Seed priming with graphene oxide improves salinity tolerance and increases productivity of peanut through modulating multiple physiological processes.

Graphene oxide (GO), beyond its specialized industrial applications, is rapidly gaining prominence as a nanomaterial for modern agriculture. However, its specific effects on seed priming for salinity tolerance and yield formation in crops remain elusive. Under both pot-grown and field-grown conditions, this study combined physiological indices with transcriptomics and metabolomics to investigate how GO affects seed germination, seedling salinity tolerance, and peanut pod yield. Peanut seeds were firstly treated with 400 mg L⁻¹ GO (termed GO priming). At seed germination stage, GO-primed seeds exhibited higher germination rate and percentage of seeds with radicals breaking through the testa. Meanwhile, omics analyses revealed significant enrichment in pathways associated with carbon and nitrogen metabolisms in GO-primed seeds. At seedling stage, GO priming contributed to strengthening plant growth, enhancing photosynthesis, maintaining the integrity of plasma membrane, and promoting the nutrient accumulation in peanut seedlings under 200 mM NaCl stress. Moreover, GO priming increased the activities of antioxidant enzymes, along with reduced the accumulation of reactive oxygen species (ROS) in response to salinity stress. Furthermore, the differentially expressed genes (DEGs) and differentially accumulated metabolites (DAMs) of peanut seedlings under GO priming were mainly related to photosynthesis, phytohormones, antioxidant system, and carbon and nitrogen metabolisms in response to soil salinity. At maturity, GO priming showed an average increase in peanut pod yield by 12.91% compared with non-primed control. Collectively, our findings demonstrated that GO plays distinguish roles in enhancing seed germination, mitigating salinity stress, and boosting pod yield in peanut plants via modulating multiple physiological processes.

Effect of peanut butter supplementation on physical and cognitive functions in community-dwelling older adults: study protocol for a 6-month randomised controlled trial.

INTRODUCTION: Ageing is associated with physical and cognitive declines, which may be further exacerbated by poor nutrition. Nuts are energy and nutrient dense, and their consumption is associated with better physical and cognitive functions in older adults, but data from interventional studies are limited. This 6-month randomised controlled trial is designed to investigate the effects of consuming 43 g/day of peanut butter (equivalent to 1.5 servings of nuts) on physical function, including walking speed (primary outcome), standing and dynamic balance, upper and lower body strength, lower body power and endurance, and associated factors including muscle mass, cognitive function and DNA telomere length in community-dwelling older adults. METHOD AND ANALYSIS: A total of 120 participants aged ≥65 years will be recruited and randomly allocated (1:1 ratio) to either the intervention group (n=60) that will receive individually packaged sealed containers containing 43 g of peanut butter to be consumed once daily for 6 months alongside habitual diet, or the control group (n=60) that will maintain their habitual diet. Primary and secondary outcomes will be assessed at baseline and at 6 months. The primary outcome is walking speed assessed using the 4 m usual gait speed test. Secondary outcomes include other physical function assessments: standing balance, chair stand time, timed-up-and-go test and four-square step test; and hand grip and knee extensor muscle strength; cognitive function assessed using the Montreal Cognitive Assessment and trail making tests; body composition; nutritional status; and DNA telomere length from participants' buccal cell samples. Linear mixed models will be used to compare changes in outcomes between intervention and control groups. ETHICS AND DISSEMINATION: The study protocol is approved by the Deakin University Human Research Ethics Committee. The trial is registered with the Australian New Zealand Clinical Trials Registry (ANZCTR): ACTRN12622001291774. The results will be disseminated through peer-reviewed journals, conference presentations and media. TRIAL REGISTRATION NUMBER: ANZCTR12622001291774.

Sustainable management of peanut damping-off and root rot diseases caused by Rhizoctonia solani using environmentally friendly bio-formulations prepared from batch fermentation broth of chitinase-producing Streptomyces cellulosae.

