Involvement of three annexin genes in the ripening of strawberry fruit regulated by phytohormone and calcium signal transduction.

KEY MESSAGE: Three annexin genes may be involved in the ripening progress of strawberry fruit. Phytohormones and calcium regulate the expressions of three annexin genes during strawberry fruit ripening. Plant annexins are multi-functional membrane- and Ca(2+)-binding proteins that are involved in various developmental progresses and stress responses. Three annexins FaAnn5a, FaAnn5b and FaAnn8 cDNA obtained from strawberry fruit encode amino acid sequences of approximately 35 kDa containing four annexin repeats, Ca(2+)-binding site, GTP-binding motif, peroxidase residue, and conserved amino acid residues of tryptophan, arginine and cysteine. During fruit development, the transcript levels of FaAnn5a and FaAnn5b increased while FaAnn5b declined after 3/4R stage. The expression patterns of annexins suggested their potential roles in strawberry fruit development and ripening. Expressions of annexin genes were also highly correlated with hormone levels. In addition, exogenous abscisic acid (ABA) enhanced the expressions of FaAnn5a and FaAnn8 while exogenous auxin (IAA) retarded it. However, both ABA and IAA promoted the transcript levels of FaAnn5b, indicating the independent regulation of annexins in fruit likely due to multi-functions of their large family. The responses of annexin genes to exogenous ABA and IAA inhibitors verified the involvement of annexins in plant hormone signaling. Besides, calcium restrained the expressions of FaAnn5s (FaAnn5a and FaAnn5b) but promoted the expression of FaAnn8. Effects of calcium and ethylene glycol tetraacetic acid (EGTA) on the transcript levels of annexins confirmed that calcium likely mediated hormone signal transduction pathways, which helped to elucidate the mechanism of calcium in fruit ripening. Therefore, FaAnn5s and FaAnn8 might be involved in plant hormones' regulation in the development and ripening of strawberry fruit through calcium signaling in the downstream.