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Alvarez-Gerding, X., Cortes-Bullemore, R., Medina, C., Romero-Romero, J. L., Inostroza-Blancheteau, C., Aquea, F., et al. (2015). Improved Salinity Tolerance in Carrizo Citrange Rootstock through Overexpression of Glyoxalase System Genes. Biomed Res. Int., 827951, 7 pp.
Abstract: Citrus plants are widely cultivated around the world and, however, are one of the most salt stress sensitive crops. To improve salinity tolerance, transgenic Carrizo citrange rootstocks that overexpress glyoxalase I and glyoxalase II genes were obtained and their salt stress tolerance was evaluated. Molecular analysis showed high expression for both glyoxalase genes (BjGlyI and PgGlyII) in 5H03 and 5H04 lines. Under control conditions, transgenic and wild type plants presented normal morphology. In salinity treatments, the transgenic plants showed less yellowing, marginal burn in lower leaves and showed less than 40% of leaf damage compared with wild type plants. The transgenic plants showed a significant increase in the dry weight of shoot but there are no differences in the root and complete plant dry weight. In addition, a higher accumulation of chlorine is observed in the roots in transgenic line 5H03 but in shoot it was lower. Also, the wild type plant accumulated around 20% more chlorine in the shoot compared to roots. These results suggest that heterologous expression of glyoxalase system genes could enhance salt stress tolerance in Carrizo citrange rootstock and could be a good biotechnological approach to improve the abiotic stress tolerance in woody plant species.
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de la Guardia, A. R. H., Ugalde, M. B., Lobos-Diaz, V., Romero-Romero, J. L., Meyer-Regueiro, C., Inostroza-Blancheteau, C., et al. (2021). Isolation and molecular characterization of MYB60 in Solanum lycopersicum. Mol. Biol. Rep., 48(2), 1579–1587.
Abstract: Stomatal closure is a common adaptation response of plants to the onset of drought condition and its regulation is controlled by transcription factors. MYB60, a transcription factor involved in the regulation of light-induced stomatal opening, has been characterized in arabidopsis and grapevine. In this work, we studied the role of MYB60 homolog SIMYB60 in tomato plants. We identified, isolated, and sequenced the SIMYB60 coding sequence, and found domains and motifs characteristic of other MYB60 proteins. We determined that SlMYB60 is mainly expressed in leaves, and its expression is repressed by abscisic acid. Next, we isolated a putative promoter region containing regulatory elements responsible for guard cell expression and other putative regulatory elements related to ABA repression and vascular tissue expression. Protein localization assays demonstrated that SlMYB60 localizes to the nucleus. Finally, SlMYB60 is able to complement the mutant phenotype of atmyb60-1 in Arabidopsis. Together, these results indicate that SlMYB60 is the homologous gene in tomato and potentially offer a molecular target to improve crops.
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Romero-Romero, J. L., Inostroza-Blancheteau, C., Orellana, D., Aquea, F., Reyes-Diaz, M., Gil, P. M., et al. (2018). Stomata regulation by tissue-specific expression of the Citrus sinensis MYB61 transcription factor improves water-use efficiency in Arabidopsis. Plant Physiol. Biochem., 130, 54–60.
Abstract: Water-use efficiency (WUE) is a quantitative measurement of biomass produced per volume of water transpired by a plant. WUE is an important physiological trait for drought response to mitigate the water deficiency. In this work, a cisgenic construction from Citrus sinensis was developed and its function in the improvement of WUE was evaluated in Arabidopsis. Sequences of the CsMYB61 coding region, a transcription factor implicated in the closure of stomata, together with a putative stomata-specific promoter from CsMYB.1.5, were identified and cloned. The protein encoded in the CsMYB61 locus harbors domains and motifs characteristic of MYB61 proteins. In addition, a 1.2 kb promoter region of the gene CsMYB15 (pCsMYB15) containing regulatory elements for expression in guard cells and in response to Abscisic Acid (ABA) and light was isolated. In Arabidopsis, pCsMYB15 directs the expression of the reporter gene GUS in stomata in the presence of light. In addition, transgenic lines expressing the CsMYB61 coding region under transcriptional control of pCsMYB15 have a normal phenotype under in vitro and greenhouse conditions. These transgenic lines exhibited a smaller opening of the stomata pore, lower stomatal conductance and respiration rate, enhanced sensitivity to exogenous ABA, and high drought stress tolerance. Our results indicate that stomata-specific expression of CsMYB61 enhances water use efficiency under drought conditions in Arabidospis.