BACKGROUND: Soil-borne plant diseases represent a severe problem that negatively impacts the production of food crops. Actinobacteria play a vital role in biocontrolling soil-borne fungi. AIM AND OBJECTIVES: The target of the present study is to test the antagonistic activity of chitinase-producing Streptomyces cellulosae Actino 48 (accession number, MT573878) against Rhizoctonia solani. Subsequently, maximization of Actino 48 production using different fermentation processes in a stirred tank bioreactor. Finally, preparation of bio-friendly formulations prepared from the culture broth of Actino 48 using talc powder (TP) and bentonite in a natural as well as nano forms as carriers. Meanwhile, investigating their activities in reducing the damping-off and root rot diseases of peanut plants, infected by R. solani under greenhouse conditions. RESULTS: Actino 48 was found to be the most significant antagonistic isolate strain at p ≤ 0.05 and showed the highest inhibition percentage of fungal mycelium growth, which reached 97%. The results of scanning electron microscope (SEM) images analysis showed a large reduction in R. solani mycelia mass. Additionally, many aberrations changes and fungal hypha damages were found. Batch fermentation No. 2, which was performed using agitation speed of 200 rpm, achieved high chitinase activity of 0.1163 U mL- 1 min- 1 with a yield coefficient of 0.004 U mL- 1 min- 1 chitinase activity/g chitin. Nano-talc formulation of Actino 48 had more a significant effect compared to the other formulations in reducing percentages of damping-off and root rot diseases that equal to 19.05% and 4.76% with reduction percentages of 60% and 80%, respectively. The healthy survival percentage of peanut plants recorded 76.19%. Furthermore, the nano-talc formulation of Actino 48 was sufficient in increasing the dry weight of the peanut plants shoot, root systems, and the total number of peanut pods with increasing percentages of 47.62%, 55.62%, and 38.07%, respectively. CONCLUSION: The bio-friendly formulations of actinobacteria resulting from this investigation may play an active role in managing soil-borne diseases.

Comparison of intestinal absorption of soybean protein isolate-, glutenin- and peanut protein isolate-bound Nε-(carboxymethyl) lysine after in vitro gastrointestinal digestion.

Advanced glycation end products (AGEs), a heterogeneous compound existed in processed foods, are related to chronic diseases when they are accumulated excessively in human organs. Protein-bound Nε-(carboxymethyl) lysine (CML) as a typical AGE, is widely determined to evaluate AGEs level in foods and in vivo. This study investigated the intestinal absorption of three protein-bound CML originated from main food raw materials (soybean, wheat and peanut). After in vitro gastrointestinal digestion, the three protein-bound CML digests were ultrafiltered and divided into four fractions: less than 1 kDa, between 1 and 3 kDa, between 3 and 5 kDa, greater than 5 kDa. Caco-2 cell monolayer model was further used to evaluate the intestinal absorption of these components. Results showed that the absorption rates of soybean protein isolate (SPI)-, glutenin (Glu)-, peanut protein isolate (PPI)-bound CML were 30.18%, 31.57% and 29.5%, respectively. The absorption rates of components with MW less than 5 kDa accounted for 19.91% (SPI-bound CML), 22.59% (Glu-bound CML), 23.64% (PPI-bound CML), respectively, and these samples were absorbed by paracellular route, transcytosis route and active route via PepT-1. Taken together, these findings demonstrated that all three protein-bound CML digests with different MW can be absorbed in diverse absorption pathways by Caco-2 cell monolayer model. This research provided a theoretical basis for scientific evaluation of digestion and absorption of AGEs in food.

Effects of enzymatic hydrolysis combined with glycation on the emulsification characteristics and emulsion stability of peanut protein isolate.

