Can Changes in Starch Content and Peroxidase Activity Be Used as Rooting Phase Markers for Rhododendron Leaf Bud Cuttings?
We examined whether peroxidase activity in cutting bases and leaves and starch content in cutting bases can be used as rooting phase markers in the elepidote rhododendron cv. ‘Babites Baltais’ (Rhododendron L.). Changes in peroxidase activity in cutting leaves and bases, as well as starch content in cutting bases, were determined in relation to anatomical stages of rhizogenesis in leaf bud cuttings treated with 1% indole-3-butyric acid (IBA+) or without IBA (IBA-). The pattern of change of peroxidase activity was similar in cutting bases and leaves of IBA- leaf bud cuttings. Three phases of adventitious root formation were identified: induction, initiation and expression. During the induction phase peroxidase activity decreased, but no anatomical changes were observed in the cuttings. Peroxidase activity increased in the initiation phase when adventitious root initials were formed. Peroxidase activity decreased during the expression phase when adventitious root primordia developed. The starch content of IBA- leaf bud cuttings decreased during the first few days and then gradually rose to maximum, followed by a sharp reduction and another increase at the end of the experiment. The changes of starch content did not coincide with rooting phases as peroxidase activity did, and cannot be used as a rooting phase marker in rhododendrons. Adventitious root formation did not occur in IBA+ leaf bud cuttings, so distinct rooting phases could not be observed. There was a significant correlation between peroxidase activity in cutting bases and leaves of IBA- leaf bud cuttings. Peroxidase activity in leaves of rhododendron leaf bud cuttings are potentially useful as a marker for rooting phases, but that requires further anatomical and physiological study of rooting in leaf bud cuttings.
Aghmir A, Kevers C, Hausmann JF, and Gaspar T. 1991. Peroxidases, compartimentation cellulaire et enracinement in vitro de pousses de Rhododendron catawbiense Michaux cv. album. Archives Internationales de Physiologie et de Biochimie 99: 9.
Ahkami AH, Lischewski S, Haensch KT, Porfirova S, Hofmann J, Rolletschek H, Melzer M, Franken P, Hause B, Druege U, and Hajirezaei MR. 2009. Molecular physiology of adventitious root formation in Petunia hybrida cuttings: involvement of wound response and primary metabolism. New Phytologist 181: 613-625.
Andersone U, and Ievinsh G. 2002. Changes of morphogenic competence in mature Pinus sylvestris L. buds in vitro. Annals of Botany 90: 293-298.
Apine I, and Kondratovics U. 2005. Effect of environmental factors on the propagation of deciduous azalea by cuttings. II. Influence of an extended growth period on budbreak, overwinter survival and carbohydrate levels of rooted cuttings. Acta Universitatis Latviensis 691: 41-50.
Arena ME, Pastur GM, Benavides MP, Zappacosta D, Eliasco E, and Curvetto N. 2003. Peroxidase and polyamine activity during the in vitro rooting of Berberis buxifolia. New Zealand Journal of Botany 41: 475-485.
Braune W, Leman A, and Taubert H. 1999. Pflanzen-anatomisches Praktikum I. Spektrum Akademischer Verlag Heidelberg, Berlin.
Davies FT, JR, Lazarte JE, and Joiner JN. 1982. Initiation and development of roots in juvenile and mature leaf bud cuttings of Ficus pumila L. Amererican Journal of Botany 69: 804-811.
De Klerk GJ, Brugge JT, and Marinova S. 1997. Effectiveness of indoleacetic acid, indolebutyric acid and naphthaleneacetic acid during adventitious root formation in vitro in Malus ‘Jork 9’. Plant Cell, Tissue and Organ Culture 49: 39-44.
Dirr MA, and Heuser Jr CW. 1987. The Reference Manual of Woody Plant Propagation: From Seed to Tissue Culture. Varsity Press, Athens.
Fekete S, Mándy A, and Stefanovits-Bányai E. 2002. Change of peroxidase enzyme activities in annual cuttings during rooting. Proceedings of the 7th Hungarian Congress on Plant Physiology. Acta Biologica Szegediensis 46: 29-31.
Gaspar T, Kevers C, Hausman JF, Berthon JY, and Ripetti V. 1992. Practical use of peroxidase activity as a predictive marker of rooting performance of micropropagated shoots. Agronomie 12: 757-65.