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Romero-Romero, J. L., Inostroza-Blancheteau, C., Reyes-D?az, M., Matte, J. P., Aquea, F., Espinoza, C., et al. (2020). Increased Drought and Salinity Tolerance in Citrus aurantifolia (Mexican Lemon) Plants Overexpressing Arabidopsis CBF3 Gene. J. Soil Sci. Plant Nutr., 20(1), 244–252.
Abstract: Citrus are a globally important fruit crop. Abiotic stressors such as drought and salinity adversely affect physiological citrus performance and survival. With the aim of improving drought tolerance in citrus plants, we constructed transgenic lines of Citrus lemon overexpressing the Arabidopsis transcription factor CBF3. Molecular, physiological, and quantitative real-time analyses showed high expression of AtCBF3 in three selected transgenic lines. During a 15-day treatment of water deficit by cessation of irrigation, the transgenic lines LM2 and LM14 showed lower stomatal conductance and transpiration paired with lower photosynthesis, whereas transgenic line LM7 maintained its photosynthesis, declining stomatal conductance, and transpiration compared to WT plants, which is manifested into more efficient water use. The genes CsRafS1 and CsGolS1 showed similar or greater expression in one of the transgenic lines with respect to control plants. Moreover, transgenic lines were more tolerant to saline stress and presented a greener phenotype with increased chlorophyll content in leaf discs compared to WT plants. In addition, a lower electrical conductivity in solution was observed in transgenic lines. Furthermore, all transgenic lines exhibited significantly less accumulation of reactive oxygen species than WT plants. Together, these results suggest the potential for heterologous expression of the AtCBF3 gene to mediate tolerance to hydric and saline stress in citrus plants.
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Vera-Maldonado, P., Aquea, F., Reyes-Díaz, M., Cárcamo-Fincheira, P., Soto-Cerda, B., Nunes-Nesi, A., et al. (2024). Role of boron and its interaction with other elements in plants. Front. Plant Sci., 15, 1332459.
Abstract: Boron (B) is an essential microelement for plants, and its deficiency can lead to impaired development and function. Around 50% of arable land in the world is acidic, and low pH in the soil solution decreases availability of several essential mineral elements, including B, magnesium (Mg), calcium (Ca), and potassium (K). Plants take up soil B in the form of boric acid (H3BO3) in acidic soil or tetrahydroxy borate [B(OH)4]- at neutral or alkaline pH. Boron can participate directly or indirectly in plant metabolism, including in the synthesis of the cell wall and plasma membrane, in carbohydrate and protein metabolism, and in the formation of ribonucleic acid (RNA). In addition, B interacts with other nutrients such as Ca, nitrogen (N), phosphorus (P), K, and zinc (Zn). In this review, we discuss the mechanisms of B uptake, absorption, and accumulation and its interactions with other elements, and how it contributes to the adaptation of plants to different environmental conditions. We also discuss potential B-mediated networks at the physiological and molecular levels involved in plant growth and development.
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Viveros, M. F. A., Inostroza-Blancheteau, C., Timmermann, T., Gonzalez, M., & Arce-Johnson, P. (2013). Overexpression of GlyI and GlyII genes in transgenic tomato (Solanum lycopersicum Mill.) plants confers salt tolerance by decreasing oxidative stress. Mol. Biol. Rep., 40(4), 3281–3290.
Abstract: The glyoxalase system plays an important role in various physiological processes in plants, including salt stress tolerance. We report the effects of overexpressing glyoxalase I and glyoxalase II genes in transgenic tomato (Solanum lycopersicum Mill.) cv. Ailsa Craig. Stable expression of both transgenes was detected in the transformed tomato plants under salt stress. The transgenic lines overexpressing GlyI and GlyII under a high NaCl concentration (800 mM) showed reduced lipid peroxidation and the production of H2O2 in leaf tissues. A greater decrease in the chlorophyll a+b content in wild-type (WT) compared with transgenic lines was also observed. These results suggest that the over expression of two genes, GlyI and GlyII, may enhance salt stress tolerance by decreasing oxidative stress in transformed tomato plants. This work will help our understanding of the putative role of the glyoxalase system in the tolerance to abiotic stress in tomato plants.
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