Peanut protein isolate (PPI) has high nutritional value, but its poor function limits its application in the food industry. In this study, peanut protein isolate was modified by enzymatic hydrolysis combined with glycation. The structure, emulsification and interface properties of peanut protein isolate hydrolysate (HPPI) and dextran (Dex) conjugate (HPPI-Dex) were studied. In addition, the physicochemical properties, rheological properties, and stability of the emulsion were also investigated. The results showed that the graft degree increased with the increase of Dex ratio. Fourier transform infrared spectroscopy (FTIR) confirmed that the glycation of HPPI and Dex occurred. The microstructure showed that the structure of HPPI-Dex was expanded, and the molecular flexibility was enhanced. When the ratio of HPPI to Dex was 1:3, the emulsifying activity and the interface pressure of glycated HPPI reached the highest value, and the emulsifying activity (61.08 m2/g) of HPPI-Dex was 5.28 times that of PPI. The HPPI-Dex stabilized emulsions had good physicochemical properties and rheological properties. In addition, HPPI-Dex stabilized emulsions had high stability under heat treatment, salt ion treatment and freeze-thaw cycle. According to confocal laser scanning microscopy (CLSM), the dispersion of HPPI-Dex stabilized emulsions was better after 28 days of storage. This study provides a theoretical basis for developing peanut protein emulsifier and further expanding the application of peanut protein in food industry.

Peanut photosynthesis response to drought can include diffusive and biochemical limitations depending on cultivar.

Photosynthesis, understood as the photosynthetic carbon assimilation rate, is one of the key processes affected by drought stress. The effects can be via decreased CO2 diffusion and biochemical constraints. However, there is still no unified consensus about the contribution of each mechanism to the drought response. This research assessed the underlying limitations to photosynthesis in nine peanut genotypes (Arachis hypogaea L.) with different water strategies (water savers vs water spenders) under progressive drought. Water saver cultivars close the stomata earlier during drought, resulting in decreased transpiration and photosynthesis, which results in less water depletion in the soil, while water spenders maintain the stomata open during drought. In order to test the performance of these genotypes, growth, transpiration per plant, gas exchange measurements, chlorophyll fluorescence and A/Ci response curves were analyzed under drought and well-watered conditions. In general, drought first affected photosynthesis (at the leaf and canopy level) via stomatal closure and then by impacts on chlorophyll fluorescence in all genotypes, but at different intensity levels. The maximum rate of carboxylation and the maximum rate of electron transport, physiological characteristics related to biochemical constraints, were not affected during the onset of drought, but they were decreased at the end of the drought period, with the exception of the PI 493329 genotype that showed higher stomatal conductance due to a bigger root system. The findings presented here highlight the importance of genetic variation in the photosynthetic response of peanut to drought, which should be considered when breeding for future climates.

Optimized nitrogen application ameliorates the photosynthetic performance and yield potential in peanuts as revealed by OJIP chlorophyll fluorescence kinetics.

BACKGROUND: Nitrogen (N) is a crucial element for increasing photosynthesis and crop yields. The study aims to evaluate the photosynthetic regulation and yield formation mechanisms of different nodulating peanut varieties with N fertilizer application. METHOD: The present work explored the effect of N fertilizer application rates (N0, N45, N105, and N165) on the photosynthetic characteristics, chlorophyll fluorescence characteristics, dry matter, N accumulation, and yield of four peanut varieties. RESULTS: The results showed that N application increased the photosynthetic capacity, dry matter, N accumulation, and yield of peanuts. The measurement of chlorophyll a fluorescence revealed that the K-phase, J-phase, and I-phase from the OJIP curve decreased under N105 treatment compared with N0, and WOI, ET0/CSM, RE0/CSM, ET0/RC, RE0/RC, φPo, φEo, φRo, and Ψ0 increased, whereas VJ, VI, WK, ABS/RC, TR0/RC, DI0/RC, and φDo decreased. Meanwhile, the photosystem activity and electron transfer efficiency of nodulating peanut varieties decreased with an increase in N (N165). However, the photosynthetic capacity and yield of the non-nodulating peanut variety, which highly depended on N fertilizer, increased with an increase in N. CONCLUSION: Optimized N application (N105) increased the activity of the photosystem II (PSII) reaction center, improved the electron and energy transfer performance in the photosynthetic electron transport chain, and reduced the energy dissipation of leaves in nodulating peanut varieties, which is conducive to improving the yield. Nevertheless, high N (N165) had a positive effect on the photosystem and yield of non-nodulating peanut. The results provide highly valuable guidance for optimizing peanut N management and cultivation measures.

