Indica Rice (Oryza sativa, BR29 and IR64)
Part of the
Methods in Molecular Biology
book series (MIMB, volume 343)
Rice is the world’s most important food crop. Indica-type rice provides the staple food for more than half of the world population. To satisfy the growing demand of the ever-increasing population, more sustained production of indica-type rice is needed. In addition, because of the high per capita consumption of indica rice, improvement of any traits including its nutritive value may have a significant positive health outcome for the rice-consuming population. Rice yield productivity is greatly affected by different biotic stresses, like diseases and insect pests, and abiotic stresses like drought, cold, and salinity. Attempts to improve resistance in rice to these stresses by conventional breeding through introgression of traits have limited success owing to a lack of resistance germplasm in the wild relatives. Gene transfer technology with genes from other sources can be used to make rice plants resistant or tolerant to insect pests, diseases, and different environmental stresses. For improving the nutritional value of the edible endosperm part of the rice, genes for increasing iron, β-carotene, or better quality protein can be introduced in rice plants by genetic engineering. Different crops have been transformed using various gene transfer methods, such as protoplast transformation, biolistic, and Agrobacterium-mediated transformation. This chapter describes the Agrobacterium-mediated transformation protocol for indica-type rice. The selectable marker genes used are hygromycin phosphotransferase (hpt), neomycin phosphotransferase (nptII), or phosphomannose isomerase (pmi), and, accordingly, the selection agents are hygromycin, kanamycin (G418), or mannose, respectively.
Key WordsIndica-type rice Agrobacterium-mediated transformation Agrobacterium tumefaciens genetic engineering marker gene
Datta, S. K., Peterhans, A., Datta, K., and Potrykus, I. (1990) Genetically engineered fertile Indica-rice plants recovered from protoplasts. Bio/Technology
, 736–740.CrossRefGoogle Scholar
Lin, W., Anuratha, C. S., Datta, K., Potrykus, I., Muthukrishnan, S., and Datta, S. K. (1995) Genetic engineering of rice for resistance to sheath blight. Bio/Technology
, 686–691.CrossRefGoogle Scholar
Datta, K., Vasquez, A., Tu, J., et al. (1998) Constitutive and tissue-specific differential expression of cryIA(b)
gene in transgenic rice plants conferring resistance to rice insect pest. Theor. Appl. Genet.
, 20–30.CrossRefGoogle Scholar
Garg, A. K., Kim, J. K., Owens, T. G., et al. (2002) Trehalose accumulation in rice plants confers high tolerance levels to different abiotic stresses. Proc. Natl. Acad. Sci. USA
, 15898–15903.PubMedCrossRefGoogle Scholar
Sakamoto, A. and Murata, N. (2000) Genetic engineering of glycinebetaine synthesis in plants, current status and implications for enhancement of stress tolerance. J. Exp. Bot.
, 81–88.PubMedCrossRefGoogle Scholar
Datta, S. K. (2004) Rice biotechnology: a need for developing countries. AgBioForum
, 30–34.Google Scholar
de Vasconcelos, M., Datta, K., Oliva, N., et al. (2003) Enhanced iron and zinc accumulation in transgenic rice with the ferritin gene. Plant Sci.
(3), 371–378.CrossRefGoogle Scholar
Datta, K., Baisakh, N., Oliva, N., et al. (2003) Bioengineered ‘golden’ indica rice cultivars with beta-carotene metabolism in the endosperm with hygromycin and mannose selection systems. Plant Biotechnol. J.
, 81–90.PubMedCrossRefGoogle Scholar
Ye, X., Al-Babili, S., Klöti, A., et al. (2000) Engineering the provitamin A (β-carotene) biosynthetic pathway into (carotenoid free) rice endosperm. Science
, 303–305.PubMedCrossRefGoogle Scholar
Baisakh N, and Datta, S. K. (2004) Metabolic pathway engineering for nutrition enrichment, in Molecular Biology and Biotechnology of Plant Organelles
(Daniell, H. and Chase, C. D., eds.), Springer, The Netherlands, pp. 527–542.CrossRefGoogle Scholar
Peterhans, A., Datta, S. K., Datta, K., Godall, G. H., Potrykus, I., and Paszkowski, J. (1990) Recognition of efficiency of Dicotylendoneae-specific promoter and RNA processing signals in rice. Mol. Gen. Genet.
, 362–368.Google Scholar
Datta, S. K., Datta, K., Soltanifar, N., Donn, G., and Potrykus, I. (1992) Herbicide-resistant Indica rice plants from IRRI breeding line IR72 after PEG-mediated transformation of protoplasts. Plant Mol. Biol.
, 619–629.PubMedCrossRefGoogle Scholar
Parkhi, V., Rai, M., Tan, J., et al. (2005) Molecular characterization of markerfree transgenic events accumulating differential expression of carotenoids in seed endosperm of indica rice. Mol. Genet. Genomics
, 325–336.PubMedCrossRefGoogle Scholar
Hiei, Y., Ohta, S., Komari, T., and Kumashiro, T. (1994) Efficient transformation of rice (Oryza sativa
) mediated by Agrobacterium
and sequence analysis of the boundaries of the T-DNA. Plant J.
, 271–282.PubMedCrossRefGoogle Scholar
Datta, K., Oliva, N., Torrizo, L., Abrigo, E., Khush, G. S., and Datta, S. K. (1996) Genetic transformation of Indica and Japonica rice by Agrobacterium tumefaciens
. Rice Genet. Newslett.
, 36–139.Google Scholar
Datta, K., Koukolíkovà-Nicola, Z., Baisakh, N., Oliva, N., and Datta, S. K. (2000) Agrobacterium
-mediated engineering for sheath blight resistance of indica rice cultivars from different ecosystems. Theor. Appl. Genet.
, 832–839.CrossRefGoogle Scholar
Chilton, M. D., Currier, T. C., Farrand, S. K., Bendich, A. J., and Nester, E. W. (1974) Agrobacterium
tumefaciens DNA and PS8 bacteriophage DNA not deleted in crown gall tumors. Proc. Natl. Acad. Sci. USA
, 3672–3676.PubMedCrossRefGoogle Scholar
Murashige, T. and Skoog, F. (1962) A revised medium for rapid growth and bioassay with tobacco tissue cultures. Physiol. Plant.
, 473–497.CrossRefGoogle Scholar
Chu, C. C., Wang, C. S., Sun, C. C., Hsu, C., Yin, K. C., and Chu, C. Y. (1975) Establishment of an efficient medium for anther culture of rice through comparative experiments on the nitrogen sources. Sci. Sinica
, 659–668.Google Scholar
Yoshida, S., Forno, D. A., Cock, J. H., and Gomez, K. A. (1976) Laboratory manual for physiological studies of rice. The International Rice Research Institute, Los Baños, Philippines.Google Scholar
Ho, N. H., Baisakh, N., Oliva, N., et al. (2005) Translational fusion hybrid Bt genes confer resistance against yellow stem borer (Scirpophaga incertulas
Walker) in transgenic elite Vietnamese rice (Oryza sativa
L.) (communicated).Google Scholar