Summary
Somatic embryogenesis from nucellus-derived callus cultures of five cultivars, including three (Caipira, Seleta Vermelha, and Valencia) of sweet oranges (C. sinesis L. Osbeck), Rangpur lime (C. limonia L. Osbeck), and Cleopatra mandarin (C. reticulata Blanco) (lines I and II), were studied. Callus lines maintained on MT medium supplemented with 50 g l−1 sucrose were transferred to MT medium supplemented with different carbohydrate sources: galactose, glucose, lactose, maltose, or sucrose at 18, 37, 75, 110, or 150 mM, or glycerol at 6, 12, 24, 36, or 50 mM. Globular embryos were observed after approximately 4 wk, in several treatments. Cultures of Valencia and Caipira sweet oranges and Cleopatra mandarin (line I) showed high numbers of embryos on medium containing galactose, lactose, and maltose. Histological studies showed somatic embryos in all developmental stages with a normal histodiffeentiation pattern. The other two cultivars (Rangpur lime and Cleopatra mandarin, line II) formed very few embryos, which did not develop further following the globular stage. Some of the abnormalities observed were lack or dedifferentiation of protoderm and absence of apical meristems and procambial strands. Embryos that followed the normal sequence of development were easily converted into plants. Non-embryogenic cultures continued as proliferating callus cultures, eventually forming a few embryos which did not convert into plants. Statistical analyses of the callus response to carbohydrate treatments was done using an overdispersion Poisson model.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Alemanno, L.; Berthouly, M.; Michaux-Ferrière, N. Histology of somatic embryogenesis from floral tissues of cocoa. Plant Cell Tiss. Organ Cult. 46:187–194; 1996.
Cabasson, C.; Ollitrault, P.; Côte, F.; Michaux-Ferrière, N.; Dambier, D.; Dalnic, R.; Teisson, C. Characteristics of citrus cell cultures during undifferentiated growth on sucrose and somatic embryogenesis on galactose. Physiol. Plant. 93:464–470; 1995.
Coutos-Trevenot, P.; Goebel-Tourand, L.; Mauro, M.; Jouanneau, J.; Boulay, M.; Deloire, A.; Guern, J. Somatic embryogenesis from grapevine cells. I. Improvement of embryo development by changes in culture conditions. Plant Cell Tiss Organ Cult. 29:125–133; 1992.
Demétrio, C. G. B.; Hinde, J. Half-normal plots and overdispersion. GLIM Newsletter 27:19–26; 1997.
Feder, N.; O'Brien, T. P. Plant microtechnique: some principles and new methods. Am. J. Bot. 55:123–142; 1968.
Gavish, H.; Vardi, A.; Fluhr, R. Suppression of somatic embryogenesis in Citrus cell cultures by extracellular proteins. Planta 186:511–517; 1992.
Gmitter, F. G.; Grosser, J. W.; Moore, G. A.; Citrus. In: Hammerschlag, F. A.; Litz, R. E., eds. Biotechnology of perennial fruit crops. Wallingford: CAB International; 1992:335–369.
Gmitter, F. G.; Moore, G. A. Plant regeneration from undeveloped ovules and embryogenic calli of Citrus: embryo production, germination, and plant survival. Plant Cell Tiss. Organ Cult. 6:139–147; 1986.
Grosser, J. W.; Gmitter, F. G. Protoplast fusion and citrus improvement. Plant Breed. Rev. 8:339–374; 1990.
Hidaka, T.; Omura, M. Control of embryogenesis in Citrus cell culture: regeneration from protoplasts and attempts to callus bank. Bull. Fruit Tree Res. Stn. B. 16:1–17; 1989.
Hinde, J.; Demétrio, C. G. B. Overdispersion: models and estimation. Comput. Statist. Data Anal. 27:151–170; 1998a.
Hinde, J.; Demétrio, C. G. B. Overdispersion: models and estimation. São Paulo: Associação Brasileira de Estatística; 1998b.
Jumin, H. B.; Nito, N. Plant regeneration via somatic embryogenesis from protoplasts of six species related to Citrus. Plant Cell Rep. 15:332–336; 1996.
