Plant Systematics and Evolution

, Volume 284, Issue 1–2, pp 33–39 | Cite as

Morphological features and isoenzyme characterization of endosymbiotic algae from green hydra

  • Goran Kovačević
  • Sandra Radić
  • Biserka Jelenčić
  • Mirjana Kalafatić
  • Hrvoje Posilović
  • Branka Pevalek-Kozlina
Original Article
First Online:
Received:
Accepted:
  • 117 Downloads

Abstract

Symbiotic associations are of a wide significance in evolution and biodiversity. Green hydra is a typical example of endosymbiosis. In its gastrodermal myoepithelial cells, it harbors individuals of unicellular green algae. Morphological characteristics of isolated algae determined by light and electron microscopy are presented. Cytological morphometric parameters (cell area, cell radius, chloroplast area) of isolated algae from green hydra (Cx), as well as from reference species Chlorella kessleri (Ck) and Chlorella vulgaris (Cv), revealed similarity between the isolated endosymbiont and C. kessleri. Isoenzyme patterns of esterase (EST), peroxidase (POX), and catalase (CAT) were used for the investigation of genetic variability in endosymbiotic algae isolated from green hydra. Out of 14 EST isoenzymes observed in Cx species, 9 were expressed in the Cx sample. Results of the EST isoenzyme analysis indicated a higher degree of similarity between Cx and Cv than between Cx and Ck. Due to much higher heterogeneity, EST isoenzymes seem to be more suitable genetic markers for identification of different Chlorella species than CAT and POX isoenzymes. Results obtained suggest that symbiogenesis in green hydra has probably not been terminated yet.

Keywords

Endosymbiotic alga Green hydra Morphometry Isoenzymes 
This is a preview of subscription content, log in to check access.

Notes

Acknowledgments

The presented results are a product of the scientific project “Molecular phylogeny, evolution and symbiosis of freshwater invertebrates” and project 119-1191196-1202 carried out with the support of the Ministry of Science, Education and Sport of the Republic of Croatia.

