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Journal of Integrative Agriculture  2019, Vol. 18 Issue (7): 1604-1612    DOI: 10.1016/S2095-3119(19)62594-3
Horticulture Advanced Online Publication | Current Issue | Archive | Adv Search |
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
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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.
 
Keywords:  apple       E3 ubiquitin ligase              MdPUB29       salt stress  
Received: 23 July 2018   Accepted: Online: 11 December 2018  
Fund:  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).
Corresponding Authors:  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;

Cite this article: 

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

Amador V, Monte E, Garc??a-Mart??nez J L, Prat S. 2001. Gibberellins signal nuclear import of PHOR1, a photoperiod-responsive protein with homology to Drosophila armadillo. Cell, 106, 343–354.
Azevedo C, Santos-Rosa M J, Shirasu K. 2001. The U-box protein family in plants. Trends in Plant Science, 6, 354–358.
Bergler J, Hoth S. 2011. Plant U-box armadillo repeat proteins AtPUB18 and AtPUB19 are involved in salt inhibition of germination in Arabidopsis. Plant Biology, 13, 725–730.
Berndsen C E, Wolberger C. 2014. New insights into ubiquitin E3 ligase mechanism. Nature Structural & Molecular Biology, 21, 301.
Cho S K, Ryu M Y, Song C, Kwak J M, Kim W T. 2008. Arabidopsis PUB22 and PUB23 are homologous U-Box E3 ubiquitin ligases that play combinatory roles in response to drought stress. The Plant Cell, 20, 1899–1914.
Ciechanover A, Finley D, Varshavsky A. 1984. Ubiquitin dependence of selective protein degradation demonstrated in the mammalian cell cycle mutant ts85. Cell, 37, 57–66.
Duplan V, Rivas S. 2014. E3 ubiquitin-ligases and their target proteins during the regulation of plant innate immunity. Frontiers in Plant Science, 5, 42.
Farmer L M, Book A J, Lee K H, Lin Y L, Fu H, Vierstra R D. 2010. The RAD23 family provides an essential connection between the 26S proteasome and ubiquitylated proteins in Arabidopsis. The Plant Cell, 22, 124–142.
Hatakeyama S, Kei-ichi I N. 2003. U-box proteins as a new family of ubiquitin ligases. Biochemical and Biophysical Research Communications, 302, 635–645.
Henriques R, Jang I C, Chua N H. 2009. Regulated proteolysis in light-related signaling pathways. Current Opinion in Plant Biology, 12, 49–56.
Hu D G, Sun M H, Sun C H, Liu X, Zhang Q Y, Zhao J, Hao Y J. 2015. Conserved vacuolar H+-ATPase subunit B1 improves salt stress tolerance in apple calli and tomato plants. Scientia Horticulturae, 197, 107–116.
Ji W, Cong R, Li S, Li R, Qin Z, Li Y, Li J. 2016. Comparative proteomic analysis of soybean leaves and roots by iTRAQ provides insights into response mechanisms to short-term salt stress. Frontiers in Plant Science, 7, 573.
Lecker S H, Goldberg A L, Mitch W E. 2006. Protein degradation by the ubiquitin-proteasome pathway in normal and disease states. Journal of the American Society of Nephrology, 17, 1807–1819.
Li R, An J P, You C X, Wang X F, Hao Y J. 2017. Molecular cloning and functional characterization of the α-subunit of heterotrimeric G protein gene MdGPA1 of apple. Scientia Agricultura Sinica, 50, 537–544. (in Chinese)
Liu L, Cui F, Li Q, Yin B, Zhang H, Lin B, Wu Y, Xia R, Tang S, Xie Q. 2011. The endoplasmic reticulum-associated degradation is necessary for plant salt tolerance. Cell Research, 21, 957–969.
Livak K J, Schmittgen T D. 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods, 25, 402–408.
Lu D, Lin W, Gao X, Wu S, Cheng C, Avila J, Shan L. 2011. Direct ubiquitination of pattern recognition receptor FLS2 attenuates plant innate immunity. Science, 332, 1439–1442.
