Silencing of FRO1 gene affects iron homeostasis and nutrient balance in tomato plants
Florinda Gama
MED—Mediterranean Institute for Agriculture, Environment and Development and CHANGE—Global Change and Sustainability Institute, Faculty of Science and Technology, University of Algarve, Faro, Portugal
GreenCoLab—Associação Oceano Verde, University of Algarve, Faro, Portugal
Search for more papers by this authorTeresa Saavedra
MED—Mediterranean Institute for Agriculture, Environment and Development and CHANGE—Global Change and Sustainability Institute, Faculty of Science and Technology, University of Algarve, Faro, Portugal
Search for more papers by this authorSusana Dandlen
MED—Mediterranean Institute for Agriculture, Environment and Development and CHANGE—Global Change and Sustainability Institute, Faculty of Science and Technology, University of Algarve, Faro, Portugal
Search for more papers by this authorPedro García-Caparrós
Agronomy Department of Superior School Engineering, University of Almeria, Almeria, Spain
Search for more papers by this authorAmarilis de Varennes
Instituto Superior de Agronomia, University of Lisbon, Lisbon, Portugal
Search for more papers by this authorGustavo Nolasco
Faculty of Science and Technology, University of Algarve, Faro, Portugal
Search for more papers by this authorPedro José Correia
MED—Mediterranean Institute for Agriculture, Environment and Development and CHANGE—Global Change and Sustainability Institute, Faculty of Science and Technology, University of Algarve, Faro, Portugal
Search for more papers by this authorCorresponding Author
Maribela Pestana
MED—Mediterranean Institute for Agriculture, Environment and Development and CHANGE—Global Change and Sustainability Institute, Faculty of Science and Technology, University of Algarve, Faro, Portugal
Correspondence
Maribela Pestana, MED—Mediterranean Institute for Agriculture, Environment and Development and CHANGE—Global Change and Sustainability Institute, Faculty of Science and Technology, University of Algarve, Building 8, Campus of Gambelas, 8005-139 Faro, Portugal.
Email: [email protected]
Search for more papers by this authorFlorinda Gama
MED—Mediterranean Institute for Agriculture, Environment and Development and CHANGE—Global Change and Sustainability Institute, Faculty of Science and Technology, University of Algarve, Faro, Portugal
GreenCoLab—Associação Oceano Verde, University of Algarve, Faro, Portugal
Search for more papers by this authorTeresa Saavedra
MED—Mediterranean Institute for Agriculture, Environment and Development and CHANGE—Global Change and Sustainability Institute, Faculty of Science and Technology, University of Algarve, Faro, Portugal
Search for more papers by this authorSusana Dandlen
MED—Mediterranean Institute for Agriculture, Environment and Development and CHANGE—Global Change and Sustainability Institute, Faculty of Science and Technology, University of Algarve, Faro, Portugal
Search for more papers by this authorPedro García-Caparrós
Agronomy Department of Superior School Engineering, University of Almeria, Almeria, Spain
Search for more papers by this authorAmarilis de Varennes
Instituto Superior de Agronomia, University of Lisbon, Lisbon, Portugal
Search for more papers by this authorGustavo Nolasco
Faculty of Science and Technology, University of Algarve, Faro, Portugal
Search for more papers by this authorPedro José Correia
MED—Mediterranean Institute for Agriculture, Environment and Development and CHANGE—Global Change and Sustainability Institute, Faculty of Science and Technology, University of Algarve, Faro, Portugal
Search for more papers by this authorCorresponding Author
Maribela Pestana
MED—Mediterranean Institute for Agriculture, Environment and Development and CHANGE—Global Change and Sustainability Institute, Faculty of Science and Technology, University of Algarve, Faro, Portugal
Correspondence
Maribela Pestana, MED—Mediterranean Institute for Agriculture, Environment and Development and CHANGE—Global Change and Sustainability Institute, Faculty of Science and Technology, University of Algarve, Building 8, Campus of Gambelas, 8005-139 Faro, Portugal.
Email: [email protected]
Search for more papers by this authorThis article has been edited by Michael Frei.
Abstract
Background
Iron chlorosis is an abiotic stress of worldwide importance affecting several agronomic crops. It is important to understand how plants maintain nutrient homeostasis under Fe deficiency and recovery.
