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A putative E3 ubiquitin ligase substrate receptor degrades transcription factor SmNAC to enhance bacterial wilt resistance in eggplant
Saturday, 2024/11/23 | 07:19:54

Shuangshuang Yan, Yixi Wang, Bingwei Yu, Yuwei Gan, Jianjun Lei, Changming Chen, Zhangsheng Zhu, Zhengkun Qiu, Bihao Cao

Horticulture Research; Volume 11, Issue 1, January 2024, uhad246, https://doi.org/10.1093/hr/uhad246

Abstract

Bacterial wilt caused by Ralstonia solanacearum is a severe soil-borne disease globally, limiting the production in Solanaceae plants. SmNAC negatively regulated eggplant resistance to Bacterial wilt (BW) though restraining salicylic acid (SA) biosynthesis. However, other mechanisms through which SmNAC regulates BW resistance remain unknown. Here, we identified an interaction factor, SmDDA1b, encoding a substrate receptor for E3 ubiquitin ligase, from the eggplant cDNA library using SmNAC as bait. SmDDA1b expression was promoted by R. solanacearum inoculation and exogenous SA treatment. The virus-induced gene silencing of the SmDDA1b suppressed the BW resistance of eggplants; SmDDA1b overexpression enhanced the BW resistance of tomato plants. SmDDA1b positively regulates BW resistance by inhibiting the spread of R. solanacearum within plants. The SA content and the SA biosynthesis gene ICS1 and signaling pathway genes decreased in the SmDDA1b-silenced plants but increased in SmDDA1b-overexpression plants. Moreover, SmDDB1 protein showed interaction with SmCUL4 and SmDDA1b and protein degradation experiments indicated that SmDDA1b reduced SmNAC protein levels through proteasome degradation. Furthermore, SmNAC could directly bind the SmDDA1b promoter and repress its transcription. Thus, SmDDA1b is a novel regulator functioning in BW resistance of solanaceous crops via the SmNAC-mediated SA pathway. Those results also revealed a negative feedback loop between SmDDA1b and SmNAC controlling BW resistance.

 

See https://academic.oup.com/hr/article/11/1/uhad246/7452860?login=false

 

Figure 1. Interaction between SmDDA1b and SmNAC and the subcellular localization analysis of SmDDA1b. (A) Yeast two-hybrid (Y2H) assays of SmNAC and SmDDA1b. The co-transformed BD-53 and AD-T in the Y2H Gold strain were used as the positive control, while co-transformed BD-Lam and AD-T in the Y2H Gold strain were used as the negative control. SmNAC1–139 indicates the N-terminal 139 aa of SmNAC. (B) Bimolecular fluorescence complementation (BiFC) assays between SmDDA1b and SmNAC. YFP indicates the interaction between two proteins. NLS represents the nucleus location. (C) The subcellular localization analysis of SmDDA1b. GFP and NLS indicate the subcellular location of SmDDA1b in the nucleus. (D) Co-Immunoprecipitation (CoIP) analysis of SmDDA1b and SmNAC. Scale bar in (B-C) represents 50 μm.

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