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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The authors thank Valeria M. Borjas Mendez and Sandy Rojas for technical assistance. We thank Alyssa Grube for assistance in language support.
Authors and Contributors
JLRR conceived the study and designed the experiments with PAJ. JLRR, CIB, MRD, CE, and JPM performed the experiments and analyzed the data. JLRR, CIB, FA, and PAJ drafted the paper. All authors contributed to the revision of the manuscript and approved the final version.
Jesús L. Romero-Romero was supported by PhD fellowship CONACYT (SC392465), COTEBAL: 1865 and CONICYT Doctoral Scholarship for foreigners (63130094). This work was supported by FONDECYT 1150220 to PAJ, CAPES FB-002-2014 to FA, and CONICYT PAI 82140040 to JPM.
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Romero-Romero, J.L., Inostroza-Blancheteau, C., Reyes-Díaz, M. et al. Increased Drought and Salinity Tolerance in Citrus aurantifolia (Mexican Lemon) Plants Overexpressing Arabidopsis CBF3 Gene. J Soil Sci Plant Nutr 20, 244–252 (2020). https://doi.org/10.1007/s42729-019-00130-y