Acetylsalicylic acid enhance tolerance of Phaseolus vulgaris L. to chilling stress, improving photosynthesis, antioxidants and expression of cold stress responsive genes

Abstract

Background: High and low temperatures constitute the most damaging type of abiotic stress and limit the survival, and productivity of plants. The present study aimed to evaluate the role of exogenous applications of acetylsalicylic acid (ASA) in reducing the deleterious effects of cold stress. Phaseolus vulgaris L. seedlings were treated with foliar-sprayed ASA at concentrations of 0-3 mM and then subjected to chilling stress at 4 °C for 2 or 4 days.

Results: Growth, photosynthesis, biochemical alterations, oxidative damage and antioxidant enzyme activities as well as the expression of cold-responsive genes (CBF3-COR47), were monitored during the experiment. ASA applications substantially improved several growth and photosynthetic parameters, including shoot biomass, dry weight, and photosynthetic pigments, of P. vulgaris seedlings exposed to different durations of chilling stresses. The ASA foliar spray treatments significantly (p < 0.05) rescued the growth and photosynthetic pigments of P. vulgaris seedlings under different chilling stresses. The total soluble sugars markedly increased during 0-4 days of chilling stress following ASA foliar spraying. The exogenous application of ASA significantly (p < 0.05) increased the accumulation of proline in P. vulgaris seedlings under chilling stress. At the gene expression level, ASA significantly (p < 0.05) upregulated the cold-responsive genes CBF3 and COR47.

Conclusions: As a result, we speculate that, the application of exogenous ASA alleviated the adverse effects of chilling stress on all measured parameters, and 1 and 2 mM ASA exhibited the greatest effects.

Keywords: ASA; Acetylsalicylic acid; Antioxidants enzymes; Catalase; Chilling stress; Peroxidase; Superoxide dismutase.

Fig. 1

Total soluble sugars (mg g⁻¹ FW) of P. vulgaris L. after foliar application of different concentration of acetylsalicylic acid (ASA; 0, 0.1, 0.5, 1, 2, 3 mM) and exposed to different level of chilling stress (0, 2, 4 days). Data represented are mean of two replicates ± standard deviation. Variations between different chilling stresses and foliar ASA concentration were assessed by univariate analyses followed by post hoc analysis. Means with the same letters are not significantly different according to Duncan’s multiple comparisons

Fig. 2

a Protein content (mg g⁻¹ FW), b total free amino acids (mg g⁻¹ FW), c proline (mg g⁻¹ FW) of P. vulgaris L. after foliar application of different concentration of acetylsalicylic acid (ASA; 0, 0.1, 0.5, 1, 2, 3 mM) and exposed to different level of chilling stress (0, 2, 4 days). Data represented are mean of two replicates ± standard deviation. Variations between different chilling stresses and foliar ASA concentration were assessed by univariate analyses followed by post hoc analysis. Means with the same letters are not significantly different according to Duncan’s multiple comparisons

Fig. 3

Cellular lipid peroxidation in terms of MDA (μmol g⁻¹ FW) of P. vulgaris L. after foliar application of different concentration of acetylsalicylic acid (ASA; 0, 0.1, 0.5, 1, 2, 3 mM) and exposed to different level of chilling stress (0, 2, 4 days). Data represented are mean of two replicates ± standard deviation. Variations between different chilling stresses and foliar ASA concentration were assessed by univariate analyses followed by post hoc analysis. Means with the same letters are not significantly different according to Duncan’s multiple comparisons

Fig. 4

a Ascorbic acid content (mg g⁻¹ FW) and b ascorbic peroxidase (APX; mmol ascorbate min⁻¹ g⁻¹ FW) in leaves of P. vulgaris L. after foliar application of different concentration of acetylsalicylic acid (ASA; 0, 0.1, 0.5, 1, 2, 3 mM) and exposed to different level of chilling stress (0, 2, 4 days). Data represented are mean of two replicates ± standard deviation. Variations between different chilling stresses and foliar ASA concentration were assessed by univariate analyses followed by post hoc analysis. Means with the same letters are not significantly different according to Duncan’s multiple comparisons

Fig. 5

a Catalase (unit mg⁻¹ protein min⁻¹) and b peroxidase activities (unit mg⁻¹ protein min⁻¹) of P. vulgaris L. after foliar application of different concentration of acetylsalicylic acid (ASA; 0, 0.1, 0.5, 1, 2, 3 mM) and exposed to different level of chilling stress (0, 2, 4 days). Data represented are mean of two replicates ± standard deviation. Variations between different chilling stresses and foliar ASA concentration were assessed by univariate analyses followed by post hoc analysis. Means with the same letters are not significantly different according to Duncan’s multiple comparisons

Fig. 6

Superoxide dismutase activities (unit mg⁻¹ protein min⁻¹) of P. vulgaris L. after foliar application of different concentration of acetylsalicylic acid (ASA; 0, 0.1, 0.5, 1, 2, 3 mM) and exposed to different level of chilling stress (0, 2, 4 days). Data represented are mean of two replicates ± standard deviation. Variations between different chilling stresses and foliar ASA concentration were assessed by univariate analyses followed by post hoc analysis. Means with the same letters are not significantly different according to Duncan’s multiple comparisons

Fig. 7

Relative expression of genes a CBF3, and b COR47 of P. vulgaris L. after foliar application of different concentration of acetylsalicylic acid (ASA; 0, 0.1, 0.5, 1, 2, 3 mM) and exposed to a different level of chilling stress (0, 2, 4 days). Data represented are mean of two replicates ± standard deviation. Variations between different chilling stresses and foliar ASA concentration were assessed by univariate analyses followed by post hoc analysis. Means with the same letters are not significantly different according to Duncan’s multiple comparisons