Enhancement of vitamin B6 levels in rice expressing Arabidopsis vitamin B6 biosynthesis de novo genes.

[en] Vitamin B6 (pyridoxine) is vital for key metabolic reactions and reported to have antioxidant properties in planta. Thus, enhancement of vitamin B6 content has been hypothesized to be a route to improve resistance to biotic and abiotic stresses. Most of the current studies on vitamin B6 in plants are on eudicot species, with monocots remaining largely unexplored. In this study, we investigated vitamin B6 biosynthesis in rice, with a view to examining the feasibility and impact of enhancing vitamin B6 levels. Constitutive expression in rice of two Arabidopsis thaliana genes from the vitamin B6 biosynthesis de novo pathway, AtPDX1.1 and AtPDX2, resulted in a considerable increase in vitamin B6 in leaves (up to 28.3-fold) and roots (up to 12-fold), with minimal impact on general growth. Rice lines accumulating high levels of vitamin B6 did not display enhanced tolerance to abiotic stress (salt) or biotic stress (resistance to Xanthomonas oryzae infection). While a significant increase in vitamin B6 content could also be achieved in rice seeds (up to 3.1-fold), the increase was largely due to its accumulation in seed coat and embryo tissues, with little enhancement observed in the endosperm. However, seed yield was affected in some vitamin B6 -enhanced lines. Notably, expression of the transgenes did not affect the expression of the endogenous rice PDX genes. Intriguingly, despite transgene expression in leaves and seeds, the corresponding proteins were only detectable in leaves and could not be observed in seeds, possibly pointing to a mode of regulation in this organ. This article is protected by copyright. All rights reserved.

Disciplines :

Biotechnology

Author, co-author :

Mangel, Nathalie

Fudge, Jared B.

Li, Kuan-Te

Wu, Ting-Ying

Tohge, Takayuki

Fernie, Alisdair R.

Szurek, Boris

Fitzpatrick, Teresa B.

Gruissem, Wilhelm

Vanderschuren, Hervé ; Université de Liège - ULiège > Agronomie, Bio-ingénierie et Chimie (AgroBioChem) > Ingénierie des productions végétales et valorisation

Language :

English

Title :

Enhancement of vitamin B6 levels in rice expressing Arabidopsis vitamin B6 biosynthesis de novo genes.

Publication date :

2019

Journal title :

The Plant journal : for cell and molecular biology

ISSN :

0960-7412

eISSN :

1365-313X

Peer reviewed :

Peer reviewed

Commentary :

Available on ORBi :

since 29 June 2019

Statistics

Number of views

62 (5 by ULiège)

Number of downloads

0 (0 by ULiège)

