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Agronomy
Volume 15
Issue 4
10.3390/agronomy15040959
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Article

Identification of PIF Gene Family and Functional Study of PbPIF3a/PbPIF4 in Anthocyanin Biosynthesis of Pear

by
Haiyan Wang
,
Diya Lei
,
Xuan Zhou
,
Shangyun Li
,
Yunting Zhang
,
Yuanxiu Lin
,
Qing Chen
,
Ya Luo
,
Haoru Tang
* and
Yong Zhang
*
College of Horticulture, Sichuan Agricultural University, Chengdu 611130, China
*
Authors to whom correspondence should be addressed.
Agronomy 2025, 15(4), 959; https://doi.org/10.3390/agronomy15040959
Submission received: 24 March 2025 / Revised: 10 April 2025 / Accepted: 11 April 2025 / Published: 15 April 2025
(This article belongs to the Special Issue Smart Strategies and Technologies for Sustainability and Biodiversity in Herbaceous and Horticultural Crops—2nd Edition)
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Abstract

Anthocyanins are key metabolites that determine red pigmentation in pear skin (Pyrus spp.) and their biosynthesis is controlled by multiple transcription factors. Although phytochrome-interacting factors (PIFs) of the bHLH family have been shown to regulate anthocyanin biosynthesis in Arabidopsis thaliana, their genome-wide identification and regulatory mechanisms in pear (Pyrus spp.) anthocyanin synthesis remain unclear. Here, we characterized PIFs family in pear, identifying eight PbPIF proteins. Promoter cis-elements and expression patterns analysis suggested that PbPIF3a and PbPIF4 might be involved in anthocyanin biosynthesis. Subcellular localization confirmed nuclear enrichment of PbPIF3a and PbPIF4. Functional studies demonstrated that overexpression of PbPIF3a and PbPIF4 significantly suppressed anthocyanin accumulation in fruit skins, downregulating key biosynthetic genes such as PbDFR and PbUFGT. In contrast, the silencing of related genes led to an enhancement of anthocyanin accumulation. Dual-luciferase reporter assays and yeast one-hybrid assays confirmed that PbPIF3a directly bound to the promoters of PbDFR and PbUFGT and repressed their transcriptional activation, while PbPIF4 specifically inhibited the activity of the PbDFR promoter. Taken together, we demonstrated that PbPIF3a and PbPIF4 negatively regulated pear fruit coloration by directly repressing the transcriptional activity of key anthocyanin biosynthesis genes, providing novel insights into PIF-mediated regulation of anthocyanin biosynthesis.
Keywords: pear; PIFs; anthocyanins; PbPIF3a; PbPIF4 pear; PIFs; anthocyanins; PbPIF3a; PbPIF4

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MDPI and ACS Style

Wang, H.; Lei, D.; Zhou, X.; Li, S.; Zhang, Y.; Lin, Y.; Chen, Q.; Luo, Y.; Tang, H.; Zhang, Y. Identification of PIF Gene Family and Functional Study of PbPIF3a/PbPIF4 in Anthocyanin Biosynthesis of Pear. Agronomy 2025, 15, 959. https://doi.org/10.3390/agronomy15040959

AMA Style

Wang H, Lei D, Zhou X, Li S, Zhang Y, Lin Y, Chen Q, Luo Y, Tang H, Zhang Y. Identification of PIF Gene Family and Functional Study of PbPIF3a/PbPIF4 in Anthocyanin Biosynthesis of Pear. Agronomy. 2025; 15(4):959. https://doi.org/10.3390/agronomy15040959

Chicago/Turabian Style

Wang, Haiyan, Diya Lei, Xuan Zhou, Shangyun Li, Yunting Zhang, Yuanxiu Lin, Qing Chen, Ya Luo, Haoru Tang, and Yong Zhang. 2025. "Identification of PIF Gene Family and Functional Study of PbPIF3a/PbPIF4 in Anthocyanin Biosynthesis of Pear" Agronomy 15, no. 4: 959. https://doi.org/10.3390/agronomy15040959

APA Style

Wang, H., Lei, D., Zhou, X., Li, S., Zhang, Y., Lin, Y., Chen, Q., Luo, Y., Tang, H., & Zhang, Y. (2025). Identification of PIF Gene Family and Functional Study of PbPIF3a/PbPIF4 in Anthocyanin Biosynthesis of Pear. Agronomy, 15(4), 959. https://doi.org/10.3390/agronomy15040959

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