Passion fruit (Passiflora edulis) is an important fruit crop with high economic value. Genetic engineering plays an important role in crop improvement with desired traits and gene functional studies. The lack of a simple, efficient, and stable transformation system for passion fruit has greatly limited gene functional studies. In this study, a simple and efficient Agrobacterium-mediated in planta transformation system for passion fruit was established, using Agrobacterium virulent strain EHA105 harboring the binary vectors pCAMBIA1301 and pCAMBIA1302 with GUS and GFP reporter genes. The system requires less time and labor costs than conventional transformation systems, and no additional phytohormones and sterile conditions are required. Regeneration efficiency of 86% and transformation efficiency of 29% were achieved, when the wounds were wrapped with Parafilm and the plants were kept in darkness for 15 days. Approximately 75% of the regenerated plants had a single shoot and 26% multiple shoots. The transformation was confirmed at the DNA and RNA levels as well as by GUS staining and GFP fluorescent measurements. The developed protocol will contribute to the genetic improvement of passion fruit breeding.

The DUF668 gene performs a critical role in mitigating the impact of abiotic stress factors. In this study, we identified 30 DUF668 genes in a soybean genome, distributed across fifteen chromosomes. The phylogenetic analysis classified the DUF668 genes into three groups (group I, group II, and group III). Interestingly, gene structure analysis illustrated that several GmDUF668 genes were without introns. Furthermore, the subcellular localization results suggested that GmDUF668 proteins were present in the nucleus, mitochondria, cytoplasm, and plasma membrane. GmDUF668 promoters were analyzed in silico to gain insight into the presence of regulatory sequences for TFs binding. The expression profiling illustrated that GmDUF668 genes showed expression in leaves, roots, nodules, and flowers. To investigate their response to salt stress, we utilized the RNA sequencing data of GmDUF668 genes. The results unveiled that GmDUF668-8, GmDUF668-20, and GmDUF668-30 genes were upregulated against salt stress treatment. We further validated these findings using qRT-PCR analysis. These findings provide a scientific basis to explore the functions of GmDUF668 genes against different stress conditions.

Peanut (Arachis hypogaea L.) is an important food and feed crop worldwide and is affected by various biotic and abiotic stresses. The cellular ATP levels decrease significantly during stress as ATP molecules move to extracellular spaces, resulting in increased ROS production and cell apoptosis. Apyrases (APYs) are the nucleoside phosphatase (NPTs) superfamily members and play an important role in regulating cellular ATP levels under stress. We identified 17 APY homologs in A. hypogaea (AhAPYs), and their phylogenetic relationships, conserved motifs, putative miRNAs targeting different AhAPYs, cis-regulatory elements, etc., were studied in detail. The transcriptome expression data were used to observe the expression patterns in different tissues and under stress conditions. We found that the AhAPY2-1 gene showed abundant expression in the pericarp. As the pericarp is a key defense organ against environmental stress and promoters are the key elements regulating gene expression, we functionally characterized the AhAPY2-1 promoter for its possible use in future breeding programs. The functional characterization of AhAPY2-1P in transgenic Arabidopsis plants showed that it effectively regulated GUS gene expression in the pericarp. GUS expression was also detected in flowers of transgenic Arabidopsis plants. Overall, these results strongly suggest that APYs are an important future research subject for peanut and other crops, and AhPAY2-1P can be used to drive the resistance-related genes in a pericarp-specific manner to enhance the defensive abilities of the pericarp.

Subtilases (SBTs), known as serine proteases or phytoproteases in plants, are crucial enzymes involved in plant development, growth, and signaling pathways. Despite their recognized importance in other plant species, information regarding their functional roles in cultivated peanut (Arachis hypogea L.) remains sparse. We identified 122 AhSBT genes in the STQ peanut genome, classifying them into six subgroups based on phylogenetic analysis. Detailed structural and motif analyses revealed the presence of conserved domains, highlighting the evolutionary conservation of AhSBTs. The collinearity results indicate that the A. hypogea SBT gene family has 17, 5, and 1 homologous gene pairs with Glycine max, Arabidopsis thaliana, and Zea mays, respectively. Furthermore, the prediction of cis-elements in promoters indicates that they are mainly associated with hormones and abiotic stress. GO and KEGG analyses showed that many AhSBTs are important in stress response. Based on transcriptome datasets, some genes, such as AhSBT2, AhSBT18, AhSBT19, AhSBT60, AhSBT102, AhSBT5, AhSBT111, and AhSBT113, showed remarkably higher expression in diverse tissues/organs, i.e., embryo, root, and leaf, potentially implicating them in seed development. Likewise, only a few genes, including AhSBT1, AhSBT39, AhSBT53, AhSBT92, and AhSBT115, were upregulated under abiotic stress (drought and cold) and phytohormone (ethylene, abscisic acid, paclobutrazol, brassinolide, and salicylic acid) treatments. Upon inoculation with Ralstonia solanacearum, the expression levels of AhSBT39, AhSBT50, AhSBT92, and AhSBT115 were upregulated in disease-resistant and downregulated in disease-susceptible varieties. qRT-PCR-based expression profiling presented the parallel expression trends as generated from transcriptome datasets. The comprehensive dataset generated in the study provides valuable insights into understanding the functional roles of AhSBTs, paving the way for potential applications in crop improvement. These findings deepen our understanding of peanut molecular biology and offer new strategies for enhancing stress tolerance and other agronomically important traits.

Tomato (Solanum lycopersicum) production is increasingly threatened by Tuta absoluta, a destructive pest that compromises yield and quality. To explore sustainable alternatives to conventional insecticides, we investigated the jasmonate-mediated defense pathway by performing a genome-wide characterization of the JAZ gene family in S. lycopersicum. A total of 39 SlJAZ genes were identified and mapped to 12 chromosomes. Detailed analysis revealed conserved motifs, diverse exon–intron structures, four major phylogenetic groups, and the presence of multiple MeJA- and stress-responsive cis-elements. Synteny analysis indicated gene duplication events and evolutionary conservation with Arabidopsis and potato. Small RNA predictions suggested that 33 SlJAZ genes are targeted by 69 microRNAs, implying multilayered regulation. Transcriptome analysis under four treatment conditions—mesoporous silica nanoparticles (MSNs) ± pest infestation—revealed 21 differentially expressed SlJAZ genes. SlJAZ1, SlJAZ19, SlJAZ20, and SlJAZ22 were notably upregulated under the combined MSN and pest treatment, with expression patterns validated by qRT-PCR (R² = 0.92). Phenotypic assessment of leaf damage index, larval survival rate, and number of leaf mines showed reduced pest activity in MSN-treated plants. Regression analysis demonstrated significant negative correlations between expression levels of SlJAZ20, SlJAZ26, and SlJAZ29 and pest-related damage traits. These findings indicate that MSNs function as effective elicitors of JA-responsive defense in tomato and modulate the expression of specific JAZ genes linked to enhanced resistance. The study provides a valuable foundation for integrating nanotechnology with molecular defense strategies to promote sustainable pest management.