Forthcoming

The first success in utilizing the RUBY reporter gene for rice transformation in Vietnam

Thi Mai Anh Pham, Khanh Linh Ly, Khanh Linh Chu, Quyen Phan, Phuong Thao Bui, Hong Diep Le, Xuan Cuong Nguyen, Hoang Ha Chu, Tien Phat Do
Author affiliations

Authors

  • Thi Mai Anh Pham \(^1\) Institute of Biology, Vietnam Academy of Science and Technology, 18 Hoang Quoc Viet, Nghia Do, Hanoi, Vietnam
  • Khanh Linh Ly \(^1\) Institute of Biology, Vietnam Academy of Science and Technology, 18 Hoang Quoc Viet, Nghia Do, Hanoi, Vietnam
    \(^5\) Agricultural Biotechnology Research Center, Academia Sinica, Taipei 11529, Taiwan
  • Khanh Linh Chu \(^1\) Institute of Biology, Vietnam Academy of Science and Technology, 18 Hoang Quoc Viet, Nghia Do, Hanoi, Vietnam
    \(^6\) Graduate School of Green-Bio Science and Crop Biotech Institute, Kyung Hee University, Yongin 17104, Republic of Korea
    https://orcid.org/0009-0003-7304-9067
  • Quyen Phan \(^1\) Institute of Biology, Vietnam Academy of Science and Technology, 18 Hoang Quoc Viet, Nghia Do, Hanoi, Vietnam
  • Phuong Thao Bui \(^1\) Institute of Biology, Vietnam Academy of Science and Technology, 18 Hoang Quoc Viet, Nghia Do, Hanoi, Vietnam
  • Hong Diep Le \(^3\) Hanoi University of Science, Vietnam National University, 334 Nguyen Trai, Thanh Xuan, Hanoi, Vietnam
  • Xuan Cuong Nguyen \(^1\) Institute of Biology, Vietnam Academy of Science and Technology, 18 Hoang Quoc Viet, Nghia Do, Hanoi, Vietnam
    \(^4\) Department of Genetics, University of Georgia, 120 East Green Street, Athens, GA 30602, USA
  • Hoang Ha Chu \(^1\) Institute of Biology, Vietnam Academy of Science and Technology, 18 Hoang Quoc Viet, Nghia Do, Hanoi, Vietnam
    \(^2\) Graduate University of Science and Technology, Vietnam Academy of Science and Technology, 18 Hoang Quoc Viet, Nghia Do, Hanoi, Vietnam
  • Tien Phat Do \(^1\) Institute of Biology, Vietnam Academy of Science and Technology, 18 Hoang Quoc Viet, Nghia Do, Hanoi, Vietnam
    \(^2\) Graduate University of Science and Technology, Vietnam Academy of Science and Technology, 18 Hoang Quoc Viet, Nghia Do, Hanoi, Vietnam
    https://orcid.org/0000-0002-4227-7396

DOI:

https://doi.org/10.15625/vjbt-23946

Keywords:

Betalains, Kitaake, reporter gene, rice, RUBY.

Abstract

To date, several reporter genes have been developed, such as GFP, GUS, and luciferase. However, GFP and luciferase genes are costly and require modern equipment, while GUS requires sample destruction. To overcome these limitations, the RUBY gene is considered a promising alternative. RUBY is an artificial open-reading frame, encoding CYP76AD1, L-DOPA 4,5-dioxygenase, and glucosyltransferase, which are key enzymes involved in the biosynthesis of betalain, which displays a red-colored pigment, providing a visual marker for transformation events. In this study, the RUBY gene was successfully transformed into the model rice variety Kitaake via the Agrobacterium-mediated method. The activity of the RUBY gene was visually detected by red spots in calli and somatic embryos through the rice transformation and regeneration procedure. Moreover, the presence and expression of this reporter gene were confirmed by PCR and the red coloration in the regenerated rice lines. The RUBY gene was stably expressed in transgenic rice lines under both in vitro and greenhouse conditions and was heritable in the T1 generation, without exhibiting any adverse effects on plant growth and development. This result demonstrates the potential of RUBY as a visual selection marker for rice transformation in Vietnam and opens the door to further applications in other important crops.

Downloads

Download data is not yet available.

References

Blochlinger, K., and Diggelmann, H. (1984). Hygromycin B phosphotransferase as a selectable marker for DNA transfer experiments with higher eucaryotic cells. Molecular and Cellular Biology, 4 (12), 2929-2931. https://doi.org/10.1128/mcb.4.12.2929-2931.1984

Phuong Thao, B., Thi Linh, N., Van Manh, N., Khanh Linh, L., Hoang Ha, C., Tien Phat, D., & Bich Ngoc, P. (2022). Optimization of Agrobacterium-mediated transformation procedure for an indica rice variety-Khang Dan 18. Vietnam Journal of Biotechnology, 20(1), 53-62. https://doi.org/10.15625/1811-4989/17200

Chalfie, M., Tu, Y., Euskirchen, G., Ward, W.W., and Prasher, D.C. (1994). Green Fluorescent Protein as a Marker for Gene Expression. Science, 263(5148), 802-805. https://doi.org/10.1126/science.8303295

Chen, L., Cai, Y., Liu, X., Yao, W., Wu, S., and Hou, W. (2024). The RUBY reporter for visual selection in soybean genome editing. aBIOTECH, 5(2), 209-213. https://doi.org/10.1007/s42994-024-00148-6

Chu, C.C., Chu, W.C., San, S.C., and Chen, H. (1975). Establishment of an efficient medium for another culture of rice through comparative experiments on the nitrogen sources. Scientia Sinica, 18(5), 659-668. https://doi.org/10.1360/ya1975-18-5-659

Davey, M.R., and Anthony, P. (2010). Plant Cell Culture: Essential Methods (1st ed.). Wiley, https://doi.org/10.1002/9780470686522

Doyle, J.J. and J.L. Doyle (1990). Isolation of plant DNA from fresh tissue. Focus, 12:13-15.

