Application of Genetic Engineering in Citrus

Yıl 2006, Cilt: 1 Sayı: 1, 74 - 86, 31.01.2006

Bayram Çevik Mehtap Şahin Çevik

Öz

As in other plants, application of genetic engineering in Citrus requires availability of genes for
desired traits and development of genetic transformation methods. Once these conditions are met, traits
that can not be integrated into the Citrus genome by conventional breeding could be introduced to citrus
by genetic engineering. Hence, a number of studies were conducted in Citrus and transgenic citrus plants
were produced from different types of plant material using various transformation methods in the last
decade. Initially, citrus plants were transformed only with reporter genes. As the transformation methods
were optimized and genes for agriculturally important traits become availible by cloning from different
sources, genetic engineering was applied for developing resistance to diseases, pests and abiotic stresses as
well as improving fruit quality in Citrus. As a result, many transgenic plants transformed with these genes
were produced and molecular and biological characteristization of these transgenic plants are still
underway. In this study, genetic transformation studies in Citrus were reviewed and its prospects in the
near future and its importance for Turkey were discussed.

Anahtar Kelimeler

Citrus, genetic engineering, genetic transformation, tissue culture, resistance to abiotic and biotic stresses,

Turunçgillerde Genetik Mühendisliği Uygulamaları

Öz

Diğer bitkilerde olduğu gibi turunçgillerde de genetik mühendisliği uygulamalarının yapılması ve
yaygınlaşması, istenilen özellikleri sağlayacak genlerin bulunmasına ve bunları bitkiye aktaracak
transformasyon yöntemlerinin geliştirilmesine bağlıdır. Bu iki temel koşulun sağlanmasıyla turunçgillere
ıslah çalışmalarıyla kazandırılamayan bazı özellikler genetik mühendisliği yoluyla kazandırılabilir. Bu
amaçla turunçgillerde son on yılda çok sayıda çalışma yapılmış olup farklı bitki materyali ve farklı gen
aktarım yöntemleri kullanılarak transgenik bitkiler üretilmiştir. Başlangıçta transformasyon çalışmaları
sadece raportör genler kullanılarak yapılmıştır. Daha sonra transformasyon yöntemlerinin optimizasyonu
ve tarımsal açıdan önemli genlerin çeşitli kaynaklardan klonlanmasıyla turunçgillerde hastalık ve zararlılar
yanında abiyotik streslere dayanıklılık ve meyve kalitesinin artırılmasına yönelik genetik mühendisliği
uygulamaları da başlamıştır. Bu uygulamalar sonucunda birçok transgenik bitki üretilmiş olup halen bu
transgenik bitkilerin moleküler ve biyolojik karakterizasyonları yapılmaktadır. Bu çalışmada dünyada
turunçgillerde yapılan genetik transformasyon çalışmaları ve bunların sonuçları derlenerek yakın
gelecekteki durumu ve ülkemiz açısından önemi tartışılmıştır.

Anahtar Kelimeler

Turunçgiller, genetik mühendisliği, genetik transformasyon, doku kültürü, Biyotik ve abiyotik streslere dayanıklılık,

