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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 W OI , ET 0 /CS M , RE 0 /CS M , ET 0 /RC, RE 0 /RC, φPo, φEo, φRo, and Ψ0 increased, whereas V J , V I , W K , ABS/RC, TR 0 /RC, DI 0 /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., (© 2024. The Author(s).)

Waterlogging stress (WS) hinders kernel development and directly reduces peanut yield; however, the mechanism of kernel filling in response to WS remains unknown. The waterlogging-sensitive variety Huayu 39 was subjected to WS for 3 days at 7 days after the gynophores touched the ground (DAG). We found that WS affected kernel filling at 14, 21, and 28 DAG. WS decreased the average filling rate and kernel dry weight, while transcriptome sequencing and widely targeted metabolomic analysis revealed that WS inhibited the gene expression in starch and sucrose metabolism, which reduced sucrose input and transformation ability. Additionally, genes related to ethylene and melatonin synthesis and the accumulation of tryptophan and methionine were upregulated in response to WS. WS upregulated the expression of the gene encoding tryptophan decarboxylase (AhTDC), and overexpression of AhTDC in Arabidopsis significantly reduced the seed length, width, and weight. Therefore, WS reduced the kernel-filling rate, leading to a reduction in the 100-kernel weight. This survey informs the development of measures that alleviate the negative impact of WS on peanut yield and quality and provides a basis for exploring high-yield and high-quality cultivation, molecular-assisted breeding, and waterlogging prevention in peanut farming., (© 2024 John Wiley & Sons Ltd.)

Photosynthesis, understood as the photosynthetic carbon assimilation rate, is one of the key processes affected by drought stress. The effects can be via decreased CO 2 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/C i 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., (© 2024 Scandinavian Plant Physiology Society.)

Background: WRKY proteins are important transcription factors (TFs) in plants, involved in growth and development and responses to environmental changes. Although WRKY TFs have been studied at the genome level in Arachis genus, including oil crop and turfgrass, their regulatory networks in controlling flowering time remain unclear. The aim of this study was to predict the molecular mechanisms of WRKY TFs regulation flowering time in Arachis genus at the genome level using bioinformatics approaches., Results: The flowering-time genes of Arachis genus were retrieved from the flowering-time gene database. The regulatory networks between WRKY TFs and downstream genes in Arachis genus were predicted using bioinformatics tools. The results showed that WRKY TFs were involved in aging, autonomous, circadian clock, hormone, photoperiod, sugar, temperature, and vernalization pathways to modulate flowering time in Arachis duranensis, Arachis ipaensis, Arachis monticola, and Arachis hypogaea cv. Tifrunner. The WRKY TF binding sites in homologous flowering-time genes exhibited asymmetric evolutionary pattern, indicating that the WRKY TFs interact with other transcription factors to modulate flowering time in the four Arachis species. Protein interaction network analysis showed that WRKY TFs interacted with FRUITFULL and APETALA2 to modulate flowering time in the four Arachis species. WRKY TFs implicated in regulating flowering time had low expression levels, whereas their interaction proteins had varying expression patterns in 22 tissues of A. hypogaea cv. Tifrunner. These results indicate that WRKY TFs exhibit antagonistic or synergistic interactions with the associated proteins., Conclusions: This study reveals complex regulatory networks through which WRKY TFs modulate flowering time in the four Arachis species using bioinformatics approaches., (© 2024. The Author(s).)

Abiotic stress is a limiting factor in peanut production. Peanut is an important oil crop and cash crop in China. Peanut yield is vulnerable to abiotic stress due to its seeds grown underground. Jasmonic acid (JA) is essential for plant growth and defense against adversity stresses. However, the regulation and mechanism of the jasmonic acid biosynthesis pathway on peanut defense against abiotic stresses are still limitedly understood. In this study, a total of 64 genes encoding key enzymes of JA biosynthesis were identified and classified into lipoxygenases (AhLOXs), alleno oxide synthases (AhAOSs), allene oxide cyclases (AhAOCs), and 12-oxo-phytodienoic acid reductases (AhOPRs) according to gene structure, conserved motif, and phylogenetic feature. A cis -regulatory element analysis indicated that some of the genes contained stress responsive and hormone responsive elements. In addition to proteins involved in JA biosynthesis and signaling, they also interacted with proteins involved in lipid biosynthesis and stress response. Sixteen putative Ah-miRNAs were identified from four families targeting 35 key genes of JA biosynthesis. A tissue expression pattern analysis revealed that AhLOX2 was the highest expressed in leaf tissues, and AhLOX32 was the highest expressed in shoot, root, and nodule tissues. AhLOX16, AhOPR1, and AhOPR3 were up-regulated under drought stress. AhLOX16, AhAOS3, AhOPR1, and AhAOC4 had elevated transcript levels in response to cold stress. AhLOX5, AhLOX16, AhAOC3, AhOPR1, and AhOPR3 were up-regulated for expression under salt stress. Our study could provide a reference for the study of the abiotic stress resistance mechanism in peanut.