Phosphorus and selenium compounding mitigates Cr stress in peanut seedlings by enhancing growth homeostasis and antioxidant properties.

Peanut is an economically important crop, but it is susceptible to Cr contamination. In this study, we used peanut as experimental material to investigate the effects of exogenous P, Se interacting with Cr on the nutrient growth and antioxidant system of peanut seedlings by simulating Cr (0 µM, 50 µM, and 100 µM) stress environment. The results showed that exogenous P, Se supply could mitigate irreversible damage to peanut seedlings by altering the distribution of Cr in roots and aboveground, changing root conformation, and repairing damaged cells to promote growth. When the Cr concentration is 100 µM, it exhibits the highest toxicity. Compared to the control group P and Se (0 MM), the treatment with simultaneous addition of P + Se (0.5 + 6.0) resulted in a significant increase in root length and root tip number by 248.7% and 127.4%, respectively. Additionally, there was a 46.9% increase in chlorophyll content, a 190.2% increase in total surface area of the seedlings, and a respective increase of 149.1% and 180.3% in soluble protein content in the shoot and roots. In addition, by restricting the absorption of Cr and reducing the synthesis of superoxide dismutase SOD (Superoxide dismutase), CAT (Catalase), POD (Peroxidase), and MDA (Malonaldehyde), it effectively alleviates the oxidative stress on the antioxidant system. Therefore, the exogenous addition of P (0.5 MM) and Se (6.0 MM) prevented the optimal concentration of chromium toxicity to peanuts. Our research provides strong evidence that the exogenous combination of P and Se reduces the risk of peanut poisoning by Cr, while also exploring the optimal concentration of exogenous P and Se under laboratory conditions, providing a basis for further field experiments.

Employing conductive porous hydrogen-bonded organic framework for ultrasensitive detection of peanut allergen Ara h1.

Peanut allergy has garnered worldwide attention due to its high incidence rate and severe symptoms, stimulating the demand for the ultrasensitive detection method of peanut allergen. Herein, we successfully developed a novel electrochemical aptasensor for ultrasensitive detection Ara h1, a major allergenic protein present in peanuts. A conductive nickel atoms Anchored Hydrogen-Bonded Organic Frameworks (PFC-73-Ni) were utilized as excellent electrocatalysts toward hydroquinone (HQ) oxidation to generate a readable current signal. The developed electrochemical aptasensor offers wide linear range (1-120 nM) and low detection limit (0.26 nM) for Ara h1. This method demonstrated a recovery rate ranging from 95.00% to 107.42% in standard addition detection of non-peanut food samples. Additionally, the developed electrochemical method was validated with actual samples and demonstrated good consistency with the results obtained from a commercial ELISA kit. This indicates that the established Ara h1 detection method is a promising tool for peanut allergy prevention.

Changes in soil properties and microbial activity unveil the distinct impact of polyethylene and biodegradable microplastics on chromium uptake by peanuts.

Microplastics (MPs) are emerging persistent pollutants, and heavy metals are typical environmental pollutants, with their coexistence potentially compounding pollution and ecological risks. However, the interactive impacts and the relevant mechanisms of heavy metal and different types of MPs in plant-soil systems are still unclear. This study investigated the differential impacts of polyethylene MPs (PE MPs) and biodegradable polybutylene adipate MPs (PBAT MPs) on chromium (Cr) uptake in peanuts, focusing on plant performance and rhizosphere soil microenvironment. Compared with nondegradable PE-MPs, biodegradable PBAT MPs produced less significant influences on plant phytotoxicity, soil Cr bioavailability, and soil properties such as pH, CEC, DOC, and MBC, with the exception of MBN in Cr-contaminated soils. Compared to the control, soil pH and cation exchange capacity (CEC) decreased by MPs, while soil-soluble carbon (DOC), microbial biomass carbon, and nitrogen (MBC and MBN) increased by MPs. Compared to the control, soil-bioavailable Cr increased by 11.8-177.8% under PE MPs treatments, while increased by 5.1-156.9% under PBAT MPs treatments. The highest Cr content in shoots and roots was observed at 500.0 mg·kg-1 Cr level, which increased by 53.1% and 79.2% under 5% PE MPs treatments, respectively, as well as increased by 38.3% and 60.4% under 5% PBAT MPs treatments, respectively, compared with the control. The regression path analysis indicated that pH, MBC, MBN, and soil-bioavailable Cr played a vital role in the changes of soil properties and Cr uptake by peanuts induced by MPs. Soil bacterial community analysis revealed that Nocardioides, Proteobacteria, and Sphingomonas were reduced by the inhibition of MPs, which affected Cr uptake by peanuts. These results indicated that the peanut soil microenvironment was affected by PBAT and PE MPs, altering the Cr bioavailability and plant Cr uptake in Cr-contaminated soil.