More statistics

Scopus citations^®

Scopus citations^®
without self-citations

OpenCitations

Bibliography

Bagri, D.S., Upadhyaya, D.C. and Kumar, A. (2018) Overexpression of PDX-II gene in potato (Solanum tuberosum L.) leads to the enhanced accumulation of vitamin B6 in tuber tissues and tolerance to abiotic stresses. Plant Sci. 272, 267–275.
Bhullar, N.K. and Gruissem, W. (2013) Nutritional enhancement of rice for human health: the contribution of biotechnology. Biotechnol. Adv. 31, 50–57.
Bilski, P., Li, M.Y., Ehrenshaft, M., Daub, M.E. and Chignell, C.F. (2000) Vitamin B6 (pyridoxine) and its derivatives are efficient singlet oxygen quenchers and potential fungal antioxidants. Photochem. Photobiol. 71, 129–134.
Blancquaert, D., Van Daele, J., Strobbe, S., Kiekens, F., Storozhenko, S., De Steur, H., Gellynck, X., Lambert, W., Stove, C. and Van Der Straeten, D. (2015) Improving folate (vitamin B9) stability in biofortified rice through metabolic engineering. Nat. Biotechnol. 33, 1076–1078.
Burns, K.E., Xiang, Y., Kinsland, C.L., McLafferty, F.W. and Begley, T.P. (2005) Reconstitution and biochemical characterization of a new pyridoxal-5′-phosphate biosynthetic pathway. J. Am. Chem. Soc. 127, 3682–3683.
Chang, S., Puryear, J. and Cairney, J. (1993) A simple and efficient method for isolating RNA from pine trees. Plant Mol. Biol. Rep. 11, 113–116.
Chen, H. and Xiong, L. (2005) Pyridoxine is required for post-embryonic root development and tolerance to osmotic and oxidative stresses. Plant J. 44, 396–408.
Chen, H. and Xiong, L. (2009) Enhancement of vitamin B6 levels in seeds through metabolic engineering. Plant Biotechnol. J. 7, 673–681.
Colinas, M. and Fitzpatrick, T.B. (2015) Natures balancing act: examining biosynthesis de novo, recycling and processing damaged vitamin B metabolites. Curr. Opin. Plant Biol. 25, 98–106.
Colinas, M. and Fitzpatrick, T.B. (2016) Interaction between vitamin B6 metabolism, nitrogen metabolism and autoimmunity. Plant Signal. Behav. 11, e1161876.
Colinas, M., Eisenhut, M., Tohge, T., Pesquera, M., Fernie, A.R., Weber, A.P. and Fitzpatrick, T.B. (2016) Balancing of B6 vitamers is essential for plant development and metabolism in Arabidopsis. Plant Cell, 28, 439–453.
Danon, A., Miersch, O., Felix, G., Camp, R.G. and Apel, K. (2005) Concurrent activation of cell death-regulating signaling pathways by singlet oxygen in Arabidopsis thaliana. Plant J. 41, 68–80.
Dell'Aglio, E., Boycheva, S. and Fitzpatrick, T.B. (2017) The pseudoenzyme PDX1.2 sustains vitamin B6 biosynthesis as a function of heat stress. Plant Physiol. 174, 2098–2112.
Denslow, S.A., Walls, A.A. and Daub, M.E. (2005) Regulation of biosynthetic genes and antioxidant properties of vitamin B6 vitamers during plant defense responses. Physiol. Mol. Plant Pathol. 66, 244–255.
Denslow, S.A., Rueschhoff, E.E. and Daub, M.E. (2007) Regulation of the Arabidopsis thaliana vitamin B6 biosynthesis genes by abiotic stress. Plant Physiol. Biochem. 45, 152–161.
Di Salvo, M., Safo, M. and Contestabile, R. (2012) Biomedical aspects of pyridoxal 5′-phosphate availability. Front. Biosci. (Elite Ed.) 4, 897–913.
Dong, W., Thomas, N., Ronald, P.C. and Goyer, A. (2016) Overexpression of thiamin biosynthesis genes in rice increases leaf and unpolished grain thiamin content but not resistance to Xanthomonas oryzae pv. oryzae. Front. Plant Sci. 7, 616.
