Keywords
chlorophyll
phosphorothioate
CHLD subunit Antisense Oligonucleotides (ASOs)
chlorophyll
phosphorothioate
tiểu đơn vị CHLD
How to Cite
Abstract
Antisense oligonucleotides (ASOs) are emerging as a promising tool for functional genomics research, particularly in the regulation of gene expression. In Viet Nam, however, studies on the application of ASOs in plant systems remains limited. This study aimed to evaluate the effects of ASOs on rice (Oryza sativa) by targeting the CHLD gene, which encodes a subunit of Mg–chelatase involved in chlorophyll biosynthesis. ASOs were synthesized in both unmodified and chemically modified phosphorothioate (PS) forms. Rice seedlings at four days after germination were treated by foliar spraying with ASOs at a concentration of 100 pmol/mL. The effects of ASO treatments were assessed at 3, 7, and 14 days after application based on leaf color, plant morphology, plant height, root length, and the contents of chlorophyll a, chlorophyll b, and carotenoids. The results showed that ASO-treated plants exhibited a marked reduction in leaf pigment contents and plant height at 3 and 7 days after treatment. Specifically, chlorophyll a content decreased by 52–72%, chlorophyll b by 47–73%, and carotenoids by 50–71%, while plant height was reduced by 17–32% compared with the control. In contrast, root development was not affected by ASO application. These findings demonstrate that ASOs can effectively suppress CHLD gene–associated phenotypes, highlighting their potential as a tool for gene expression control and functional gene analysis in crop plants.
References
- Dhuri K, Bechtold C, Quijano E, et al. Antisense Oligonucleotides: An Emerging Area in Drug Discovery and Development. J Clin Med. 2020;9(6):1-24.
- Hunter WB, Cooper WR, Sandoval-Mojica AF, McCollum G, Aishwarya V, Pelz-Stelinski KS. Improving Suppression of Hemipteran Vectors and Bacterial Pathogens of Citrus and Solanaceous Plants: Advances in Antisense Oligonucleotides (FANA). Front Agron. 2021;Volume 3 - 2021.
- Lima LFF de, Carvalho IGB, Souza-Neto RR de, et al. Antisense Oligonucleotide as a New Technology Application for CsLOB1 Gene Silencing Aiming at Citrus Canker Resistance. Phytopathology. 2024;114(8):1802-1809.
- Gruber C, Gursinsky T, Gago-Zachert S, Pantaleo V, Behrens SE. Effective Antiviral Application of Antisense in Plants by Exploiting Accessible Sites in the Target RNA. Int J Mol Sci. 2023;24(24).
- Krasnodębski C, Sawuła A, Kazmierczak U, Żuk MJIJoMS. Oligo—Not Only for Silencing: Overlooked Potential for Multidirectional Action in Plants. Int J Mol Sci. 2023;24(5):4466.
- Ayub R, Guis M, Amor MB, Gillot L, Roustan J-P, Latché A, et al. Expression of ACC oxidase antisense gene inhibits ripening of cantaloupe melon fruits. Nature Biotechnology. 1996;14(7):862-6.
- Sheehy RE, Kramer M, Hiatt WR. Reduction of polygalacturonase activity in tomato fruit by antisense RNA. Proc Natl Acad Sci. 1988;85(23):8805-9.
- Smith CJS, Watson CF, Ray J, Bird CR, Morris PC, Schuch W, et al. Antisense RNA inhibition of polygalacturonase gene expression in transgenic tomatoes. Nature. 1988;334(6184):724-6.
- Gräfe S, Saluz H-P, Grimm B, Hänel F. Mg-chelatase of tobacco: The role of the subunit CHL D in the chelation step of protoporphyrin IX. Journal of Plant Growth Regulation. 1999;96(5):1941-6.
- Zhang H, Li J, Yoo J-H, Yoo S-C, Cho S-H, Koh H-J, et al. Rice Chlorina-1 and Chlorina-9 encode ChlD and ChlI subunits of Mg-chelatase, a key enzyme for chlorophyll synthesis and chloroplast development. Plant Molecular Biology. 2006;62(3):325-37.
- Du S-Y, Zhang X-F, Lu Z, Xin Q, Wu Z, Jiang T, et al. Roles of the different components of magnesium chelatase in abscisic acid signal transduction. Plant Molecular Biology. 2012;80(4):519-37.
- Wu C-j, Wang J, Zhu J, Ren J, Yang Y-x, Luo T, et al. Molecular Characterization of Mg-Chelatase CHLI Subunit in Pea (Pisum sativum L.). Front Plant Sci. 2022;13(January):1-13.
- Yang X, Cai J, Xue J, Luo X, Zhu W, Xiao X, et al. Magnesium chelatase subunit D is not only required for chlorophyll biosynthesis and photosynthesis, but also affecting starch accumulation in Manihot esculenta Crantz. BMC Plant Biology. 2023;23(1):258.
- Jensen PE, Gibson LCD, Henningsen KW, Hunter CN. Expression of the chlI, chlD, and chlH Genes from the Cyanobacterium Synechocystis PCC6803 in Escherichia coli and Demonstration That the Three Cognate Proteins Are Required for Magnesium-protoporphyrin Chelatase Activity. Journal of Biological Chemistry. 1996;271(28):16662-7.
- Walker CJ, Willows RD. Mechanism and regulation of Mg-chelatase. Biochemical Journal. 1997;327(2):321-33.
- Willows RD. Biosynthesis of chlorophylls from protoporphyrin IX. Natural Product Reports. 2003;20(3):327-41.
- Nayek S, Choudhury I, Haque, Jaishee N, Roy S. Spectrophotometric Analysis of Chlorophylls and Carotenoids from Commonly Grown Fern Species by Using Various Extracting Solvents. Research Journal of Chemical Sciences. 2014;4:2231-606.
- Shaw J-P, Kent K, Bird J, Fishback J, Froehler B. Modified deoxyoligonucleotides stable to exonuclease degradation in serum. Nucleic Acids Research. 1991;19(4):747-50.
- Dinç E, Tóth SZ, Schansker G, Ayaydin F, Kovács L, Dudits D, et al. Synthetic Antisense Oligodeoxynucleotides to Transiently Suppress Different Nucleus- and Chloroplast-Encoded Proteins of Higher Plant Chloroplasts Plant Physiology. 2011;157(4):1628-41.
- Pleskot R, Potocký M, Pejchar P, Linek J, Bezvoda R, Martinec J, et al. Mutual regulation of plant phospholipase D and the actin cytoskeleton. The Plant Journal. 2010;62(3):494-507.
- Moutinho A, Hussey PJ, Trewavas AJ, Malhó R. cAMP acts as a second messenger in pollen tube growth and reorientation. Plant Science. 2001;98(18):10481-6.
- Mizuta Y, Higashiyama T. Antisense gene inhibition by phosphorothioate antisense oligonucleotide in Arabidopsis pollen tubes. The Plant Journal. 2014;78(3):516-26.
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