle Aplidium californicum Aplidium solidum Aplidium californicum Archidistoma diaphanes Aplidium californicum Archidistoma ritteri Aplidium californicum Distaplia occidentalis Archidistoma psammion Archidistoma molle Archidistoma psammion Aplidium solidum Archidistoma psammion Archidistoma diaphanes 11 Rene Toolson p. 13 arde MOUNDS + MOUNDS Archidistoma molle Archidistoma molle Archidistoma molle Aplidium solidum Archidistoma molle Archidistoma diaphanes Aplidium solidum Aplidium solidum Aplidium solidum Archidistoma diaphanes Archidistoma diaphanes Archidistoma diaphanes LOBES + SHEE Polyclinum planum Archidistoma psammion Synoicum parfustis Aplidium californicum Synoicum parfustis Archidistoma psammion Aplidium propinquum Aplidium californicum Aplidium propinquum Archidistoma psammion 19 10 1 3 2-3 — a Rene Toolson 7 p.14 alle Aplidium arenatum Aplidium californicum Aplidium arenatum Archidistoma psammion — — LOBES + MOUNDS Synoicum parfustis Archidistoma molle Synoicum parfustis Archidistoma diaphanes Synoicum parfustis Archidistoma ritteri Aplidium propinguum Archidistoma molle Aplidium propinquum Archidistoma diaphanes Aplidium propinquum Aplidium solidum Aplidium arenatum Archidistoma molle Aplidium arenatum Aplidium solidum Aplidium arenatum Archidistoma diaphanes 1 . 8 Rene Toolson p.15 lable SOCIAL COL + LOBES Clavelina huntsmani Synoicum parfustis Clavelina huntsmani Aplidium propinquum Clavelina huntsmani Aplidium arenatum Euherdmania claviforme: Synoicum parfustis SOCIAL + SHEETS Clavelina huntsmani Aplidium californicum Clavelina huntsmani Archidistoma psammion Euherdmanis claviformes Archidistoma psammion SOCIAL + MOUNDS Clavelina huntsmani Archidistoma molle Clavelina huntsmani Aplidium solidum Clavelina huntsmani Archidistoma diaphanes Euherdmania claviforme: Archidistoma molle Rene Toolson p.16 Euherdmania claviformes Aplidium solidum - Euherdmania claviformes Archid stoma diaphanes IAL + SOCIAL Clavelina huntsmani Euherdmania claviforme Perophora annectens all, except A. molle - 4 10 Rene Toolson Jable Species Pairs — Aplidium californicum Aplidium californicum Aplidium californicum Archidistoma psammion Archidistoma psammion Archidistoma psammion Aplidium californicum Archidistoma molle Aplidium californicum Aplidium solidum Archidistoma psammion Archidistoma molle — Archidistoma molle Aplidium solidum — — Synoicum parfustis Aplidium californicum Perophora annectens Archidistoma psammion 8 — Rene Toolson Talle Field: Glass Plates Isolated Species Aplidium californicum * Aplidium solidum * Archidistoma psammion * Archidistoma molle Archidistoma diaphanes Synoicum parfustis Clavelina huntsmani Aplidium californicum Aplidium solidum Archidistoma psammion Archidistoma molle Archidistoma diaphanes Aplidium propinquum Clavelina huntsmani Increase ir Colony Area 16% 15% 69 14% 12-19% 117 1% -9% Rene Toolson Observed Rearrangement Time of Systems (days) no no no no yes (3 days) yes 12 (7 days) no 10 no 10 no 10 no 10 budding 10 labl e OO a- (5. 5. (5. 5. 88 Rene Toolson 80 88 5. . 0 8 (lole 5 — 4. 4. 8 — 85 22.4. — + —+— O + O 4 Rene Toolson Aplidium californicum Archidistoma psammion — Archidistoma molle — Aplidium solidum Archidistoma diaphanes — Archidistoma ritteri a- Distaplia occidentalis Synoicum parfustis —1 Aplidium propinquum Aplidium arenatun Polyclinum planum Perophora annectens Clavelina huntsmani — Euherdmania claviformes p.21 Rene Toolson TABLE LEGENDS Table 1: Responses of species combinations of colonial ascidians based upon field observations and sections of interactive borders. Numbers indicate the proportion of a response occurring for that interaction. Blanks indicate that the response was not observed. Thick test = given species had a layer of thick test with a conspicuous lack of zooids at the border of interaction; sand encrusted = bordering test was sand encrusted; loose sand - sand in between the two colonies, not encrusted; displacement = given specieswas displaced by neighbor species; projection - given species, tunic and zooids, was leaning