In-vitro cormel formation and changes of biochemical composition in calli during morphogenesis in gladiolus (Gladiolus × grandiflorus)
287 / 155
Authors
-
KOUSHIK DUTTA
Uttar Banga Krishi Viswavidhyalaya, Pundibari, Cooch Behar West Bengal 736 165
-
SUBHENDU S GANTAIT
Uttar Banga Krishi Viswavidhyalaya, Pundibari, Cooch Behar West Bengal 736 165
Keywords:
Callus, Cormel tip, Gladiolus × grandiflorus, In-vitro cormel formation, Metabolitic variation, RegenerationAbstract
The present investigation was carried out for in-vitro cormel formation and to study the changes of biochemical composition during shoot and root differentiation from callus in gladiolus (Gladiolus × grandifforus L.). Cormel tip was used as explant for in-vitro regeneration on MS media supplemented with different plant growth regulators. Maximum callus formation was observed on MS medium supplemented with 4.0 mg/l 2,4-D with excellent callus growth. The inducted calli were subcultured on MS medium supplemented with 4.0 mg/l BAP and 0.5 mg/l NAA for best shooting followed by on ½ MS medium fortified with 3.0 mg/l IAA for best rooting. After two weeks of root initiation, the whole cluster of rooted plantlets was equally divided into two halves and the divided clusters were subcultured on cormel induction media. The different stages of morphogenesis, viz. non-differentiating callus, differentiating green callus, shoot differentiating callus and root differentiating callus were used for biochemical analysis. Maximum (55.0 %) and earliest (56.0 days) formation of cormels was observed on MS medium supplemented with 4.0 mg/l NAA elevated with 6% sucrose with good cormel size of up to 7.0 mm diameter. Decrease in metabolities like starch, reducing sugars and total phenols while, increase in total soluble proteins was found during shoot and root differentiation. Activities of enzymes, viz. polyphenol oxidase and peroxidase increased during shoot and root differentiation. From the present investigation, it was proved that metabolic and enzymatic activity during in-vitro morphogenesis help gladiolus for rapid organogenesis and multiplication through cormel formation.
Downloads
References
Ahmad T, Ahmad M S, Nasir I A and Riazuddin S. 2000. In vitro production of cormels in gladiolus. Pakistan Journal of Biological Science 3(5): 819–21. DOI: https://doi.org/10.3923/pjbs.2000.819.821
Amorim H A, Dougall D K and Sharp W R. 1977. The effect of carbohydrate and nitrogen concentration on phenol synthesis in Paul’s Scarlet Rose cells grown in tissue culture. Physiologia Plantarum 39: 91–5. DOI: https://doi.org/10.1111/j.1399-3054.1977.tb09291.x
Beura S, Singh R and Jagadev PN. 2003. In vitro multiplication studies in gladiolus cv. American Beauty. Orissa Journal of Horticulture, 31: 101–5.
Bradford M M. 1976. A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72: 248–54. DOI: https://doi.org/10.1016/0003-2697(76)90527-3
Chawla H S. 1991. Regeneration potentiality and isoenzyme variation during morphogenesis of barely callus. Biologia Plantarum 33: 175–80. DOI: https://doi.org/10.1007/BF02897878
Carrillo-Castaneda G and Mata A. 2000. Succession of esterase and peroxidase isozymes associated with the in vitro sugarcane tissue dedifferentiation and shoot induction. Biotechnologia Aplicada. 17: 225–30.
Dharmasena P A I U, Karunananda D P and Eeswara J P. 2011. Effect of gibberellic acid (GA3) and sugar on in vitro cormlet formation, multiplication and ex vitro sprouting of sladiolus hybrida variety princess Lee. Tropical Agricultural Research. 23: 1–10. DOI: https://doi.org/10.4038/tar.v23i1.4626
Dutta Gupta S and Datta S. 2003. Antioxidant enzyme activities during in vitro morphogenesis of gladiolus and the effect of application of antioxidants on plant regeneration. Biologia Plantarum 47: 179–83. DOI: https://doi.org/10.1023/B:BIOP.0000022248.62869.c7
Goo D.H., Joung H.Y. and Kim K.W., 2003. Differentiation of gladiolus plantlets from callus and subsequent flowering. Acta Horticultural. 620: 339–42. DOI: https://doi.org/10.17660/ActaHortic.2003.620.42
Handa A K, Bressan R A, Handa S and Hasegawa P M. 1982. Characteristics of cultured tomato cells after prolonged exposure to medium containing polyethylene glycol. Plant Physiology. 69: 514–21. DOI: https://doi.org/10.1104/pp.69.2.514
Hasbullah N A, Saleh A and Taha R M. 2011. Establishment of somatic embryogenesis from Gerbera jamesonii Bolus EX. Hook F. through suspension culture. African Journal of Biotechnology. 10(63): 13 762–8 DOI: https://doi.org/10.5897/AJB11.1653
Hammerschmidt R, Nuckles E M and Kuae J. 1982. Association of enhanced peroxidase activity with induced systemic resistance of cucumber to Colletorichum lagenarium. Physiological Plant Pathology, 20: 73–82. DOI: https://doi.org/10.1016/0048-4059(82)90025-X
Hassid W Z and Neufeld B F. 1964. Quantitative determination of starch in plant tissue. Methods in Carbohydrate Chemiistry. 4: 33.
Jala A. 2013. Potential of benzyl adenine, naphthalene acetic acid and sucrose concentration on growth, development, and regeneration of new shoot and cormel on gladiolus. American Transactions on Engineering & Applied Sciences 2: 277–85.