Background: Peanut (Arachis hypogaea), a vital oil and food crop globally, is susceptible to web blotch which is a significant foliar disease caused by Phoma arachidicola Marasas Pauer&Boerema leading to substantial yield losses in peanut production. Calcium treatment has been found to enhance plant resistance against pathogens., Results: This study investigates the impact of exogenous calcium on peanut resistance to web blotch and explores its mechanisms. Greenhouse experiments revealed that exogenous calcium treatment effectively enhanced resistance to peanut web blotch. Specifically, amino acid calcium and sugar alcohol calcium solutions demonstrated the best induced resistance effects, achieving reduction rates of 61.54% and 60% in Baisha1016, and 53.94% and 50% in Luhua11, respectively. All exogenous calcium treatments reduced malondialdehyde (MDA) and relative electrical conductivity (REC) levels in peanut leaves, mitigating pathogen-induced cell membrane damage. Exogenous calcium supplementation led to elevated hydrogen peroxide (H 2 O 2 ) content and superoxide anion (O 2 ∙- ) production in peanut leaves, facilitating the accumulation of reactive oxygen species (ROS) crucial for plant defense responses. Amino acid calcium and sugar alcohol calcium treatments significantly boosted activities of peroxidase (POD), superoxide dismutase (SOD), catalase (CAT), and ascorbate peroxidase (APX) in peanut leaves. Activation of these antioxidant enzymes effectively scavenged excess ROS, maintaining ROS balance and mitigating cellular damage., Conclusions: In summary, exogenous calcium treatment triggered ROS production, which was subsequently eliminated by the activation of antioxidant enzymes, thereby reducing cell membrane damage and inducing defense responses against peanut web blotch., (© 2024. The Author(s).)

Water stress is a major cause of yield loss in peanut cultivation. Melatonin seed priming has been used to enhance stress tolerance in several crops, but not in peanut. We investigated the impact of seed priming with melatonin on the growth, development, and drought tolerance of two peanut cultivars, TUFRunner™ '511', a drought tolerant cultivar, and New Mexico Valencia A, a drought sensitive cultivar. Peanut seed priming tests using variable rates of melatonin (0-200 μM), indicated that 50 μM of melatonin resulted in more uniform seed germination and improved seedling growth in both cultivars under non stress conditions. Seed priming with melatonin also promoted vegetative growth, as evidenced by higher whole-plant transpiration, net CO 2 assimilation, and root water uptake under both well-watered and water stress conditions in both cultivars. Higher antioxidant activity and protective osmolyte accumulation, lower reactive oxygen species accumulation and membrane damage were observed in primed compared with non-primed plants. Seed priming with melatonin induced a growth promoting effect that was more evident under well-watered conditions for TUFRunnner™ '511', whereas for New Mexico Valencia A, major differences in physiological responses were observed under water stress conditions. New Mexico Valencia A primed plants exhibited a more sensitized stress response, with faster down-regulation of photosynthesis and transpiration compared with non-primed plants. The results demonstrate that melatonin seed priming has significant potential to improve early establishment and promote growth of peanut under optimal conditions, while also improve stress tolerance during water stress., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Masson SAS. All rights reserved.)

Peanut production is threatened by climate change. Damage to seedlings from low temperatures in early spring can limit yield. Plant adaptations to chilling stress remain unclear in peanut seedlings. It is essential to understand how peanut acquires chilling tolerance. We evaluated effects of chilling stress on growth and recovery of peanut seedlings. We compared and analysed biological characteristics, antioxidants, photosynthesis, biochemical and physiological responses, and nutrient absorption at varying levels of chilling. Compared with chilling-sensitive FH18, the reduced impact of chilling stress on chilling-tolerant NH5 was associated with reduced ROS accumulation, higher ascorbate peroxidase activity and soluble sugar content, lower soluble protein content, and smaller reductions in nutrient content during stress. After removal of chilling stress, FH18 had significant accumulation of O 2 •- and H 2 O 2 , which decreased photosynthesis, nutrient absorption, and transport. ROS-scavenging reduced damage from chilling stress, allowed remobilization of nutrients, improved chilling tolerance, and restored plant functioning after chilling stress removal. These findings provide a reference for targeted research on peanut seedling tolerance to chilling and lay the foundation for bioinformatics-based research on peanut chilling tolerance mechanisms., (© 2024 Wiley‐VCH GmbH. Published by John Wiley & Sons Ltd.)

