Isolation and expression analysis of proline metabolism-related genes in Chrysanthemum lavandulifolium |
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Authors: | Mi Zhang He HuangSilan Dai |
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Affiliation: | Beijing Key Laboratory of Ornamental Plants Germplasm Innovation & Molecular Breeding, Beijing 100083, China; National Engineering Research Centre for Floriculture, Beijing 100083, China; College of Landscape Architecture, Beijing Forestry University,Beijing 100083, China |
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Abstract: | Proline plays a significant role in plant resistance to abiotic stresses, and its level is determined by a combination of synthesis, catabolism and transport. The primary proteins involved are Δ1-pyrroline-5-carboxylate synthetase (P5CS), proline dehydrogenase (PDH) and proline transporter (ProT). To utilise proline metabolism to improve the stress resistance of Chrysanthemum × morifolium, we isolated two P5CS-homologous genes (ClP5CS1 and ClP5CS2), one PDH gene (ClPDH) and four ProT-homologous genes (ClProT1-4) (GenBANK accession numbers: KF743136–KF743142) from Chrysanthemum lavandulifolium, which is closely related to chrysanthemums and exhibits strong resistance to stresses. Expression analysis of these genes in different organs and under various stresses indicated that ClP5CSs showed substantial constitutive expression, while ClPDH was only strongly expressed in the capitulum and was inhibited under most stresses. The expression patterns of four ClProT genes presented characteristics of organ specificity and disparity under stresses. Above all, the expression of ClProT2 was restricted to above-ground organs, especially strong in the capitulum and could be obviously induced by various stress conditions. Promoters of ClPDH and ClProTs contained many cis-acting regulatory elements involved in stress responses and plant growth and development. High levels of free proline were found in flower buds, the capitulum under the non-stress condition and later periods of stress conditions except cold treatment. Interestingly, organ specificity and disparity also exist in the level of free proline under different stress conditions. Our study indicates that ClProTs play significant roles in proline accumulation and stress responses, and that ClProT2 could be used to genetically modify the stress resistance of chrysanthemums. In addition, proline metabolism might be closely related to plant flowering and floral development. |
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Keywords: | 3&prime RACE, 3&prime rapid amplification of cDNA ends AAAP, amino acid/auxin permease ABA, abscisic acid APC, amino acid-polyamine-choline ATF, amino acid transporter family CAT, cationic amino acid transporter CBF, C-repeat binding factor ClTUA, α-tubulin gene in Chrysanthemum lavandulifolium ESTs, expressed sequence tags GSA, glutamate-semialdehyde LHT, lysine&ndash histidine transporter ORF, open reading frame P5C, Δ1-pyrroline-5-carboxylate P5CDH, Δ1-pyrroline-5-carboxylate dehydrogenase P5CR, Δ1-pyrroline-5-carboxylate reductase P5CS, Δ1-pyrroline-5-carboxylate synthetase PCR, polymerase chain reaction PDH, proline dehydrogenase PRE, proline- or hypoosmolarity-responsive element ProT, proline transporter RT-PCR, reverse transcription-polymerase chain reaction RT-qPCR, real-time quantitative PCR δOAT, ornithine-δ-aminotransferase |
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