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| Cloning and expression anysis of Dehydrin gene in Tea Plant(Camellia sinensis (L.) O. Kuntz) |
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Abstract Dehydrins (DHNs), known as a group of late embryogenesis abundant proteins (LEA) in plants, are related to various environment stresses. In this study, CsDHN (GenBank accession No. FJ436978), a full length cDNA of DHN in tea plant(Camellia sinensis) was amplified via rapid-amplification of cDNA ends (RACE) technology. It was shown that the full-length cDNA was 960 bp, encoding 201 amino acids by sequence analysis. The deduced molecular weight of DHN protein was 21 kD, together with isoelectric point 8.3, which belonged to the Y3SK2 type of dehydrins. There were 2 exons and 1 intron in CsDHN. The bioinformatics prediction revealed that CsDHN located in the cytoplasm with no signal peptide or transmembrane domain. It was predicted to show strong hydrophilicity. The analysis of characteristic expression showed that CsDHN was up-regulated in tea plant exposed to low temperature, dehydration, abscisic acid(ABA) treatment and salt stress. The expression of CsDHN was increased not satisfactorily at 3 and 6 h under low temperature stress at 4 ℃, while the expression level was increased greatly (two times higher than control, P＜0.05) at 12 h. And the highest expression, 4.2-fold than that of control, was found at 18 h treatment, then decreased to its original level as control at 24 h. After the treatment of dehydration, the expression of CsDHN was increased dramatically. The expression level was 7.5-fold than that of control at 3 h and continuously increased to the highest level at 18 h, with the expression level was 93.4-fold (P＜0.05) than that of control. After that, the expression was decreased to some extent, not maintained a high level. In accordance to ABA treatment, the expression amount was markedly increased at 3 h, and reached to its greatest amount at 6 h (4.3-fold than that of control, P＜0.05), then began to decline. A little bit higher than its original level was found at 18 h to 24 h treatment. When treated with the solution of 300 mmol/L NaCl, the expression level of CsDHN was increased all the time till to 24 h, which was 2.5-fold and 17.8-fold than that of the original level at 3 h and 24 h, respectively. In the normal growth conditions, the expression of CsDHN existed in all organs in tea plant, which CsDHN was a constitutive expression gene. While, the expression patterns in all organs were obviously different. The highest expression was found in mature seeds (the expression was 11.2-fold than that of mature leaves); similar expression level were found in bud and leaves. But the expression levels in young stems, flowers and young roots were low with no significance, which were 0.24, 0.26, 0.32 -folds compared with leaves, respectively. This study showed that the CsDHN might participates with the defense against abiotic stress for tea plants and dehydration vitality protection during tea seeds maturation, which provides a certain theory foundation for understanding the molecular mechanism to tea stress resistance.
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Received: 04 November 2015
Published: 05 February 2016
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