Kobayashi, S.; Ohgawara, T.; Ohgawara, E.; Oiyama I.; Ishii, S. A somatic hybrid plant obtained by protoplast fusion between navel orange (Citrus sinesis) and satsuma mandarin (C. unshiu). Plant Cell Tiss. Organ Cult. 14:63–69; 1988.
Kobayashi, S.; Uchimiya, H. Expression and integration of a foreign gene in orange (Citrus sinensis Osb.) protoplast by direct DNA transfer. Jpn. J. Genet. 64:91–97; 1989.
Kochba, J.; Button, J. The stimulation of embryogenesis and embryoid development in habituated ovular callus from the ‘Shamouti’ orange (Citrus sinensis) as affected by tissue age and sucrose concentration. Z. Pflanzenphysiol. 73:415–421; 1974.
Kochba, J.; Spiegel-Roy, P. The effects of auxins, cytokinins and inhibitors on embryogenesis in habituated ovular callus of the ‘Shamouti’ orange (Citrus sinensis). Z. Pflanzenphysiol. 81:283–288; 1977.
Kochba, J.; Spiegel-Roy, P.; Neumann, H.; Saad, S. Stimulation of embryogenesis in citrus ovular callus by ABA, ethephon, CCC and Alar and its suppression by GA3. Z. Pflanzenphysiol. 89:427–432; 1978a.
Kochba, J.; Spiegel-Roy, P.; Neumann, H.; Saad, S. Effect of carbohydrates on somatie embryogenesis in subcultured nucellar callus of Citrus cultivars. Z. Pflanzenphysiol. 105:359–368; 1982.
Kochba, J.; Spiegel-Roy, P.; Saad, S.; Neumann, H. Stimulation of embryogenesis in Citrus tissue culture by galactose. Naturwissenschaften 65:261–262 1978b.
Komai, F.; Okuse, I.; Saga, K.; Harada, T. Improvement on the efficiency of somatic embryogenesis from spinach root tissues by applying various sugars. J. Jpn Soc. Sci. 65:67–72; 1996.
Kunitake, H.; Kagami, H.; Mii, M. Somatic embryogenesis and plant regeneration from protoplasts of ‘Satsuma’ mandarin (Citrus unshiu Marc). Scient. Hortic. 47:27–33; 1991.
Kunitake, H.; Mii, M. Somatic embryogenesis in Citrus species. In: Bajaj Y. P. S., ed. Somatic embryogenesis and synthetic seed I. Biotechnology in agricultural and forestry, vol. 30. Berlin: Springer-Verlag; 1995:280–298.
Ling, J.; Iwamasa, M. Plant regeneration from embryogenic calli of six Citrus related genera. Plant Cell Tiss. Organ Cult. 49:145–148; 1997.
Mendes-da-Glória, F. J.; Mourão Filho, F. A. A.; Demétrio, C. G. B.; Mendes, B. M. J. Embryogenic calli induction from nucellar tissue of citrus cultivars. Scient. Agric. 56:1111–1115; 1999.
Merkle, S. A.; Parrot, W. A.; Flinn, B. S. Morphogenic aspects of somatic embryogenesis. In: Thorpe, T. A., ed. In vitro embryogenesis in plant. Dordrecht: Kluwer Academic Publishers; 1995:155–203.
Merkle, S. A.; Wiecko A. T.; Sotak, R. J.; Sommer, H. E. Maturation and conversion of Liriodendron tulipifera somatic embryos. In Vitro Cell. Dev. Biol. 26:1086–1093; 1990.
Murashige, T.; Tucker, D. P. H. Growth factor requirements of citrus tissue culture. Proc. Int. Soc. Citricult. 1:1155–1161; 1969.
Oliveira, R. P.; Mendes, B. M. J.; Tulmann Neto, A., Obtenção e cultura de calos nucelares de limão Cravo, tangerinaCleopatra e Poncirus trifoliata. Rev. Bras. Fisiol. Veg. 6:115–119; 1994.