Aims
We used the virus-induced gene silencing (VIGS) method to elucidate the role of the FRO1 gene in tomato plants and identify the impact on regulation of the root ferric-chelate reductase (FCR) activity and nutritional homeostasis.
Methods
Tomato plantlets cv. “Cherry” were transferred into half-strength Hoagland's nutrient solution containing 0.5 µM of Fe (Fe0.5). In phase I, two treatments were established: control (Fe0.5) plants and VIGS-0.5 plants corresponding to plants with the FRO1 gene silenced. In phase II, plants from Fe0.5 and VIGS-0.5 were transferred to new nutrient solution and then grown for a further 14 days under 0 and 10 µM of Fe (as 0.5 µM would not be enough for the larger plants during phase II). Therefore, four treatments were imposed: Fe0, Fe10, VIGS-0, and VIGS-10.
Results
VIGS-0.5 plants had significantly lower chlorophyll (Chl) and root FCR activity compared to the respective non-silenced plants and retained more Cu and Zn in the roots at the expense of stems (Cu) or young leaves (Zn). Iron concentration in roots and stems decreased in FRO1 gene-silenced plants, compared to control plants, but the allocation to different organs was similar in both treatments.
Conclusions
There was a partial recovery of leaf Chl in the VIGS-10 plants and a higher concentration of Fe in all organs. In contrast, the allocation of Cu to roots decreased in the VIGS-10 plants.
Open Research
DATA AVAILABILITY STATEMENT
The data that support the findings of this study are available from the corresponding author upon reasonable request.
REFERENCES
- Abadía, J., & Abadía, A. (1993). Iron and pigments. In L. L. Barton & B. C. Hemming (Eds.), Iron chelation in plants and soil microorganisms (pp. 327–343). Academic Press Inc.
10.1016/B978-0-12-079870-4.50020-X Google Scholar
- Abadía, J., Vázquez, S., Rellán-Álvarez, R., El-Jendoubi, H., Abadía, A., Álvarez-Fernández, A., & López-Millán, A. F. (2011). Towards a knowledge-based correction of iron chlorosis. Plant Physiology and Biochemistry, 49(5), 471–482.
- Agüero, J., del Carmen Vives, M., Velázquez, K., Pina, J. A., Navarro, L., Moreno, P., & Guerri, J. (2014). Effectiveness of gene silencing induced by viral vectors based on Citrus leaf blotch virus is different in Nicotiana benthamiana and citrus plants. Virology, 460, 154–164.
- AOAC. (1990). Official methods of analysis ( 12th ed.). Springer Nature.
- Arce-Rodriguez, M. L., & Ochoa-Alejo, N. (2015). Silencing AT3 gene reduces the expression of pAmt, BCAT, Kas, and Acl genes involved in capsaicinoid biosynthesis in chili pepper fruits. Biologia Plantarum, 59(3), 477–484.
- Barberon, M., Dubeaux, G., Kolb, C., Isono, E., Zelazny, E., & Vert, G. (2014). Polarization of Iron-Regulated Transporter 1 (IRT1) to the plant-soil interface plays crucial role in metal homeostasis. Proceedings of the National Academy of Sciences, 111(22), 8293–8298.
- Becker, A., & Lange, M. (2010). VIGS-genomics goes functional. Trends in Plant Science, 15(1), 1–4.
- Bernal, M., Casero, D., Singh, V., Wilson, G. T., Grande, A., Yang, H., Dodani, S. C., Pellegrini, M., Huijser, P., Connolly, E. L., Mechant, S. S., & Krämer, U. (2012). Transcriptome sequencing identifies SPL7-regulated copper acquisition genes FRO4/FRO5 and the copper dependence of iron homeostasis in Arabidopsis. The Plant Cell, 24(2), 738–761.
- Bienfait, H. F., Bino, R. J., Van der Bliek, A. M., Duivenvoorden, J. F., & Fontaine, J. M. (1983). Characterization of ferric reducing activity in roots of Fe-deficient Phaseolus vulgaris. Physiologia Plantarum, 59(2), 196–202.
- Brigneti, G., Martín-Hernández, A. M., Jin, H., Chen, J., Baulcombe, D. C., Baker, B., & Jones, J. D. (2004). Virus-induced gene silencing in Solanum species. The Plant Journal, 39(2), 264–272.
- Burch-Smith, T. M., Anderson, J. C., Martin, G. B., & Dinesh-Kumar, S. P. (2004). Applications and advantages of virus-induced gene silencing for gene function studies in plants. The Plant Journal, 39(5), 734–746.