Fitzpatrick, T.B. (2011) Vitamin B6 in plants: more than meets the eye. Adv. Bot. Res. 59, 1–38.
Fitzpatrick, T.B., Amrhein, N., Kappes, B., Macheroux, P., Tews, I. and Raschle, T. (2007) Two independent routes of de novo vitamin B6 biosynthesis: not that different after all. Biochem. J. 407, 1–13.
Fudge, J., Mangel, N., Gruissem, W., Vanderschuren, H. and Fitzpatrick, T.B. (2017) Rationalising vitamin B6 biofortification in crop plants. Curr. Opin. Biotechnol. 44, 130–137.
Furtado, A., Henry, R.J. and Takaiwa, F. (2008) Comparison of promoters in transgenic rice. Plant Biotechnol. J. 6, 679–693.
Gonzalez, C., Szurek, B., Manceau, C., Mathieu, T., Sere, Y. and Verdier, V. (2007a) Molecular and pathotypic characterization of new Xanthomonas oryzae strains from West Africa. Mol. Plant Microbe Interact. 20, 534–546.
Gonzalez, E., Danehower, D. and Daub, M.E. (2007b) Vitamer levels, stress response, enzyme activity, and gene regulation of Arabidopsis lines mutant in the pyridoxine/pyridoxamine 5′-phosphate oxidase (PDX3) and the pyridoxal kinase (SOS4) genes involved in the vitamin B6 salvage pathway. Plant Physiol. 145, 985–996.
von Grebmer, K., Saltzman, A., Birol, E., Wiesmann, D., Prasai, N., Yin, S., Yohannes, Y., Menon, P., Thompson, J. and Sonntag, A. (2014) 2014 Global hunger index: The challenge of hidden hunger. Welthungerhilfe, International Food Policy Research Institute, and Concern Worldwide. Bonn, Washington, DC, and Dublin.
Gregory, J.F. III (1998) Nutritional properties and significance of vitamin glycosides. Annu. Rev. Nutr. 18, 277–296.
Gregory, J.F. III (2012) Accounting for differences in the bioactivity and bioavailability of vitamers. Food Nutr. Res. 56, 5809.
Gregory, J.F. III and Ink, S.L. (1987) Identification and quantification of pyridoxine beta-glucoside as a major form of vitamin B6 in plant-derived foods. J. Agric. Food Chem. 35, 76–82.
Hanson, A.D., Beaudoin, G.A., McCarty, D.R. and Gregory, J.F. (2016) Does abiotic stress cause functional B vitamin deficiency in plants? Plant Physiol. 172, 2082–2097.
Havaux, M., Ksas, B., Szewczyk, A., Rumeau, D., Franck, F., Caffarri, S. and Triantaphylides, C. (2009) Vitamin B6 deficient plants display increased sensitivity to high light and photo-oxidative stress. BMC Plant Biol. 9, 130.
Hellmann, H. and Mooney, S. (2010) Vitamin B6: a molecule for human health? Molecules, 15, 442–459.
Herrero, S. and Daub, M.E. (2007) Genetic manipulation of vitamin B6 biosynthesis in tobacco and fungi uncovers limitations to up-regulation of the pathway. Plant Sci. 172, 609–620.
Herrero, S., Gonzalez, E., Gillikin, J.W., Velez, H. and Daub, M.E. (2011) Identification and characterization of a pyridoxal reductase involved in the vitamin B6 salvage pathway in Arabidopsis. Plant Mol. Biol. 76, 157–169.
Hood, E., Gelvin, S., Melchers, L. and Hoekema, A. (1993) New Agrobacterium helper plasmids for gene-transfer to plants. Transgenic Res. 2, 208–218.
Hruz, T., Laule, O., Szabo, G., Wessendorp, F., Bleuler, S., Oertle, L., Widmayer, P., Gruissem, W. and Zimmermann, P. (2008) Genevestigator v3: a reference expression database for the meta-analysis of transcriptomes. Adv. Bioinformatics, 2008, 420747.