Peanut (Arachis hypogaea L.) is an important oilseed crop worldwide. However, soil salinization becomes one of the main limiting factors of peanut production. Therefore, developing salt-tolerant varieties and understanding the molecular mechanisms of salt tolerance is important to protect peanut yield in saline areas. In this study, we selected four peanut varieties with contrasting response to salt challenges with T1 and T2 being tolerance and S1 and S2 being susceptible. High-throughput RNA sequencing resulted in more than 314.63 Gb of clean data from 48 samples. We identified 12,057 new genes, 7,971of which have functional annotations. KEGG pathway enrichment analysis of uniquely expressed genes in salt-tolerant peanut revealed that upregulated genes in the root are involved in the MAPK signaling pathway, fatty acid degradation, glycolysis/gluconeogenesis, and upregulated genes in the shoot were involved in plant hormone signal transduction and the MAPK signaling pathway. Na + content, K + content, K + / Na + , and dry mass were measured in root and shoot tissues, and two gene co-expression networks were constructed based on weighted gene co-expression network analysis (WGCNA) in root and shoot. In this study, four key modules that are highly related to peanut salt tolerance in root and shoot were identified, plant hormone signal transduction, phenylpropanoid biosynthesis, starch and sucrose metabolism, flavonoid biosynthesis, carbon metabolism were identified as the key biological processes and metabolic pathways for improving peanut salt tolerance. The hub genes include genes encoding ion transport (such as HAK8, CNGCs, NHX, NCL1) protein, aquaporin protein, CIPK11 (CBL-interacting serine/threonine-protein kinase 11), LEA5 (late embryogenesis abundant protein), POD3 (peroxidase 3), transcription factor, and MAPKKK3. There were some new salt-tolerant genes identified in peanut, including cytochrome P450, vinorine synthase, sugar transport protein 13, NPF 4.5, IAA14, zinc finger CCCH domain-containing protein 62, beta-amylase, fatty acyl-CoA reductase 3, MLO-like protein 6, G-type lectin S-receptor-like serine/threonine-protein kinase, and kinesin-like protein KIN-7B. The identification of key modules, biological pathways, and hub genes in this study enhances our understanding of the molecular mechanisms underlying salt tolerance in peanuts. This knowledge lays a theoretical foundation for improving and innovating salt-tolerant peanut germplasm., (© 2024. The Author(s).)

Globally, drought stress poses a significant threat to crop productivity. Improving the drought tolerance of crops with microbial biostimulants is a sustainable strategy to meet a growing population's demands. This research aimed to elucidate microbial biostimulants' (Plant Growth Promoting Rhizobacteria) role in alleviating drought stress in oil-seed crops. In total, 15 bacterial isolates were selected for drought tolerance and screened for plant growth-promoting (PGP) attributes like phosphate solubilization and production of indole-3-acetic acid, siderophore, hydrogen cyanide, ammonia, and exopolysaccharide. This research describes two PGPR strains: Acinetobacter calcoaceticus AC06 and Bacillus amyloliquefaciens BA01. The present study demonstrated that these strains (AC06 and BA01) produced abundant osmolytes under osmotic stress, including proline (2.21 and 1.75 µg ml - 1 ), salicylic acid (18.59 and 14.21 µg ml - 1 ), trehalose (28.35 and 22.74 µg mg - 1 FW) and glycine betaine (11.35 and 7.74 mg g - 1 ) respectively. AC06 and BA01 strains were further evaluated for their multifunctional performance by inoculating in Arachis hypogaea L. (Groundnut) under mild and severe drought regimes (60 and 40% Field Capacity). Inoculation with microbial biostimulants displayed distinct osmotic-adjustment abilities of the groundnut, such as growth parameters, plant biomass, photosynthetic pigments, relative water content, proline, and soluble sugar in respective to control during drought. On the other hand, plant sensitivity indexes such as electrolyte leakage and malondialdehyde (MDA) contents were decreased as well as cooperatively conferred plant drought tolerance by induced alterations in stress indicators such as catalase (CAT), ascorbate peroxidase (APX), and superoxide dismutase (SOD). Thus, Acinetobacter sp. AC06 and Bacillus sp. BA01 can be considered as osmolyte producing microbial biostimulants to simultaneously induce osmotic tolerance and metabolic changes in groundnuts under drought stress., (© 2024. The Author(s).)

Drought stress threatens the productivity of numerous crops, including chilli pepper (Capsicum annuum). DnaJ proteins are known to play a protective role against a wide range of abiotic stresses. This study investigates the regulatory mechanism of the chloroplast-targeted chaperone protein AdDjSKI, derived from wild peanut (Arachis diogoi), in enhancing drought tolerance in chilli peppers. Overexpressing AdDjSKI in chilli plants increased chlorophyll content, reflected in the maximal photochemical efficiency of photosystem II (PSII) (Fv/Fm) compared with untransformed control (UC) plants. This enhancement coincided with the upregulated expression of PSII-related genes. Our subsequent investigations revealed that transgenic chilli pepper plants expressing AdDjSKI showed reduced accumulation of superoxide and hydrogen peroxide and, consequently, lower malondialdehyde levels and decreased relative electrolyte leakage percentage compared with UC plants. The mitigation of ROS-mediated oxidative damage was facilitated by heightened activities of antioxidant enzymes, including superoxide dismutase, catalase, ascorbate peroxidase, and peroxidase, coinciding with the upregulation of the expression of associated antioxidant genes. Additionally, our observations revealed that the ectopic expression of the AdDjSKI protein in chilli pepper plants resulted in diminished ABA sensitivity, consequently promoting seed germination in comparison with UC plants under different concentrations of ABA. All of these collectively contributed to enhancing drought tolerance in transgenic chilli plants with improved root systems when compared with UC plants. Overall, our study highlights AdDjSKI as a promising biotechnological solution for enhancing drought tolerance in chilli peppers, addressing the growing global demand for this economically valuable crop., (© 2024 Scandinavian Plant Physiology Society.)

