Abstract

Abscisic acid (ABA) is a phytohormone that is necessary for stress adaptation. Recent studies have reported that attenuated levels of ABA improved grain yield and seedling growth under low temperature in cereals. To improve plant growth under low temperature, we attempted to generate ABA-insensitive transgenic rice by expressing a clade A type 2C protein phosphatase (OsPP2C), OsABIL2, with or without the mutation equivalent to the Arabidopsis abi1-1 mutation. A yeast two-hybrid assay revealed that the interaction between OsABIL2 and a putative rice ABA receptor, OsPYL1, was ABA-dependent, and the interaction was lost with amino acid substitution from glycine to aspartic acid at the 183rd amino acid of the OsABIL2 protein, corresponding to abi1-1 mutation. The constitutive expression of OsABIL2 or OsABIL2G183D in Arabidopsis or rice decreased ABA sensitivity to differing degrees. Moreover, the transgenic rice expressing OsABIL2G183D exhibited improved seedling growth under low temperature, although the transgenic lines showed unfavorable traits, such as viviparous germination and elongated internodes. These results indicated that the introduction of abi1-1 type dominant mutation was also effective in OsABIL2 at decreasing ABA sensitivity in plants, and the attenuation of ABA sensitivity could be an alternative parameter to improve rice performance under low temperatures.

Highlights

  • Abscisic acid (ABA) possesses two opposite effects related to stress adaptation in plants

  • AtPYR1 is a member of the dimeric ABA receptors that interact with clade A PP2C in an ABA-dependent manner[30]

  • The mutation of abi[1] resulted in substitution of the 180th amino acid from glycine to aspartic acid in Arabidopsis ABI1 protein and homogenous amino acid substitution in ABI2 and HAB1 led to ABA insensitivity[27,31,32]

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Summary

Introduction

ABA possesses two opposite effects related to stress adaptation in plants. On the one hand, genetic enhancements of ABA metabolism or sensitivity effectively improve drought stress tolerance in various plant species at the vegetative stage[16,17,18]. On the other hand, when crops were subjected at the reproductive stage to environmental stresses such as high temperatures, drought, or cold, stress-induced pollen sterility led to the loss of grain or fruit. HAB1W300A does not interact with ABA receptor in yeast, and the ectopic expression of HAB1W300A in Arabidopsis presented ABA-resistant germination and wilty phenotypes as a result of ABA insensitivity, indicating that ABA-receptor-insensitive PP2C could reduce ABA sensitivity in plants[25]. Several studies have reported that the ectopic expression of clade A PP2Cs harboring the abi[1] type dominant mutation showed a stronger ABA insensitivity than that of regular clade A PP2C in dicotyledonous plants such as Arabidopsis and poplar[26,27]. We investigated whether transgenic rice with reduced ABA sensitivity presented improved growth under low temperature conditions or not

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