Main Article Content
Beta-glucan in cereal crops is known as a functional food, which can reduce cardiovascular diseases by lowering blood cholesterol levels. However, beta-glucan content is relatively low in rice grains, despite being relatively abundant in barley and oat grains. Taking advantage of rice as the staple food for Asians, increasing beta-glucan content in rice for their consumption may help to reduce cardiovascular-related diseases among them. Previous attempts in increasing beta-glucan content in rice via transgene expression of beta-glucan synthase genes from barley into rice were unsuccessful due to the use of non-tissue specific as well as constitutively expressing promoter. The current transgenic expression study was performed to characterise the promoter of beta-glucan synthase gene in barley using beta-glucuronidase (GUS) reporter gene. Two fragments of HvCslF6 promoter (2771 bp and 1257 bp) were successfully fused with GUS reporter gene and integrated into rice plants, demonstrated that the promoter was functional in the heterologous plant system. The presence of blue GUS staining was observed on the leaf, root, stem, and grain of the transgenic rice regardless of the promoter length used and stayed functional up to the next generation. GUS qualitative analysis confirmed that the shorter promoter length generated a stronger GUS activity in comparison to the longer one. This indicated that the presence of repressor elements in between the -2771 bp and -1257 bp regions. The preliminary results shed light on the strong promoter activity in the rice endosperm tissue. It can become an alternative to the collection of plant promoters that can be used for grain quality improvement and biofortification.
Transfer of Copyrights
- In the event of publication of the manuscript entitled [INSERT MANUSCRIPT TITLE AND REF NO.] in the Malaysian Journal of Science, I hereby transfer copyrights of the manuscript title, abstract and contents to the Malaysian Journal of Science and the Faculty of Science, University of Malaya (as the publisher) for the full legal term of copyright and any renewals thereof throughout the world in any format, and any media for communication.
Conditions of Publication
- I hereby state that this manuscript to be published is an original work, unpublished in any form prior and I have obtained the necessary permission for the reproduction (or am the owner) of any images, illustrations, tables, charts, figures, maps, photographs and other visual materials of whom the copyrights is owned by a third party.
- This manuscript contains no statements that are contradictory to the relevant local and international laws or that infringes on the rights of others.
- I agree to indemnify the Malaysian Journal of Science and the Faculty of Science, University of Malaya (as the publisher) in the event of any claims that arise in regards to the above conditions and assume full liability on the published manuscript.
Copyright: Rights of the Author(s)
- Effective 2007, it will become the policy of the Malaysian Journal of Science (published by the Faculty of Science, University of Malaya) to obtain copyrights of all manuscripts published. This is to facilitate:
(a) Protection against copyright infringement of the manuscript through copyright breaches or piracy.
(b) Timely handling of reproduction requests from authorized third parties that are addressed directly to the Faculty of Science, University of Malaya.
- As the author, you may publish the fore-mentioned manuscript, whole or any part thereof, provided acknowledgement regarding copyright notice and reference to first publication in the Malaysian Journal of Science and Faculty of Science, University of Malaya (as the publishers) are given.
You may produce copies of your manuscript, whole or any part thereof, for teaching purposes or to be provided, on individual basis, to fellow researchers.
- You may include the fore-mentioned manuscript, whole or any part thereof, electronically on a secure network at your affiliated institution, provided acknowledgement regarding copyright notice and reference to first publication in the Malaysian Journal of Science and Faculty of Science, University of Malaya (as the publishers) are given.
- You may include the fore-mentioned manuscript, whole or any part thereof, on the World Wide Web, provided acknowledgement regarding copyright notice and reference to first publication in the Malaysian Journal of Science and Faculty of Science, University of Malaya (as the publishers) are given.
- In the event that your manuscript, whole or any part thereof, has been requested to be reproduced, for any purpose or in any form approved by the Malaysian Journal of Science and Faculty of Science, University of Malaya (as the publishers), you will be informed. It is requested that any changes to your contact details (especially e-mail addresses) are made known.
Copyright: Role and responsibility of the Author(s)
- In the event of the manuscript to be published in the Malaysian Journal of Science contains materials copyrighted to others prior, it is the responsibility of current author(s) to obtain written permission from the copyright owner or owners.
- This written permission should be submitted with the proof-copy of the manuscript to be published in the Malaysian Journal of Science
Arena, M. P., Graziano C., Fiocco D., Russo, P., Torelli, M., Spano, G., Capozzi, V. (2014) Barley β-Glucans-Containing Food Enhances Probiotic Performances of Beneficial Bacteria, International Journal of Molecular Sciences 15 (2): 3025–3039.
