Developments of Rice Cell Suspension Culture and A Novel Strategy for Screening New Resistant Lines to Rice Blight Disease Caused by Xanthomonas oryzae pv. oryzae

Restu Rizkyta Kusuma, Liliek Sulistyowati, Chiu-Chsiung Cheng, Yi-Hsien Lin

Abstract


The research aimed to develop a rice cell culture system with high proliferation and screening resistant cell lines of rice to bacterial blight disease caused by Xanthomonas oryza pv. oryzae (Xoo). The culture cells obtained from the callus, cultured on CS-1 medium containing 3 % sucrose and 2 mg L-1 2,4-D for 4 weeks. The results showed that proliferation cell was signifcantly increased 1-fold in 3 weeks of primary culture in CS-1 conditioned medium (fresh/spent medium ratio 1:1) containing 3 % sucrose, 0.5 % glucose, 0.05 % fructose and 2 mg L-1 2,4-D. This medium was used to screen the cell lines through applying culture filtrate of Xoo. The method was to find a novel cell line which could produce high amounts of reactive oxygen species (ROS). Screening results showed 33 % cell lines were strong ROS-producing, two cell lines were selected and cultured for second round screening. The ratio of strong ROS-producing cell lines was increased up to 67 % in the third round screening. The strong ROS-producing cell lines in third round screening can be further cultured for plant regeneration. The rice cell lines with high ROS production may have potential of resistant cell lines against Xoo.

Keywords


Bacterial blight; Cell culture; Reactive oxygen species; Resistant cell lines; Screening

Full Text:

PDF

References


Ashraf, M., & Akram, N. A. (2009). Improving salinity tolerance of plants through conventional breeding and genetic engineering: An analytical comparison. Biotechnology Advances, 27(6), 744–752. crossref

Ben Amar, A., Cobanov, P., Boonrod, K., Krczal, G., Bouzid, S., Ghorbel, A., & Reustle, G. M. (2007). Efficient procedure for grapevine embryogenic suspension establishment and plant regeneration: Role of conditioned medium for cell proliferation. Plant Cell Reports, 26(9), 1439–1447. crossref

Bi, Y., Chen, W., Zhang, W., Zhou, Q., Yun, L., & Xing, D. (2009). Production of reactive oxygen species, impairment of photosynthetic function and dynamic changes in mitochondria are early events in cadmium-induced cell death in Arabidopsis thaliana. Biology of the Cell, 101(11), 629–643. crossref

Botha, F. C., & O’Kennedy, M. M. (1998). Carbohydrate utilisation by cell suspension cultures of Phaseolus vulgaris. Physiologia Plantarum, 102(3), 429–436. crossref

Chen, H., Lin, Y., & Zhang, Q. (2009). Review and prospect of transgenic rice research. Chinese Science Bulletin, 54, 4049. crossref

Daub, M. E. (1986). Tissue culture and the selection of resistance to pathogens. Annual Review of Phytopathology, 24(1), 159–186. crossref

Eruslanov, E., & Kusmartsev, S. (2010). Identification of ROS using oxidized DCFDA and flow-cytometry. In Armstrong, D. (Ed.), Advanced Protocols in Oxidative Stress II (vol. 594, pp. 57–72). Totowa, NJ: Humana Press. crossref

Gomes, A., Fernandes, E., & Lima, J. L. F. C. (2005). Fluorescence probes used for detection of reactive oxygen species. Journal of Biochemical and Biophysical Methods, 65(2–3), 45–80. crossref

Hayashi, T., Ohsumi, C., Kato, Y., Yamanouchi, H., Toriyama, K., & Hinata, K. (1994). Effects of amino acid medium on cell aggregation in suspension-cultured rice cells. Bioscience, Biotechnology, and Biochemistry, 58(2), 256–260. crossref

Htwe, N. N., Maziah, M., Ling, H. C., Zaman, F. Q., & Zain, A. M. (2011). Regeneration capacity of cell suspension culture in Malaysian rice genotypes under salinity stress. Asian Journal of Biotechnology, 3(4), 357–367. crossref

Jones, J. D. G., & Dangl, J. L. (2006). The plant immune system. Nature, 444, 323–329. crossref

Lee, T. J., Shultz, R. W., Hanley-Bowdoin, L., & Thompson, W. F. (2004). Establishment of rapidly proliferating rice cell suspension culture and its characterization by fluorescence-activated cell sorting analysis. Plant Molecular Biology Reporter, 22(3), 259–267. crossref

Mew, T. W., Alvarez, A. M., Leach, J. E., & Swings, J. (1993). Focus on bacterial blight of rice. Plant Disease, 77(1), 5–12. crossref

Niño-Liu, D. O., Ronald, P. C., & Bogdanove, A. J. (2006). Xanthomonas oryzae pathovars: Model pathogens of a model crop. Molecular Plant Pathology, 7(5), 303–324. crossref

Ohira, K., Ikeda, M., & Ojima, K. (1976). Thiamine requirements of various plant cells in suspension culture. Plant and Cell Physiology, 17(3), 583–590. crossref

Ohira, K., Ojima, K., & Fujiwara, A. (1973). Studies on the nutrition of rice cell culture I. A simple, defined medium for rapid growth in suspension culture. Plant & Cell Physiology, 14(6), 1113–1121. crossref

Ong, S., Ling, A., Poospooragi, R., & Moosa, S. (2011). Production of Flavonoid compounds in cell cultures of Ficus deltoidea as influenced by medium composition. International Journal of Medicinal and Aromatic Plants, 1(2), 62–74. Retrieved from PDF

Ryba, W. M., Jean-Loup, N., & Leach, J. E. (1995). Comparison of Xanthomonas oryzae pv. oryzae strains from Africa, North America, and Asia by restriction fragment length polymorphism analysis. International Rice Research Notes, 20(1), 25–26. Retrieved from website

Sharp, W. R., Evans, D. A., Ammirato, P. V., & Yamada, Y. (Eds.). (1984). Handbook of plant cell culture (Vol. 2 Crop species). New York: Macmillan Publishing Co.

Song, F., & Goodman, R. M. (2001). Molecular biology of disease resistance in rice. Physiological and Molecular Plant Pathology, 59(1), 1–11. crossref

Taji, A. M., Kumar, P. P., & Lakshmanan, P. (2002). In vitro plant breeding. Abingdon, UK: Taylor & Francis. Retrieved from PDF

Torres, M. A., Jones, J. D. G., & Dangl, J. L. (2006). Reactive oxygen species signaling in response to pathogens. Plant Physiology, 141(2), 373–378. crossref

van Hengel, A. J., Guzzo, F., van Kammen, A., & de Vries, S. C. (1998). Expression pattern of the carrot EP3 endochitinase genes in suspension cultures and in developing seeds. Plant Physiology, 117(1), 43–53. crossref

Wieczorek, A. M., & Wright, M. G. (2012). History of agricultural biotechnology: How crop development has evolved. Nature Education Knowledge, 3(10), 9. Retrieved from website




DOI: http://doi.org/10.17503/agrivita.v40i3.1779

Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.