Kaynakça

• Almeida, W.A.B, Mourão-Filho, F.A.A., Pino, L.E., Boscariol, R.L., Rodríguez A.P.M., Mendes, B.M.J. 2003. Genetic transformation and plant recovery from mature tissues of Citrus sinensis L. Osbeck. Plant Science 164: 203–211.
• Bond, J.E. and Roose, M.L. 1998. Agrobacterium-mediated transformation of the commercially important citrus cultivar Washington navel orange. Plant Cell Rep. 18: 229–234.
• Boscariol R. L., Almeida, W.A.B., Derbyshire, M.T.V.C., Mourao-Filho, F.A.A. and Mendes, B.M.J. 2003. The use of the PMI/mannose selection system to recover transgenic sweet orange plants (Citrus sinensis L. Osbeck). Plant Cell Rep. 22:122–128.
• Ceccardi, T.L., Barthe, G.A. and Derrick, K.S. 1998. A novel protein associated with citrus blight has sequence similarities to expansin. Plant Mol. Biol. 38: 775–783.
• Cervera, M., Ortega C., Navarro, A., Navarro, L. and Pena, L. 2000. Generation of transgenic citrus plants with the tolerance-tosalinity gene HAL2 from yeast, J. Hort. Sci. Biotech. 75 26–30.
• Cervera, M., Pina, J.A., Juarez, J., Navarro, L. and Pena, L. 1998. Agrobacterium-mediated transformation of citrange: factors affecting transformation and regeneration. Plant Cell Rep. 18: 271–278.
• Cevik, B., Lee, R.F., Moore, G.A. and Niblett, C.L. 2000. Genetic Transformation of Citrus paradisi with the RNA-Dependent RNA Polymerase Gene of Citrus Tristeza Closterovirus. In: Proceedings of the 9th International Citrus Congress. (Ed.:L. G. Albrigo),Vol. 1, CREC-UF/IFAS, Lake Alfred, pp. 205.
• Cevik, B. 2001. Characterization of the RNA-dependent RNA polymerase gene of citrus tristeza closterovirus. Ph.D Thesis. University of Florida, Gainesville, Florida, 138 pp.
• Cevik, B., Lee, R.F. and Niblett, C.L. 2006. Genetic Transformation of Citrus paradisi with antisense and untranslatable constructs of the RNA-Dependent RNA Polymerase Gene of Citrus Tristeza Closterovirus. T. J. Agric. Forest. In press.
• Champ, K. 2004. Isolation and characterization of components of low temperature-induced signal transduction pathways in Poncirus trifoliata (L.) Raf. and Citrus paradisi Macf. . Ph.D Thesis. University of Florida, Gainesville, Florida,125 pp.
• Chávez-Vela, N.A., Lucía, I., Chávez-Ortiz, E. And Pérez-Molphe B. 2003. Genetic transformatıon of sour orange using Agrobacterium rhizogenes. Agrociencia. 37: 629-639.
• Chiba, H., Uehara, M., Wu, J., Wang, X., Masuyama, R., Suzuki, K., Kanazawa, K. and Ishimi, Y. 2003. Hesperidin, a citrus flavonoid, inhibits bone loss and decreases serum and hepatic lipids in ovaryectomized mice. J. Nutr. 133:1892–1897.
• Costa, M.G.C., Otoni, W.C. and Moore, G.A. 2002. An evaluation of factors affecting the efficiency of Agrobacteriummediated transformation of Citrus paradisi (Macf.) and production of transgenic plants containing carotenoid biosynthetic genes. Plant Cell Rep. 21: 365–373.
• Derrick, K.S., Lee, R.F., Brlansky, R.H., Timmer, L.W., Hewitt, B.G. and Barthe, G.A. 1990. Proteins associated with citrus blight. Plant Dis. 74:168–170. Derrick, K.S. and Timmer, L.W. 2000. Citrus blight and other diseases of recalcitrant etiology. Annu. Rev. Phytopathol. 38:181–201.