From agro-food waste to nanoparticles: green synthesis of copper nanoparticles with lignin peroxidase enzyme produced by Anoxybacillus rupiensis using peanut shells.

This study presents a novel approach for the eco-friendly green synthesis of copper nanoparticles (Cu NPs) using enzymatic mediation which is an environmentally benign alternative to conventional methods, offering control over particle size and shape. Anoxybacillus rupiensis BS1 thermophilic bacterium was isolated from Erzurum's Pasinler hot spring and lignin peroxidase enzyme production conditions (incubation time 96 h, 40 g/L shell amount, pH 8.5, 150 rpm, and 60 °C temperature) were used in the production of peroxidase enzyme using peanut waste which has been optimized. The characterization of the synthesized Cu NPs was performed using various analytical techniques, including UV-vis spectroscopy, Fourier-transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), and scanning electron microscopy (SEM), confirming the successful production of stable and well-defined nanoparticles. Furthermore, the biological activities of the synthesized Cu NPs were explored, revealing their potential for antimicrobial applications. The antibacterial efficacy of the Cu NPs against some pathogens such as Escherichia coli, Klebsiella pneumoniae, Staphylococcus aureus, Streptococcus pyogenes, and Bacillus cereus was examined. It was determined that Cu NPs were effective on all pathogens and had the highest effectiveness against the S. pyogenes pathogen (19.0 mm). This study not only presents an innovative and sustainable approach for the synthesis of Cu NPs but also highlights the multifaceted biological activities of these nanoparticles, opening avenues for diverse applications in the fields of medicine, agriculture, and environmental remediation. The utilization of peanut shell wastes as a substrate for enzyme production adds value to agricultural by-products, contributing to the development of a circular and sustainable economy.

Root Endophyte-Manipulated Alteration in Rhizodeposits Stimulates Claroideoglomus in the Rhizosphere to Enhance Drought Resistance in Peanut.

Drought dramatically affects plant growth and yield. A previous study indicated that endophytic fungus Phomopsis liquidambaris can improve the drought resistance of peanuts, which is related with the root arbuscular mycorrhizal fungi (AMF) community; however, how root endophytes mediate AMF assembly to affect plant drought resistance remains unclear. Here, we explored the mechanism by which endophytic fungus recruits AMF symbiotic partners via rhizodeposits to improve host drought resistance. The results showed that Ph. liquidambaris enhanced peanut drought resistance by enriching the AMF genus Claroideoglomus of the rhizosphere. Furthermore, metabolomic analysis indicated that Ph. liquidambaris significantly promoted isoformononetin and salicylic acid (SA) synthesis in rhizodeposits, which were correlated with the increase in Claroideoglomus abundance following Ph. liquidambaris inoculation. Coinoculation experiments confirmed that isoformononetin and SA could enrich Claroideoglomus etunicatum in the rhizosphere, thereby improving the drought resistance. This study highlights the crucial role of fungal consortia in plant stress resistance.

Prevalence of Clostridium botulinum in Retail Peanut Butters from a 2007 Survey in Ottawa, Canada.