- Chai, Y. M., Jia, H. F., Li, C. L., Dong, Q. H., & Shen, Y. Y. (2011). FaPYR1 is involved in strawberry fruit ripening. Journal of Experimental Botany, 62(14), 5079–5089.
- Connolly, E. L., Campbell, N. H., Grotz, N., Prichard, C. L., & Guerinot, M. L. (2003). Overexpression of the FRO2 ferric chelate reductase confers tolerance to growth on low iron and uncovers posttranscriptional control. Plant Physiology, 133(3), 1102–1110.
- Divte, P. R., Yadav, P., Pawar, A. B., Sharma, V., Anand, A., Pandey, R., & Singh, B. (2021). Crop response to iron deficiency is guided by cross-talk between phytohormones and their regulation of the root system architecture. Agricultural Research, 10, 347–360.
- Du, J., Huang, Z., Wang, B., Sun, H., Chen, C., Ling, H. Q., & Wu, H. (2015). SlbHLH068 interacts with FER to regulate the iron-deficiency response in tomato. Annals of Botany, 116(1), 23–34.
- Fu, D. Q., Meng, L. H., Zhu, B. Z., Zhu, H. L., Yan, H. X., & Luo, Y. B. (2016). Silencing of the SlNAP7 gene influences plastid development and lycopene accumulation in tomato. Scientific Reports, 6(1), 38664.
- Gama, F., Saavedra, T., Da Silva, J. P., Miguel, M. G., de Varennes, A., Correia, P. J., & Pestana, M. (2016). The memory of iron stress in strawberry plants. Plant Physiology and Biochemistry, 104, 36–44.
- Gama, F., Saavedra, T., Dandlen, S., de Varennes, A., Correia, P. J., Pestana, M., & Nolasco, G. (2017). Silencing of the FRO1 gene and its effects on iron partition in Nicotiana benthamiana. Plant Physiology and Biochemistry, 114, 111–118.
- Ghoshal, B., & Sanfaçon, H. (2015). Symptom recovery in virus-infected plants: Revisiting the role of RNA silencing mechanisms. Virology, 479, 167–179.
- Gogorcena, Y., Abadía, J., & Abadía, A. (2000). Induction of in vivo root ferric chelate reductase activity in fruit tree rootstock. Journal of Plant Nutrition, 23(1), 9–21.
- Graziano, M., & Lamattina, L. (2007). Nitric oxide accumulation is required for molecular and physiological responses to iron deficiency in tomato roots. The Plant Journal, 52(5), 949–960.
- Grotz, N., & Guerinot, M. L. (2006). Molecular aspects of Cu, Fe and Zn homeostasis in plants. Biochimica et Biophysica Acta (BBA)—Molecular Cell Research, 1763(7), 595–608.
- He, X., Jin, C., Li, G., You, G., Zhou, X., & Zheng, S. (2008). Use of the modified viral satellite DNA vector to silence mineral nutrition-related genes in plants: Silencing of the tomato ferric chelate reductase gene, FRO1, as an example. Science in China Series C: Life Sciences, 51(5), 402–409.
- Ivanov, R., Brumbarova, T., & Bauer, P. (2012). Fitting into the harsh reality: Regulation of iron-deficiency responses in dicotyledonous plants. Molecular Plant, 5(1), 27–42.
- Jain, A., Wilson, G. T., & Connolly, E. L. (2014). The diverse roles of FRO family metalloreductases in iron and copper homeostasis. Frontiers in Plant Science, 5, 100. https://doi.org/10.3389/fpls.2014.00100
- Jeong, J., & Connolly, E. L. (2009). Iron uptake mechanisms in plants: Functions of the FRO family of ferric reductases. Plant Science, 176(6), 709–714.
- Jeong, J., & Guerinot, M. L. (2009). Homing in on iron homeostasis in plants. Trends in Plant Science, 14(5), 280–285.
- Jiménez González, M. R., Casanova Lerma, L., Saavedra, T., Gama, F., Suárez García, M. P., Correia, P. J., & Pestana, M. (2019). Responses of tomato (Solanum lycopersicum L.) plants to iron deficiency in the root zone. Folia Horticulturae, 31(1), 223–234.