• Dominguez, A., Guerri, J., Cambra, M., Navarro, L., Moreno, P. and Peña L. 2000. Efficient production of citrus transgenic plants expressing the coat protein gene of Citrus Tristeza Virus. Plant Cell Rep. 19: 427–433.
• Febres, V.J., Niblett, C.L., Lee, R.F. and Moore, G.A. 2003. Characterization of grapefruit plants (Citrus paradisi Macf.) transformed with citrus tristeza closterovirus genes. Plant Cell Rep. 21:421–428.
• Frydman, A., Weisshaus, O., Bar-Peled, M., Huhman, D.V., Sumner, L.W., Marin, FR., Lewinsohn, E., Fluhr, R., Gressel, J. and Eyal, Y. 2004. Citrus fruit bitter flavors: Isolation and functional characterization of the gene Cm1,2RhaT encoding a 1,2 rhamnosyltransferase, a key enzyme in the biosynthesis of the bitter flavonoids of citrus. The Plant Journal 40: 88–100.
• Gmitter, F.G.J., Grosser, J.W., and Moore, G.A. 1992 Citrus. (ed.): Hammerschlag, F.A., and Litz, R.E. Biotechnology in agriculture, vol 8. CAB International. pp 335-336, Wallingford, Oxford.
• Ghorbel, R., Dominguez, A., Navarro, L. and Peña, L. 2000. High efficiency genetic transformation of sour orange Citrus aurantium L. and production of transgenic trees containing the coat protein gene of Citrus Tristeza Virus. Tree Physiol. 20: 1183–1189.
• Ghorbel, R., Lopez, C., Moreno, P., Navarro, L., Flores, R. and Peña L. 2001. Transgenic citrus plants expressing the Citrus Tristeza Virus p23 protein exhibit viral-like symptoms. Mol. Plant Pathol. 2: 27–36.
• Gross, J. 1987. Carotenoids: Pigments in Fruits. Academic Press, London.
• Gutierrez M.A., Luth, E.D. and Moore. G.A. 1997. Factors affecting Agrobacterium-mediated transformation in citrus and production of sour orange (Citrus aurantium L.) plants expressing the coat protein gene of citrus tristeza virus. Plant Cell Rep. 16745–753
• Harborne, J. Plant Flavonoids in Biology and Medicine, Biochemical, Pharmacological,and Structure-Activity Alan R. Liss. pp. 163–175, New York.
• Hidaka, T., Omura, M., Ugaki, M., Tomiyama, M., Kato, A., Ohshima, M. and Motoyoshi, F., 1990. Agrobacteriummediated transformation and regeneration of Citrus from suspension cells. Jpn. J. Breed. 40, 199–207.
• Hidaka, T. and Omura, M. 1993. Transformation of citrus protoplasts by electroporation. J. Jpn. Soc. Hortic. Sci.62:371–376.
• Horowitz, R.M. 1986. Taste effects of flavonoids. (ed.): Cody, V., Middleton, E. Jr. and Kaneyoshi, J., Kobayashi, S. Nakamura, Y., Shigemoto, N. and Doi, Y. 1994. A simple and efficient gene transfer system of trifoliate orange (Poncirus trifoliata (L.) Raf.), Plant Cell Rep. 13: 541–545.
• Kato, M., Ikoma, Y., Matsumoto, H., Sugiura, M., Hyodo, H. and Masamichi, Y. 2004. Accumulation of Carotenoids and Expression of Carotenoid Biosynthetic Genes during Maturation in Citrus Fruit. Plant Physiol. 134: 824–837.
• Kayım, M. 1997. Genetic transformation of Kutdiken lemon [Citrus limon (L.) Burm. f] variety and regeneration of transgenic plants. PhD thesis, University of Cukurova, Adana.
• Kayım, M., Grosser, J.W., Barthe, G.A. and Derrick, K. S. 2003. Shoot regeneration from epicotyl segments of rough lemon (Citrus jambhiri Lush.). In: Proc. Int. Soc. Citrus. Int. Congr. 2000. Int. Soc. Citriculture. Orlando, Fla., p 212.