The spore-forming, anaerobic bacterium, Clostridium botulinum, can cause intestinal toxemia (colonization) botulism in adults and infants by colonizing the gut and producing botulinum neurotoxin in situ. In 2006, peanut butter was identified as a lab-confirmed source of C. botulinum spores for an adult colonization botulism case in Canada. It is recommended for infants to be exposed to peanut butter at an early age to help prevent the development of a peanut allergy, yet the prevalence of C. botulinum in retail peanut butters is currently unknown. This report details a survey that was conducted in 2007 for the presence of viable C. botulinum spores in 92 peanut butters and 12 other nut butter spreads obtained from retail grocery stores in Ottawa, Canada. Samples were tested for viable C. botulinum spores by detecting botulinum neurotoxin in enrichment cultures by mouse bioassay. Three of the peanut butters from the entire survey of nut butter spreads (3/104, 3%) produced cultures containing botulinum neurotoxin. Whole genome sequencing performed on one isolate from this survey, as well as a clinical isolate and peanut butter isolates associated with the 2006 adult colonization case revealed that all C. botulinum isolates contained a full-length chromosomal bont/A1 gene within an ha-orf + cassette. This study identifies retail peanut butters as a potential source of viable C. botulinum spores at the time of sampling. Whether peanut butter represents a food category that may be contributing to the incidence of infant botulism has yet to be determined.

Combining Hyperspectral Techniques and Genome-Wide Association Studies to Predict Peanut Seed Vigor and Explore Associated Genetic Loci.

Seed vigor significantly affects peanut breeding and agricultural yield by influencing seed germination and seedling growth and development. Traditional vigor testing methods are inadequate for modern high-throughput assays. Although hyperspectral technology shows potential for monitoring various crop traits, its application in predicting peanut seed vigor is still limited. This study developed and validated a method that combines hyperspectral technology with genome-wide association studies (GWAS) to achieve high-throughput detection of seed vigor and identify related functional genes. Hyperspectral phenotyping data and physiological indices from different peanut seed populations were used as input data to construct models using machine learning regression algorithms to accurately monitor changes in vigor. Model-predicted phenotypic data from 191 peanut varieties were used in GWAS, gene-based association studies, and haplotype analyses to screen for functional genes. Real-time fluorescence quantitative PCR (qPCR) was used to analyze the expression of functional genes in three high-vigor and three low-vigor germplasms. The results indicated that the random forest and support vector machine models provided effective phenotypic data. We identified Arahy.VMLN7L and Arahy.7XWF6F, with Arahy.VMLN7L negatively regulating seed vigor and Arahy.7XWF6F positively regulating it, suggesting distinct regulatory mechanisms. This study confirms that GWAS based on hyperspectral phenotyping reveals genetic relationships in seed vigor levels, offering novel insights and directions for future peanut breeding, accelerating genetic improvements, and boosting agricultural yields. This approach can be extended to monitor and explore germplasms and other key variables in various crops.

System-wide analysis of groundnut's salinity resilience: Integrating plant-cell interactions with environmental stress dynamics through cutting-edge transcriptomics.

Salinity stress is a major concern in regions where irrigation relies on saline water. This study aimed to investigate the relative water content (RWC), electrolytic leakage (EL), total chlorophyll content, free amino acid content, and total soluble sugar content were analyzed in different groundnut species subjected to various salinity treatments. The results showed that salinity stress significantly reduced the RWC in groundnut leaves, with A. duranensis (wild type) exhibiting higher RWC values compared to the Arachis hypogaea species. RNA sequencing was performed to identify differentially expressed genes (DEGs) during salt stress. A total of 9079 DEGs were identified, with 1372 genes upregulated and 2509 genes downregulated. Genes belonging to transcription factor families, such as WRKY, MYB, bHLH, E2F, and Auxin efflux carrier proteins, were induced under salt stress in the tolerant genotype. Conversely, genes encoding NADH dehydrogenase, glutathione S-transferase, protein kinases, UDP-glycosyltransferase, and peroxidase were downregulated. Gene ontology and pathway analyses revealed several enriched categories and metabolic pathways associated with salt stress response, including catalytic activity, response to salt stress, ATP-dependent activity, and oxidative phosphorylation. The findings of this study provide insights into the physiological and molecular responses of groundnut to salinity stress. A. duranensis exhibited better salinity tolerance than Arachis hypogaea, as indicated by higher RWC values, lower electrolytic leakage, and differential gene expression patterns. These results contribute to our understanding of the mechanisms underlying salt stress tolerance in groundnut and may guide future efforts to develop salinity-tolerant groundnut species, ultimately improving crop yield in saline-affected regions.

