Abstract
We established an efficient plant regeneration system for Catharanthus roseus L. (G.) Don through somatic embryogenesis. Embryogenic callus was induced from hypocotyl of seed germinated in vitro. Somatic embryogenesis in Catharanthus has been categorized into three distinct stages: (1) initiation and proliferation of embryo; (2) maturation, and; (3) germination or plantlet conversion. Beside plant growth regulators, various stages of embryogenesis were screened for their response to a wide variety of factors (pH, gelrite, light, sugar alcohols, polyethyleneglycol and amino acids), which affect embryogenesis. All of the tested factors had a small to marked influence on embryogeny and eventual conversion to plantlets. The plantlets were acclimatized successfully in a greenhouse. To our knowledge, this is the first report describing a detailed study of various cultural factors which regulate embryogenesis in C. roseus. The results discussed in this paper may be used in mass propagation to produce medicinal raw material, and the embryo precursor cells could be used in genetic modification programmes that aim to improve the alkaloid yield as well.
Similar content being viewed by others
Abbreviations
- BA:
-
N 6-benzyladenine
- 2,4-D:
-
2,4-Dichlorophenoxyacetic acid
- NAA:
-
α-Naphthalene acetic acid
- GA3 :
-
Gibberellic acid
- PEG:
-
Polyethylene glycol
- MS:
-
Murashige and Skoog’s medium
- SE:
-
Somatic embryo
- DMRT:
-
Duncan’s multiple range test
References
Bajaj YPS (1995) Somatic embryogenesis and its applications for crop improvement. In: Bajaj YPS (ed) Biotechnology in agriculture and forestry, somatic embryogenesis and synthetic seed. I. Springer, Berlin, pp 105–125
Bhansali RR, Driver JA, Durzan DJ (1990) Rapid multiplication of adventitious somatic embryos in peach and nectarine by secondary embryogenesis. Plant Cell Rep 9:280–284
Blanc G, Michaux-Ferriere N, Teisson C, Lardet L, Carron MP (1999) Effect of carbohydrate addition on the induction of somatic embryogenesis in Hevea brasiliensis. Plant Cell Tissue Organ Cult 59:103–112
Bozhkov PV, Von Arnold S (1998) Polyethylene glycol promotes maturation but inhibits further development of Picea abies somatic embryos. Physiol Plant 104:211–224
Brown DCW, Finstad KI, Watson EM (1995) Somatic embryogenesis in herbaceous dicots. In: Thorpe TA (ed) In vitro embryogenesis in plants. Kluwer, Dordrecht, pp 345–415
Cabasson C, Ollitrault P, Cote FX, Michaux-Ferriére N, Dambier D, Dalnic R, Teisson C (1995) Characteristic of citrus cell cultures during undifferentiated growth on sucrose and somatic embryogenesis on galactose. Physiol Plant 93:464–470
Chang YF, Wong JR (1994) Regeneration of plants from protoplasts of Triticum aestivum L. (Wheat). In: Bajaj YPS (ed) Plant protoplasts and genetic engineering V (Biotechnology in agriculture and forestry, vol 29). Springer, Berlin, pp 161–171
Cheong EJ, Pooler MR (2004). Factors affecting somatic embryogenesis in Prunus incisa cv. February Pink. Plant Cell Rep 22:810–815
Choi YE, Yang DC, Choi KT (1998) Induction of somatic embryos by macrosalt stress from mature zygotic embryos of Panax ginseng. Plant Cell Tissue Organ Cult 52:177–181
De March G, Grenier E, Miannay N, Sulmont G, David H, David A (1993) Potential of somatic embryogenesis in Prunus avium immature zygotic embryos. Plant Cell Tissue Organ Cult 34:209–215
Druart P (1981) Embryogenese somatique et obtention de plantules chez Prunus incise x serrula (GM 9) cultive in vitro. Bull Rech Agron Gembloux 16:205–220
Fiore MC, Trabace T, Sunseri F (1997) High frequency of plant regeneration in sunflower from cotyledons via somatic embryogenesis. Plant Cell Rep 16:295–298
Gaj MD (2004) Factors influencing somatic embryogenesis induction and plant regeneration with particular reference to Arabidopsis thaliana (L.) Heynh. Plant Growth Reg 43:27–47
Gray DJ, Compton ME, Harrell RC, Cantliffe DJ (1995) Somatic embryogenesis and the technology of synthetic seed. In: Bajaj YPS (ed) Biotechnology in agriculture and forestry, somatic embryogenesis and seed I, vol 30. Springer, Berlin, pp 126–151
