Gene Cloning and Functional Analysis of Heat Shock Protein VvDnaJ05 and VvDnaJ26 in Grape (Vitis vinifera)
CHEN Tian-Chi1,*, XU Tao1,*, SHEN Le-Yi1, LUO Ying-Jie1, LIU Ting-Ting2, JIA Yong-Hong1, WU Yue-Yan1**
1 College of Biological and Environmental Sciences, Zhejiang Wanli University, Ningbo 315100, China; 2 Institute of Plant Virology, Ningbo University, Ningbo 315211, China
Abstract：Grape (Vitis vinifera) often suffers various environmental stresses. The mechanism of tolerance to stresses has been a hot topic of cultivation techniques in grape. Plant stress resistance known to rely on special proteins. DnaJ protein (HSP40) serves as a co-chaperone of heat shock protein 70 (HSP70), playing crucial roles in various stress responses. This study focused on the VvDnaJ05 (GenBank No. OQ102602) and VvDnaJ26 (GenBank No. OQ108519) cloning from DnaJ gene family. The expression patterns of the VvDnaJ05 and VvDna26 in different tissues and under various hormones and stresses were determined by qPCR. Meanwhile, the overexpression and interaction vectors were constructed to explore the protein function and regulatory mechanism of the VvDnaJ05 and VvDnaJ26. The results indicated that the full-length of VvDnaJ05 was 822 bp, encoding 273 amino acids. The full-length of VvDnaJ26 was 1 254 bp, encoding 417 amino acids. VvDnaJ05 and VvDnaJ26 showed the highest expression levels in skin and seed, respectively. Hormones responsive cis-acting elements were found in the promoter region of VvDnaJ05 and VvDna26, significantly changes were observed in the expression level of both under methyl jasmonate and salicylic acid treatment (P<0.05). VvDnaJ05 and VvDnaJ26 may play an essential role when the grape plants were exposed to salt, shade and hot stresses, especially in heat stress, the expression level of both under heat stress (35 ℃) was significantly increased in 6 h after the treatment (P<0.05). Transient expression results showed that the VvDnaJ05 and VvDnaJ26 were able to improve the heat tolerance ability of the tobacco (Nicotiana tabacum), and the malondialdehyde and H2O2 significantly decreased (P<0.05) compared with that of the control. The subcellular localization results showed that the VvDnaJ05 was localized in the cytoplasm and nuclear, and the VvDnaJ26 was localized in the cytoplasm. Additionally, the VvDnaJ05 and VvDnaJ26 did not present interaction relationship. The results above indicated that VvDnaJ05 and VvDnaJ26 respectively exert their biological functions and play a key role in stress resistance. The findings of this study could provide a theoretical foundation for the further investigation on the protein functions of the VvDnaJs and breeding of new grape varieties resisted stresses.
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