Bradford, Marion M. 1976. “A Rapid and Sensitive Method for the Quantitation of Microgram Quantities of Protein Utilizing the Principle of Protein-Dye Binding.” Analytical Biochemistry 72 (1): 248–54.
Burton, R. A., Collins, H.M., Kibble, N. A. J., Smith, J. A., Shirley, N. J., Jobling, S. A., Henderson, M. (2011) Over-Expression of Specific HvCslF Cellulose Synthase-like Genes in Transgenic Barley Increases the Levels of Cell Wall (1,3;1,4)-β-D-Glucans and Alters Their Fine Structure, Plant Biotechnology Journal 9 (2): 117–135.
Burton, R. A., Jobling, S. A., Harvey, A. J., Shirley, N. J., Mather, D. E., Bacic, A., Fincher, G. B. (2008) The Genetics and Transcriptional Profiles of the Cellulose Synthase-Like HvCslF Gene Family in Barley, Plant Physiology 146 (4): 1821–33.
Burton, R. A., Wilson, S. M., Hrmova, M., Harvey, A. J., Shirley, N. J., Medhurst, A., Stone, B. A., Newbigin, E. J., Bacic, A., Fincher, G. B. (2006) Cellulose Synthase-like CslF Genes Mediate the Synthesis of Cell Wall (1,3;1,4)-Beta-D-Glucans, Science 311 (5769): 1940–1942.
Dimitroff, G. (2016) Investigating the Synthesis and Regulation of (1,3;1,4)-Beta-Glucan Biosynthesis, Australia: University of Adelaide.
Ding, J., Jia, J., Yang, L., Wen, H., Zhang, C., Liu, W., Zhang, D. (2004) Validation of a Rice Specific Gene, Sucrose Phosphate Synthase, Used as the Endogenous Reference Gene for Qualitative and Real-Time Quantitative PCR Detection of Transgenes, Journal of Agricultural and Food Chemistry 52 (11): 3372–77.
Ermawar, R. A., Collin, H. M., Byrt, C. S., Henderson, M., O’Donovan, L. A., Shirley, N. J., Schwerdt, J. G., Lahnstein, J., Fincher, G. B., Burton R.A. (2015) Genetics and Physiology of Cell Wall Polysaccharides in the Model C4 Grass, Setaria Viridis Spp, BMC Plant Biology 15 (236)
Fu, C., Donovan, W. P., Shikapwashya-Hasser, O., Ye, X., Cole, R. H. (2014) Hot Fusion: An Efficient Method to Clone Multiple DNA Fragments as Well as Inverted Repeats without Ligase, PLOS ONE 9 (12): e115318
Garcia-Gimenez, G., Russell, J., Aubert, M. K., Fincher, G. B., Burton, R. A., Waugh, R., Tucker, M. R., Houston, K. (2019) Barley Grain (1,3;1,4)-β-Glucan Content: Effects of Transcript and Sequence Variation in Genes Encoding the Corresponding Synthase and Endohydrolase Enzymes, Scientific Reports 9 (1): 1–11.
Joyce, S. A., Kamil, A., Fleige, L., Gahan, C. G. M. (2019) The Cholesterol-Lowering Effect of Oats and Oat Beta Glucan: Modes of Action and Potential Role of Bile Acids and the Microbiome, Nutrition, 171
Juhász, A., Makai, S.,Sebestyén, E., Tamás, L., Balázs, E. (2011). Role of Conserved Non-Coding Regulatory Elements in LMW Glutenin Gene Expression, PLOS ONE 6 (12): e29501
Higo, K., Ugawa, Y., Iwamoto, M., Higo, H. (1998) PLACE: A Database of Plant Cis -Acting Regulatory DNA Elements, Nucleic Acids Research 26 (1): 358–59.
Hiei, Yukoh, Toshihiko Komari, and Tomoaki Kubo. 1997. “Transformation of Rice Mediated by Agrobacterium Tumefaciens.” Plant Molecular Biology 35 (1–2): 205–18.
Hobbs, S. L., Kpodar, P., DeLong. C. M. (1990) The Effect of T-DNA Copy Number, Position and Methylation on Reporter Gene Expression in Tobacco Transformants, Plant Molecular Biology 15 (6): 851–64.
Huang, J., Wei, Z., An, H., & Zhu, Y. (2001). Agrobacterium tumefaciens -mediated transformation of rice with the spider insecticidal gene conferring resistance to leaf folder and striped stem borer. Cell Research, 11, 149–155.
Kim, S., Zemelis-Durfee, S., Jensen, J. K., Wilkerson, C.G., Keegstra, K., Brandizzi, F. (2018) In the Grass Species Brachypodium Distachyon, the Production of Mixed-Linkage (1,3;1,4)-β-Glucan (MLG) Occurs in the Golgi Apparatus, The Plant Journal 93 (6): 1062–1075.
