Abstract
As one of the most important phytohormones, the abscisic acid (ABA) is often used to breed stress-tolerant crop lines with both higher yields and active ingredient contents. In higher plants, the 9-cis-epoxycarotenoid dioxygenase (NCED) has been found to be a regulatory enzyme involved in ABA biosynthesis. In research, the novel gene SmNCED3 was isolated from S. miltiorrhiza. The open reading frame of SmNCED3 was 1725-bp, and it was encoding 574 amino acids with a calculated molecular mass of 63,822 kDa, which was verified by the expression of SmNCED3 in E. coli. The deduced SmNCED3 amino acid sequence had high sequence homology with NCED sequences from other plants and contained a putative chloroplast transit targeting signal peptide at its N terminus. Phylogenetic analysis demonstrated that SmNCED3 had a closer affinity to NCED3 in Arabidopsis thaliana (AtNCED3). The 1732-bp 5′ flanking sequence of SmNCED3 was also cloned. It contained several phytohormone response elements, biotic or abiotic stress-related elements, and plant development-related elements. Real-time PCR revealed that SmNCED3 was highly expressed in leaves, and was strongly induced by exogenous ABA. A subcellular localization experiment indicated that SmNCED3 was located in chloroplast stroma, chloroplast membranes, and thylakoid membranes. The overexpression of SmNCED3 promoted ABA accumulation. These results indicated that SmNCED3 might be a rate-limiting gene regulating ABA biosynthesis, and improving abiotic stresses tolerance and active ingredient contents in plants.
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Acknowledgements
This work was partly supported by the National Nature Science Foundation of China (Grant nos. 81403033, 81673536 and 81373908) and partly supported by the Zhejiang Provincial Natural Science Key Project (Grant No. LZ16H280001). We also thank Prof. Weibo Jin (Zhejiang Sci-Tech University, China) for his assistance in this work.
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Communicated by H. Peng.
Yanyan Jia and Jingling Liu are co-first authors.
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Jia, Y., Liu, J., Bai, Z. et al. Cloning and functional characterization of the SmNCED3 in Salvia miltiorrhiza. Acta Physiol Plant 40, 133 (2018). https://doi.org/10.1007/s11738-018-2704-x
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DOI: https://doi.org/10.1007/s11738-018-2704-x