- Kobayashi, T., & Nishizawa, N. K. (2012). Iron uptake, translocation, and regulation in higher plants. Annual Review of Plant Biology, 63, 131–152.
- Lange, M., Yellina, A. L., Orashakova, S., & Becker, A. (2013). Virus-induced gene silencing (VIGS) in plants: An overview of target species and the virus-derived vector systems. In A. Becker (Ed.), Virus-induced gene silencing. Methods in molecular biology (Vol. 975, pp. 1–14). Humana Press.
10.1007/978-1-62703-278-0_1 Google Scholar
- Li, L., Ye, L., Kong, Q., & Shou, H. (2019). A vacuolar membrane ferric-chelate reductase, OsFRO1, alleviates Fe toxicity in rice (Oryza sativa L.). Frontiers in Plant Science, 10, 700.
- Lichtenthaler, H. K. (1987). Chlorophylls and carotenoids: Pigments of photosynthetic biomembranes. Methods in Enzymology, 148, 350–382.
- López-Millán, A. F., Morales, F., Gogorcena, Y., Abadía, A., & Abadía, J. (2001). Iron resupply-mediated deactivation of Fe-deficiency stress responses in roots of sugar beet. Functional Plant Biology, 28(3), 171–180.
- Lu, R., Martin-Hernandez, A. M., Peart, J. R., Malcuit, I., & Baulcombe, D. C. (2003). Virus-induced gene silencing in plants. Methods, 30(4), 296–303.
- Marschner, H., Römheld, V., & Kissel, M. (1986). Different strategies in higher plants in mobilization and uptake of iron. Journal of Plant Nutrition, 9(3–7), 695–713.
- Mukherjee, I., Campbell, N. H., Ash, J. S., & Connolly, E. L. (2006). Expression profiling of the Arabidopsis ferric chelate reductase (FRO) gene family reveals differential regulation by iron and copper. Planta, 223, 1178–1190.
- Murgia, I., Marzorati, F., Vigani, G., & Morandini, P. (2022). Plant iron nutrition: The long road from soil to seeds. Journal of Experimental Botany, 73(6), 1809–1824.
- Pfaffl, M. W. (2001). A new mathematical model for relative quantification in real-time RT–PCR. Nucleic Acids Research, 29(9), e45–e45. https://doi.org/10.1093/nar/29.9.e45
- Pestana, M., Correia, P. J., David, M., Abadía, A., Abadía, J., & de Varennes, A. (2011). Response of five citrus rootstocks to iron deficiency. Journal of Plant Nutrition and Soil Science, 174(5), 837–846.
- Pestana, M., Correia, P. J., Saavedra, T., Gama, F., Abadía, A., & de Varennes, A. (2012). Development and recovery of iron deficiency by iron resupply to roots or leaves of strawberry plants. Plant Physiology and Biochemistry, 53, 1–5.
- Nozoye, T., Aung, M. S., Masuda, H., Nakanishi, H., & Nishizawa, N. K. (2017). Bioenergy grass [Erianthus ravennae (L.) Beauv.] secretes two members of mugineic acid family phytosiderophores which involved in their tolerance to Fe deficiency. Soil Science and Plant Nutrition, 63(6), 543–552.
- Rai, S., Singh, P. K., Mankotia, S., Swain, J., & Satbhai, S. B. (2021). Iron homeostasis in plants and its crosstalk with copper, zinc, and manganese. Plant Stress, 1, 100008. https://doi.org/10.1016/j.stress.2021.100008
- Rajniak, J., Giehl, R. F., Chang, E., Murgia, I., von Wirén, N., & Sattely, E. S. (2018). Biosynthesis of redox-active metabolites in response to iron deficiency in plants. Nature Chemical Biology, 14(5), 442–450.
- Ratcliff, F., Martin-Hernandez, A. M., & Baulcombe, D. C. (2001). Technical advance: Tobacco rattle virus as a vector for analysis of gene function by silencing. The Plant Journal, 25(2), 237–245.
- Rengel, Z. (2001). Xylem and phloem transport of micronutrients. In W. J. Horst, M. K. Schenk, A. Bürkert, N. Claasen, H. Flessa, W. B. Frommer, H. Goldbach, H.-W. Olfs, V. Römheld, B. Sattelmacher, U. Schmidhalter, S. Schubert, N. Wirén, & L. Wittenmayer (Eds.), Plant nutrition. Developments in plant and soil sciences (pp. 628–629). Springer.