Jana S and Shekhawat G S. 2012. In vitro regeneration of Anethum graveolens, antioxidative enzymes during organogenesis and RAPD analysis for clonal fidelity. Biologia Plantarum 56: 9– 14. DOI: https://doi.org/10.1007/s10535-012-0009-2
Mader M and Fussel R. 1982. Role of peroxidise in lignification of tobacco cells. Plant Physiology 70: 1 132–4. DOI: https://doi.org/10.1104/pp.70.4.1132
Mangat B S, Pelekis M K and Cassells A C. 2006. Changes in the starch content during organogenesis in in vitro cultured Begonia rex stem explants. Physiologia Plantarum 79: 267– 74. DOI: https://doi.org/10.1034/j.1399-3054.1990.790209.x
Mayer A M Harel E and Shaul R B. 1965. Assay of catechol oxidase -a critical comparison of methods. Phytochemistry, 5: 783–9. DOI: https://doi.org/10.1016/S0031-9422(00)83660-2
Memon N, Qasim M, Jaskani M J and Ahmad R. 2010. In vitro cormel production of gladiolus. Pakistan Journal of Agricultural Science 47: 115–23.
Misra P, Pandey V and Kochhar S. 2010. Study of antioxidant enzymes activity during organogenesis and in vitro propagation of Asiatic hybrid lily. Journal of Plant Biochemistry and Biotechnology 19: 119–22. DOI: https://doi.org/10.1007/BF03323449
Misra R L. 1994. Propagation of gladiolus through sprouts-An entirely new method in gladiolus propagation. (In) Prakash J and Bhandary K R (Eds). Floriculture- Technology, Trades and Trends pp 67–70. Oxford & IBH Pub Co. Pvt Ltd.
Mohapatra H P and Rath S P. 2005. In vitro studies of Bacopa monnieri- An important medicinal plant with reference to its biochemical variations. Indian Journal of . Experimental Biology 43: 373–6.
Panigrahi J, Behera M, Maharana S and Mishra R R. 2007. Biomolecular changes during in vitro organogenesis of Asteracantha longifolia (L.) Nees ¯# A medicinal herb. Indian Journal. of Experimental Biology 45: 911–9.
Prasad V S S and Gupta S D. 2006. In vitro shoot regeneration of gladiolus in semisolid agar versus liquid cultures with support systems. Plant Cell Tissue and Organ Culture 87: 263–71. DOI: https://doi.org/10.1007/s11240-006-9160-9
Rathod D, Patel A and Shrimali G. 2014. Biochemical changes during in vitro organogenesis of Tylophora indica (Burm. F.) Merrill. Indian Journal of Applied Research 4: 274–7. DOI: https://doi.org/10.15373/2249555X/JAN2014/79
Samantaray S, Rout G R and Das P. 1999. Changes in isozyme patterns during in vitro regeneration from cotyledon explants of Brassica Species, Biologia Plantarum 42: 169. DOI: https://doi.org/10.1023/A:1002196231904
Sharma R K, Ahujaa A M, Sharada and Govilb C M. 2009. Biochemical changes associated with morphogenesis in saffron. (In) Plant Tissue Culture and Molecular Marker, pp 531–47. Kumar A and Shekhawat N S (Eds). I. K. International Publisher, New Delhi.
Shaheenuzzaman M, Haque M S, Karim M M and Noor Z U. 2011. In vitro shoot proliferation and development of micropropagation protocol from leaf disc of gladiolus. Journal of. Bangladesh Agrilcultural University 9: 21–6. DOI: https://doi.org/10.3329/jbau.v9i1.8739
Sharifi G. and Ebrahimzadeh H. 2010. Changes of antioxidant enzyme activities and isoenzyme profiles during in vitro shoot formation in saffron (Crocus sativus L.). Acta biol. hung. 61: 73–89. DOI: https://doi.org/10.1556/ABiol.61.2010.1.8
Singh R, Singal H R and Goyal S C. 2006. Biochemical changes during shoot differentiation in callus cultures of Chlorophytum borivilianum Sant. Et Fernand. Indian Journal Plant Physiology 11: 130–5.
Thorpe T A. 1990. The current status of plant tissue culture. (In) Bhojwani S S (Eds). Plant Tissue Culture: Applications and Limitations, pp 1–33. Elsevier Science Publisher, New York. DOI: https://doi.org/10.1016/B978-0-444-88883-9.50005-4
Vanhoof P and Gaspar T. 1976. Peroxidase and isoperoxidase changes in relation to root initiation of Asparagus cultures in vitro. Acta Horticultural. 4: 27. DOI: https://doi.org/10.1016/0304-4238(76)90061-3
Verghese T M and Kour V. 1991. Micropropagation of Albizzia lebbeck Benth. Acta Horticultural. 289: 161–2. DOI: https://doi.org/10.17660/ActaHortic.1991.289.41
Downloads
Submitted
Published
Issue
Section
License
Copyright (c) 2016 The Indian Journal of Agricultural Sciences
This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License.
The copyright of the articles published in The Indian Journal of Agricultural Sciences is vested with the Indian Council of Agricultural Research, which reserves the right to enter into any agreement with any organization in India or abroad, for reprography, photocopying, storage and dissemination of information. The Council has no objection to using the material, provided the information is not being utilized for commercial purposes and wherever the information is being used, proper credit is given to ICAR.
How to Cite