Fifteen peanut varieties at the pod filling stage were exposed to waterlogging stress for 7 days, the enzyme activities and fluorescence parameters were measured after 7 days of waterlogging and drainage. The waterlogging tolerance and recovery capability of varieties were identified. After waterlogging, waterlogging tolerance coefficient (WTC) of relative electrolyte linkage (REL), malondialdehyde (MDA) content, superoxide dismutase (SOD) activity, and catalase (CAT) activity, non-photochemical quenching (NPQ) and photochemical quenching (qL) of leaves of most peanut varieties were increased, while the WTC of the soil and plant analysis development (SPAD) value, PS II actual quantum yield (Φ PS   II ), maximum photochemical efficiency (Fv/Fm) were decreased. After drainage, the WTC of REL, MDA content, SOD and CAT activity of leaves were decreased compared with that of after waterlogging, but these indicators of a few cultivars were increased. Φ PS   II , Fv/Fm and qL can be used as important indexes to identify waterlogging recovery capability. There was a significant negative correlation between recovery capability and the proportion of reduction in yield, while no significant correlation was found between waterlogging tolerance and the proportion of reduction in yield. Therefore, it is recommended to select varieties with high recovery capability and less pod number reduction under waterlogging in peanut breeding and cultivation., Competing Interests: The authors declare there are no competing interests., (©2022 Zeng et al.)

The frequency and severity of extreme climatic conditions such as drought, salinity, cold, and heat are increasing due to climate change. Moreover, in the field, plants are affected by multiple abiotic stresses simultaneously or sequentially. Thus, it is imperative to compare the effects of stress combinations on crop plants relative to individual stresses. This study investigated the differential regulation of physio-biochemical and metabolomics parameters in peanut ( Arachis hypogaea L.) under individual (salt, drought, cold, and heat) and combined stress treatments using multivariate correlation analysis. The results showed that combined heat, salt, and drought stress compounds the stress effect of individual stresses. Combined stresses that included heat had the highest electrolyte leakage and lowest relative water content. Lipid peroxidation and chlorophyll contents did not significantly change under combined stresses. Biochemical parameters, such as free amino acids, polyphenol, starch, and sugars, significantly changed under combined stresses compared to individual stresses. Free amino acids increased under combined stresses that included heat; starch, sugars, and polyphenols increased under combined stresses that included drought; proline concentration increased under combined stresses that included salt. Metabolomics data that were obtained under different individual and combined stresses can be used to identify molecular phenotypes that are involved in the acclimation response of plants under changing abiotic stress conditions. Peanut metabolomics identified 160 metabolites, including amino acids, sugars, sugar alcohols, organic acids, fatty acids, sugar acids, and other organic compounds. Pathway enrichment analysis revealed that abiotic stresses significantly affected amino acid, amino sugar, and sugar metabolism. The stress treatments affected the metabolites that were associated with the tricarboxylic acid (TCA) and urea cycles and associated amino acid biosynthesis pathway intermediates. Principal component analysis (PCA), partial least squares-discriminant analysis (PLS-DA), and heatmap analysis identified potential marker metabolites (pinitol, malic acid, and xylopyranose) that were associated with abiotic stress combinations, which could be used in breeding efforts to develop peanut cultivars that are resilient to climate change. The study will also facilitate researchers to explore different stress indicators to identify resistant cultivars for future crop improvement programs.

Storage is an important step after peanut harvest and drying. Many factors could affect the peanut quality during storage. The quality change differences of peanut after being dried by solar radiation and at 35°C, 40°C, 45°C, 50°C during later storage were investigated, including moisture content (MC) and germination percentage (GP) of peanut kernels, acid value (AV), peroxide value (PV), iodine value (IV), vitamin E (VE) content and fatty acid composition (FAC) of extracted peanut oil. And the impact of four storage conditions, air-room temperature (A-RT), air-low temperature (A-LT), vacuum-room temperature (V-RT) and nitrogen-room temperature (N-RT) on peanut quality after 10 months' storage were also studied in this paper. The results revealed that drying conditions had only a little influence on peanut quality during later storage. Peanut dried by solar radiation was more easily oxidized than that dried under other drying conditions. The effects of storage time were much greater. The GP, AV, PV, VE content and FAC, showed significantly changes along with storage. GP and VE content decreased, AV and PV increased, and some linoleic acid was oxidized to oleic acid after 10 months' storage. In addition, A-LT exhibited best performance in keeping peanut quality than A-RT, V-RT and N-RT, which demonstrated that low temperature was more advantageous for peanut storage than controlled atmosphere. These results above would provide useful information and reference for the peanut storage to apply in food industry.

