Overexpression of RcOLEO1 Enhanced the Tolerance of Arabidopsis thaliana toDrought and High-temperature Stress

doi:10.13560/j.cnki.biotech.bull.1985.2025-1043

Abstract

Abstract:

Objective Oleosin (OLEO, OLE) play crucial roles in oil body formation and lipid storage in plants, while also contributing significantly to the stress resisting process of plants. Racomitrium canescens is a typical xerophytic moss exhibiting extreme tolerance to desiccation and high-temperature. This study aimed to investigate whether the stress-responsive oleosin gene RcOLEO1 from R. canescens can enhance plant resistance to stress, providing a theoretical foundation for elucidating the molecular mechanisms of underlying drought and high-temperature in R. canescens and identifying potential drought-resistant genetic resources. Method The expression patterns of RcOLEO1 under rehydration, dehydration and high-temperature stress treatments were analyzed using real-time quantitative PCR (RT-qPCR). The coding sequence of RcOLEO1 was cloned and used to construct overexpressing A. thaliana lines, the transgenic A. thaliana plants were subjected to have drought and high-temperature stress treatments, respectively, for phenotypic observations and physiological and biochemical indexes measurements. Result RT-qPCR results revealed that RcOLEO1 was responsive to rehydration, dehydration and high-temperature stress treatments in R. canescens. The coding sequence of RcOLEO1 was successfully cloned, and T₂ generation transgenic A. thaliana overexpressing RcOLEO1 was obtained. Under drought and high-temperature stress, RcOLEO1-overexpresion lines demonstrated enhanced tolerance compared to wild-type plants. Physiological analyses, including measurements of osmoregulatory substances content, malondialdehyde content, and chlorophyll content, preliminarily elucidated the mechanisms by which the overexpression of RcOLEO1 improved drought and high-temperature tolerance in A. thaliana. Conclusion The overexpression of RcOLEO1 likely enhances plant tolerance to drought and high-temperature stress by modulating the accumulation of osmoregulatory compounds and antioxidant enzyme activity.

Key words: Racomitrium canescens, RcOLEO1, oleosin gene, tolerance to drought stress, tolerance to high-temperature stress, gene expression, bioinformatics analysis, detection of physiological and biochemical indexes

GUO Miao, XU Jia-jia, SUN Tian-guo, CAI Can, CAO Wan-di, BAO Ji-xing, SHA Wei, ZHANG Mei-juan, PENG Yi-fang, MA Tian-yi. Overexpression of RcOLEO1 Enhanced the Tolerance of Arabidopsis thaliana toDrought and High-temperature Stress[J]. Biotechnology Bulletin, 2026, 42(5): 312-322.

Figures/Tables 7

Fig. 1 Expression analysis of RcOLEO1 during rehydration and dehydration treatments of Racomitrium canescensThe error bars indicates the standard deviation, and the letters on the error bars indicate different significance (P<0.05), the same below

Fig. 2 Phylogenic analysis (A) and sequence alignment (B) of RcOLEO1 and homologous proteinsThe red boxes indicate the conserved proline knot motif

Fig. 3 Screening and characterization of RcOLEO1 overexpression transgenic A. thalianaA: Screening of T1 generation plants of transgenic A. thaliana overexpressing RcOLEO1.B: Identification of T2RcOLEO1 overexpression transgenic A. thaliana

Fig. 4 Phenotypes of RcOLEO1 overexpression transgenic A. thaliana under drought stress

Fig. 5 Changes in physiological and biochemical indexes of wild-type and RcOLEO1 overexpression transgenic A. thaliana under drought treatment

Fig. 6 Phenotypes of RcOLEO1 overexpression transgenic A. thaliana plants under 45 ℃ high-temperature treatment

Fig. 7 Changes in physiological and biochemical indexes of wild-type and RcOLEO1 overexpression transgenic A. thaliana under 45 °C high-temperature treatment

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