References

  1. Aebi H (1984) Catalase in vitro. Methods Enzymol 105:121–126CrossRefPubMedGoogle Scholar
  2. Agostinucci K, Manfredi TG, Cosmas A, Martin K, Han SN, Wu D, Sastre J, Meydani SN, Meydani M (2002) Vitamin E and age alter liver mitochondrial morphometry. J Anti-Aging Med 5:173–178CrossRefGoogle Scholar
  3. Arkhipchuk W, Blaise C, Malinovskaya MV (2005) Use of hydra for chronic toxicity assessment of waters intended for human consumption. Environ Pollut 142:200–211CrossRefPubMedGoogle Scholar
  4. Balen B, Krsnik-Rasol M, Simeon-Rudolf V (2003) Isoenzymes of peroxidase and esterase related to morphogenesis in Mammillaria gracilis Pfeiff. tissue culture. J Plant Physiol 160:1401–1406CrossRefPubMedGoogle Scholar
  5. Baron PJ, Real LE, Ciocco NF, Re ME (2004) Morphometry, growth and reproduction of an Atlantic population of the razor clam Ensis macha. Sci Mar 68:211–217CrossRefGoogle Scholar
  6. Beach MJ, Pascoe D (1998) The role of Hydra vulgaris (Pallas) in assessing the toxicity of freshwater pollutants. Wat Res 32:101–106CrossRefGoogle Scholar
  7. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254CrossRefPubMedGoogle Scholar
  8. Brown AHD, Weir RS (1983) Measuring genetic variability in plant populations. In: Tankley SD, Orton TI (eds) Isozymes in plant genetics and breeding. Development in plant genetics and breeding 1. Elsevier, Amsterdam, pp 219–240Google Scholar
  9. Burlina A, Galzigna L (1972) A new and simple procedure for serum arylesterase. Clin Chim Acta 39:255–257CrossRefGoogle Scholar
  10. Burnett AL (1973) Biology of hydra. Academic Press, New YorkGoogle Scholar
  11. Chance B, Maehly AC (1955) Assay of catalases and peroxidases. In: Colowick SP, Kaplan NO (eds) Methods in enzymology. Academic Press, New York, pp 764–775CrossRefGoogle Scholar
  12. Cubadda R, Quattrucci E (1974) Separation by gel electrofocussing and characterization of wheat esterses. J Sci Food Agric 25:417–422CrossRefPubMedGoogle Scholar
  13. Cummins I, Burnet M, Edwards R (2001) Biochemical characterisation of esterases active in hydrolysing xenobiotics in wheat and competing weeds. Physiol Plant 113:477–485CrossRefGoogle Scholar
  14. De Carvalho VM, Marques RM, Lapenta AS, Machado MFPS (2003) Functional classification of esterases from leaves of Aspidosperma polyneuron M. Arg. (Apocynaceae). Gen Mol Biol 26:195–198Google Scholar
  15. Diaz-Pulido G, Villamil L, Almanza V (2007) Herbivory effects on the morphology of the brown alga Padina boergesenii (Phaeophyta). Phycologia 46:131–136CrossRefGoogle Scholar
  16. Douglas AE, Smith DC (1984) The green hydra symbiosis. VIII. Mechanisms in symbiont regulation. Proc R Soc Lond B 221:298–319CrossRefGoogle Scholar
  17. Dunn K (1987) Growth of endosymbiotic algae in the green hydra, Hydra viridissima. J Cell Sci 88:571–578Google Scholar
  18. Falconer DS (1986) Introducción a la genética cuantitativa. CECSA, MéxicoGoogle Scholar
  19. Fawley MW, Fawley KP, Buchheim MA (2004) Molecular diversity among communities of freshwater microchlorophytes. Microb Ecol 48:489–499CrossRefPubMedGoogle Scholar
  20. Friedl T (1997) Evolution of the green algae. In: Bhattacharya D (ed) Origins of algae and their plastids. Berlin, Springer, pp 87–101Google Scholar
  21. Gershater MC, Edwards R (2007) Regulating biological activity in plants with carboxylesterases. Plant Sci 173:579–588CrossRefGoogle Scholar
  22. Habetha M, Bosch TCG (2005) Symbiotic Hydra express a plant-like peroxidase gene during oogenesis. J Exp Biol 208:2157–2165CrossRefPubMedGoogle Scholar
  23. Habetha M, Anton-Erksleben F, Neumann K, Bosch TCG (2003) The Hydra viridis/Chlorella symbiosis. Growth and sexual differentiation in polyps without symbionts. Zoology 106:1–8CrossRefGoogle Scholar
  24. Handa S, Nakahara M, Tsuboa H, Deguchi H, Masuda Y, Nakano T (2006) Choricystis minor (Trebouxiophyceae, Chlorophyta) as a symbiont of several species of freshwater sponge. Hikobia 14:365–373Google Scholar
  25. Hegewald E, Hanagata N (2000) Phylogenetic studies on Scenedesmaceae (Chlorophyta). Algol Stud 100:29–49Google Scholar