• Kayım, M., Ceccardi ,T.L., Berretta, M.J.G., Barthe, G.A. and Derrick, K.S. 2004. Introduction of a citrus blight-associated gene into Carrizo citrange [Citrus sinensis (L.) Osbc. Poncirus trifoliata (L.) Raf.] by Agrobacterium-mediated transformation Plant Cell Rep. 23:377–385.
• Kayım,M. and Koç, N.K. 2005. Improved transformation efficiency in citrus by plasmolysis treatment. Journal of Plant Biochemistry and Biotechnology, 14 (1), 15-20.
• Kim, H.K., Jeon, W.K. and Ko, B.S. 2001. Flavanone glycosides from Citrus junos and their anti-influenza virus activity. Planta Med. 67:548–549.
• Kobayashi, S. and Uchimiya, H. 1989. Expression and integration of a foreign gene in orange (Citrus sinensis Osb.) protoplasts by direct DNA transfer. Jpn. J. Genet 64:91–97.,
• Koca, U. 2005. Manipulation of the flavanoid pathway in citrus. Ph.D Thesis. University of Florida, Gainesville, Florida, 110 pp.
• Li, D.D., Shi, W. and Deng, X.X. 2002. Agrobacterium-mediated transformation of embryogenic calluses of Ponkan mandarin and the regeneration of plants containing the chimeric ribonuclease gene. Plant Cell Rep. 21:153–156.
• Luth, D. and Moore, G.A. 1999. Transgenic grapefruit plants obtained by Agrobacterium tumefaciens-mediated transformation. Plant Cell Tiss. Org. 57: 219–222.
• Manthey, J.A., Grohmann, K. and Guthrie, N. 2001. Biological properties of citrus flavonoids pertaining to cancer and inflammation. Curr. Med. Chem. 8: 135–153.
• Manthey, J.A. and Guthrie, N. 2002. Antiproliferative activities of citrus flavonoids against six human cancer cell lines. J. Agric. Food Chem. 50: 5837–5843. Middleton, Jr.E., Kandaswami, C. and Theoharides, T.C. 2000. The effects of plant flavonoids on mammalian cells: implications for inflammation,heart disease, and cancer. Pharmacol. Rev. 52: 673–751
• Molinari, H.B.C., Marur, C.J., Bespalhok, J.C., Kobayashi, A.K., Pileggi, M., Leite, R.P., Protasio-Pereira, L.P. and Esteves-Vieira, L.G. 2004. Osmotic adjustment in transgenic citrus rootstock Carrizo citrange (Citrus sinensis Osb. x Poncirus trifoliata L. Raf.) overproducing proline. Plant Science. 167: 1375-1381.
• Moore, G.A., Jacono, C.C., Neidigh, J.L., Lawrence, S.D. and Cline, K. 1992. Agrobacterium-mediated transformation of citrus stem segments and regeneration of transgenic plants, Plant Cell Rep.11: 238–242.
• Moore, G.A., Jacono, C.C., Neidigh, J.L., Lawrence, S.D. and Cline, K. 1993. Transformation in citrus. In: Bajaj YPS (ed.) Biotechnology in agriculture and forestry, vol 23, plant protoplasts and genetic engineering IV. Springer, Berlin Heidelberg New York, pp 194–208.
• Peña, L., Cervera, M., Navarro, A., Pina, J.A., Duran-Vila, N. and Navarro, L. 1995a. Agrobacterium-mediated transformation of sweet orange and regeneration of transgenic plants. Plant Cell Rep.14:616-619.
• Peña, L., Cervera, M., Juarez, J., Ortega, C., Pina, J.A., Durán-Vila, N. and Navarro, L. 1995b. High efficiency Agrobacterium-mediated transformation and regeneration of Citrus. Plant Sci. 104:183-191.
• Peña, L., Cervera, M., Juarez, J., Navarro, A., Pina, J.A. and Navarro, L. 1997. Genetic transformation of lime (Citrus aurantifolia Swing): factors affecting transformation and regeneration. Plant Cell Rep. 16:731-737.
• Pérez-Molphe-Balch, E. and Ochoa-Alejo, N. 1998. Regeneration of transgenic plants of Mexican lime from Agrobacterium rhizogenes-transformed tissues. Plant Cell Rep. 17 :591–596.
