Glycine betaine involvement in freezing tolerance and water stress in Arabidopsis thaliana

https://doi.org/10.1016/S0098-8472(01)00078-8Get rights and content

Abstract

Levels of endogenous glycine betaine in the leaves were measured in response to cold acclimation, water stress and exogenous ABA application in Arabidopsis thaliana. The endogenous glycine betaine level in the leaves increased sharply during cold acclimation treatment as plants gained freezing tolerance. When glycine betaine (10 mM) was applied exogenously to the plants as a foliar spray, the freezing tolerance increased from −3.1 to −4.5°C. In addition, when ABA (1 mM) was applied exogenously, the endogenous glycine betaine level and the freezing tolerance in the leaves increased. However, the increase in the leaf glycine betaine level induced by ABA was only about half of that by the cold acclimation treatment. Furthermore, when plants were subjected to water stress (leaf water potential of ∼−1.6 MPa), the endogenous leaf glycine betaine level increased by about 18-fold over that in the control plants. Water stress lead to significant increase in the freezing tolerance, which was slightly less than that induced by the cold acclimation treatment. The results suggest that glycine betaine is involved in the induction of freezing tolerance in response to cold acclimation, ABA, and water stress in Arabidopsis plants.

Introduction

Many plants accumulate low-molecular-weight compounds with cryoprotectant activity in response to low temperatures (Guy, 1990, Rajashekar, 2000). Glycine betaine is one of several such compatible solutes (Dorffling et al., 1990, Koster and Lynch, 1992, Stushnoff et al., 1997) that has osmoprotection function, and is known to protect proteins and enzyme activities under water deficits, and even stabilize membranes during freezing (Coughlan and Heber, 1982, Anchordoguy et al., 1987, Zhao et al., 1992, Rhodes and Hanson, 1993). The cryoprotective effects of glycine betaine appear to come from its compatibility with macromolecular structure and function. It has been suggested that glycine betaine can help stabilize protein tertiary structure and prevent or reverse the disruption of the tertiary structure of proteins caused by non-compatible (perturbing) solutes (Bateman et al., 1992). Glycine betaine has been shown to protect spinach thylakoids against freezing stress, and it was proposed that this protection may be due to a weak interaction between the positive quaternary ammonium cation and the anionic carboxyl groups of the exposed membrane proteins (Coughlan and Heber, 1982).

Accumulation of glycine betaine in response to low temperatures has been reported in wheat (Naidu et al., 1991, Allard et al., 1998), Puma rye (Koster and Lynch, 1992), barley (Kishitani et al., 1994), and strawberry (Rajashekar et al., 1999). The accumulation of endogenous glycine betaine was closely related to the development of freezing tolerance (Kishitani et al., 1994). In addition, significant increase in freezing tolerance has been observed after exogenous application of glycine betaine in many plants (Zhao et al., 1992, Allard et al., 1998, Rajashekar et al., 1999). These observations suggest that accumulation of glycine betaine may play a role in the induction of freezing tolerance during cold acclimation.

Water stress has long been known to induce freezing tolerance in a number of plant species including Arabidopsis thaliana (Chen et al., 1975, Cloutier and Siminovitch, 1982, Guy et al., 1992, Mantyla et al., 1995). In many of these studies, water stress was as effective as the cold acclimation treatment in inducing freezing tolerance (Cloutier and Siminovitch, 1982, Mantyla et al., 1995) and by and large elicited similar responses in plants as do low temperatures (Shinozaki and Yamaguchi-Shinozaki, 1996, Guy et al., 1992). Activation of a number of genes has been reported in response to both cold acclimation and water stress (Curry et al., 1991, Holappa and Walker-Simmons, 1995, Mantyla et al., 1995). In addition, a cis- acting promoter element responsive to both low temperature and water stress in Arabidopsis has been identified (Yamaguchi-Shinozaki and Shinozaki, 1994). Abscisic acid is known to accumulate in response to low temperatures and water stress (Chen et al., 1983, Cornish and Zeevaart, 1984) and can induce freezing tolerance in a number of plant species, when applied exogenously (Chen and Gusta, 1983, Lang et al., 1989). Many genes responsive to cold and drought are also induced by ABA, which suggests that its pathway in inducing freezing tolerance may overlap or share with that in response to low temperatures.