In agroecosystems, drought stress severely threatens crops development. Although potassium (K) is required in amounts by crops under drought stress, the mobilization and availablity of K are limited by the soil water status. Arbuscular mycorrhizal (AM) fungi can form mutualistic associations with most crops and play direct or indirect roles in the host drought resistance. Considering that the glomalin generated by living AM fungal hyphae can sequester multiple minerals, however, the function of mineral-sequestering glomalin in the crop drought resistance remains unclear. In this study, peanuts cultivated in the sterilized soil with a history of AM fungi inoculation showed significantly enhanced leaf K accumulation, drought resistance and pod yield under drought stress. Through the collection of different types of mineral-sequestering glomalin from living AM fungal hyphae, the peanut drought resistance was improved only when K-sequestering glomalin was added. Moreover, we found that peanut root exudates could prime the dissociation of glomalin-bound K and further satisfy the K requirement of crops. Our study is the first report that K-sequestering glomalin could improve drought performance and peanut pod yield, and it helps us to understand the ecological importance of improving AM symbiosis to face agricultural challenges., (Copyright © 2021 Elsevier GmbH. All rights reserved.)

Nitric oxide-mediated S-nitrosation through S-nitrosoglutathione reductase (GSNOR) plays important roles in cellular processes and signaling of plants; however, the regulatory mechanism of programmed cell death (PCD) by S-nitrosation remains unclear. In this study, the S-nitrosated proteomic and functions of GSNOR during Al-induced PCD in peanut were investigated. Al stress induced an increase of S-nitrosothiol (SNO) content and GSNOR activity in Al-induced PCD. There was significant positive correlation between SNO content and hydrogen peroxide content. The S-nitrosated proteomic analysis identified 402 S-nitrosated proteins containing 551 S-nitrosated sites during Al-induced PCD in the root tips of peanut. These S-nitrosated proteins were involved in regulation of various biological processes including energy metabolism, maintenance of cell wall function and organic acid secretion. Among them, 128 S-nitrosated proteins were up-regulated and one was down-regulated after Al stress. Experiments with recombinant AhGSNOR revealed that activity of the enzyme was inhibited by its S-nitrosation, with a moderate decrease of 17.9 % after 100 μM GSNO incubation. These data provide novel insights to understanding the functional mechanism of NO-mediated S-nitrosation during plant PCD., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Peanut is one of the important oil and economic crops, among which the variegated testa peanut is a unique member. The molecular mechanisms underlying the pigment synthesis in variegated testa are still unclear. Differentially expressed genes (DEGs) in the flavonoid metabolism pathway in pigmented areas indicated that there were 27 DEGs highly related to the synthesis of variegated testa color among 1,050 DEGs. Of these 27, 13 were up-regulated and 14 were down-regulated, including 3 PALs, 1 C4H, 2 CHSs, 1 F3H, 1 F3'H, 2 DFRs, 2 LARs, 2 IAAs, 4 bHLHs, and 9 MYBs. GO (Gene Ontology) analysis indicated that DEGs were similarly enriched in three branches. KEGG (Kyoto Encyclopedia of Genes and Genomes) analysis suggested flavonoid biosynthesis is the most direct metabolic pathway for the synthesis of testa variegation. The liquid chromatography-tandem mass spectrometry (LC-MS/MS) results showed that cyanidin and delphinidin were the primary metabolites that caused the color differences between the pigmented and the non-pigmented areas. Through the verification of 20 DEGs via qPCR, the results were consistent with transcriptome sequencing in four comparison groups. The results in this study lay the foundation for revealing the molecular regulation mechanisms of flavonoid synthesis in variegated testa peanut.

The scarcity of information on the maturation physiology of the peanut seed (Arachis hypogaea L.; Virgínia group) makes harvesting high quality seeds a challenge for the seed industry. During two consecutive crop seasons, we studied the acquisition of physiological quality of peanut seeds during maturation in tropical conditions. We bring new insights about the period of late maturation of seeds and the influence of the maternal environment on physiological quality. We monitored water content, dry weight, ability of germination, desiccation tolerance, vigor and longevity. In addition, we monitored temperature and precipitation throughout plant growth. We demonstrate that the physiological quality of peanut seeds is acquired during development, with a maximum between 57 and 76 days after flowering in the late stage of maturation. This final period represents about 25% of the development, considered the best time to harvest peanut seeds with the highest quality. Our findings also support the idea that the adequate proportion of rainfall and thermal sum in the maternal environment are factors that favor the acquisition of peanut seed longevity., Competing Interests: The authors have declared that no competing interests exist.

