Abstract
Studies were undertaken in order to improve the micropropagation ofCymbidiun ensifolium 'Yuh Hwa’. It is easy toinduce rhizomes in vitro in this cultivar but not upright shoots, which areneeded for eventual production of a flowering plant. Exposure ofCymbidium ensifolium 'Yuh Hwa’ rhizomeexplants in vitro to greater than 2.5 μM BA for 12weeks or longer inhibited both elongation of upright, aerial shoots and rootgrowth. Through transfer experiments, it was determined that the commitment toinduction of upright shoots occurred after 10–14 days of exposure to 2.5μM BA. BA supplied at 20 μM during the first 14days of culture was sufficient to induce upright shoot formation, but shootelongation did not continue under these conditions. Using radiolabelled BA,adenine was found to be a major metabolite found in the rhizome tissue at day14. BA comprised 37% of the total DPM recovered in rhizomes grown in 20μM BA compared to that of 61% of the total DPM recoveredfrom rhizomes grown in 2.5 μM BA. The total amount of BA takenup per gram fresh weight was similar for both green and etiolated rhizomesgrownin 2.5 μM BA. Rhizomes grown in 20 μM BA tookupapproximately five times as much BA as those grown in 2.5 μMBA.Free IAA levels were highest in the rhizome tip (12.7 ng/g FW) anddecreased to one fourth the value at 1 cm behind the tip. Free IAAlevels increased initially in rhizornes transferred to medium with or without20μM BA (although at different amounts) and then decreased. Whenactivated charcoal was added to the medium as well as BA there was no initialincrease in IAA. When 20 μM BA was added to the medium, solubleperoxidase activity increased while IAA concentrations remained at about thesame level over a 7-day period. Peroxidase activity in rhizomes eitherdecreasedor remained constant when activated charcoal was added to the medium containingBA.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Auer C.A., Cohen J.D., Lalou M. and Cooke T.J. 1992. Comparison of benzyl adenine metabolism in two Petunia hybrida lines differing in shoot organogenesis. Plant Physiol 98: 1035–1041.
Auer C.A., Laloue M., Cohen J.D. and Cooke T.J. 1992. Uptake and metabolism of benzyladenine during shoot organogenesis in Petunia leaf rhizomes. Plant Growth Regul. 11: 105–114.
Auer C.A., Motyka V., Brezinova A. and Kaminek M. 1999. Endogenous cytokinin accumulation and cytokinin oxidase activity during shoot organogenesis of Petunia hybrida. Physiol Plant 105: 141–147.
Biondi S.L., Canciani S.L. and Bagni N. 1984. Uptake and translocation of benzyladenine by elm shoots cultured in vitro. Can. J. Bot. 62: 2385–2390.
Bond S. and Alderson P.G. 1993. The effect of explant density, temperature and light on rhizome growth in vitro of Alstroemeria. J. Hort. Sci. 68: 855–859.
Bradford K.J. and Trewavas K.J. 1994. Sensitivity thresholds and variable time scales in plant hormone action. Plant Physiology 105: 1029–1036.
Chen K.-H., Miller A.N., Patterson G.W. and Cohen J.D. 1988. A rapid and simple procedure for purification of indole-3-acetic acid prior to GC-SIM-MS analysis. Plant Physiol 86: 822–825.
Faivre-Rampant O., Kevers C. and Gaspar T. 2000. IAA-oxidase activity and auxin protectors in nonrooting, rac, mutant shoots of tobacco in vitro. Plant Science 153: 73–80.
Gabryszewska E. 1996. The influence of temperature, daylength and sucrose concentation on the growth and development of Alstroemeria “Zebra” in vitro. Acta. Agrobotanica. 49: 131–140.
Gaspar T., Kevers C., Hausman J.F., Berthon J.Y. and Ripetti V. 1992. Practical uses of peroxidase activity as a predictive marker of rooting performance of micropropagated shoots. Agronomie. 12: 757–765.
