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Plant Biol (Stuttg) 2006; 8(6): 813-820
DOI: 10.1055/s-2006-924177
Research Paper

Georg Thieme Verlag Stuttgart KG · New York

Fluorescence Labelling of Phosphatase Activity in Digestive Glands of Carnivorous Plants

B. J. Płachno1 , L. Adamec2 , I. K. Lichtscheidl3 , M. Peroutka3 , W. Adlassnig3 , J. Vrba4
  • 1Department of Plant Cytology and Embryology, The Jagiellonian University, Grodzka 52, 31-044 Cracow, Poland
  • 2Institute of Botany, Academy of Sciences of the Czech Republic, Section of Plant Ecology, Dukelská 135, 379 82 Třeboň, Czech Republic
  • 3Cell Imaging and Ultrastructure Research Unit, University of Vienna, Althanstraße 14, 1090 Vienna, Austria
  • 4Hydrobiological Institute, Academy of Sciences of the Czech Republic, Na Sádkách 7, 370 05 České Budějovice, Czech Republic
Further Information

Publication History

Received: September 29, 2005

Accepted: April 5, 2006

Publication Date:
24 July 2006 (online)

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Abstract

A new ELF (enzyme labelled fluorescence) assay was applied to detect phosphatase activity in glandular structures of 47 carnivorous plant species, especially Lentibulariaceae, in order to understand their digestive activities. We address the following questions: (1) Are phosphatases produced by the plants and/or by inhabitants of the traps? (2) Which type of hairs/glands is involved in the production of phosphatases? (3) Is this phosphatase production a common feature among carnivorous plants or is it restricted to evolutionarily advanced species? Our results showed activity of the phosphatases in glandular structures of the majority of the plants tested, both from the greenhouse and from sterile culture. In addition, extracellular phosphatases can also be produced by trap inhabitants. In Utricularia, activity of phosphatase was detected in internal glands of 27 species from both primitive and advanced sections and different ecological groups. Further positive reactions were found in Genlisea, Pinguicula, Aldrovanda, Dionaea, Drosera, Drosophyllum, Nepenthes, and Cephalotus. In Utricularia and Genlisea, enzymatic secretion was independent of stimulation by prey. Byblis and Roridula are usually considered as “proto-carnivores”, lacking digestive enzymes. However, we found high activity of phosphatases in both species. Thus, they should be classified as true carnivores. We suggest that the inflorescence of Byblis and some Pinguicula species might also be an additional “carnivorous organ”, which can trap a prey, digest it, and finally absorb available nutrients.

Key words

Carnivorous plants - digestion - enzyme-labelled fluorescence - glandular cells - mutualism - enzymatic activity - trap functioning - Lentibulariaceae.

