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Jinhui Lin

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Open access Jul 2026

Mesophyll protoplasts: a key tool for deciphering plant nutrient starvation pathways

The mesophyll protoplast transient expression system is an essential and robust methodology for examining gene expression regulation. Despite its potential, it has not been effectively employed to elucidate the genetic regulatory pathways and networks underlying plant responses to nutrient starvation, such as those involving phosphorus (Pi) and iron (Fe). In this study, we identified differentially expressed genes in response to Pi starvation in Arabidopsis using transcriptome analysis and RT-qPCR validation. Our findings revealed that Pi starvation significantly upregulated the expression of Pi-starvation induced (PSI) genes, including SPX1, SPX3, IPS1, and PS2, while simultaneously downregulating the expression of Fe starvation-responsive genes, such as FIT, IRT1, and FRO2. Additionally, through a dual luciferase transient expression assay in mesophyll protoplasts, we demonstrated that the transcription factor PHR1 serves as a crucial transcriptional regulator, modulating the expression of phosphate starvation response (PSR) genes. This regulation significantly enhances the transcriptional activity of the SPX1, SPX2, SPX3, IPS1, PS2, and PHT1;4 promoters. Upon the addition of the SPX1 protein, the activation of these gene promoters by PHR1 were alleviated. Concurrently, the transcriptional regulator FIT, which governs the expression of genes responsive to Fe starvation, markedly increased the transcriptional activity of the IRT1 and FRO2 promoters. Based on these findings, we propose the mesophyll protoplast transient expression system as a rapid and reliable method for investigating complex genetic networks. Overall, our study provides substantial evidence for understanding the role of the mesophyll protoplast transient expression system in elucidating the genetic regulatory pathways and networks involved in plant responses to nutrient starvation.

Yashan Tian, Jian-Ju She, Jin-Hui Lin et al. · Frontiers in Plant Science · 0 citations

Open access Aug 2026

Transcription Factor TCP9 Enhances Arabidopsis Tolerance to Cadmium Toxicity via ZAT6 and ZAT10 Activation

Cadmium (Cd), a highly toxic and mobile heavy metal, has emerged as a severe environmental concern in global agroecosystems, posing a substantial threat to human health. Although prior studies have established that ZAT6 and ZAT10 positively regulate Arabidopsis tolerance to Cd toxicity, the underlying molecular mechanisms remain largely elusive. The present study provides evidence that a class I TCP transcription factor, TCP9, significantly enhances Arabidopsis tolerance to Cd toxicity through the direct activation of ZAT6 and ZAT10 expression. The real-time quantitative PCR (RT-qPCR) analysis indicates that the expression of TCP9 was induced under Cd toxicity. Meanwhile, the tcp9 mutant exhibited heightened sensitivity to Cd toxicity, accompanied by elevated Cd accumulation in both shoots and roots. Notably, the complemented lines exhibited phenotypic characteristics analogous to those observed in the wild-type (WT) plants. Further physiological and biochemical analyses revealed that, in comparison to WT, the tcp9 mutant displayed elevated hydrogen peroxide (H2O2) accumulation and reduced contents of catalase (CAT), ascorbate peroxidase (APX), and peroxidase (POD) under Cd toxicity. Furthermore, TCP9 directly interacted with the promoters of ZAT6 and ZAT10 in vitro, facilitating their transcription and consequently enhancing plant tolerance to Cd toxicity. Overall, our findings showed that TCP9 enhances Cd tolerance via modulating ZAT6 and ZAT10, thereby identifying TCP9 as a potential key target for improving plant tolerance to Cd toxicity.

Jianju She, Feng Chen, Jia-Yi Liu et al. · Plants · 0 citations