Huang, S., Zhang, J., Wang, L. and Huang, L. (2013) Effect of abiotic stress on the abundance of different vitamin B6 vitamers in tobacco plants. Plant Physiol. Biochem. 66, 63–67.
Jain, M., Nijhawan, A., Tyagi, A.K. and Khurana, J.P. (2006) Validation of housekeeping genes as internal control for studying gene expression in rice by quantitative real-time PCR. Biochem. Biophys. Res. Commun. 345, 646–651.
Kauffman, H.E., Reddy, A.P.K., Hsieh, S.P.Y. and Merca, S.D. (1973) An improved technique for evaluating resistance of rice varieties to Xanthomonas oryzae. Plant Dis. Rep. 57, 537–541.
Kennedy, G., Burlingame, B. and Nguyen, V.N. (2003) Nutritional contribution of rice and impact of biotechnology and biodiversity in rice-consuming countries. Proceedings of the 20th Session of the International Rice Commission: FAO corporate document repository. Bangkok.
Kobayashi, T., Suzuki, M., Inoue, H., Itai, R.N., Takahashi, M., Nakanishi, H., Mori, S. and Nishizawa, N.K. (2005) Expression of iron-acquisition-related genes in iron-deficient rice is co-ordinately induced by partially conserved iron-deficiency-responsive elements. J. Exp. Bot. 56, 1305–1316.
Kuwano, M., Masumura, T. and Yoshida, K.T. (2011) A novel endosperm transfer cell-containing region-specific gene and its promoter in rice. Plant Mol. Biol. 76, 47–56.
Lamesch, P., Berardini, T.Z., Li, D. et al. (2012) The Arabidopsis Information Resource (TAIR): improved gene annotation and new tools. Nucleic Acids Res. 40, D1202–1210.
Leach, J.E., Rhoads, M.L., Vera Cruz, C.M., White, F.F., Mew, T.W. and Leung, H. (1992) Assessment of genetic diversity and population structure of Xanthomonas oryzae pv. oryzae with a repetitive DNA element. Appl. Environ. Microbiol. 58, 2188–2195.
Li, K.T., Moulin, M., Mangel, N., Albersen, M., Verhoeven-Duif, N.M., Ma, Q., Zhang, P., Fitzpatrick, T.B., Gruissem, W. and Vanderschuren, H. (2015) Increased bioavailable vitamin B6 in field-grown transgenic cassava for dietary sufficiency. Nat. Biotechnol. 33, 1029–1032.
Livak, K.J. and Schmittgen, T.D. (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods, 25, 402–408.
Long, X., Liu, Q., Chan, M., Wang, Q. and Sun, S.S. (2013) Metabolic engineering and profiling of rice with increased lysine. Plant Biotechnol. J. 11, 490–501.
Lucca, P., Poletti, S. and Sautter, C. (2006) Genetic engineering approaches to enrich rice with iron and vitamin A. Physiol. Plant. 126, 291–303.
Lum, H.K., Kwok, F. and Lo, S.C. (2002) Cloning and characterization of Arabidopsis thaliana pyridoxal kinase. Planta, 215, 870–879.
Meng, F., Wei, Y. and Yang, X. (2005) Iron content and bioavailability in rice. J. Trace Elem. Med. Biol. 18, 333–338.
Moccand, C., Kaufmann, M. and Fitzpatrick, T.B. (2011) It takes two to tango: defining an essential second active site in pyridoxal 5′-phosphate synthase. PLoS One, 6, e16042.
Moccand, C., Boycheva, S., Surriabre, P., Tambasco-Studart, M., Raschke, M., Kaufmann, M. and Fitzpatrick, T.B. (2014) The pseudoenzyme PDX1.2 boosts vitamin B6 biosynthesis under heat and oxidative stress in Arabidopsis. J. Biol. Chem. 289, 8203–8216.
Nishimura, A., Aichi, I. and Matsuoka, M. (2006) A protocol for Agrobacterium-mediated transformation in rice. Nat. Protoc. 1, 2796–2802.
