Cloning and expression analysis of PmCBL 3 from Pinus massoniana

doi:10.3969/j.issn.1000-2006.201609007

Cloning and expression analysis of PmCBL 3 from Pinus massoniana

CAI Qiong, DING Guijie,DING Bo, ZHANG Renbo, DU Mingfeng

JOURNAL OF NANJING FORESTRY UNIVERSITY ›› 2017, Vol. 41 ›› Issue (04) : 30-36.

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JOURNAL OF NANJING FORESTRY UNIVERSITY ›› 2017, Vol. 41 ›› Issue (04) : 30-36. DOI: 10.3969/j.issn.1000-2006.201609007

Cloning and expression analysis of PmCBL 3 from Pinus massoniana

CAI Qiong, DING Guijie^*,DING Bo, ZHANG Renbo, DU Mingfeng

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Abstract

【Objective】CBLs participates in plants response to abiotic stress. The objectives of this study were to investigate biological function of CBL, and to provide a candidate gene and theoretical basis for the drought tolerant breeding for Pinus massoniana, by cloning the CBL from P. massoniana, and analyzing its sequence features, evolutionary relationships and expression characteristics. 【Method】The full-length cDNA of CBL had been isolated from P. massoniana by RT-PCR and RACE, and bioinformatics analysis was conducted to analyze the properties and phylogenetic tree. In addition, the expression profile of CBL was analyzed by qRT-PCR under drought stress.【Result】A full-length cDNA, designated PmCBL3, was obtained. Sequence analysis revealed that PmCBL3 was 1 035 bp in length, including a 681 bp open reading frame that encodes 226 amino acids. PmCBL3 comprised four EF-Hand functional domains, and the distance between the EF-hand motifs was conservative. Evolutionary analysis further revealed that PmCBL3 had the closest relationship with Picea sitchensis, and CBL exhibited 62%-98% homologies with other plants. qRT-PCR results showed that PmCBL3 expression was the highest in the roots, then was in the stems and leaves. PmCBL3 expression was in the peak at 15 d after treatment, but decreased with drought stress aggravation. 【Conclusion】The results indicated that PmCBL3 belonged to the member of CBLs family genes, and had the closest relationship with CBL of P. sitchensis. PmCBL3 responded drought stress, which suggested that it was involved in drought stress regulation

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CAI Qiong, DING Guijie,DING Bo, ZHANG Renbo, DU Mingfeng. Cloning and expression analysis of PmCBL 3 from Pinus massoniana[J]. JOURNAL OF NANJING FORESTRY UNIVERSITY. 2017, 41(04): 30-36 https://doi.org/10.3969/j.issn.1000-2006.201609007

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