Hasegawa A., Ohashi H. and Goi M. 1985. Effects of BA, rhizome Iength, mechanical treatment and liquid shaking culture on the shoot formation from rhizome in Cymbidium ensifolium faberi Rolfe. Acta Horticulturae. 166: 25–40.
Hausman J.F. 1993. Changes in peroxidase activity, auxin level and ethylene production during root formation by poplar shoots raised in vitro. Plant Growth Regul. 13: 263–268.
Lagrimini L.M., Gingas F., Finger F., Rothstein S. and Liu T.T.Y. 1997. Characterization of antisense transformed plants deficient in the tobacco anionic peroxidase. Plant Physiol 114: 1187–1196.
Lu I.-L. 1988. In vitro seed germination and organogenesis of Cymbidium ensifolium var misericors and Cymbidiun ensifolium dayanum. Master, National Taiwan University.
Lu I.-L. 1993. The roles of various growth regulators in the organogenesis and gravitropism of Cymbidium ensifolium cv Yuh Hwa rhizomes. Ph D dissertation, University of California, Davis.
Murashige T. and Skoog F. 1962. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15: 473–497.
Nissen S. and Sutter E.G. 1990. Stability of IAA and IBA in nutrient medium to severa] tissue culture procedures. HortScience 25: 800–802.
Normanly J., Slovin J.P. and Cohen J.D. 1995. Rethinking auxin biosynthesis and metabolism. Plant Physiol 107: 323–329.
Paek K.Y. and Yeung E.C. 1991. The effects of 1-naphthaleneacetic acid and N6-benzyladenine on the growth of Cymbidium eusifolium forrestii rhizomes in vitro. Plant Cell Tissue and Organ Culture 24: 65–71.
Peres L.E.P. and Kerbauy G.B. 1999. High cytokinin accumulation following root tip excision changes the endogenous auxin-tocytokinin ratio during root-to-shoot conversion in Catasetum fimbriatum Lindl (Orchidaceae). Plant Cell Rep. 18: 1002–1006.
Podwyszynska M., Gabryszewska E. and Przybyla A. 1998. Effect of growth retardants, cytokinins and auxins on the multiplication and rooting in vitro of Alstroemeria x hybrida “Juanita”. Acta Agrobotanica. 51: 23–31.
Quesada M.A., Sanchez-Roldan C., Heredia A., Valpuesta V. and Bukovac M.J. 1992. Peroxidase and IAA oxidase activities and peroxidase isoenzymes in the pericarp of seeded and seedless “Redhaven” peach fruit. J. Plant Growth Regul. 11: 1–6.
Shimasaki K. and Uemoto S. 1991. Rhizome induction and plantlet regeneration of Cymbidium ensifolium goeringii from flower bud cultures in vitro. Plant Cell, Tissue and Organ Culture 25: 49–52.
Skoog F. and Miller C.O. 1957. Chemical regulation of growth and organ formation in plant tissues cultured in vitro. Symp. Soc. Exp. Biol. 11: 118–131.
Strnad M. 1997. The aromatic cytokinins. Physiol Plant 101: 674–688.
Ueda H. and Torikata H. 1969. Organogenesis in the meristem tissue cultures of Cymbidium ensifolium II Effects of growth substances on the organogenesis in dark culture. J. Japan Soc. Hort. Sci. 38: 188–193.
Van Staden J. and Crouch N.R. 1996. Benzyladenine and derivatives - their significance and interconversion in plants. Plant Growth Regulation 19: 153–175.
Yip W.-K. and Yang S.F. 1986. Effect of thidiazuron, a cytokininactive urea derivative, in cytokinin-dependent ethylene production systems. Plant Physiol 80: 515–519.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Lu, Il., Sutter, E. & Burger, D. Relationships between benzyladenine uptake, endogenous free IAA levels and peroxidase activities during upright shoot induction of Cymbidium ensifoilum cv. Yuh Hwa rhizomes in vitro . Plant Growth Regulation 35, 161–170 (2001). https://doi.org/10.1023/A:1014430715746
Issue Date:
DOI: https://doi.org/10.1023/A:1014430715746