Peanut is an important oilseed crop whose production is threatened by various abiotic and biotic stresses. Study of the molecular mechanism of salt tolerance could provide important information for the salt tolerance of this crop. WRKY transcription factors (TFs) are one of the largest TF families in plants and are involved in growth and development, defense regulation and the stress response. Here, we cloned a novel WRKY transcription factor gene belonging to the WRKY IIc subfamily, AhWRKY75, from the salt-tolerant mutant M34. The expression of AhWRKY75 was induced by NaCl stress treatment. After salt treatment, AhWRKY75-overexpressing peanuts grew better than wild-type plants. Furthermore, several genes related to the reactive oxygen species (ROS) scavenging system were up-regulated; the activities of superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT) were significantly higher in transgenic lines than in non-transgenic control plants; and the malondialdehyde (MDA) and superoxide anion contents were significantly lower in transgenic lines than in control plants. The net photosynthetic rate (Pn), stomatal conductance (GS) and transpiration rate (Tr) of transgenic lines were significantly higher in transgenic plants than in control plants, and the intercellular CO 2 concentration (Ci) was significantly lower in transgenic plants than in control plants. These results demonstrated that the AhWRKY75 gene conferred salt tolerance in transgenic peanut lines by improving the efficiency of the ROS scavenging system and photosynthesis under stress treatment. This study identifies a novel WRKY gene for enhancing the tolerance of peanut and other plants to salt stress., (Copyright © 2021 Elsevier Masson SAS. All rights reserved.)

Microbial symbiosis in legumes is achieved through nitrogen-fixing root nodules, and these are important for sustainable agriculture. The molecular mechanisms underlying development of root nodules in polyploid legume crops are largely understudied. Through map-based cloning and QTL-seq approaches, we identified a pair of homoeologous GRAS transcription factor genes, Nodulation Signaling Pathway 2 (AhNSP2-B07 or Nb) and AhNSP2-A08 (Na), controlling nodulation in cultivated peanut (Arachis hypogaea L.), an allotetraploid legume crop, which exhibited non-Mendelian and Mendelian inheritance, respectively. The segregation of nodulation in the progeny of Nananbnb genotypes followed a 3:1 Mendelian ratio, in contrast to the 5:3~1:1 non-Mendelian ratio for nanaNbnb genotypes. Additionally, a much higher frequency of the nb allele (13%) than the na allele (4%) exists in the peanut germplasm collection, suggesting that Nb is less essential than Na in nodule organogenesis. Our findings reveal the genetic basis of naturally occurred non-nodulating peanut plants, which can be potentially used for nitrogen fixation improvement in peanut. Furthermore, the results have implications for and provide insights into the evolution of homoeologous genes in allopolyploid species., (© The Author(s) 2020. Published by Oxford University Press on behalf of the Society for Experimental Biology. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Peanut is an important crash crop worldwide, and it is often threatened by drought stress due to unexpected extreme weather events. In this work, NH5 and FH18 were selected as drought-tolerant and drought-sensitive varieties, respectively. Comparison of their physiological responses revealed that NH5 showed less wilting, higher relative water content and lower water loss rate of detached leaves, lower electrolyte leakage, and stronger antioxidant ability under drought stress than did FH18. Based on comparative transcriptomic analysis, 5376 differentially expressed mRNAs were commonly identified in the two varieties, and 2993 genes specifically changed in the drought-tolerant variety and were mainly enriched in photosynthesis-antenna proteins and photosynthetic pathways. Furthermore, 73 microRNAs (miRNAs) were differentially expressed in the drought tolerance variety specifically under drought stress; of these, two key candidate miRNAs, novel miR_416 and novel miR_73, were identified, and the majority of their target genes were enriched in phenylpropanoid biosynthesis, linoleic acid metabolism, and cutin, suberine, and wax biosynthesis. This study lays the foundation for the analysis of the molecular mechanism of drought tolerance and promotes the genetic improvement of peanut drought tolerance.

Background: The peanut is one of the most important oil crops worldwide. Qualities and yields of peanut can be dramatically diminished by abiotic stresses particularly by drought. Therefore, it would be beneficial to gain a comprehensive understanding on peanut drought-responsive transcriptional regulatory activities, and hopefully to extract critical drought-tolerance-related molecular mechanism from it., Results: In this study, two peanut Arachis hypogaea L. varieties, NH5 (tolerant) and FH18 (sensitive), which show significantly differential drought tolerance, were screened from 23 main commercial peanut cultivars and used for physiological characterization and transcriptomic analysis. NH5 leaves showed higher water and GSH contents, faster stomatal closure, and lower relative conductivity (REC) than FH18. Under the time-course of drought-treatments 0 h (CK), 4 h (DT1), 8 h (DT2) and 24 h (DT3), the number of down-regulated differential expressed genes (DEGs) increased with the progression of treatments indicating repressive impacts on transcriptomes by drought in both peanut varieties., Conclusions: Nevertheless, NH5 maintained more stable transcriptomic dynamics than FH18. Furthermore, annotations of identified DEGs implicate signal transduction, the elimination of reactive oxygen species, and the maintenance of cell osmotic potential which are key drought-tolerance-related pathways. Finally, evidences from the examination of ABA and SA components suggested that the fast stomatal closure in NH5 was likely mediated through SA rather than ABA signaling. In all, these results have provided us a comprehensive overview of peanut drought-responsive transcriptomic changes, which could serve as solid foundation for further identification of the molecular drought-tolerance mechanism in peanut and other oil crops.

