Biology and Fertility of Soils

, Volume 15, Issue 2, pp 127–131 | Cite as

Effect of earthworm humic substances on esterase and peroxidase activity during growth of leaf explants of Nicotiana plumbaginifolia

  • A. Muscolo
  • M. Felici
  • G. Concheri
  • S. Nardi


Leaf explants of Nicotiana plumbaginifolia were compared in cultures supplemented with hormones or humic substances (extracted from faeces of Allolobophora caliginosa) of various molecular complexity and concentration. The results showed that the humic substances (F1, F2, and T) at the concentration of 1 mg C l-1 produced greater leaf explants than those grown in the control. Furthermore, humic fractions like gibberellic alone induced a rhizogenic activity in leaf explants. Quantitative differences were also observed in the peroxidase activity induced in Nicotiana plumbaginifolia by humic matter (F2, F2, and T). In addition, the Nicotiana sp. tissue treated with humic fractions revealed, in the esterase enzyme pattern, the appearance of the 2a band, which was attributable to indoleacetic acid, since its profile was consistent with those obtained from tissues treated with indoleacetic acid. These differences demonstrate that humic substances exhibited a hormone-like behaviour, but no evidence of a relationship between biological activities and chemical characteristics of humus substances was found.

Key words

Nicotiana plumbaginifolia Tissue culture Esterase Peroxidase Humic fractions Earthworms 


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  1. Bradford MM (1976) A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254Google Scholar
  2. Coppens L, Dewitte B (1990) Esterase and peroxidase zymograms from barley (Hordeum vulgare L.) callus as a biochemical marker system of embryogenesis and organogenesis. Plant Sci 67:97–105Google Scholar
  3. Davis BJ (1964) Disc electrophoresis. II. Method and application to human serum proteins. Ann NY Acad Sci 121:404–427Google Scholar
  4. Dell'Agnola G, Ferrari G (1971) Effect of humic acids on anion uptake by excised barley roots. In: Novak B, Macura J, Kutilek M, Pakorna-Kozova J, Tichy V (eds) Studies about humus. Trans Int Symp Humus et Planta V, 13–17 September, Prague, pp 567–570Google Scholar
  5. Dell'Agnola G, Nardi S (1987) Hormone-like effect and enhanced nitrate uptake induced by depolycondensed humic fractions obtained from Allolobophora rosea and A. caliginosa faeces. Biol Fertil Soils 4:115–118Google Scholar
  6. Iunge W, Klees H (1984) Peroxidases. In: Bergmeyer HV (ed) Methods of enzymatic analysis IV, 3rd edn. Academic Press, New York, pp 8–14Google Scholar
  7. Kay LE, Basile DV (1987) Specific peroxidase isoenzymes are correlated with organogenesis. Plant Physiol 84:99–105Google Scholar
  8. Krsnik-Rasol M, Jalaska S, Sermann D (1982) Isoperoxidases: Early indicators of somatic embryoid differentiation in pumpkin tissue. Acta Bot Croat 41:33–39Google Scholar
  9. Ladd JN, Butler JHA (1971) Inhibition by soil humic acids of native and acetylated proteolytic enzymes. Soil Biol Biochem 3:157–160Google Scholar
  10. Lavce S, Galston AW (1968) Structural, physiological and biochemical gradients in tobacco pith tissue. Plant Physiol 43:1760–1768Google Scholar
  11. Lee TT (1972) Interaction of cytokinin, auxin, and gibberellin on peroxidase isoenzymes in tobacco tissues cultured in vitro. Can J Bot 50:2471–2477Google Scholar
  12. Lee TT (1973) On extraction and quantitation of plant peroxidase isoenzymes. Physiol Plant 29:198–203Google Scholar
  13. Malcolm RE, Vaughan D (1979) Humic substances and phosphatase actities in plant tissues. Soil Biol Biochem 11:253–259Google Scholar
  14. Murashige T, Skoog K (1962) A revised medium for rapid growth and bioassay with tobacco tissue cultures. Physiol Plant 15:473–497Google Scholar
  15. Nardi S, Arnoldi G, Dell'Agnola G (1988) Release of the hormone-like activities from Allolobophora rosea (Sav.) and Allolobophora caliginosa (Sav.) feces. Can J Soil Sci 68:563–567Google Scholar
  16. Piccolo A, Nardi S, Concheri G (1992) Structural characteristics of humus and biological activity. Soil Biol Biochem 24:373–380Google Scholar
  17. Putter J (1974) Peroxidases. In: Bergmeyer HV (ed) Methods of enzymatic analysis II. Academic Press, New York, pp 685–690Google Scholar
  18. Shaw CR (1970) Starch gel electrophoresis of enzymes: A compilation of recipes. Biochem Genet 4:297–370Google Scholar
  19. Stevenson FJ (1982) Humus chemistry: Genesis, composition, reactions. Wiley, New York Chichester Brisbane Toronto Singapore, pp 26–42Google Scholar
  20. Van Huystee RB, Cairns WL (1982) Progress and prospects in the use of peroxidase to study cell development. Phytochemistry 21: 1843–1847Google Scholar
  21. Vaughan D, Malcolm RE (1979) Effect of humic acid on invertase synthesis in roots of higher plants. Soil Biol Biochem 11:247–252Google Scholar
  22. Vaughan D, Malcolm RE (1985) Influence of humic substances on growth and physiological processes. In: Vaughan D, Malcolm RE (eds) Soil organic matter and biological activity. Martinus Nijhoff/Dr W Junk Publishers. Dordrecht Boston Lancaster, pp 37–76Google Scholar
  23. Vaughan D, Ord BG, Malcolm RE (1978) Effect of soil organic matter on some root surface enzymes and uptakes into winter wheat. J Exp Bot 29:1337–1344Google Scholar
  24. Vaughan D, Malcolm RE, Ord BG (1985) Influence of humic substances on biochemical processes in plants. In: Vaughan D, Malcolm RE (eds) Soil organic matter and biological activity. Martinus Nijhoff/Dr W Junk Publishers, Dordrecht Boston Lancaster, pp 77–108Google Scholar
  25. Verma DPS, Van Huystee RB (1970) Cellular differentiation and peroxidase isozymes in cell culture of peanut cotyledons. Can J Bot 48:429–431Google Scholar
  26. Wochok SZ, Burleson B (1974) Isoperoxidase activity and induction in culture tissue of wild carrot: A comparison of proembryos and embryos. Physiol Plant 31:73–75Google Scholar
  27. Wolter KE, Gordon JC (1975) Peroxidases as indicators of growth and differentiation in aspen callus cultures. Physiol Plant 33:219–223Google Scholar

Copyright information

© Springer-Verlag 1993

Authors and Affiliations

  • A. Muscolo
    • 1
  • M. Felici
    • 1
  • G. Concheri
    • 2
  • S. Nardi
    • 2
  1. 1.Istituto di Chimica Agraria e ForestaleUniversità degli Studi di Reggio CalabriaItaly
  2. 2.Dipartimento di Biotecnologie AgrarieUniversità degli Studi di PadovaPadovaItaly

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