Ollilainen, V.M. (1999) HPLC analysis of vitamin B6 in foods. Agri. Food Sci. 8, 519–618.
Ouyang, S., Zhu, W., Hamilton, J. et al. (2007) The TIGR rice genome annotation resource: improvements and new features. Nucleic Acids Res. 35, D883–887.
Percudani, R. and Peracchi, A. (2003) A genomic overview of pyridoxal-phosphate-dependent enzymes. EMBO Rep. 4, 850–854.
le Qu, Q. and Takaiwa, F. (2004) Evaluation of tissue specificity and expression strength of rice seed component gene promoters in transgenic rice. Plant Biotechnol. J. 2, 113–125.
Raschke, M., Boycheva, S., Crevecoeur, M., Nunes-Nesi, A., Witt, S., Fernie, A.R., Amrhein, N. and Fitzpatrick, T.B. (2011) Enhanced levels of vitamin B6 increase aerial organ size and positively affect stress tolerance in Arabidopsis. Plant J. 66, 414–432.
Raschle, T., Amrhein, N. and Fitzpatrick, T.B. (2005) On the two components of pyridoxal 5′-phosphate synthase from Bacillus subtilis. J. Biol. Chem. 280, 32291–32300.
Raschle, T., Arigoni, D., Brunisholz, R., Rechsteiner, H., Amrhein, N. and Fitzpatrick, T.B. (2007) Reaction mechanism of pyridoxal 5′-phosphate synthase. Detection of an enzyme-bound chromophoric intermediate. J. Biol. Chem. 282, 6098–6105.
Raschle, T., Speziga, D., Kress, W., Moccand, C., Gehrig, P., Amrhein, N., Weber-Ban, E. and Fitzpatrick, T.B. (2009) Intersubunit cross-talk in pyridoxal 5′-phosphate synthase, coordinated by the C terminus of the synthase subunit. J. Biol. Chem. 284, 7706–7718.
Ristila, M., Strid, H., Eriksson, L.A., Strid, A. and Savenstrand, H. (2011) The role of the pyridoxine (vitamin B6) biosynthesis enzyme PDX1 in ultraviolet-B radiation responses in plants. Plant Physiol. Biochem. 49, 284–292.
Ruiz, A., Garcia-Villoria, J., Ormazabal, A., Zschocke, J., Fiol, M., Navarro-Sastre, A., Artuch, R., Vilaseca, M.A. and Ribes, A. (2008) A new fatal case of pyridox(am)ine 5′-phosphate oxidase (PNPO) deficiency. Mol. Genet. Metab. 93, 216–218.
Sang, Y., Barbosa, J.M., Wu, H., Locy, R.D. and Singh, N.K. (2007) Identification of a pyridoxine (pyridoxamine) 5′-phosphate oxidase from Arabidopsis thaliana. FEBS Lett. 581, 344–348.
Sang, Y., Locy, R.D., Goertzen, L.R., Rashotte, A.M., Si, Y., Kang, K. and Singh, N.K. (2011) Expression, in vivo localization and phylogenetic analysis of a pyridoxine 5′-phosphate oxidase in Arabidopsis thaliana. Plant Physiol. Biochem. 49, 88–95.
Savenstrand, H., Brosche, M. and Strid, A. (2004) Ultraviolet-B signalling: Arabidopsis brassinosteroid mutants are defective in UV-B regulated defence gene expression. Plant Physiol. Biochem. 42, 687–694.
Seck, P.A., Diagne, A., Mohanty, S. and Wopereis, M.C.S. (2012) Crops that feed the world 7: Rice. Food Security, 4, 7–24.
Sheu, J.J., Yu, T.S., Tong, W.F. and Yu, S.M. (1996) Carbohydrate starvation stimulates differential expression of rice alpha-amylase genes that is modulated through complicated transcriptional and posttranscriptional processes. J. Biol. Chem. 271, 26998–27004.
Shi, H. and Zhu, J.K. (2002) SOS4, a pyridoxal kinase gene, is required for root hair development in Arabidopsis. Plant Physiol. 129, 585–593.