Understanding the changes in peanut (Arachis hypogaea L.) anther lipidome under heat stress (HT) will aid in understanding the mechanisms of heat tolerance. We profiled the anther lipidome of seven genotypes exposed to ambient temperature (AT) or HT during flowering. Under AT and HT, the lipidome was dominated by phosphatidylcholine (PC), phosphatidylethanolamine (PE), and triacylglycerol (TAG) species (> 50% of total lipids). Of 89 lipid analytes specified by total acyl carbons:total carbon-carbon double bonds, 36:6, 36:5, and 34:3 PC and 34:3 PE (all contain 18:3 fatty acid and decreased under HT) were the most important lipids that differentiated HT from AT. Heat stress caused decreases in unsaturation indices of membrane lipids, primarily due to decreases in highly-unsaturated lipid species that contained 18:3 fatty acids. In parallel, the expression of Fatty Acid Desaturase 3-2 (FAD3-2; converts 18:2 fatty acids to 18:3) decreased under HT for the heat-tolerant genotype SPT 06-07 but not for the susceptible genotype Bailey. Our results suggested that decreasing lipid unsaturation levels by lowering 18:3 fatty-acid amount through reducing FAD3 expression is likely an acclimation mechanism to heat stress in peanut. Thus, genotypes that are more efficient in doing so will be relatively more tolerant to HT.

Plants encounter diverse stressors simultaneously with changing environmental factors. The combined effect of different types of stresses can have a wide range of effects on plants. The present study demonstrated that various stress factors such as the combination of chemical elicitors, namely paraquat (PQ), methyl jasmonate (MeJA) and methyl-β-cyclodextrin (CD), light exposure versus darkness, and mechanical shearing stress affected the defence response in peanut hairy root culture. The antioxidant activities were dramatically increased at all time points after hairy roots were subjected to elicitation with PQ + MeJA + CD under root cutting in both light and dark conditions. The stilbene compounds were highly increased in the culture medium after elicitor treatment of uncut hairy roots under dark conditions. In contrast to the high stilbene contents detected in culture medium under dark conditions, the transcription of the stilbene biosynthesis genes PAL, RS and RS3 was enhanced by the effect of light in uncut hairy root tissues. The antioxidant enzyme genes APX, GPX and CuZn-SOD of uncut and cut hairy roots were more highly expressed in light conditions than in dark conditions. The pathogenesis-related protein (PR)-encoding genes chitinase, PR4A, PR5 and PR10 of uncut hairy roots were highly expressed in response to light conditions compared to dark conditions at all time points. Recent evidence of the production of antioxidant stilbene compounds and defence response genes has implicated plant protective functions through defence responses under different stress challenges. Plant responses might therefore be regulated by the coordination of different signal responses through dynamic pathways., (Copyright © 2020 Elsevier Masson SAS. All rights reserved.)

Peanut is widely grown and provides protein and edible oil for millions of people. Peanut growth and productivity are frequently negatively affected by abiotic and biotic environmental factors. However, the research on improving peanut germplasm resources by genetic transformation is very limited. Here, the novel R2R3-MYB repressor GmMYB3a was introduced into peanut plants by Agrobacterium-mediated transformation for the first time for thorough evaluation of the function of GmMYB3a in drought stress plant responses. We generated GmMYB3a-transgenic peanut plants. The GmMYB3a-overexpressing lines showed significantly improved physiological responses and no yield loss non-transgenic plants, in terms of survival rates. Thus, the GmMYB3a-overexpressing plants showed better photosynthetic performance, higher relative water content, and greater water use efficiency, demonstrating their adaptive capacity to water deficit. We conclude that overexpression of GmMYB3a can improve drought tolerance and productivity in peanut.