over neighbor species; overgrowth = given species was growing over the other species, but was also attaded to primary substrate; 2° substrate = given species was using other species as substrate; other zooids = presence of zooids of given species in the test of the other species; budding = buds were attached to the species. Damage was rated on the following scale : O = no damage; 1 = slight damage (lack of zooids, very thick test); 2 - significant damage (no zooids, deterioration of test); 3 = death. No response = interaction had no visible effect on either colony. *The tracings of these colonies appear in the appendix. Table 2: Responses of species combinations of colonial ascidians removed from the field and tied to glass plates. + = given response was observed; - = opposite of given response was observed; blank indicates that the response was not observed. Expansion = size of the colony increased; overgrowth = given species grew over neighbor species; projection = given species leaned over other species; spreading = systems within the p.22 Rene Toolson colony spread out, bec me less dense within the test. *The tracings of these colonies appear in the appendix. Table 3 : A summary of the responses of colonial ascidians to isolation, both by clearing borders in the field and tying colonies on glass plates. Colony area was determined by tracing photographs and calculating the area within the tracing. + = the change in colony size of these species was positive, yet not determinable by tracing. *The tracings of these isolated colonies appear in the appendix. Table 4: List of all species classified according to growth forms with compet¬ itive strategies summarized for each growth form. Table 5: A matrix of dominance of ascidian species observed in the field. Dominance, indicated by the direction of the arrow, was determined through observations and sectioning interactive borders.— - direction of dominance;+ - both share dominance, but empty arrow less often than blackened arrow;dominance is approximately equalrelation- ship was only observedon glass plates;almost total dominance, occasionally, the other species won;()-no observable effects of the interaction; blank indicates that the interaction was not seen. Rene Toolson LITERATURE CITED Buss, L.W. 1979. Habitat selection, directional growth and spatial refuges: why colonial animals have more hiding places. in B. Rosen and G. Larwood, eds. Biology and Systematics of Colonial Animals. Academic Press. London. 598pp. Buss, L.W. and J.B.C. Jackson. 1979. Competitive networks: nontransitive competitive relationships in cryptic coral reef environments. Am. Nat, 113:223-233 Connell, J.H. 1961. The influence of interspecific competition and other factors on the distribution of the barnacle Cthamalus stellatus. Ecology 42:710-723 Dayton, P.K. 1971. Competition, disturbance, and community organization: the provision and subsequent utilization of space in a rocky intertidal community. Ecol. Monogr. 41:351-389 Jackson, J.B.C. 1977. Competition on marine hard substrata: the adaptive significance of solitary and colonial strategies. Am. Nat. 111:743-768 Jackson, J.B.C. 1979. Morphological strategies of sessile animals. in B. Rosen and G. Larwood, eds. Biology and Systematics of Colonial Animals. Academic Press. London. 598pp. Morris, R.H., Donald P. Abbott and Eugene C. Haderlie. Intertidal Invert- ebrates of California. Stanford University Press. Stanford, California. 1980. 69opp. Paine, R.T. 1974. Intertidal community structure: experimental studies on the relationship between a dominant competitor and its principal predator. Oecologia. 15:93-120 APPENDIX: A. californicum + A. propinquum (5 days) - - - - Ac Ap A. californicum + A. solidum (5 days) - - - - - Ac (All colonies 1X) Rene Toolson A. californicum + A. molle — — - (5 days) - - - Ac Am A. psammion + A. molle (10 days)- - - p.2. ) N S Rene Toolson A. solidum (2X) (6 days) A. psammion (2X) (6 days) alifornicum (1X) (5 days) Rene Toolson A. molle (2X) (6 days) A. propinquum (1X) (10 days) p.27