• Rahbe, Y. and Febvay, G. 1993. Protein toxicity to aphids: an in vitro test on Acyrthosiphon pisum. Entomol Exp. Appl. 67: 149–160.
• Rock, C.D. and Zeevaart, J.A.D. 1991. The aba mutant of Arabidopsis thaliana is impaired in epoxy-carotenoid biosynthesis. Proc. Natl. Acad. Sci. 88:7496–7499.
• Rao, K.V., Rathore, K.S., Hodges, T.K., Fu, X., Stoger, E., Sudhakar, D., Williams, S., Christou, P., Bharathi, M., Bown, D.P., Powell, K.S, Spence, J., Gatehouse, A.M, and Gatehouse J.A. 1998. Expression of snowdrop lectin (GNA) in transgenic rice plants confers resistance to rice brown planthopper. Plant J. 15:469-477.
• Gatehouse, J.A., Gatehouse A, and Fitches, E. 1997. Effects of snowdrop lectin (GNA) delivered via artificial diet and transgenic plants on the development of tomato moth (Lacanobia oleracea) larvae in laboratory and glasshouse trials. J Insect Physiol. 43:727-739.
• Silalahi, J. 2002. Anticancer and health protective properties of citrus fruit components. Asia. Pac. J. Clin. Nutr. 11(1),79–84.
• Şahin-Çevik, M. 2003. Identification and Characterization of cold-regulated genes in cold-hardy Citrus relative Poncirus trifoliata (L.) Raf. Ph.D Thesis. University of Florida, Gainesville, Florida, 130 pp.
• Sahin-Çevik M, and Moore GA. 2006a. Identification and expression analysis of cold-regulated genes from the cold-hardy Citrus relative Poncirus trifoliata (L.) Raf. Plant Mol Biol. 262: 83-97.
• Şahin-Çevik, M. and Moore, G.A. 2006b. Isolation and characterization of a novel RING-H2 finger gene induced in response to cold and drought in the interfertile Citrus relative Poncirus trifoliata. Physiologia Plantarum. 126: 153-161.
• Şahin-Çevik, M. and Moore, G.Aç 2006c. Two AP2 domain containing genes isolated from the cold-hardy Citrus relative Poncirus trifoliata are induced in response to cold. Functional Plant Biology 33: 863–875
• Sauvion, N., Rahbe, Y., Peumans, W.J., van Damme, E., Gatehouse, J.A. and Gatehouse, A.M.R. 1996. Effects of GNA and other mannosebinding lectins on development and fecundity of the peachpotato aphid. Entomol. Exp. Appl. 79: 285–293. Wong, W.S., Li, G.G., Ning, W., Xu, Z.F., Hsiao, W.L.W., Zhang, L.Y. and Li, N. 2001. Repression of chilling-induced ACC accumulation in transgenic citrus by over-production of antisense1-aminocyclopropa ne-1-carboxylate synthase RNA. Plant Science 161: 969–977.
• Van Damme, E.J.M., DeClerq, N., Claessens, F., Hemschoote, K., Peeters, B. and Peumans, W. 1991. Molecular cloning and characterisation of multiple isoforms of the snowdrop (Galanthus nivalis L.) lectin. Planta. 186: 35–43.
• Vardi, A., Bleichman, S. and Aviv, D. 1990. Genetic transformation of Citrus protoplasts and regeneration of transgenic plants. Plant Sci. 69:199–206.
• Yang, Z.N., Ingelbrecht, I.L., Louzada, E., Skaria, M. and Mirkov, T.E. 2000. Agrobacterium-mediated transformation of the commercially important grapefruit cultivar Rio Red (Citrus paradisi Macf.). Plant Cell Rep. 19:1203-1211.
• Yao, J.L., Wu, J.H., Gleave, A.P. and Morris, B.A.M. 1996. Transformation of citrus embryogenic cells using particle bombardment and production of transgenic embryos. Plant Sci. 113; 175–183.