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Updated: Jun 7, 2025

Measuring Gene Expression in Bombarded Barley Aleurone Layers with Increased Throughput
Published on: March 30, 2018
TRX h2-PP2AC2 module serves as a convergence node for aluminum stress and leaf senescence signals, regulating cell
Xia Li1,2, Guijun Su1,2, Chunliu Pan2
1Guangxi Key Laboratory for Agro-Environment and Agro-Product Safety, Nanning, 530004, People's Republic of China.
Abstract:
ROS/redox signaling plays an important role in the regulation of signal transduction and acclimation pathways activated by multiple abiotic stresses and leaf senescence. However, the regulatory events that produce ROS under different stimuli are far from clear. Here, we report the elucidation of the molecular mechanism of an h type thioredoxin, AhTRX h2, positively regulates Al sensitivity and leaf senescence by promoting ROS. AhTRX h2 transcript levels increased greatly during both natural senescence and Al stress condition in peanut. Ectopic expression of AhTRX h2 in Arabidopsis conferred Al sensitivity as well as premature leaf senescence, manifested by multiple indices, including inhibiting root elongation, severe cell death, and accelerated expression of MC1 and CEX17. AhTRX h2 exhibited similar functions to AtTRX h2, as AhTRX h2 was able to restore the phenotypes of the AtTRX h2 defective mutant (trxh2-4) which showed Al tolerant and late senescence phenotypes. The knock down of AhTRX h2 markedly suppressed Al- and senescence-induced cell death in peanut. AhTRX h2 could recruit catalytic subunit of protein phosphatase 2A (PP2AC2) to form a stable complex. The interaction between AhTRX h2 and AtPP2AC2, as well as AhPP2AC2 and AtTRX h2 was also proved. Overexpression of AhPP2AC2 significantly enhanced Al sensitivity and leaf senescence in Arabidopsis. Protein stability assay revealed that AhTRX h2 was more stable during aging or aluminum stress. Moreover, PP2AC2 could greatly enhance the stability of AhTRX h2 in vivo. Consistent with these observations, overexpression of AhPP2AC2 effectively enhanced AhTRX h2-induced Al sensitivity and precocious leaf senescence. AhTRX h2 and AhPP2AC2 required ABA and ROS in response to cell death under Al stress and senescence, and it was evidence to suggest that ABA acted upstream of ROS in this process. Together, AhTRX h2 and AhPP2AC2 constitute a stable complex that promotes the accumulation of ABA and ROS, effectively regulate cell death. These findings suggest that TRX h2-PP2AC2-mediated pathway may be a widespread mechanism in regulating Al stress and leaf senescence.
Insights
Thioredoxin h2 (TRX h2) in peanut promotes aluminum (Al) sensitivity and leaf senescence by increasing reactive oxygen species (ROS). This TRX h2 protein interacts with protein phosphatase 2A (PP2AC2), forming a complex that enhances Al stress and senescence responses.
Area of Science:
- Plant Biology
- Molecular Biology
- Biochemistry
Background:
- Reactive oxygen species (ROS) are crucial for plant acclimation to abiotic stresses and leaf senescence.
- The precise mechanisms generating ROS under various stimuli remain unclear.
Purpose of the Study:
- To elucidate the molecular mechanism by which peanut thioredoxin h2 (AhTRX h2) regulates aluminum (Al) sensitivity and leaf senescence.
- To investigate the role of AhTRX h2 in promoting ROS accumulation.
Main Methods:
- Investigated AhTRX h2 transcript levels under natural senescence and Al stress.
- Performed ectopic expression of AhTRX h2 in Arabidopsis and analyzed phenotypes.
- Utilized knock-down of AhTRX h2 in peanut.
- Examined protein-protein interactions between AhTRX h2 and PP2AC2.
- Assessed protein stability and the effect of PP2AC2 on AhTRX h2 stability.
Main Results:
- AhTRX h2 expression increased during senescence and Al stress in peanut.
- Ectopic expression of AhTRX h2 in Arabidopsis induced Al sensitivity and premature leaf senescence.
- AhTRX h2 restored phenotypes of the Arabidopsis trxh2-4 mutant.
- Knock-down of AhTRX h2 reduced Al- and senescence-induced cell death.
- AhTRX h2 forms a stable complex with PP2AC2, enhancing its stability and promoting Al sensitivity and senescence.
- The TRX h2-PP2AC2 complex promotes ABA and ROS accumulation, regulating cell death.
Conclusions:
- AhTRX h2 positively regulates Al sensitivity and leaf senescence by promoting ROS accumulation.
- The AhTRX h2-PP2AC2 complex is a key mediator of Al stress and senescence.
- This pathway involving ABA and ROS is critical for regulating cell death under stress conditions.
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