Gupta PK, Grobe JA (1995) Somatic embryogenesis in conifer. In: Jain SM, Gupta PK, Newton RJ (eds) Somatic embryogenesis in woody plants, vol 1. Kluwer, Dordrecht, pp 81–98
Holme IB, Krogstrup P, Hansen J (1997) Embryogenic callus formation, growth and regeneration in callus and suspension cultures of Miscanthus x ogiformis Honda Giganteus as affected by proline. Plant Cell Tissue Organ Cult 50:203–210
Huang LC, Vits H, Staba EJ, Cooke TJ, Hu WS (1992) Effect of cultivated age and embryo size on specific oxygen uptake rate in developing somatic embryos of Daucus carota L. Biotechol Lett 14:701–706
Ikeda-Iwai M, Umehara M, Satoh S, Kamada H (2003) Stress-induced somatic embryogenesis in vegetative tissues of Arabidopsis thaliana. Plant J 34:107–114
Jimenez VM (1996) El Cultivo de protoplastos en citricos y su potencial para el mejoramiento genetico. Agron Costarric 20:187–204
Junaid A, Bhat MA, Mujib A, Sharma MP (2004). Somatic embryogenesis study in Catharanthus roseus (L.) G. Don: an anticancerous plant. In: Khan IA, Khanum A (eds) Role of biotechnology in medicinal and aromatic plants, vol XI. Ukaaz, Hyderabad, pp 98–106
Junaid A, Bhatt MA, Mujib A, Sharma MP (2006) Somatic embryo proliferation maturation and germination in Catharanthus roseus. Plant Cell Tissue Organ Cult 84:325–332
Junaid A, Mujib A, Sharma MP, Tang W (2007a) Growth regulators affect primary and secondary somatic embryogenesis in Madagaskar periwinkle (Catharanthus roseus (L) G. Don) at morphological and biochemical levels. Plant Growth Regul 51:271–281
Junaid A, Mujib A, Bhat MA, Sharma MP, Šamaj J (2007b) Somatic embryogenesis and plant regeneration in Catharanthus roseus. Biol Plant 51(4):641–646
Kevei E, Nagy F (2003) Phytochrome controlled signaling cascades in higher plants. Physiol Plant 117:305–313
Koh WL, Loh CS (2000) Direct somatic embryogenesis, plant regeneration and in vitro flowering in rapid-cycling Brassica napus. Plant Cell Rep 19:1177–1183
Krikorian AD, Berquam D (1969) Plant cell and tissue culture: the role of Haberlandt. Bot Rev 35:58–88
Linossier L, Veisseire P, Cailloux F, Coudret A (1997) Effects of abscisic acid and high concentrations of PEG on Hevea brasiliensis somatic embryos development. Plant Sci 124:183–191
Lipavská H, Konrádová H (2004) Somatic embryogenesis in conifers: the role of carbohydrate metabolism. In Vitro Cell Dev Biol Plant 40:23–30
Machado AC, Puschmann M, Puhringer H, Kremen R, Katinger H, Machado MLC (1995) Somatic embryogenesis of Prunus subhirtella ‘autumno rosa’ and regeneration of transgenic plants after Agrobacterium-mediated transformation. Plant Cell Rep 14:335–340
Marchant R, Davey MR, Lucas JA, Power JB (1996) Somatic embryogenesis and plant regeneration in Floribunda rose (Rosa hybrida L.) cvs. Trumpeter and Glad tidings. Plant Sci 120:95–105
Maruyama E, Hosoi Y, Ishii K (2002) Somatic embryogenesis in sawara cypress (Chamaecyparis pisifera Sieb. Et Zucc.) for stable and efficient plant regeneration, propagation and protoplast culture. J For Res 7:23–34
Mujib A, Samaj J (2006) Somatic embryogenesis. Springer, Berlin
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497
Nato A, Fresneau C, Moursalimowa N, De Buyser J, Lavergne D, Henry Y (2000) Expression of auxin and light regulated arrestin-like proteins, G proteins and nucleoside diphosphate kinase during induction and development of wheat somatic embryos. Plant Physiol Biochem 38:483–490
Norgaard JV (1997) Somatic embryo maturation and plant regeneration in Abies nordmanniana LK. Plant Sci 124:211–221
Nuutila AM, Villiger C, Oksman-Caldentey KM (2002) Embryogenesis and regeneration of green plantlets from oat (Avena sativa L.) leaf-base segments: influence of nitrogen balance, sugar and auxin. Plant Cell Rep 20:1156–1161
Ochatt SJ, Pontecaille C, Rancillac M (2000) The growth regulators used for bud regeneration and shoot rooting affect the competence for flowering and seed set in regenerated plants of protein peas. In Vitro Cell Dev Biol Plant 36:188–193
Owen HR, Wengerd D, Miller AR (1991) Culture medium pH is influenced by basal medium, carbohydrate source, gelling agent, activated charcoal and medium storage method. Plant Cell Rep 10:583–586