Chloroplast and whole-genome sequencing shed light on the evolutionary history and phenotypic diversification of peanuts.

Cultivated peanut (Arachis hypogaea L.) is a widely grown oilseed crop worldwide; however, the events leading to its origin and diversification are not fully understood. Here by combining chloroplast and whole-genome sequence data from a large germplasm collection, we show that the two subspecies of A. hypogaea (hypogaea and fastigiata) likely arose from distinct allopolyploidization and domestication events. Peanut genetic clusters were then differentiated in relation to dissemination routes and breeding efforts. A combination of linkage mapping and genome-wide association studies allowed us to characterize genes and genomic regions related to main peanut morpho-agronomic traits, namely flowering pattern, inner tegument color, growth habit, pod/seed weight and oil content. Together, our findings shed light on the evolutionary history and phenotypic diversification of peanuts and might be of broad interest to plant breeders.

Molecular Cloning and Functional Identification of a Pericarp- and Testa-Abundant Gene's (AhN8DT-2) Promoter from Arachis hypogaea.

Cultivated peanut (Arachis hypogaea L.) is a key oil- and protein-providing legume crop of the world. It is full of nutrients, and its nutrient profile is comparable to that of other nuts. Peanut is a unique plant as it showcases a pegging phenomenon, producing flowers above ground, and after fertilization, the developing peg enters the soil and produces seeds underground. This geocarpic nature of peanut exposes its seeds to soil pathogens. Peanut seeds are protected by an inedible pericarp and testa. The pericarp- and testa-specific promoters can be effectively used to improve the seed defense. We identified a pericarp- and testa-abundant expression gene (AhN8DT-2) from available transcriptome expression data, whose tissue-specific expression was further confirmed by the qRT-PCR. The 1827bp promoter sequence was used to construct the expression vector using the pMDC164 vector for further analysis. Quantitative expression of the GUS gene in transgenic Arabidopsis plants showed its high expression in the pericarp. GUS staining showed a deep blue color in the pericarp and testa. Cryostat sectioning of stained Arabidopsis seeds showed that expression is only limited to seed coat (testa), and staining was not present in cotyledons and embryos. GUS staining was not detected in any other tissues, including seedlings, leaves, stems, and roots, except for some staining in flowers. Under different phytohormones, this promoter did not show an increase in expression level. These results indicated that the AhN8DT-2 promoter drives GUS gene expression in a pericarp- and testa-specific manner. The identified promoter can be utilized to drive disease resistance genes, specifically in the pericarp and testa, enhancing peanut seed defense against soil-borne pathogens. This approach has broader implications for improving the resilience of peanut crops and other legumes, contributing to sustainable agricultural practices and food security.

Risk assessment of resistance to prochloraz in Phoma arachidicola causing peanut web blotch.

