Somatic Embryogenesis and Plant Regeneration from Hypocotyl and Leaf Explants of Brassica Oleracea var. Botrytis (Cauliflower)
We investigated direct and indirect formation of somatic embryogenesis in Brassica oleracea var. botrytis (cauliflower), a very important vegetable crop worldwide. Direct somatic embryogenesis, which is rather rare, was achieved in culture of 2-week-old hypocotyl explants of Brassica oleracea var. botrytis on MS medium supplemented with 1.0 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.5; 1.0; and 1.5 mg/l kinetin. Initial induction of embryogenic callus was achieved on MS supplemented with very low concentrations of 2,4-D (0.05 mg/l and 0.1 mg/l). Indirect somatic embryogenesis from leaf sections was obtained on MS supplemented with 0.05 or 0.1 mg/l 2,4-D. We examined various stages of somatic embryos (globular, heart, torpedo, cotyledonary). More embryos per explant were produced through the indirect pathway (23-25) than through the direct pathway (14-19). The number of embryos produced was high. There is a potential for recurrent, repeated or secondary somatic embryogenesis, possibly an unlimited source for mass propagation and ideal for synthetic seed production in this species. Plant regeneration was achieved on half-strength MS medium without any hormones.
Ahmed ABA, RAO AS, RAO MV, and Taha RM. 2011. Effect of picloram, additives and plant growth regulators on somatic embryogenesis of Phyla nodiflora (L.) Greene. Brazilian Archives of Biology and Technology 54: 7-13.
Ahloowalia BS. 1991. Somatic embryos in monocots: The genesis and genetic stability. Revue de Cytolgie et de Biologie Vegetales, Le Botaniste 14: 223-235.
Ali A, Naz S, and Iqbal J. 2007. Effect of different explants and media compositions for efficient somatic embryogenesis in sugarcane (Saccharum officinarum). Pakistan Journal of Botany 39(6): 1961-1977.
Arnold VS, Sabala I, Bozhkov P, Dyachok J, and Filonova L. 2002. Developmental pathways of somatic embryogenesis. Plant Cell Tissue Organ Culture 69: 233-249.
Bhalla PL, and Weerd DN. 1999. In vitro propagation of cauliflower, Brassica oleracea var. botrytis for hybrid seed production. Plant Cell Tissue and Organ Culture 56: 89-95.
Chikkala VRN, Nugent GD, Dix PJ, and Stevenson TW. 2008. Regeneration from leaf explants and protoplasts of Brassica oleracea var. botrytis (cauliflower). Scientia Horticulturae 119: 330-334.
Duncan DB. 1955. Multiple range and multiple F-tests. Biometrics 11: 1-42.
Eimert K, and Siegemund F. 1992. Transformation of cauliflower (Brassica oleracea var. botrytis)-an experimental survey. Plant Molecular Biology 19: 485-490.
Fransz PF, Leunissen EHM, Van EFA, Keizer PLC, and Colijn-Hooymans TM. 1994. Improvement of protoplast regeneration from a recalcitrant inbred line of Brassica oleracea: a morphogenetic analysis. Plant Science 98: 87-95.
Ikeda-Iwai M, Umehara M, Satoh S, and Kamada H. 2003. Stress-induced somatic embryogenesis in vegetative tissues of Arabidopsis thaliana. The Plant Journal 34: 107-114.
Jourdan PS, Earle ED, and Mutschler MA. 1990. Improved protoplast culture and stability of cytoplasmic traits in plants regenerated from leaf protoplasts of cauliflower (Brassica oleracea ssp. botrytis). Plant Cell Tissue and Organ Culture 21: 227-236.
Karami O. 2008. Induction of embryogenic callus and plant regeneration in Carnation (Dianthus caryophyllus L.). Online Journal of Biological Sciences 8(4): 68-72.
Kieffer M, Fuller MP, and Jellings AJ. 1995. Rapid mass production of cauliflower propagules from fractionated and graded curd. Plant Science 107: 229-235.
Kieffer M, Simkins N, Fuller MP, and Jellings AJ. 2001. A cost effective protocol for in vitro mass propagation of cauliflower. Plant Science 160: 1015-1024.
Kumar A, Kumar VA, and Kumar J. 1993. Rapid in vitro propagation of cauliflower. Plant Science 90: 175-178.
Leroy XJ, Leon K, Charles G, and Branchard M. 2000. Cauliflower somatic embryogenesis and analysis of regenerant stability by ISSRs. Plant Cell Report 19: 1102-1107.
Lv LL, Lei JJ, Song M, Li LY, and Cao BH. 2005. Study on transformation of cowpea trypsin inhibitor gene into cauliflower (Brassica oleracea var. botrytis). African Journal of Biotechnology 4(1): 45-49.
Martin KP, and Madassery J. 2005. Direct and indirect somatic embryogenesis on cotyledon explants of Quassia amara L., an antileukaemic drug plant. In Vitro Cellular and Developmental Biology 41: 54-57.
Murashige T, and Skoog F. 1962. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiology Plantarum 51: 473-497.
Murthy HN, Hahn EJ, and Paek KY. 2008. Recurrent somatic embryogenesis and plant regeneration in Coriandrum sativum L. Scientia Horticulturae 118: 168-171.
Namasivayam P. 2007. Acquisition of embryogenic competence during somatic embryogenesis. Plant Cell, Tissue and Organ Culture 90: 1-8.
Pareek LK, and Chandra N. 1978. Somatic embryogenesis in leaf callus from cauliflower (Brassica oleracea var. botrytis). Plant Science Letters 11: 311-316.
Pogrebnyak N, Markley K, Smimov Y, Brodzik R, and Bandurska K. 2006. Collard and cauliflower as a base for production of recombinant antigens. Plant Science 171: 677-685.
Schumann G, Ryschika U, Schulze J, and Klocke E. 1995. Anatomy of somatic embryogenesis. In: Bajaj, YPS [ed.] Biotechnology in Agriculture and Forestry, vol. 30: Somatic Embryogenesis and Synthetic Seed 1, 71-86. Springer-Verlag, Berlin, Heidelberg.
Sharp WR, Sondahl MR, Caldas LS, and Maraffa SB. 1980. The physiology of in vitro sexual embryogenesis. Horticulture Review 2: 268-310.
Qin Y, Gap LH, Pulli S, and Guo YD. 2006. Shoot differentiation, regeneration and analysis of somaclonal variation by RAPD. Hereditas 143: 91-98.
Vatsya B, and Bhaskaran S. 1982. Plant regeneration from cotyledonary protoplasts of cauliflower (Brassica oleracea var. botrytis). Protoplasma 113: 161-163.
Vikrant, and Rashid A. 2001. Direct as well as indirect somatic embryogenesis from immature (unemerged) inflorescence of a minor millet Paspalum scrobiculatum L. Euphytica 120: 167-172.
Watts LE. 1963. Investigations into the breeding system of cauliflower Brassica oleracea var. botrytis (L.). Euphytica 12: 323-340.
William EG, and Maheswaran G. 1986. Somatic embryogenesis: factors influencing coordinate behavior of cells as an embryogenic group. Annals of Botany 57: 44-462.
Yang Q, Chauvin JE, and Herve Y. 1992. A study of factors affecting anther culture of cauliflower (Brassica oleracea var. botrytis). Plant Cell Tissue and Organ Culture 28: 289-296.
Zobayed SMA, Armstrong J, and Armstrong W. 1999. Cauliflower shoot-culture: effects of different types of ventilation on growth and physiology. Plant Science 141: 209-217.