References

  • 1 Adamec L.. Mineral nutrition of carnivorous plants: a review.  The Botanical Review. (1997);  63 273-299
    PubMedGoogle Scholar
  • 2 Albert V. A., Stephen E. W., Chase M. W.. Carnivorous plants: phylogeny and structural evolution.  Science. (1992);  257 1491-1495
    PubMedGoogle Scholar
  • 3 Anderson B.. Adaptation to foliar absorption of faeces: a pathway in plant carnivory.  Annals of Botany. (2005);  95 757-761
    CrossrefPubMedGoogle Scholar
  • 4 Anderson B., Midgley J. J.. It takes two to tango but three is a tangle: mutualists and cheaters on the carnivorous plant Roridula.  Oecologia. (2002);  132 369-373
    CrossrefPubMedGoogle Scholar
  • 5 Beaver R. A.. The communities living in Nepenthes pitcher plants: fauna and food webs. Frank, J. H. and Lounibos, L. P., eds. Phytotelmata: Terrestrial Plants as Hosts for Aquatic Insect Communities. Medford; Plexus (1983): 129-159
    Google Scholar
  • 6 Bradshaw W.. Interaction between the mosquito Wyeomyia smithii, the midge Metriocnemus knabi, and their carnivorous host Sarracenia purpurea. Frank, J. H. and Lounibos, L. P., eds. Phytotelmata: Terrestrial Plants as Hosts for Aquatic Insect Communities. Medford; Plexus (1983): 161-189
    Google Scholar
  • 7 Broussaud F., Vintéjoux C.. Etudes ultrastructurales et cytochimiques des tissues superficiels placés à l'entrée des urnes d’Utricularia (Lentibulariacée).  Bulletin de la Société Botanique de France 129, Lettres Botaniques. (1982);  3 191-201
    PubMedGoogle Scholar
  • 8 Chróst R. J.. Environmental control of the synthesis and activities of aquatic microbial ectoenzymes. Chróst, R. J., ed. Microbial Enzymes in Aquatic Environments. New York; Springer Verlag (1991): 29-59
    Google Scholar
  • 9 Clancy F. G. A., Coffey M. D.. Acid phosphatase and protease release by the insectivorous plant Drosera rotundifolia.  Canadian Journal of Botany. (1976);  56 480-488
    PubMedGoogle Scholar
  • 10 Cohn F.. Über die Function der Blasen von Aldrovanda und Utricularia.  Beiträge der Biologie der Pflanzen. (1874);  3 71-92
    PubMedGoogle Scholar
  • 11 Cox G. W., Singer V. L.. A high-resolution, fluorescence-based method for localization of endogenous alkaline phosphatase activity.  Journal of Histochemistry and Cytochemistry. (1999);  47 1443-1455
    PubMedGoogle Scholar
  • 12 Darnowski D. W.. Triggerplants. Dural, New South Wales, Australia; Rosenberg Publishers (2002)
    Google Scholar
  • 13 Darnowski D. W., Carroll D., Stielper J. M.. Are triggerplants (Stylidium; Stylidiaceae) carnivorous? Proceedings of the 4th International Carnivorous Plant Conference, Tokyo, Japan, 2002. (2002): 209-213
    Google Scholar
  • 14 Darwin C. R.. Insectivorous Plants. London; J. Murray (1875)
    Google Scholar
  • 15 Darwin F.. Experiments on the nutrition of Drosera rotundifolia.  Journal of the Linnaean Society of Botany. (1978);  17 17-32
    PubMedGoogle Scholar
  • 16 Duff S. M. G., Sarath G., Plaxton W. C.. The role of acid phosphatases in plant phosphorus metabolism.  Physiologia Plantarum. (1994);  90 791-800
    PubMedGoogle Scholar
  • 17 Ellis A. G., Midgley J. J.. A new plant-animal mutualism involving a plant with sticky leaves and resident hemipteran.  Oecologia. (1996);  106 478-481
    CrossrefPubMedGoogle Scholar
  • 18 Ellison A. M., Gotelli N. J., Brewer J. S., Cochran-Stafira D. L., Kneitel J. M., Miller T. E., Worley A. C., Zamora R.. The evolutionary ecology of carnivorous plants.  Advances in Ecological Research. (2003);  33 1-74
    PubMedGoogle Scholar
  • 19 Feder J.. The phosphatases. Griffith, E. J., ed. Environmental Phosphorus Handbook. New York; J. Willey (1973): 475-505
    Google Scholar
  • 20 Fineran B. A.. Glandular trichomes in Utricularia: a review of their structure and function.  Israel Journal of Botany. (1985);  34 295-330
    PubMedGoogle Scholar
  • 21 Fineran B. A., Lee M. S. L.. Organization of quadrifid and bifid hairs in the trap of Utricularia monanthos.  Protoplasma. (1975);  84 43-70
    CrossrefPubMedGoogle Scholar
  • 22 Fish D.. Phytotelmata: Flora and Fauna. Frank, J. H. and Lounibos, L. P., eds. Phytotelmata: Terrestrial Plants as Hosts for Aquatic Insect Communities. Medford; Plexus (1983): 1-27
    Google Scholar
  • 23 Friday L. E., Quarmby C.. Uptake and translocation of prey-derived 15N and 32P in Utricularia vulgaris L.  New Phytologist. (1994);  126 273-281
    CrossrefPubMedGoogle Scholar