Fish, N., Lindsey, K., and Jones, M.G.K. (1988). Plant Protoplast Fusion. Methods in molecular biology (Clifton, N.J.), 4, 481-498. Fish, N., Lindsey, K., and Jones, M.G.K. (1988). Plant Protoplast Fusion. Methods in molecular biology (Clifton, N.J.), 4, 481-498. https://doi.org/10.1385/0-89603-127-6:481

Gamborg, O.L., Miller, R.A., and Ojima, K. (1968). Nutrient requirements of suspension cultures of soybean root cells. Experimental Cell Research, 50(5), 151-158. https://doi.org/10.1016/0014-4827(68)90403-5

He, Y., Zhang, T., Sun, H., Zhan, H., and Zhao, Y. (2020). A reporter for noninvasively monitoring gene expression and plant transformation. Horticulture Research, 7(1), 152. https://doi.org/10.1038/s41438-020-00390-1

Jefferson, R.A., Kavanagh, T.A., and Bevan, M.W. (1987). GUS fusions: beta-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. The EMBO Journal, 6(13), 3901-3907. https://doi.org/10.1002/j.1460-2075.1987.tb02730.x

Khanh Linh, L., Van Doai, N., Thi Linh, N., Quyen, P., Hoang Ha, C., Bich Ngoc, P., and Tien Phat, D. (2023). Development of CRISPR/Cas9 systems to induce targeted mutations in the promoter region of the OsSRFP1 gene in rice. Vietnam Journal of Biotechnology, 21(3), 463-475. https://doi.org/10.15625/1811-4989/19532

Lee, K., Kang, M., Ji, Q., Grosic, S., and Wang, K. (2023). New T-DNA binary vectors with NptII selection and RUBY reporter for efficient maize transformation and targeted mutagenesis. Plant Physiology, 192 (4), 2598-2603. https://doi.org/10.1093/plphys/kiad231.

Li, L., Qu, R., De Kochko, A., Fauquet, C., and Beachy, R.N. (1993). An improved rice transformation system using the biolistic method. Plant Cell Reports, 12(5), 250-255. https://doi.org/10.1007/BF00237129

Miah, M.A.A., Pathan, M.S., and Quayum, H.A. (1996). Production of salt tolerant rice breeding line via doubled haploid. Euphytica, 91(3), 285-288. https://doi.org/10.1007/BF00033089

Murashige, T., and Skoog, F. (1962). A Revised Medium for Rapid Growth and Bioassays with Tobacco Tissue Cultures. Physiologia Plantarum, 15(3), 473-497. https://doi.org/10.1111/j.1399-3054.1962.tb08052.x

Ohira, K., Ojima, K., and 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. https://doi.org/10.1093/oxfordjournals.pcp.a074950

Ow, D.W., Wood, K.V., DeLuca, M., De Wet, J.R., Helinski, D.R., and Howell, S.H. (1986). Transient and Stable Expression of the Firefly Luciferase Gene in Plant Cells and Transgenic Plants. Science, 234(4778), 856-859. https://doi.org/10.1126/science.234.4778.856

Sallaud, C., Meynard, D., Van Boxtel, J., Gay, C., Bès, M., Brizard, J.P., et al. (2003). Highly efficient production and characterization of T-DNA plants for rice (Oryza sativa L.) functional genomics. Theoretical and Applied Genetics, 106(8), 1396-1408. https://doi.org/10.1007/s00122-002-1184-x

Slamet-Loedin, I.H., Chadha-Mohanty, P., and Torrizo, L. (2014). Agrobacterium-Mediated Transformation: Rice Transformation, in: Henry, R.J., Furtado, A. (Eds.), Cereal Genomics, Methods in Molecular Biology (Clifton, N.J.), 1099, 261-271. https://doi.org/10.1007/978-1-62703-715-0_21

Sun, H., Wang, S., Zhu, C., Yang, K., Liu, Y., and Gao, Z. (2023). A new biotechnology for in-planta gene editing and its application in promoting flavonoid biosynthesis in bamboo leaves. Plant Methods 19(1), 20. https://doi.org/10.1186/s13007-023-00993-4

Verma, D.K., and Srivastav, P.P. (2020). Bioactive compounds of rice (Oryza sativa L.): Review on paradigm and its potential benefit in human health. Trends in Food Science & Technology, 97, 355-365. https://doi.org/10.1016/j.tifs.2020.01.007

Wang, D., Zhong, Y., Feng, B., Qi, X., Yan, T., Liu, J., et al. (2023). The RUBY reporter enables efficient haploid identification in maize and tomato. Plant Biotechnology Journal, 21(8), 1707-1715. https://doi.org/10.1111/pbi.14071.

Downloads

Published

15-04-2026

How to Cite

Pham, T. M. A., Ly, K. L., Chu, K. L., Phan, Q., Bui, P. T., Le, H. D., … Do, T. P. (2026). The first success in utilizing the RUBY reporter gene for rice transformation in Vietnam. Vietnam Journal of Biotechnology. https://doi.org/10.15625/vjbt-23946

Issue

Section

Articles

Funding data

Most read articles by the same author(s)

Similar Articles

<< < 9 10 11 12 13 14 15 > >> 

You may also start an advanced similarity search for this article.