Padmanabhan K.; Cantliffe D. J.; Harrell, R. C.; McConnell, D. B. A comparison of shoot-forming and non-shoot-forming somatic embryos of sweet potato (Ipomea batatas (L.) Lam.) using computer vision and histological analyses. Plant Cell Rep. 17:685–692; 1998.
Peña, L.; Cervera, M.; Juárez, J.; Navarro, A.; Piña, J. A.; Durán-Vila, N.; Navarro, L. Agrobacterium-mediated transformation sweet orange and regeneration of transgenic plants. Plant Cell Rep. 14:616–619; 1995a.
Peña, L.; Cervera, M.; Juárez, J.; Ortega, C.; Piña, J. A.; Durán-Vila, N.; Navarro, L. High efficiency Agrobacterium-mediated transformation and regeneration of citrus. Plant Sci. 104:183–191, 1995b.
Pérez, R. M.; Galiana, A. M.; Navarro, L.; Duran-Vila, N. Embryogenesis in vitro of several Citrus species and cultivars. J. Hort. Sci. Biotechnol. 73:796–802; 1998.
Pérez, R. M.; Mas, O.; Navarro, L.; Duran-Vila, N. Production and cryoconservation of embryogenic cultures of mandarin and mandarin hybrids. Plant Cell Tiss. Organ Cult. 55:71–74; 1999.
Ranga Swamy, N. S. Culture of nucellar tissue of Citrus in vitro Experientia 14:111–112; 1958.
Ranga Swamy, N. S. Experimental studies on female reproductive structures of Citrus microcarpa Bunge. Phytomorphology 11:109–127; 1961.
Rangan, T. S.; Murashige, T.; Bitters, w. P. In vitro initiation of nucellar embryos in monoembryonic Citrus. HortScience 3:226–227; 1968.
Rangan, T.S.; Murashige, T.; Bitters, W.P. In vitro studies of zygotic and nucellar embryogenesis in citrus. Proc. Int. Soc. Citricult. 1:225–229; 1969.
Rodriguez, A. P. M.; Wetzstein, H. Y. The effect of auxin type and concentration on pecan (Carya illinoinensis) somatic embryo morphology and subsequent conversion into plants. Plant Cell Rep. 13:607–611; 1994.
Rodriguez, A. P. M.; Wetzstein, H. Y. A morphological and histological comparison of the initiation and development of pecan (Carya illinoinensis) somatic embryogenic cultures induced with naphthaleneacetic acid or 2,4-dichlorophenoxyacetic acid. Protoplasma 204:71–83; 1998.
Singh, A. K.; Nito, N.; Iwamasa, M. Influence of lactose and glycerol on growth and somatic embryogenesis of Citrus callus. Acta Hortic. 321:606–609; 1992.
Spiegel-Roy, P.; Saad, S. Effect of carbohydrates and inhibitors of GA3 biosynthesis on embryogenic potential of salt tolerant and nontolerant callus lines of orange (Citrus sinensis Osbeck). Plant Sci. 47:215–220; 1986.
Strickland, S. G.; Nichol, J. W.; McCall, C. M.; Stuart, D. A. Effect of carbohydrate source on alfafa somatic embryogenesis. Plant Sci. 48:113–121; 1987.
Vu, J. C. V.; Niedz, R. P.; Yelenosky, G. Glycerol stimulation of chlorophyll synthesis, embryogenesis, and carboxylation and sucrose metabolism enzymes in nucellar callus of Hamlin sweet orange. Plant Cell Tiss. Organ Cult. 33:75–80; 1993.
Yeung, E. C. Structural and developmental patterns in somatic embryogenesis. In: Thorpe, T. A., ed. In vitro embryogenesis in plants. Dordrecht: Kluwer Academic Publishers; 1995:205–247.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Tomaz, M.L., Januzzi Mendes, B.M., Mourão Filho, f.D.A.A. et al. Somatic embryogenesis in Citrus SPP.: Carbohydrate stimulation and histodifferentiation. In Vitro Cell.Dev.Biol.-Plant 37, 446–452 (2001). https://doi.org/10.1007/s11627-001-0078-y
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/s11627-001-0078-y