Gaspar T, Kevers C, Hausman JF, and Ripetti V. 1994. Peroxidase activity and endogenous free auxin during adventitious root formation. In: Lumsden PJ, Nicholas JR, Davies WJ [eds.], Physiology, Growth and Development of Plants in Culture, 289-298. Kluwer Academic Publishers, Dordrecht.
Goreau T. 1980. Rhododendron propagation. Proceedings of Plant Propagator Society 30: 532-537.
Grönroos R, and Von Arnold S. 1987. Initiation of roots on hypocotyl cuttings of Pinus contorta in vitro. Physiologia Plantarum 69: 227-236.
Haissig BE. 1989. Carbohydrate relations during propagation of cuttings from sexually mature Pinus banksiana trees. Tree Physiology 5: 319-328.
Hartmann HT, Kester DE, Davies FT, and Geneve RL. 2002. Hartmann and Kester's Plant Propagation: Principles and Practice, 7th edn. Prentice Hall, Upper Saddle River, New Jersey.
Hatzilazarou SP, Syros TD, Yupsanis TA, Bosabalidis AM, and Economou AS. 2006. Peroxidases, lignin and anatomy during in vitro and ex vitro rooting of gardenia (Gardenia jasminoides Ellis) microshoots. Journal of Plant Physiology 163: 827-836.
Husen A, and PAL M. 2007. Metabolic changes during adventitious root primordium development in Tectona grandis Linn. f. (teak) cuttings as affected by age of donor plants and auxin (IBA and NAA) treatment. New Forests 33: 309-323.
Klopotek Y, Haensch KT, Hause B, Hajirezaei MR, and Druege U. 2010. Dark exposure of petunia cuttings strongly improves adventitious root formation and enhances carbohydrate availability during rooting in the light. Journal of Plant Physiology 167: 547-554.
Leslie A. 2006. The International Rhododendron Register and Checklist 2004: First Supplement. Royal Horticultural Society, Great Britain.
Li M, and Leung DWM. 2000. Starch accumulation is associated with adventitious root formation in hypocotyl cuttings of Pinus radiate. Journal of Plant Growth Regulation 19: 423-428.
Ludwig-Müller J. 2003. Peroxidase isoenzimes as markers for the rooting ability of easy-to-root and difficult-to-root Grevillea species and cultivars of Protea obstusifolia (Proteaceae). In Vitro Cellular & Developmental Biology - Plant 39: 377-383.
Metaxas D, Syros T, Yupsanis T, and Economou AS. 2004. Peroxidases during adventitious rooting in cuttings of Arbutus unedo and Taxus baccata as affected by plant genotype and growth regulator treatment. Plant Growth Regulation 44: 257-66.
Naija S, Elloumi N, Jbir N, Ammar S, and Kevers C. 2008. Anatomical and biochemical changes during adventitious rooting of apple rootstocks MM 106 cultured in vitro. Comptes Rendus Biologies 331: 518-525.
Nawrocka-Grześkowiak U. 2004. Effect of growth substances on the rooting of cuttings of rhododendron species. Folia Horticulturae 16: 115-123.
Rival A, Bernard F, and Mathieu Y. 1997. Changes in peroxidase activity during in vitro rooting of oil palm (Elaeis guineensis Jacq.). Scientia Horticulturae 71: 103-112.
Rout GR, Samantaray S, and DAS P. 2000. In vitro rooting of Psoralea corylifolia Linn: peroxidase activity as a marker. Plant Growth Regulation 305: 215-219.
Ruzin SE. 1999. Plant Mictotechnique and Microscopy. University Press, New York Oxford.
Sanders CH. 1978. Some aspects of propagation of Rhododendron, Mahonia and Ilex by cuttings. Proceedings of International Plant Propagator Society 28: 228-232.
Strong FM, and Koch GH. 1974. Biochemistry Laboratory Manual, 2nd ed. Wm. C. Brown, Dubuque, IA.
Syros T, Yupsanis T, Zafiriadis H, and Economou AS. 2004. Activity and isoforms of peroxidases, lignin and anatomy, during adventitious rooting in cuttings of Ebenus cretica L. Journal of Plant Physiology 161: 69-77.
Veierskov B. 1988. Relations between carbohydrates and adventitious root formation. In: Dudley TR [ed.], Adventitious Root Formation in Cuttings, 70-78. Advances in Plant Sciences Series, vol. 2, Dioscorides Press, Portland, OR.