Molecular cloning and functional characterization of apple U-box E3 ubiquitin ligase gene MdPUB29 reveals its involvement in salt tolerance

doi:10.1016/S2095-3119(19)62594-3

Journal of Integrative Agriculture ›› 2019, Vol. 18 ›› Issue (7): 1604-1612.DOI: 10.1016/S2095-3119(19)62594-3

收稿日期:2018-07-23 修回日期:2018-12-11 出版日期:2019-07-01 发布日期:2019-07-01

Molecular cloning and functional characterization of apple U-box E3 ubiquitin ligase gene MdPUB29 reveals its involvement in salt tolerance

HAN Peng-liang, DONG Yuan-hua, JIANG Han, HU Da-gang, HAO Yu-jin

State Key Laboratory of Crop Biology, National Research Center for Apple Engineering and Technology, Shandong Agricultural University, Tai’an 271018, P.R.China

Received:2018-07-23 Revised:2018-12-11 Online:2019-07-01 Published:2019-07-01
Contact: Correspondence HU Da-gang, Tel/Fax: +86-538-8246151, E-mail: fap_296566@163.com; HAO Yu-jin, E-mail: haoyujin@sdau.edu.cn
About author:HAN Peng-liang, E-mail: 18763825273@163.com;
Supported by:
This work was supported by the grants from the National Natural Science Foundation of China (31601728, 31471854 and 31772288), the Innovation Team Support Program from the Ministry of Education of China (IRT15R42), the Shandong Natural Science Foundation, China (ZR2016CQ13), the Shandong Modern Agriculture Industry Technology System, China (SDAIT-06-03), the Shandong Agricultural University Outstanding Youth Fund, China (564024), and the Shandong Agricultural University Science and Technology Innovation Fund Project, China (24024).

摘要/Abstract

Abstract:

An E3 ubiquitin ligase gene (Genbank accession no.: MD01G1010900) was cloned from the Royal Gala apple genome (Malus×domestica Borkh.). Sequence analysis showed that the length of the MdPUB29 gene was 1 275 bp, encoding 424 amino acids. Phylogenetic tree analysis indicated that the apple E3 ubiquitin ligase exhibited the greatest sequence similarity to Pyrus×bretschneideri. The predicted protein structural domain of MdPUB29 showed that it contained a U-box domain. qRT-PCR analysis showed that MdPUB29 was expressed widely in different tissues of the Royal Gala apple species, and was highly expressed in the root, while the expression of MdPUB29 was significantly inhibited by exogenous NaCl. Immunoblotting assays revealed that MdPUB29 protein abundance in tissue cultures of the Royal Gala apple accumulated under NaCl stress conditions. Three-dimensional protein structure prediction indicated that MdPUB29 was highly homologous with AtPUB29. The growing potential of MdPUB29-expressing apple calli and Arabidopsis were much stronger than that of the control under salt stress conditions, suggesting that MdPUB29 may positively regulate salt tolerance.

Key words: apple , E3 ubiquitin ligase , , MdPUB29 , salt stress

. [J]. Journal of Integrative Agriculture, 2019, 18(7): 1604-1612.

HAN Peng-liang, DONG Yuan-hua, JIANG Han, HU Da-gang, HAO Yu-jin. Molecular cloning and functional characterization of apple U-box E3 ubiquitin ligase gene MdPUB29 reveals its involvement in salt tolerance[J]. Journal of Integrative Agriculture, 2019, 18(7): 1604-1612.

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Molecular cloning and functional characterization of apple U-box E3 ubiquitin ligase gene MdPUB29 reveals its involvement in salt tolerance

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