Introduction: Recombinant single‐chain variable fragments (scFvs) are promising antibody formats for cost‐effective and scalable production. However, their soluble expression in Escherichia coli is often limited by misfolding and aggregation, particularly for scFvs targeting small molecule haptens such as abscisic acid (ABA). To address this bottleneck, molecular chaperones can be co‐expressed to enhance folding efficiency and functional yield.Methods: An ABA‐specific scFv was expressed in E. coli BL21(DE3) using five different chaperone plasmids (pG‐KJE8, pGro7, pKJE7, pG-Tf2, and pTf16). Soluble expression was quantified by His‐tag ELISA, and protein identity was confirmed by SDS‐PAGE and Western blot. Functional characterization included competitive ELISA for IC₅₀ and specificity, while secondary structure was analyzed by FT‐IR and circular dichroism spectroscopy.Results: Trigger Factor (pTf16) significantly improved soluble scFv yield (19.65%) compared to the control (14.20%). The pKJE7 system achieved the highest sensitivity with the lowest IC₅₀, whereas the pTf16 system provided superior specificity and a broader detection range by minimizing cross‐reactivity. Structural analysis revealed that pKJE7‐assisted scFv closely matched the predicted β‐sheet content, correlating with high sensitivity, while pTf16‐assisted scFv avoided non-native α-helices, supporting enhanced specificity. Circular dichroism further demonstrated that pKJE7‐ and pTf16‐assisted scFvs exhibited conformational rigidity consistent with a lock-and-key binding mechanism.Discussion: This study highlights that molecular chaperone choice influences both structural fidelity and functional performance of ABA‐scFv in E. coli. While pKJE7 favors high sensitivity, pTf16 yields highly specific and structurally stable antibodies. These findings establish a practical basis for developing low‐cost ABA immunoassays with tailored performance for agricultural biotechnology.

Sugarcane tillering is a key determinant of crop productivity, yet the integrated roles of rhizosphere microbiome dynamics, nutrient status, and hormone signaling in regulating tiller development remain poorly understood. Here, we compared rhizosphere microbial communities, endogenous hormone profiles, and nutrient element concentrations in sugarcane cultivars with contrasting tillering capacities. High-tillering varieties exhibited significantly greater microbial diversity and more complex co-occurrence network structures in the rhizosphere, characterized by enrichment of Acidobacteriota, Chloroflexi, and Planctomycetes and functional pathways related to nitrogen fixation, phosphorus solubilization, and auxin biosynthesis. In contrast, low-tillering varieties harbored simplified, stress-adapted microbial consortia and prioritized pathways linked to oxidative stress response and heavy metal detoxification. Hormonal analysis revealed that high-tillering cultivars maintained higher levels of growth-promoting hormones—particularly auxin (IAA) and active cytokinins—in tiller buds while low-tillering cultivars accumulated elevated abscisic acid (ABA) and inactive cytokinin conjugates. Nutrient analysis indicated that high-tillering genotypes possessed higher nitrogen and phosphorus contents, supporting vigorous axillary bud activation and shoot proliferation, whereas low-tillering varieties accumulated more zinc and manganese, potentially reflecting stress adaptation. Network-level integration of microbial, hormonal, and nutrient profiles underscored genotype-specific feedback between rhizosphere microbiota and plant physiological states, highlighting modular associations that link microbial hubs with tissue-specific nutrient and hormone signatures. Our findings reveal a systems-level mechanism by which rhizosphere microbial community structure and function interact with plant-nutrient–hormonal status to regulate tillering in sugarcane. These insights provide a basis for microbiome-informed strategies to enhance sugarcane productivity through integrated nutrient–hormonal–microbe management.

Introduction: Sugarcane tillering represents a complex developmental process shaped by multiple interacting molecular networks. To outline a multi-omics regulatory framework linking hormonal, ionomic, and transcriptomic variation to contrasting tillering capacities in sugarcane, we analyzed contrasting genotypes using an integrated multi-omics approach. Methods: Four field-grown sugarcane genotypes at the peak tillering stage were analyzed using integrated transcriptomics, targeted phytohormone metabolomics, and ionomics, with three biological replicates per genotype × tissue combination. Results: High-tillering plants maintained a transcriptional and metabolic state optimized for growth, including enhanced expression of photosynthetic and carbon metabolic pathways, marked accumulation of the auxin-associated metabolite tryptamine (Log2FC = 4.97), elevated auxin- and cytokinin-associated metabolites, and preferential enrichment of micronutrients in tiller tissues. In contrast, low-tillering genotypes adopted a stress-prepared phenotype characterized by activation of defense and protein synthesis pathways, reduced 1-aminocyclopropane-1-carboxylic acid (ACC) levels in high-tillering plants relative to low-tillering plants (Log2FC = −2.26), accumulation of jasmonate-, salicylic acid-, and abscisic acid (ABA)-associated metabolites, and distinct ionomic signatures associated with nutrient imbalance. Weighted Gene Co-expression Network Analysis (WGCNA) further revealed a polarized regulatory structure, with the growth-associated module positively correlated with cis-Zeatin riboside (cZR) (yellow-labelled, r = 0.71), whereas the defense-associated module showed strong negative correlations with cZR/cis-Zeatin (cZ) (brown-labelled, r = −0.74/−0.75), and another defense-associated module showed positive correlations with ACC and Salicylic acid (SA) (turquoise-labelled, r = 0.77 and 0.79, respectively) . Discussion: This study provides an integrated multi-omics framework for understanding sugarcane tillering and highlights a coordinated growth–defense trade-off associated with contrasting tillering strategies.

Averrhoa carambola L. (family Oxalidaceae), known as star fruit, is a berry species found in tropical and subtropical regions, recognized for its unique crisp texture, high juiciness, and refreshing, slightly tangy flavor profile. A. carambola is known for its relatively high total oxalate content (fresh weight: 200–1840 mg/100 g), and excessive consumption is linked to toxicity, particularly in individuals with chronic kidney disease or other renal disorders. Therefore, understanding the mechanisms behind oxalate accumulation is crucial for developing effective strategies to manage oxalate levels in A. carambola . To our knowledge, this represents the first comprehensive transcriptomic investigation conducted across distinct fruit types of A. carambola . Results demonstrated that the downregulation of genes associated with oxalate oxidation contributes to the elevated oxalate content observed in sour-type A. carambola fruits. In addition, L-ascorbate concentration exhibits a positive correlation with oxalate levels across different varieties. These findings establish a foundational work for formulating rational and targeted strategies to regulate oxalate content in A. carambola , encompassing genetic improvement, optimized harvest timing, and carefully designed post-harvest management practices.