Peanut web blotch (PWB) caused by Phoma arachidicola, is one of the most serious foliar diseases of peanut. Although prochloraz is an active fungicide with broad anti-fungal spectrum, it has not been registered for the control of PWB in China. The activity of prochloraz against P. arachidicola and the risk of resistance to prochloraz in P. arachidicola are still unclear. In current study, the inhibitory activity of prochloraz against 96 P. arachidicola strains was determined with the average EC50 value of 1.2700 ± 0.7786 µg/mL. Prochloraz exhibited excellent protective and curative effect on detached peanut leaves, and the effect was obviously better than that of carbendazim and difenoconazole at the same concentration. After prochloraz treatment, the mycelium of P. arachidicola contorted, shrunk and ruptured, with shrinking of cell wall and membrane, enhanced cell membrane permeability, and reduced ergosterol content. Totally 80 prochloraz-resistant mutants were obtained by fungicide adaptation with the frequency of 6.7 × 10-3. All the selected 12 prochloraz-resistant mutants lost their resistance to prochloraz after 10 transfers on PDA plates. And these mutants exhibited decreased biological fitness in mycelial growth and pathogenicity. Moreover, there was positive cross-resistance between prochloraz and other demethylation inhibitor (DMI) fungicides, such as tebuconazole, triflumizole and difenoconazole, but no cross-resistance was found between prochloraz and other classes of fungicides, such as carbendazim, pydiflumetofen or fludioxonil. Overexpression of PaCYP51 and PaAtrB genes were detected in the resistant mutants. All the above results demonstrated that prochloraz has a great potential in management of PWB. The risk of P. arachidicola developing resistance to prochloraz is relatively low-to-medium. Overexpressing of PaCYP51 and PaAtrB might be linked to prochloraz resistance in P. arachidicola.

Peanut Rhizosphere Achromobacter xylosoxidans Inhibits Aspergillus flavus Development and Aflatoxin Synthesis by Inducing Apoptosis through Targeting the Cell Membrane.

Contamination of crop seeds and feed with Aspergillus flavus and its associated aflatoxins presents a significant threat to human and animal health due to their hepatotoxic and carcinogenic properties. To address this challenge, researchers have screened for potential biological control agents in peanut soil and pods. This study identified a promising candidate, a strain of the nonpigmented bacterium, Achromobacter xylosoxidans ZJS2-1, isolated from the peanut rhizosphere in Zhejiang Province, China, exhibiting notable antifungal and antiaflatoxin activities. Further investigations demonstrated that ZJS2-1 active substances (ZAS) effectively inhibited growth at a MIC of 60 µL/mL and nearly suppressed AFB1 production by 99%. Metabolomic analysis revealed that ZAS significantly affected metabolites involved in cell wall and membrane biosynthesis, leading to compromised cellular integrity and induced apoptosis in A. flavus through the release of cytochrome c. Notably, ZAS targeted SrbA, a key transcription factor involved in ergosterol biosynthesis and cell membrane integrity, highlighting its crucial role in ZJS2-1's biocontrol mechanism. Moreover, infection of crop seeds and plant wilt caused by A. flavus can be efficiently alleviated by ZAS. Additionally, ZJS2-1 and ZAS demonstrated significant inhibitory effects on various Aspergillus species, with inhibition rates ranging from 80 to 99%. These findings highlight the potential of ZJS2-1 as a biocontrol agent against Aspergillus species, offering a promising solution to enhance food safety and protect human health.

Enhanced detection of Aspergillus flavus in peanut kernels using a multi-scale attention transformer (MSAT): Advancements in food safety and contamination analysis.

In this study, a multi-scale attention transformer (MSAT) was coupled with hyperspectral imaging for classifying peanut kernels contaminated with diverse Aspergillus flavus fungi. The results underscored that the MSAT significantly outperformed classic deep learning models, due to its sophisticated multi-scale attention mechanism which enhanced its classification capabilities. The multi-scale attention mechanism was utilized by employing several multi-head attention layers to focus on both fine-scale and broad-scale features. It also integrated a series of scale processing layers to capture features at different resolutions and incorporated a self-attention mechanism to integrate information across different levels. The MSAT model achieved outstanding performance in different classification tasks, particularly in distinguishing healthy peanut kernels from those contaminated with aflatoxigenic fungi, with test accuracy achieving 98.42±0.22%. However, it faced challenges in differentiating peanut kernels contaminated with aflatoxigenic fungi from those with non-aflatoxigenic contamination. Visualization of attention weights explicitly revealed that the MSAT model's multi-scale attention mechanism progressively refined its focus from broad spatial-spectral features to more specialized signatures. Overall, the MSAT model's advanced processing capabilities marked a notable advancement in the field of food quality safety, offering a robust and reliable tool for the rapid and accurate detection of Aspergillus flavus contaminations in food.