  • 24 Givnish T., Burkhardt E., Happel R., Weintraub J.. Carnivory in the bromeliad Brocchinia reducta, with a cost/benefit model for the general restriction of carnivorous plants to sunny, moist, nutrient-poor habitats.  The American Naturalist. (1984);  124 479-497
    CrossrefPubMedGoogle Scholar
  • 25 Gonzalez J. M., Jaffe K., Michelangeli F.. Competition for prey between the carnivorous Bromeliaceae Brocchinia reducta and Sarraceniaceae Heliamphora nutans.  Biotropica. (1991);  23 602-604
    CrossrefPubMedGoogle Scholar
  • 26 Hammond J. P., Broadley M. R., White P. J.. Genetic responses to phosphorus deficiency.  Annals of Botany. (2004);  94 323-332
    CrossrefPubMedGoogle Scholar
  • 27 Hanslin H. M., Karlsson P. S.. Nitrogen uptake from prey and substrate as affected by prey capture level and plant reproductive status in four carnivorous plant species.  Oecologia. (1996);  106 370-375
    CrossrefPubMedGoogle Scholar
  • 28 Hartmeyer S.. Carnivory of Byblis revisited - a simple method for enzyme testing on carnivorous plants.  Carnivorous Plant Newsletter. (1997);  26 39-45
    PubMedGoogle Scholar
  • 29 Hartmeyer S.. Carnivory in Byblis revisited II: the phenomenon of symbiosis on insect trapping plants.  Carnivorous Plant Newsletter. (1998);  27 110-113
    PubMedGoogle Scholar
  • 30 Henry Y., Steer M. W.. Acid phosphatase localization in the digestive glands of Dionaea muscipula Ellis flytrap.  Journal of Histochemistry and Cytochemistry. (1985);  33 339-344
    PubMedGoogle Scholar
  • 31 Heslop-Harrison Y.. Enzyme release in carnivorous plants. Dingle, J. T. and Dean, R. T., eds. Lysozymes in Biology and Pathology. Amsterdam; North Holland Publishing Company (1975): 525-578
    Google Scholar
  • 32 Heslop-Harrison Y.. Enzyme secretion and digest uptake in carnivorous plants. Sunderland, N., ed. Perspectives in Experimental Biology , S.E.B. Symposial Volume 2,. Oxford; Pergamon Press (1976 a): 463-476
    Google Scholar
  • 33 Heslop-Harrison Y.. Carnivorous plants a century after Darwin.  Endeavour. (1976 b);  35 114-122
    CrossrefPubMedGoogle Scholar
  • 34 Heslop-Harrison Y., Knox R. B.. A cytochemical study of the leaf-gland enzymes of insectivorous plants of the genus Pinguicula.  Planta. (1971);  96 183-211
    CrossrefPubMedGoogle Scholar
  • 35 Istock C. A., Tanner K., Zimmer H.. Habitat selection by the pitcher plant mosquito, Wyeomyia smithii: behavioral and genetic aspects. Frank, J. H. and Lounibos, L. P., eds. Phytotelmata: Terrestrial Plants as Hosts for Aquatic Insect Communities. Medford; Plexus (1983): 191-204
    Google Scholar
  • 37 Jobson R. W., Morris E. C., Burgin S.. Carnivory and nitrogen supply affect the growth of the bladderwort Utricularia uliginosa.  Australian Journal of Botany. (2000);  48 549-560
    CrossrefPubMedGoogle Scholar
  • 38 Jobson R. W., Playford J., Cameron K. M., Albert V. A.. Molecular phylogenetics of Lentibulariaceae inferred from plastid rps16 intron and trnL‐F DNA sequences: implications for character evolution and biogeography.  Systematic Botany. (2003);  28 157-171
    PubMedGoogle Scholar
  • 39 Juniper B. E., Robins R. J., Joel D. M.. The Carnivorous Plants. London; Academic Press (1989)
    Google Scholar
  • 40 Karlsson P. S., Carlsson B.. Why does Pinguicula vulgaris L. trap insects?.  New Phytologist. (1984);  97 25-30
    CrossrefPubMedGoogle Scholar
  • 41 Karlsson P. S., Pate J. S.. Contrasting effects of supplementary feeding of insects or mineral nutrients on the growth and nitrogen and phosphorus economy of pygmy species of Drosera.  Oecologia. (1992);  92 8-13
    CrossrefPubMedGoogle Scholar
  • 42 Krafft C. C., Handel S. N.. The role of carnivory in the growth and reproduction of Drosera filiformis and D. rotundifolia.  Bulletin of the Torrey Botanical Club. (1991);  118 12-19
    CrossrefPubMedGoogle Scholar
  • 44 Larison K. D., Bremiller R., Wells K. S., Clements I., Haugland R. P.. Use of a new fluorogenic phosphatase substrate in immunohistochemical applications.  Journal of Histochemistry and Cytochemistry. (1995);  43 77-83
    PubMedGoogle Scholar
  • 45 Lollar A. Q., Coleman D. C., Boyd C. E.. Carnivorous pathway of phosphorus uptake by Utricularia inflata.  Archiv für Hydrobiologie. (1971);  69 400-404
    PubMedGoogle Scholar
  • 46 Lloyd F. E.. The Carnivorous Plants. Waltham, MS, USA; Chronica Botanica Company (1942)
    Google Scholar
  • 47 Lowrie A.. Carnivorous Plants of Australia, Vol. 3. Nedlands; University of Western Australia Press (1998)
    Google Scholar
  • 48 Mabberley W. J.. The Plant Book. Cambridge; Cambridge University Press (2000)