10.1007/0-306-47624-X_304 Google Scholar
- Ricachenevsky, F. K., & Sperotto, R. A. (2014). There and back again, or always there? The evolution of rice combined strategy for Fe uptake. Frontiers in Plant Science, 5, 189. https://doi.org/10.3389/fpls.2014.00189
- Ryu, C. M., Anand, A., Kang, L., & Mysore, K. S. (2004). Agrodrench: A novel and effective agroinoculation method for virus-induced gene silencing in roots and diverse Solanaceous species. The Plant Journal, 40(2), 322–331.
- Robinson, N. J., Procter, C. M., Connolly, E. L., & Guerinot, M. L. (1999). A ferric-chelate reductase for iron uptake from soils. Nature, 397(6721), 694–697.
- Sahu, P. P., Puranik, S., Khan, M., & Prasad, M. (2012). Recent advances in tomato functional genomics: Utilization of VIGS. Protoplasma, 249, 1017–1027.
- Schmid, N. B., Giehl, R. F., Döll, S., Mock, H. P., Strehmel, N., Scheel, D., Kong, S., Hider, R. C., & von Wirén, N. (2014). Feruloyl-CoA 6′-hydroxylase1-dependent coumarins mediate iron acquisition from alkaline substrates in Arabidopsis. Plant Physiology, 164(1), 160–172.
- Senthil-Kumar, M., Gowda, H. R., Hema, R., Mysore, K. S., & Udayakumar, M. (2008). Virus-induced gene silencing and its application in characterizing genes involved in water-deficit-stress tolerance. Journal of Plant Physiology, 165(13), 1404–1421.
- Senthil-Kumar, M., & Mysore, K. S. (2011). New dimensions for VIGS in plant functional genomics. Trends in Plant Science, 16(12), 656–665.
- Shi, G., Hao, M., Tian, B., Cao, G., Wei, F., & Xie, Z. (2021). A methodological advance of tobacco rattle virus-induced gene silencing for functional genomics in plants. Frontiers in Plant Science, 12, 671091. https://doi.org/10.3389/fpls.2021.671091
- Singh, B., Kukreja, S., Salaria, N., Thakur, K., Gautam, S., Taunk, J., & Goutam, U. (2019). VIGS: A flexible tool for the study of functional genomics of plants under abiotic stresses. Journal of Crop Improvement, 33(5), 567–604.
- Singh, A. K., Ghosh, D., & Chakraborty, S. (2022). Optimization of tobacco rattle virus (TRV)-based virus-induced gene silencing (VIGS) in tomato. In K. S. Mysore & M. Senthil-Kumar (Eds.), Plant gene silencing. Methods in molecular biology (Vol. 2408, pp. 133–145). Humana.
10.1007/978-1-0716-1875-2_9 Google Scholar
- Tian, S. L., Li, L., Chai, W. G., Shah, S. N. M., & Gong, Z. H. (2014). Effects of silencing key genes in the capsanthin biosynthetic pathway on fruit color of detached pepper fruits. BMC Plant Biology, 14(1), 1–12.
- Velásquez, A. C., Chakravarthy, S., & Martin, G. B. (2009). Virus-induced gene silencing (VIGS) in Nicotiana benthamiana and tomato. JoVE (Journal of Visualized Experiments), 28, e1292. https://doi.org/10.3791/1292
10.3791/1292 Google Scholar
- Vigani, G., Di Silvestre, D., Agresta, A. M., Donnini, S., Mauri, P., Gehl, C., Bittner, F., & Murgia, I. (2017). Molybdenum and iron mutually impact their homeostasis in cucumber (Cucumis sativus) plants. New Phytologist, 213(3), 1222–1241.
- Walker, E. L., & Connolly, E. L. (2008). Time to pump iron: Iron-deficiency-signaling mechanisms of higher plants. Current Opinion in Plant Biology, 11(5), 530–535.
- Zhao, N., Yu, G., Wang, Q., Wang, R., Zhang, J., Liu, C., & He, N. (2020). Conservative allocation strategy of multiple nutrients among major plant organs: From species to community. Journal of Ecology, 108(1), 267–278.
- Zahra, N., Hafeez, M. B., Shaukat, K., Wahid, A., & Hasanuzzaman, M. (2021). Fe toxicity in plants: Impacts and remediation. Physiologia Plantarum, 173(1), 201–222.