Lescot, M., Déhais, P., Thijs, G., Marchal, K., Moreau, Y., Peer, Y. V. D., Rouzé, P., Rombauts, S. (2002) PlantCARE, a Database of Plant Cis-Acting Regulatory Elements and a Portal to Tools for in Silico Analysis of Promoter Sequences, Nucleic Acids Research 30 (1): 325–27.
Lim, W. L., Collin, H. M., Byrt, C. S., Lahnstein, J., Shirley, N. J., Aubert, M. K., Tucker, M. R., Peukert, M., Matros, A., Burton, R. A. (2019) Overexpression of HvCslF6 in Barley Grain Alters Carbohydrate Partitioning plus Transfer Tissue and Endosperm Development, Journal of Experimental Botany 71 (1): 138–53.
Li, S., Chen, L., Peng, X., Wang, C., Qin, B., Tan, D., Han, C. (2019) Overview of the Reporter Genes and Reporter Mouse Models, Animal Models and Experimental Medicine, (8) 29–35.
Little, A., Schwerdt, J., Shirley, N. J., Khor, S. F., Neumann, K., O’Donovan, L. A., Lahnstein, J. (2018) Revised Phylogeny of the Cellulose Synthase Gene Superfamily: Insights into Cell Wall Evolution, Plant Physiology 177 (7): 1124–1141.
Liu, Q., Zhang J., Wang, Z., Hong, M., Gu, M. (1998) A highly efficient transformation system mediated by Agrobacterium tumefaciens in rice ( Oryza sativa L.), Acta Phytophysiol Sin 24 (3): 259–71.
Marković, S. M.,Đukić N. H., Knežević, D., Leković, S. V. (2017) Divergence of Barley and Oat Varieties according to the Content of β-Glucan (Short Communication), Journal of the Serbian Chemical Society 82 (4): 379–88.
Pelletier, J. M., Kwong, R. W., Park, S., Le, B. H., Baden, R., Cagliari, A., Hashimoto, M. (2017) LEC1 Sequentially Regulates the Transcription of Genes Involved in Diverse Developmental Processes during Seed Development, Proceedings of the National Academy of Sciences 114 (32): E6710–19.
Ren, Y., Xie, H, Liu, L., Jia, D., Yao, K., Chi, Y. (2018) Processing and Prebiotics Characteristics of β-Glucan Extract from Highland Barley, Applied Sciences 8 (1481): 1–11.
Wang, J. C., Xu, H., Zhu, Y., Liu, Q. Q., Cai, X. L. (2013) OsbZIP58, a Basic Leucine Zipper Transcription Factor, Regulates Starch Biosynthesis in Rice Endosperm, Journal of Experimental Botany 64 (11): 3453–3466.
Wilson, S. M., Ho, Y. Y., Lampugnani, E. R., Van de Meene a. M. L., Bain, M. P., Bacic, A., Doblin, M.S. (2015) Determining the Subcellular Location of Synthesis and Assembly of the Cell Wall Polysaccharide (1,3; 1,4)-β-D-Glucan in Grasses, The Plant Cell 27 (3): 754–71.
Wolever, T., Tosh, S., Gibbs, A., Brand-Miller, J., Duncan, A., Hart, V., Lamarche, B., Thomson, B., Duss, R., Wood, P. (2010)
Physicochemical Properties of Oat Beta-Glucan Influence Its Ability to Reduce Serum LDL Cholesterol in Humans: A Randomized Clinical Trial, The American Journal of Clinical Nutrition 92 (4): 723–32.
Wong, S. C., Shirley, N. J., Little, A., Khoo, K. H. P., Schwerdt, J., Fincher, G. B., Burton, R. A., Mather, D. E. (2015) Differential Expression of the HvCslF6 Gene Late in Grain Development May Explain Quantitative Differences in (1,3;1,4)-β-Glucan Concentration in Barley, Molecular Breeding 35 (1): 1–12.
Sambrook, J., Fritsch, E. F., Maniatis, T. (1989). Molecular Cloning: A Laboratory Manual. 2nd ed. Vol. 1, New York: Cold Spring Harbor Laboratory Press.
Vega-Sánchez, M. E., Verhertbruggen, Y., Christensen, U., Chen, X., Sharma, V., Varanasi, P., Jobling, S. A. (2012) Loss of Cellulose Synthase-Like F6 Function Affects Mixed-Linkage Glucan Deposition, Cell Wall Mechanical Properties, and Defense Responses in Vegetative Tissues of Rice, Plant Physiology 159 (1): 56–69.