    Google Scholar
  • 49 Müller K., Borsch T., Legendre L., Porembski S., Theisen I., Barthlott W.. Evolution of carnivory in Lentibulariaceae and the Lamiales.  Plant Biology. (2004);  6 477-490
    Article in Thieme ConnectPubMedGoogle Scholar
  • 50 Nedoma J., Štrojsová A., Vrba J., Komárková J., Šimek K.. Extracellular phosphatase activity of natural plankton studied with ELF97 phosphate: fluorescence quantification and labelling kinetics.  Environmental Microbiology. (2003);  5 462-472
    CrossrefPubMedGoogle Scholar
  • 51 Olczak M.. Acid phosphatases from higher plants.  Wiadomości Botaniczne. (1996);  40 39-51
    PubMedGoogle Scholar
  • 52 Otto C.. Effects of prey and turion size on the growth and turion production of the carnivorous bladderwort, Utricularia vulgaris L.  Archiv für Hydrobiologie. (1999);  145 469-478
    PubMedGoogle Scholar
  • 53 Płachno B. J., Jankun A.. Transfer cell wall architecture in secretory hairs of Utricularia intermedia Traps.  Acta Biologica Cracoviensia, Series Botanica. (2004);  46 193-200
    PubMedGoogle Scholar
  • 54 Płachno B. J., Jankun A., Faber J.. Development of the wall labyrinth in pavement epithelium hairs of some Utricularia species.  Acta Biologica Cracoviensia, Series Botanica. (2005 a);  47 109-113
    PubMedGoogle Scholar
  • 55 Płachno B. J., Adamus K., Faber J., Kozłowski J.. Feeding behaviour of carnivorous Genlisea plants in the laboratory.  Acta Botanica Gallica,. (2005 b);  152 159-164
    PubMedGoogle Scholar
  • 56 Richards J. H.. Bladder function in Utricularia purpurea (Lentibulariaceae): is carnivory important?.  American Journal of Botany. (2001);  88 170-176
    PubMedGoogle Scholar
  • 57 Robins R. J.. Studies in secretion and absorption in Dionaea muscipula Ellis. PhD Thesis, University of Oxford. (1978)
    Google Scholar
  • 58 Robins R. J., Juniper B. E.. The secretory cycle of Dionaea muscipula Ellis. IV. The enzymology of the secretion.  New Phytologist. (1980);  86 401-412
    CrossrefPubMedGoogle Scholar
  • 59 Sasago A., Sibaoka T.. Water extrusion in the trap bladders of Utricularia vulgaris. I. A possible pathway of water across the bladder wall.  Botanical Magazine (Tokyo). (1985);  98 55-66
    PubMedGoogle Scholar
  • 60 Schumacher G. J.. Further notes on the occurrence of desmids in Utricularia bladders.  Castanea. (1960);  25 62-65
    PubMedGoogle Scholar
  • 61 Sirová D., Adamec L., Vrba J.. Enzymatic activities in traps of four aquatic species of the carnivorous genus Utricularia.  New Phytologist. (2003);  159 669-675
    CrossrefPubMedGoogle Scholar
  • 62 Sydenham P. H., Findlay G. P.. Transport of solutes and water by resetting bladders of Utricularia.  Australian Journal of Plant Physiology. (1975);  2 335-351
    PubMedGoogle Scholar
  • 63 Štrojsová A., Vrba J., Nedoma J., Komárková J., Znachor P.. Seasonal study of extracellular phosphate expression in the phytoplankton of a eutrophic reservoir.  European Journal of Phycology. (2003);  38 295-306
    PubMedGoogle Scholar
  • 64 Štrojsová M., Vrba J.. Direct detection of digestive enzymes in planktonic rotifers using enzyme labelled fluorescence (ELF).  Marine and Freshwater Research. (2005);  56 189-195
    CrossrefPubMedGoogle Scholar
  • 65 Taylor P.. The Genus Utricularia - A Taxonomic Monograph. Kew Bulletin Additional Series 14. London; HMSO (1989)
    Google Scholar
  • 66 van Aarle I. M., Olsson P. A., Söderström B.. Microscopic detection of phosphatase activity of saprophytic and arbuscular mycorrhiza using a fluorogenic substrate.  Mycologia. (2001);  93 17-24
    PubMedGoogle Scholar
  • 67 Vintéjoux C.. Études des aspects ultrastructuraux de certaines cellules glandulaires en rapport avec leur activité secretive chez l’Utricularia neglecta L. (Lentibulariaceae).  Comptes Rendues. (1973);  277D 2345-2348
    PubMedGoogle Scholar
  • 68 Vintéjoux C.. Ultrastructural and cytochemical observations on the digestive glands of Utricularia neglecta L. (Lentibulariaceae). Distribution of protease and acid phosphatase activities.  Portugaliae Acta Biologica, Series A. (1974);  14 463-474
    PubMedGoogle Scholar
  • 69 Zamora R., Gómez J. M., Hódar J. A.. Responses of a carnivorous plant to prey and inorganic nutrients in a Mediterranean environment.  Oecologia. (1997);  111 443-451
    CrossrefPubMedGoogle Scholar

B. J. Płachno

Department of Plant Cytology and Embryology
The Jagiellonian University

Grodzka 52

31-044 Cracow

Poland

Email: bartek78pl@poczta.onet.pl

Guest Editor: S. Porembski