Kim Young-Cheon, Choi Eun Bin, Kim Gee Woo, Lee Jeong Hwan
Department of Life Sciences, Jeonbuk National University, 567 Baekje-daero, Deokjin-gu, Jeonju, 54896, Jeonbuk State, Korea.
HOPS Biosciences Company, Jeonbuk National University, 567 Baekje-daero, Deokjin-gu, Jeonju, 54896, Jeonbuk State, Korea.
Planta. 2025 Sep 6;262(4):97. doi: 10.1007/s00425-025-04809-8.
PHYTOCHROME INTERACTING FACTOR4 (PIF4) plays an important role in regulating plant thermomorphogenesis. In this study, two PIF4 homologous genes, BcPIF4-1 and BcPIF4-2 (Brassica rapa subsp. CHINENSIS PIF4-1 and PIF4-2), were investigated. Amino acid sequence comparison with Arabidopsis thaliana PIF4 (AtPIF4) showed that BcPIF4-1 and BcPIF4-2 had an active phytochrome B binding (APB) motif at the N-terminus and a basic helix-loop-helix (bHLH) domain at the C-terminus. Both BcPIF4-1 and BcPIF4-2 were highly expressed in the leaves of Pak choi and showed increasing expression patterns during vernalization. Subcellular localization and yeast two-hybrid analyses also showed that the three BcPIF4-1 and BcPIF4-2 proteins were localized in the nucleus and could interact with Pak choi and Arabidopsis TEOSINTE BRANCHED1/CYCLOIDEA/PROLIFERATING CELL FACTORS (BcTCPs and AtTCPs). Interestingly, transgenic approaches have shown that the overexpression of only BcPIF4-1 in Col-0 and pif4-101 plants results in early flowering phenotypes by upregulating the expression of Arabidopsis FLOWERING LOCUS T (AtFT). An important BcPIF4-1 motif that directly binds to the AtFT promoter is also suggested in this study. Furthermore, the overexpression of BcPIF4-1 in Col-0 and pif4-101 plants affected hypocotyl length by increasing the expression levels of Arabidopsis INDOLE-3-ACETIC ACID INDUCIBLE 29 (AtIAA29). Our results indicate that BcPIF4-1 isolated from Pak choi is a functional equivalent of AtPIF4 in terms of flowering time and hypocotyl elongation, suggesting that BcPIF4-1 is a candidate gene for developing high-temperature-insensitive Pak choi cultivars. MAIN CONCLUSION: Pak choi BcPIF4-1 controls flowering time and hypocotyl length by affecting AtFT and AtIAA29 expression, respectively in Arabidopsis.
光敏色素互作因子4(PIF4)在调节植物热形态建成中起重要作用。在本研究中,对两个PIF4同源基因BcPIF4-1和BcPIF4-2(小白菜PIF4-1和PIF4-2)进行了研究。与拟南芥PIF4(AtPIF4)的氨基酸序列比较表明,BcPIF4-1和BcPIF4-2在N端具有一个活性光敏色素B结合(APB)基序,在C端具有一个碱性螺旋-环-螺旋(bHLH)结构域。BcPIF4-1和BcPIF4-2在小白菜叶片中均高度表达,且在春化过程中呈现表达增加的模式。亚细胞定位和酵母双杂交分析还表明,BcPIF4-1和BcPIF4-2这三种蛋白定位于细胞核,并且可以与小白菜和拟南芥的TEOSINTE BRANCHED1/CYCLOIDEA/PROLIFERATING CELL FACTORS(BcTCPs和AtTCPs)相互作用。有趣的是,转基因方法表明,仅在Col-0和pif4-101植物中过表达BcPIF4-1会通过上调拟南芥开花位点T(AtFT)的表达导致早花表型。本研究还提出了一个直接与AtFT启动子结合的重要BcPIF4-1基序。此外,在Col-0和pif4-101植物中过表达BcPIF4-1通过增加拟南芥吲哚-3-乙酸诱导型29(AtIAA29)的表达水平影响下胚轴长度。我们的结果表明,从小白菜中分离出的BcPIF4-1在开花时间和下胚轴伸长方面与AtPIF4功能相当,这表明BcPIF4-1是培育对高温不敏感的小白菜品种的候选基因。主要结论:小白菜BcPIF4-1分别通过影响拟南芥中AtFT和AtIAA29的表达来控制开花时间和下胚轴长度。