In this study, we tested the hypothesis that glycine betaine accumulates in response to cold acclimation conditions and water stress, and is involved in the freezing tolerance of Arabidopsis thaliana.

Section snippets

Plant materials and growing conditions

Arabidopsis thaliana (Columbia) plants were grown in a greenhouse at 23/18°C day/night temperatures under natural day light in 15-cm pots containing a sterile growing medium of peat, perlite, and soil (2:2:1 v/v). Plants were irrigated once every 2 days to the field capacity and fertilized weekly with Peat-Lite Special (N–P–K; 10–10–20, Scotts-Sierra Horticultural Products Co., OH) at 250 ppm of nitrogen in irrigation water. For cold-acclimation, plants were transferred to walk-in cold chambers

Results

In response to the cold acclimation treatment, Arabidopsis plants acquired freezing tolerance rather rapidly. The freezing tolerance of leaves increased by about 2°C after the first day of cold acclimation treatment (Fig. 1). Plants reached their maximum freezing tolerance (about −7°C) by the end of 1 week of cold acclimation. As plants cold hardened, the leaves became much thicker and appeared dark green with slight wrinkling along the edges compared with the unhardened control plants.

Discussion

Glycine betaine has been known to accumulate in a wide range of plants typically in response to salt and drought stress (McCue and Hanson, 1990, Rhodes and Hanson, 1993, Xing and Rajashekar, 1999). However, glycine betaine has also been shown to accumulate in response to low temperatures during cold acclimation in barley (Kishitani et al., 1994), wheat (Naidu et al., 1991), and Puma rye (Koster and Lynch, 1992). Typically, the increase in endogenous glycine betaine levels ranged from two- to

References (36)

  • P. Chen et al.

    Induction of frost hardiness in red osier dogwood stems by water stress

    HortSci.

    (1975)
  • Y. Cloutier et al.

    Correlation between cold- and drought- induced frost hardiness in winter wheat and rye varieties

    Plant Physiol.

    (1982)
  • K. Cornish et al.

    Abscisic acid metabolism in relation to water stress and leaf age in Xanthium strumarium

    Plant Physiol.

    (1984)
  • S.J. Coughlan et al.

    The role of glycine betaine in the protection of spinach thylakoids against freezing stress

    Planta

    (1982)
  • J. Curry et al.

    Sequence analysis of a cDNA encoding a group 3 LEA mRNA inducible by ABA or dehydration stress in wheat

    Plant Mol. Biol.

    (1991)
  • K. Dorffling et al.

    Abscisic acid and proline levels in cold hardened winter wheat leaves in relation to variety-specific differences in freezing resistance

    J. Agron. Crop Sci.

    (1990)
  • C.L. Guy

    Cold acclimation and freezing stress tolerance: role of protein metabolism

    Annu. Rev. Plant Physiol. Plant Mol. Biol.

    (1990)
  • C.L. Guy et al.

    Hydration- state-responsive proteins link cold and drought stress in spinach

    Planta

    (1992)
  • Cited by (139)

    • Influence of hormonal seed priming on seedling growth, development, and potential antioxidant performance under abiotic stress

      2023, Hormonal Cross-Talk, Plant Defense and Development: Plant Biology, Sustainability and Climate Change
    • Role of glycine betaine in the protection of plants against environmental stresses

      2023, Plant Stress Mitigators: Types, Techniques and Functions
    • Evolution, family expansion, and functional diversification of plant aldehyde dehydrogenases

      2022, Gene
      Citation Excerpt :

      Precisely, ALDHs have been reported to deal with a variety of abiotic stressors, such as dehydration, high temperature, high salinity, heavy metals, and oxidative stress, implying that ALDHs may play a significant role in environmental adaptability (Gao & Han, 2009; Kotchoni et al., 2006). Aside from the stress resistance capability, ALDHs also perform many other activities, in particular, (i) regulating secondary metabolism, especially the metabolism of amino acids and retinoic acid (Zhang et al., 2012); (ii) producing osmoprotectants, such as glycine betaine, to defend against osmotic stress (Ishitani et al., 1995; Xing & Rajashekar, 2001); (iii) generating NADPH and NADH like other oxidoreductases, which help to maintain redox equilibrium (Hou & Bartels, 2015). There are also several physiological processes in which plant ALDH participates and aids plant survival and appropriate growth.

    View all citing articles on Scopus
    View full text