Pan ZG, Liu CZ, Murch SJ, El-Demerdash M, Saxena PK (2003) Plant regeneration from mesophyll protoplasts of the Egyptian medicinal plants Artemisia judaica L. and Echinops spinosissimus Turra. Plant Sci 165:681–687
Pasternak TP, Prinsen E, Ayaydin F, Miskolczi P, Potters G, Asard H, Van Onckelen HA, Dudits D, Feher A (2002) The role of auxins, pH, and stress in the activation of embryogenic cell division in leaf protoplast derived cells of alfalfa. Plant Physiol 129:1807–1819
Pinto G, Santos C, Neves L, Araújo C (2002) Somatic embryogenesis and plant regeneration in Eucalyptus globules Labill. Plant Cell Rep 21:208–213
Puigderrajols P, Jofre A, Mir G, Pla M, Verdaguer D, Huguet G, Molinas M (2002) Developmentally and stress induced small heat shock proteins in cork oak somatic embryos. J Exp Bot 53:1445–1452
Pullman GS, Johnson S, Peter G, Cairney J, Xu N (2003) Improving loblolly pine somatic embryo maturation: comparision of somatic and zygotic embryo morphology, germination and gene expression. Plant Cell Rep 21:747–758
Pullman GS, Gupta PK, Timmis R, Carpenter C, Kreitinger M, Welty E (2005) Improved Norway spruce somatic embryo development through the use of abscisic acid combined with activated carbon. Plant Cell Rep 24:271–279
Ramage CM, Williams RR (2004) Cytokinin induced abnormal shoot regeneration is associated with elevated knottedl—type homeobox gene expression in tobacco. Plant Cell Rep 22:919–924
Redenbaugh K (1993) Synseeds. Application of synthetic seeds to crop improvement. CRC Press, Boca Raton, FL
Reidiboym-Talleux L, Diemer F, Sourdioux M, Chapelain K, Grenier-DeMarch G (1998) Improvement of somatic embryogenesis in wild cherry (Prunus avium). Effect of maltose and ABA supplements. Plant Cell Tissue Organ Cult 55:199–209
Rey HY, Sansberro PA, Collavino MM, Daviña JR, Gonzàlez AM, Mroginski LA (2002) Colchicine, trifluralin and oryzalin promoted development of somatic embryos in Ilex paraguariensis (Aquifoliaceae). Euphytica 123:49–56
Robichaud RL, Lessard VC, Merkle SA (2004) Treatments affecting maturation and germination of American chestnut somatic embryos. J Plant Physiol 161:957–969
Schuller A, Reuther G (1993) Response of Abies alba embryonal- suspensor mass to various carbohydrate treatments. Plant Cell Rep 12:199–202
Scott P, Lyne RL (1994) The effect of different carbohydrate sources upon the initiation of embryogenesis from barley microspores. Plant Cell Tissue Organ Cult 36:129–133
Smith DL, Krikorian AD (1990) Somatic embryogenesis of carrot in hormone- free medium: external pH control over morphogenesis. Am J Bot 77:1634–1647
Strickland SG, Nichol JW, McCall CM, Stuart DA (1987) Effect of carbohydrate source on alfalfa somatic embryogenesis. Plant Sci 48:113–121
Taniguchi T, Kurita M, Itahana N, Kondo T (2004) Somatic embryogenesis and plant regeneration from immature zygotic embryos of Hinoki cypress (Chamaecyparis obtuse Sieb. et Zucc.). Plant Cell Rep 23:26–31
Thorpe TA (1995) In vitro embryogenesis in plants. Kluwer, Dordrecht
Torne JM, Moysset L, Santos M, Simon E (2001) Effects of light quality on somatic embryogenesis in Araujia sercifera. Physiol Plant 111:405–411
Van Winkle SC, Johnson S, Pullman GS (2003) The impact of gelrite and activated carbon on the elemental composition of two conifer embryogenic tissue initiation media. Plant Cell Rep 21:1175–1182
Verma DC, Dougall DK (1997) Influence of carbohydrates on quantitative aspects of growth and embryo formation in wild carrot suspension cultures. Plant Physiol 59:81–85
Xing Z, Powell WA, Maynard CA (1999) Development and germination of American chestnut somatic embryos. Plant Cell Tissue Organ Cult 57:47–55
Acknowledgments
The AFMI Fellowship provided by AFMI to one of us (Dr. A. Junaid) is gratefully acknowledged.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Aslam, J., Mujib, A., Fatima, S. et al. Cultural conditions affect somatic embryogenesis in Catharanthus roseus L. (G.) Don. Plant Biotechnol Rep 2, 179–189 (2008). https://doi.org/10.1007/s11816-008-0060-9
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11816-008-0060-9