Overexpression of RcFDH2 from Racomitrium canescens Enhances the Tolerance of Arabidopsis thaliana to Drought and High-temperature

doi:10.13560/j.cnki.biotech.bull.1985.2025-1056

Abstract

Abstract:

Objective To investigate whether a formate dehydrogenase (FDH) gene RcFDH2 of Racomitrium canescens possesses drought- and high-temperature-tolerant functions, and to provide theoretical support for revealing the drought- and high-temperature-tolerant mechanisms of R. canescens and for cultivating resistant crops. Method Real-time fluorescence quantitative PCR technology was used for the detection of the expression variations of RcFDH2 during the rehydration process of desiccated R. canescens. The coding sequence of RcFDH2 was successfully cloned. RcFDH2-overexpressed transgenic Arabidopsis thaliana lines were acquired, followed by drought and high-temperature treatments, the plant phenotypes were observed, and the physiological and biochemical indexes were determined. Result RcFDH2 showed differential expression during the rehydration process of R. canescens; the coding sequence of RcFDH2 was successfully cloned, and multiple RcFDH2-overexpressed transgenic A. thaliana lines were obtained. It was found that under drought and high-temperature treatments, the RcFDH2 transgenic lines showed stronger tolerances to drought and high-temperature compared to wild-type plants. The physiological and biochemical indicators of RcFDH2-overexpressed transgenic A. thaliana and wild-type plants revealed that in the transgenic plants, the contents of three osmotic regulatory substances, the total chlorophyll content, and the activities of three antioxidant enzymes were significantly higher than those in the wild-type plants, while the malondialdehyde content was lower than that in the wild-type plants. Conclusion RcFDH2 can respond to desiccation and high-temperature stress treatments in R. canescens, and its overexpression probably enhanced the drought and high-temperature tolerance of A. thaliana maybe by regulating osmotic regulatory substances and antioxidant enzyme activities.

Key words: Racomitrium canescens, formate dehydrogenase gene, RcFDH2, tolerance to desiccation, tolerance to drought, tolerance to high-temperature, abiotic stress

MA Tian-yi, SUN Ming-yu, LIANG Shu-ting, BAO Wei, QIN Xu-yang, ZHANG Mei-juan, SHA Wei, PENG Yi-fang. Overexpression of RcFDH2 from Racomitrium canescens Enhances the Tolerance of Arabidopsis thaliana to Drought and High-temperature[J]. Biotechnology Bulletin, 2026, 42(8): 229-238.

Figures/Tables 8

Fig. 1 Expression changes of RcFDH2 in desiccated R. canescens under rehydration treatmentThe error bars indicate the standard deviation, and the different letters on the error bars indicate the difference is significant (P<0.05), the same below

Fig. 2 Coding sequence of RcFDH2 and its amino acid sequences

Fig. 3 Sequence alignment of RcFDH2 with homologous proteinsRed box indicates the conserved domains of FDH, black box indicates NAD⁺ binding sites, and blue arrow indicates the catalytic sites

Fig. 4 Analysis of RcFDH2 expression in T2 generation lines of RcFDH2-overexpressed transgenic A. thaliana

Fig. 5 Phenotypesof RcFDH2-overexpressed transgenic A. thaliana under drought stress

Fig. 6 Variation in physiological and biochemical indexes of wild-type and RcFDH2-overexpressed transgenic A. thaliana under drought stress

Fig. 7 Phenotypes of RcFDH2-overexpressed transgenic A. thaliana under 45 ℃ high-temperature treatments

Fig. 8 Variation in physiological and biochemical indexes of wild-type and RcFDH2-overexpressed transgenic A. thaliana under 45 ℃ high-temperature treatments

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