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Cloning, in silico characterization, subcellular localization, and expression of a heat shock cognate 70 kDa protein/gene (EjHsc70-2) from Eriobotrya japonica

Lin, Shoukai, Wu, Ti, Li, Ming, Huang, Xiaoming, Zhang, Yanqing, Han, Linxiao, Wu, Bisha, Chen, Yu, Lin, Suying, Lin, Dahe, Wu, Minlan, Wu, Jincheng
Acta physiologiae plantarum 2019 v.41 no.7 pp. 119
Eriobotrya japonica, amino acids, bioinformatics, cold storage, complementary DNA, fruits, genes, heat-shock protein 70, loquats, open reading frames, phylogeny, quantitative polymerase chain reaction, reverse transcriptase polymerase chain reaction, stress response
Heat shock protein 70 kDa proteins (Hsp70 s) are among the most important molecular chaperone groups and play a significant role in the stress responses and development of plants. In the present study, the full-length cDNA of the heat shock cognate 70 protein 2 gene EjHsc70-2, which encodes a loquat Hsp70 s member, was cloned and characterized, and its expression and subcellular localization were also investigated. The full-length cDNA of EjHsc70-2 consists of an open reading frame (ORF) of 1950 bp, a 5′-UTR of 103 bp, and a 3′-UTR of 62 bp, and the ORF encodes 649 amino acid residues. The structure of the loquat Hsc70-2 protein was analysed using several bioinformatics tools, and the results showed that the protein was, indeed, a member of the Hsp70 s. Phylogenetic tree analysis suggested that the genetic evolution of Hsc70-2 genes conformed well to the morphology based taxonomic classification of seed plants. BLAST and multiple alignment analyses determined that the Hsc70-2 genes and Hsc70-2 proteins were both highly conserved among loquat and other seed plants, suggesting that the functions of EjHsc70-2 might be similar to those of other Hsc70-2 genes. The bioinformatics and experimental subcellular localization analyses both supported that EjHsc70-2 was a cytoplasmic and/or nuclear protein. Quantitative real-time RT-PCR (RT-qPCR) suggested its conserved functions involved in loquat organ development. Moreover, EjHsc70-2 were also inducible, which may contribute to the low-temperature adaptation of loquat fruits in cold storage. These results provide new insights into the characteristics and functions of Hsp70 s in Eriobotrya japonica.