Abstract
Wounding is a constant threat to plant survival throughout their lifespan; therefore, understanding the biological responses to wounds at the cellular level is important. The protoplast system is versatile for molecular biology, however, no wounding studies on this system have been reported. We established a new approach for wounding research using mechanically damaged Arabidopsis mesophyll protoplasts. Wounded protoplasts showed typical wounding responses, such as increased MPK6 kinase activity and upregulated JAZ1 expression. We also assessed expression profiles and protein stability of the basic helix-loop-helix transcription factor MYC2 in wounded protoplasts. Promoter activity, gene expression, and protein stability of MYC2 were compromised, but recovered in the early stage of wounding. In the late stage, the promoter activity and expression of MYC2 were increased, but the protein stability was not changed. According to the results of the present study, this new cell-based approach will be of use in various molecular studies on plant wounding.
Highlights
Wounding is defined as mechanical damage that occurs frequently in plants due to biotic and abiotic stresses [1]
The expression of numerous genes associated with phytohormones, oxidative stress, dehydration stress, and heat-shock proteins is rapidly upregulated during wounding [6,7,8], and protein turnover, transport processes, metabolism modulation, and gene expression reprogramming occur [9]
In Arabidopsis, the basic helix-loop-helix leu zipper transcription factor MYC2 is a major regulator of the JA signaling pathway and response [11]
Summary
Wounding is defined as mechanical damage that occurs frequently in plants due to biotic and abiotic stresses [1]. Wounding generates damage-associated molecular patterns and activates MPK6 [3,4,5]. The expression of numerous genes associated with phytohormones, oxidative stress, dehydration stress, and heat-shock proteins is rapidly upregulated during wounding [6,7,8], and protein turnover, transport processes, metabolism modulation, and gene expression reprogramming occur [9]. Jasmonate (JA) is a major immune phytohormone that accumulates after wounding [10]. In Arabidopsis, the basic helix-loop-helix leu zipper transcription factor MYC2 is a major regulator of the JA signaling pathway and response [11]. MYC2 is involved in various phytohormone crosstalk and several signaling pathways [12,13,14]
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