Shi, H., Xiong, L., Stevenson, B., Lu, T. and Zhu, J.K. (2002) The Arabidopsis salt overly sensitive 4 mutants uncover a critical role for vitamin B6 in plant salt tolerance. Plant Cell, 14, 575–588.
Singh, G., Kumar, S. and Singh, P. (2003) A quick method to isolate RNA from wheat and other carbohydrate-rich seeds. Plant Mol. Biol. Rep. 21, 93–93.
Singh, U., Praharaj, C.S., Singh, S.S. and Singh, N.P. (2016) Biofortification of Food Crops. New Delhi, Heidelberg, New York, Dordrecht, London: Springer India.
Storozhenko, S., De Brouwer, V., Volckaert, M., Navarrete, O., Blancquaert, D., Zhang, G.F., Lambert, W. and Van Der Straeten, D. (2007) Folate fortification of rice by metabolic engineering. Nat. Biotechnol. 25, 1277–1279.
Strobbe, S. and Van Der Straeten, D. (2018) Toward Eradication of B-Vitamin Deficiencies: Considerations for Crop Biofortification. Front. Plant Sci. 9, 443.
Svozil, J., Hirsch-Hoffmann, M., Dudler, R., Gruissem, W. and Baerenfaller, K. (2014) Protein abundance changes and ubiquitylation targets identified after inhibition of the proteasome with syringolin A. Mol. Cell Proteomics, 13, 1523–1536.
Szydlowski, N., Bürkle, L., Pourcel, L., Moulin, M., Stolz, J. and Fitzpatrick, T.B. (2013) Recycling of pyridoxine (vitamin B6) by PUP1 in Arabidopsis. Plant J. 75, 40–52.
Tambasco-Studart, M., Titiz, O., Raschle, T., Forster, G., Amrhein, N. and Fitzpatrick, T.B. (2005) Vitamin B6 biosynthesis in higher plants. Proc. Natl Acad. Sci. USA, 102, 13687–13692.
Tambasco-Studart, M., Tews, I., Amrhein, N. and Fitzpatrick, T.B. (2007) Functional analysis of PDX2 from Arabidopsis, a glutaminase involved in vitamin B6 biosynthesis. Plant Physiol. 144, 915–925.
Thompson, B. and Amoroso, L. (2011) Combating Micronutrient Deficiencies: Food-based Approaches. Rome: CAB International and FAO.
Titiz, O., Tambasco-Studart, M., Warzych, E., Apel, K., Amrhein, N., Laloi, C. and Fitzpatrick, T.B. (2006) PDX1 is essential for vitamin B6 biosynthesis, development and stress tolerance in Arabidopsis. Plant J. 48, 933–946.
Vanderschuren, H., Alder, A., Zhang, P. and Gruissem, W. (2009) Dose-dependent RNAi-mediated geminivirus resistance in the tropical root crop cassava. Plant Mol. Biol. 70, 265–272.
Vanderschuren, H., Boycheva, S., Li, K.T., Szydlowski, N., Gruissem, W. and Fitzpatrick, T.B. (2013) Strategies for vitamin B6 biofortification of plants: a dual role as a micronutrient and a stress protectant. Front. Plant Sci. 4, 143.
Yang, Y.Z., Ding, S., Wang, Y., Li, C.L., Shen, Y., Meeley, R., McCarty, D.R. and Tan, B.C. (2017) Small kernel2 encodes a glutaminase in vitamin B6 biosynthesis essential for maize seed development. Plant Physiol. 174, 1127–1138.
Zhang, Y., Liu, B., Li, X., Ouyang, Z., Huang, L., Hong, Y., Zhang, H., Li, D. and Song, F. (2014) The de novo biosynthesis of vitamin B6 is required for disease resistance against Botrytis cinerea in tomato. Mol. Plant Microbe Interact. 27, 688–699.
Zhang, Y., Jin, X., Ouyang, Z., Li, X., Liu, B., Huang, L., Hong, Y., Zhang, H., Song, F. and Li, D. (2015) Vitamin B6 contributes to disease resistance against Pseudomonas syringae pv. tomato DC3000 and Botrytis cinerea in Arabidopsis thaliana. J. Plant Physiol. 175, 21–25.
Zhao, C., Piao, S., Wang, X. et al. (2016) Plausible rice yield losses under future climate warming. Nat. Plants, 3, 16202.