Nakashima K, Yamaguchi-Shinozaki K, Shinozaki K. 2014. The transcriptional regulatory network in the drought response and its crosstalk in abiotic stress responses including drought, cold, and heat. Frontiers in Plant Science, 5, 170.
Orosa B, He Q, Mesmar J, Gilroy E M, McLellan H, Yang C, Craig A, Bailey M, Zhang C, Moore J D, Boevink P C, Tian Z, Birch P R J, Sadanandom A. 2017. BTB-BACK domain protein POB1 suppresses immune cell death by targeting ubiquitin E3 ligase PUB17 for degradation. PLoS Genetics, 13, e1006540.
Qi C H, Zhao X Y, Han P L, Jiang H, Wang Y X, Hu D G, Hao Y J. 2017. Functional identification of salt tolerance and ABA sensitivity of apple U-box E3 ubiquitin ligase MdPUB24. Acta Horticulturae Sinica, 44, 2255–2264. (in Chinese)
Sivasankaramoorthy S, Balasubramanian T T, Amuthavalli P, Sivaraman P. 2011. Effect of NaCl on growth, ion accumulation and oxidative enzymes of Suaeda nudiflora Moq. Recent Research in Science and Technology, 3, 123–127.
Smalle J, Vierstra R D. 2004. The ubiquitin 26S proteasome proteolytic pathway. Annual Review of Plant Biology, 55, 555–590.
Stegmann M, Anderson R G, Ichimura K, Pecenkova T, Reuter P, ?árský V, Trujillo M. 2012. The ubiquitin ligase PUB22 targets a subunit of the exocyst complex required for PAMP-triggered responses in Arabidopsis. The Plant Cell, 24, 4703–4716.
Stone S L, Hauksdóttir H, Troy A, Herschleb J, Kraft E, Callis J. 2005. Functional analysis of the RING-type ubiquitin ligase family of Arabidopsis. Plant Physiology, 137, 13–30.
Yang C W, González-Lamothe R, Ewan R A, Rowland O, Yoshioka H, Shenton M, Sadanandom A. 2006. The E3 ubiquitin ligase activity of Arabidopsis plant U-box 17 and its functional tobacco homolog ACRE276 are required for cell death and defense. The Plant Cell, 18, 1084–1098.
Yang J X, Zhang H, Wang Z L, Wang X L, Wang G L. 2015. Rencent progresses in the regulation mechanism of E3 ligases in plant disease resistance. Plant Protection, 41, 1–8. (in Chinese)
Yee D. 2010. The expanding diversity of plant U-box E3 ubiquitin ligases in Arabidopsis: Identifying AtPUB18 and AtPUB19 function during abiotic stress responses. Ph D thesis, University of Toronto, Toronto, Canada.
Yee D, Goring D R. 2009. The diversity of plant U-box E3 ubiquitin ligases: From upstream activators to downstream target substrates. Journal of Experimental Botany, 60, 1109–1121.
Zeng L R, Qu S, Bordeos A, Yang C, Baraoidan M, Yan H, Wang G L. 2004. Spotted leaf11, a negative regulator of plant cell death and defense, encodes a U-box/armadillo repeat protein endowed with E3 ubiquitin ligase activity. The Plant Cell, 16, 2795–2808.
Zhang M, Fang Y, Ji Y, Jiang Z, Wang L. 2013. Effects of salt stress on ion content, antioxidant enzymes and protein profile in different tissues of Broussonetia papyrifera. South African Journal of Botany, 85, 1–9.
Zhang M, Zhao J, Li L, Gao Y, Zhao L, Patil S B, Li X. 2017. The Arabidopsis U-box E3 ubiquitin ligase PUB30 negatively regulates salt tolerance by facilitating BRI1 kinase
inhibitor 1 (BKI1) degradation. Plant, Cell & Environment, 40, 2831–2843.
Zhang Q Y, Yu J Q, Wang J H, Hu D G, Hao Y J. 2017. Functional characterization of MdMYB73 reveals its involvement in cold stress response in apple calli and Arabidopsis. Journal of Integrative Agriculture, 16, 2215–2221.
Zhao S, Xu C C, Zou Q, Meng Q W. 1994. Improvements of method for measurement of malondialdehyde in plant tissues. Plant Physiol Commun, 30, 207–210.
Zheng Q. 2013. Preliminary identification of U-box protein PUB53 from Arabidopsis thaliana. MSc thesis, Nanjing Agricultural University, China. (in Chinese)
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