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. 2019 Dec;17(12):2231-2233.
doi: 10.1111/pbi.13196. Epub 2019 Jul 10.

MdBBX22 regulates UV-B-induced anthocyanin biosynthesis through regulating the function of MdHY5 and is targeted by MdBT2 for 26S proteasome-mediated degradation

Jian-Ping An  1 , Xiao-Fei Wang  1 , Xiao-Wei Zhang  1 , Si-Qi Bi  1 , Chun-Xiang You  1 , Yu-Jin Hao  1
Affiliations

MdBBX22 regulates UV-B-induced anthocyanin biosynthesis through regulating the function of MdHY5 and is targeted by MdBT2 for 26S proteasome-mediated degradation

Jian-Ping An et al. Plant Biotechnol J. 2019 Dec.
No abstract available

Keywords: B-box; BT2; HY5; UV-B; anthocyanin biosynthesis; ubiquitination.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
MdBBX22 positively regulates UV‐B‐induced anthocyanin biosynthesis by enhancing the binding of MdHY5 to its target genes, and MdBT2 decreases MdBBX22‐promoted anthocyanin biosynthesis by degrading the MdBBX22 protein. (a) Phenotypes and (b) anthocyanin levels of apple calli in the dark (Dark) or in the light but with (+UV‐B) or without (‐UV‐B) UV‐B treatment. WT: wild‐type; MdHY5‐Anti: MdHY5 antisense suppression. ‐UV‐B: white fluorescent light; +UV‐B: white fluorescent light plus UV‐B. Apple calli of 15‐day‐old were treated with dark, ‐UV‐B, and +UV‐B for 10 days. Anthocyanin content of ‐UV‐B‐treated wild‐type apple calli was used as the reference and set to 1. Different letters above the bars indicated significant difference (P < 0.05) as obtained by one‐way ANOVA and LSD test. Asterisks denoted t‐test significance: *P < 0.05, **P < 0.01. (c) Detection of MdBBX22 transcription in response to UV‐B treatment using qRT‐PCR. ‘Fuji’ apple fruits were moved from white fluorescent light (‐UV‐B) to white fluorescent light plus UV‐B (+UV‐B). (d) Detection of the MdBBX22‐HIS fusion protein after UV‐B and MG132 treatments. ‘Fuji’ apple fruits were treated with white fluorescent light (‐UV‐B, Control) and white fluorescent light plus UV‐B (+UV‐B, UV‐B) for 5 days. Total proteins extracted from apple peel were incubated with the purified MdBBX22‐HIS protein. For MG132 treatment, total proteins of ‐UV‐B‐treated apple fruits were pre‐treated using 100 µM MG132 for 0.5 h. (e) Phenotypes and (f) anthocyanin levels of apple calli with (+UV‐B) or without (‐UV‐B) UV‐B treatments. WT: wild‐type; MdBBX22‐OX: MdBBX22‐overexpression; MdBBX22‐Anti: MdBBX22 antisense suppression. Apple calli of 15‐day‐old were treated with ‐UV‐B and +UV‐B for 3 days. (g) MdHY5 interacts with MdBBX22. MdHY5 (MDP0000586302) and MdBBX22 (MDP0000298804). (h) Phenotypes and (i) anthocyanin levels of apple calli with (+UV‐B) or without (‐UV‐B) UV‐B treatments. WT: wild‐type; MdBBX22‐OX: MdBBX22‐overexpression; MdHY5‐Anti: MdHY5 antisense suppression; MdBBX22‐OX/MdHY5‐Anti: antisense suppression of MdHY5 in the background of MdBBX22‐overexpression. Apple calli of 15‐day‐old were treated with ‐UV‐B and +UV‐B for 3 days. (j) and (k) EMSA showing that MdBBX22 enhances the transcriptional activity of MdHY5 on MdMYB10 and MdCHS. (l) MdHY5 interacts with MdBBX proteins. MdBBX10 (MDP0000733075), MdBBX20 (MDP0000177126), MdBBX22 (MDP0000298804), MdBBX23 (MDP0000222881), MdBBX24 (MDP0000800387), MdBBX25 (MDP0000901915), MdBBX33 (MDP0000697407), MdBBX37 (MDP0000157816), MdBBX43 (MDP0000140484), and MdBBX48 (MDP0000759984). (m) MdBBX22 interacts with MdBBX37 and MdBBX48 proteins. (n) MdBBX22 interacts with MdBT2. MdBT2 (MDP0000151000). (o) Expression of MdBT2 in response to UV‐B treatment using qRT‐PCR. (p) Detection of the MdBT2‐GST fusion protein after UV‐B and MG132 treatments. (q) Phenotypes and (r) anthocyanin contents of wild‐type (WT) and MdBT2 transgenic apple calli in response to UV‐B treatments. WT: wild‐type; MdBT2‐OX: MdBT2‐overexpression; MdBT2‐Anti: MdBT2 antisense suppressing apple calli. Apple calli of 15‐day‐old were treated with ‐UV‐B and +UV‐B for 4 days. (s) Phenotypes and (t) anthocyanin contents of wild‐type (WT) and transgenic apple calli. WT: wild‐type; MdBBX22‐OX: MdBBX22‐overexpression; MdBBX22‐OX/MdBT2‐OX: overexpression of MdBT2 in the background of MdBBX22‐overexpression; MdBBX22‐OX/MdBT2‐Anti: antisense suppression of MdBT2 in the background of MdBBX22‐overexpression. Apple calli of 15‐day‐old were treated with ‐UV‐B for 4 days. (u) MdBT2 promoted the degradation of the MdBBX22 protein in vitro. (v) MdBT2 promotes the ubiquitination of the MdBBX22 protein in vivo. (w) MdBT2 interacts with MdBBX proteins. (x) A working model of MdBBX22 functioning in UV‐B‐induced anthocyanin biosynthesis. The blue line represents post‐translational regulation; the red line represents transcriptional regulation, and the black line represents transcriptional and post‐translational regulations.
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References

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