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Auxin response factor gene MdARF2 is involved in ABA signaling and salt stress response in apple
WANG Chu-kun, ZHAO Yu-wen, HAN Peng-liang, YU Jian-qiang, HAO Yu-jin, XU Qian, YOU Chun-xiang, HU Da-gang
2022, 21 (8): 2264-2274.   DOI: 10.1016/S2095-3119(21)63843-1
Abstract239)      PDF in ScienceDirect      

Auxin response factors (ARFs) play key roles throughout the whole process of plant growth and development, and mediate auxin response gene transcription by directly binding with auxin response elements (AuxREs).  However, their functions in abiotic stresses are largely limited, especially in apples.  Here, the auxin response factor gene MdARF2 (HF41569) was cloned from apple cultivar ‘Royal Gala’ (Malus×domestica Borkh.).  Phylogenetic analysis showed that ARF2 proteins are highly conserved among different species and MdARF2 is the closest relative to PpARF2 of Prunus persica, but they differ at the DNA level.  MdARF2 contains three typical conserved domains including the B3 DNA-binding domain, Auxin_resp domain and AUX_IAA domain.  The subcellular localization demonstrated that MdARF2 is localized in the nucleus.  The three-dimensional structure prediction of the proteins showed that MdARF2 is highly similar with AtARF2, and they contain helices, folds, and random coils.  The promoter of MdARF2 contains cis-acting elements which respond to various stresses, as well as environmental and hormonal signals.  Expression analysis showed that MdARF2 is widely expressed in all tissues of apple, with the highest expression of MdARF2 in root.  Functional analysis with a series of MdARF2 transgenic apple calli indicated that MdARF2 can reduce the sensitivity to ABA signaling and enhance salt tolerance in apple.  In summary, the results of this research provide a new basis for studying the regulation of abiotic stresses by ARFs.

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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
2019, 18 (7): 1604-1612.   DOI: 10.1016/S2095-3119(19)62594-3
Abstract227)      PDF in ScienceDirect      
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.
 
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