  26. Holstein T, Emschermann P (1995) Cnidaria: Hydrozoa Kamptozoa. Gustav Fischer, StuttgartGoogle Scholar
  27. Horvatić J, Vidaković-Cifrek Ž, Regula I (2000) Trophic level, bioproduction and toxicity of the water of lake Sakadaš (Nature Park Kopački rit, Croatia). Limnol Report 33:89–94Google Scholar
  28. Huss VAR, Holweg C, Seidel B, Reich V, Rahat M, Kessler E (1993/1994) There is an ecological basis for host/symbiont specificity in Chlorella/Hydra symbioses. Endocyt Cell Res 10:35–46Google Scholar
  29. Kalafatić M, Kopjar N (1995) Response of green hydra to pirimicarb. Biol Bratislava 50:289–292Google Scholar
  30. Kaltenpoth M, Göttler W, Herzne G, Strohm (2005) Symbiotic bacteria protect wasp larvae from fungal infections. Curr Biol 15:475–479CrossRefPubMedGoogle Scholar
  31. Kessler E, Kauer G, Rahat M (1991) Excretion of sugars by Chlorella species capable and incapable of symbiosis with Hydra viridis. Bot Acta 104:58–63Google Scholar
  32. Kovačević G, Kalafatić M, Ljubešić N, Šunjić H (2001) The effect of chloramphenicol on the symbiosis between alga and hydra. Biol Bratislava 56:605–610Google Scholar
  33. Kovačević G, Ljubešić N, Kalafatić M (2005) Newly described mechanims in hydra-alga symbiosis. In: Bosch TGC, Holstein TW, David CN (eds) Abstract book of the international workshop hydra and the molecular logic of regeneration. DFG, Tutzing,p 95Google Scholar
  34. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685CrossRefPubMedGoogle Scholar
  35. Lewontin RC (1974) The genetic basis of evolutionary change. Columbia University Press, New YorkGoogle Scholar
  36. Mädgefrau K, Ehrendorfer F (1988) Botanika: Sistematika, evolucija i geobotanika. Školska knjiga, ZagrebGoogle Scholar
  37. McAuley PJ, Smith DC (1982) The green hydra symbiosis. V. Stages in the intracellular recognition of algal symbionts by digestive cells. Proc R Soc Lond B 216:7–23CrossRefGoogle Scholar
  38. Mittler R, Zilinskas B (1993) Detection of ascorbate peroxidase activity in native gels by inhibition of the ascorbate-dependent reduction of nitroblue tetrazolium. Anal Biochem 212:540–546CrossRefPubMedGoogle Scholar
  39. Nakahara M, Handa S, Nakano T, Deguchi H (2003) Culture and pyrenoid structure of a symbiotic Chlorella species isolated from Paramecium bursaria. Symbiosis 34:203–214Google Scholar
  40. O’Brien TL (1982) Inhibition of vacuolar membrane fusion by intracellular symbiotic algae in Hydra viridis (Florida strain). J Exp Zool 223:211–218CrossRefPubMedGoogle Scholar
  41. Pratt R (1941) Studies on Chlorella vulgaris IV. Am J Bot 28:492–497CrossRefGoogle Scholar
  42. Rahat M (1991) An ecological approach to hydra-cell colonization by algae-algae/hydra symbioses. Oikos 62:381–388CrossRefGoogle Scholar
  43. Reisser W, Wiessner W (1984) Autotrophic eukaryotic freshwater symbionts. In: Linskens HF, Heslop-Harrison J (eds) Cellular interactions (encyclopedia of plant physiology). Springer, Berlin, pp 59–90Google Scholar
  44. Scandalios JG (1990) Response of plant antioxidant defense genes to environmental stress. Adv Genet 28:1–41CrossRefPubMedGoogle Scholar
  45. Siegel BZ (1993) Plant peroxidases: an organismic perspective. Plant Growth Regul 12:303–312CrossRefGoogle Scholar
  46. Tonar Z, Markos A (2004) Microscopy and morphometry of integument of the foot of pulmonate gastropods Arion rufus and Helix pomatia. Acta Vet Brno 73:3–7Google Scholar
  47. Vianello A, Zancani M, Nagy G, Macrì F (1997) Guaiacol peroxidase associated to soybean root plasma membranes oxidizes ascorbate. J Plant Physiol 150:573–577Google Scholar
  48. Woodbury W, Spencer AK, Stahmann MA (1971) An improved procedure using ferrycyanide for detecting catalase isozymes. Anal Biochem 44:301–305CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Goran Kovačević
    • 1
  • Sandra Radić
    • 1
  • Biserka Jelenčić
    • 1
  • Mirjana Kalafatić
    • 1
  • Hrvoje Posilović
    • 2
  • Branka Pevalek-Kozlina
    • 1
  1. 1.Division of Biology, Faculty of ScienceUniversity of ZagrebZagrebCroatia
  2. 2.Department of Geology, Faculty of ScienceUniversity of ZagrebZagrebCroatia

Personalised recommendations