N4-acetylcytidine (ac4C) modification is the only known RNA acetylation modification identified in eukaryotes, which plays critical roles in the regulation of plant development and the responses to biotic and abiotic stresses. In this study, 15 members of the RNA N-acetyltransferase (NAT) gene family were identified across nine plant species, including Arabidopsis thaliana, Zea mays, Oryza sativa, Brachypodium distachyon, Solanum lycopersicum, Solanum tuberosum, Sorghum bicolor, Glycine max, and Brassica napus. Physicochemical property analysis revealed that most NAT proteins were hydrophilic proteins with highly conserved amino acid sequence lengths. Phylogenetic analysis, conserved motif and gene structure analysis, conserved protein domain analysis, and synteny analysis demonstrated that the NAT gene family exhibited strong homology and conservation in both evolutionary relationship and functional architecture. Prediction of cis-acting elements indicated that NAT genes may function critically in regulating plant growth, development and in response to abiotic stresses. Tissue expression profiling of Brassica napus NAT genes showed that BnaNAT1, BnaNAT2, BnaNAT3 and BnaNAT4 were highly expressed in all detected tissues. Expression pattern analysis under drought and salt stresses showed that BnaNAT1, BnaNAT2, BnaNAT3 and BnaNAT4 responded to drought and salt stresses to varying degrees, suggesting their essential roles in plant stress response. Collectively, the findings of this study lay a solid foundation for the in-depth elucidation of the biological functions of NAT genes and for the identifying of stress-tolerance genes and the improvement of stress tolerance traits in Brassica napus.
Virus-induced gene silencing (VIGS) is a common method using modified viral vectors to decipher the functions of plant genes. Therefore, developing an effective transformation method for peanut plants is of interest. Here, two pTRV-VIGS-based systems were developed for peanut: leaf–injected agroinoculation (LIA) and seed-soaked agroinoculation (SSA). With the LIA-VIGS method, stable gene silencing was achieved in approximately 32–34 days with an average efficiency of 42.31–92.61%. In contrast, with the SSA-VIGS method performed by scratching naked peanut seeds and soaking them in bacterial solution, approximately 23–25 days were required to induce gene silencing with an efficiency of 49.93–95.40%. Overall, under the conditions tested, the SSA-VIGS strategy features shorter experimental cycles, easier operation, and faster phenotypic observation, allowing earlier evaluation of gene function during seedling establishment.