Arachis hypogaea L. (peanut) is a major oil yielding crop and its productivity is largely affected by the availability of nitrogen and phosphorus. The present study aims to elucidate the differential physiological and biochemical mechanisms involved in two contrasting genotypes of peanut for mitigation of N and/or P deficiency. The plants of two contrasting genotypes of peanut (GG7 and TG26) were subjected to N and/or P deficiency under hydroponic culture condition. After 15 d of N and/or P deficiency, various growth parameters, mineral nutrient status, nutrient use efficiency, photosynthesis, transpiration, water use efficiency, chlorophyll fluorescence, ROS level, and changes in enzymatic and non-enzymatic antioxidative components were measured in control and nutrient deficient plants. Our results showed that GG7 is fast-growing genotype than TG26 under control condition, whereas under N and/or P deficiency growth performance of GG7 was significantly declined as compared to TG26. The levels of photosynthetic pigments, net photosynthesis activity (P N ), and stomatal conductance (g s ) declined in N and/or P deficient plants of both the genotypes. However, quantum efficiency of photosystem II (Fv/Fm) did not change significantly under N and/or P starvation in both the genotypes. In the present investigation, most of the antioxidative enzymes either remained in steady state or downregulated in both the genotypes of peanut under N and/or P deficiency condition. N and/or P deficiency did not influence the levels of ROS and oxidative stress indicators such as O 2 ·- , H 2 O 2 , and MDA in both the genotypes. In the present investigation, the decline in growth in both the genotypes under N and/or P deficiency might be due to the reduced photosynthetic performance. Our results suggest that TG26 is more resistant to N and P deficiency than GG7 genotype. Higher NUE value of GG7 as compared to TG26 suggests that GG7 can utilize N more efficiently to promote biomass production than TG26 under sufficient nutrient condition. On the other hand, mineral resource allocation to leaf and higher PUE are key adaptive features of the TG26 genotype under N, and P deficiency conditions. The differential regulations of various enzymatic and non-enzymatic antioxidative components in peanut genotypes maintain the cellular redox homeostasis under mineral deficiency conditions and prevent the peanut plants from oxidative stress, thereby maintaining PSII efficiency. The information from the present study can be useful for the improvement of traits in peanut that can maintain the productivity under N and P deficient environment with minimum input of fertilizers., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Plants counteract Cd toxicity by activating cellular stress responses. The simultaneous exogenous application of methyl jasmonate (MeJA) and methyl-β-cyclodextrin (CD) before Cd exposure improved the response of Arachis hypogaea hairy root culture to the unfavourable effects of Cd toxicity. At 24 h after elicitation, genes that encode key enzymes in the phenylpropanoid biosynthesis pathway (i.e., PAL and RS3) were up-regulated to 3.2- and 5.4-fold changes respectively, thereby inducing stilbene production. The up-regulation of genes that encode transcription factors (i.e., ERF1 and ERF6) significantly increased the expression of several genes (PR4A, PR5, PR10, and chitinase) that encode the pathogenesis-related (PR) proteins to 25.8-, 45-, 5- and 12.6-fold changes, respectively. The more dramatic up-regulation of PR protein-encoding genes demonstrated the significant role of defence proteins in plant protective mechanisms. The prolonged (i.e., 72-h) treatment with MeJA + CD_Cd triggered adaptive responses by substantially increasing the levels of antioxidants, stilbenes, and other phenolic substances. These findings suggest that the interaction between signalling elicitors (MeJA and CD) and Cd modulates a complex signalling network for plant defence system., (Copyright © 2020 Elsevier GmbH. All rights reserved.)

Arsenic (As) can be present naturally in groundwater from peanut fields, constituting a serious problem, as roots can accumulate and mobilize the metalloid to their edible parts. Understanding the redox changes in the legume exposed to As may help to detect potential risks to human health and recognize tolerance mechanisms. Thirty-days old peanut plants inoculated with Bradyrhizobium sp. strains (SEMIA6144 or C-145) were exposed to a realistic arsenate concentration, in order to unravel the redox response and characterize the oxidative stress indexes. Thus, root anatomy, reactive oxygen species detection by fluorescence microscopy and, ROS histochemical staining along with the NADPH oxidase activity were analyzed. Besides, photosynthetic pigments and damage to lipids and proteins were determined as oxidative stress indicators. Results showed that at 3 μM As V , the cross-section areas of peanut roots were augmented; NADPH oxidase activity was significantly increased and O 2 ˙¯and H 2 O 2 accumulated in leaves and roots. Likewise, an increase in the lipid peroxidation and protein carbonyls was also observed throughout the plant regardless the inoculated strain, while chlorophylls and carotenes were increased only in those inoculated with Bradyrhizobium sp. C-145. Interestingly, the oxidative burst, mainly induced by the NADPH oxidase activity, and the consequent oxidative stress was strain-dependent and organ-differential. Additionally, As modifies the root anatomy, acting as a possibly first defense mechanism against the metalloid entry. All these findings allowed us to conclude that the redox response of peanut is conditioned by the rhizobial strain, which contributes to the importance of effectively formulating bioinoculants for this crop., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Flavonol is an important functional bioactive substance in peanut seeds, and plays important roles responding to abiotic stress. The flavonol content is closely related to the activity and regulation of gene expression patterns of flavonol synthase (FLS). In this study, eight FLS genes, AhFLSs were cloned and their expression characterization in different peanut organ and seedling under different abiotic stress were conducted. The results showed that the expressions levels of AhFLSs were differed in all assayed peanut organs and seedlings under abiotic stress treatments. Expression levels of AhFLS2, AhFLS3, AhFLS4, and AhFLS6 were higher than those of other AhFLSs. The flavonol contents of peanut organs and seedlings under different abiotic stress were also determined using high performance liquid chromatography (HPLC). Dried mature peanut seeds were the organ tissue with the highest flavonol content, and flavonol content increased with seed development. Under abiotic stress treatments, the types of flavonols induced differed among stress treatments. Correlation analysis results suggested that eight AhFLS genes may have different functions in peanut. Moreover, changes in the expression of the eight genes appear to has substrate preference. These results can lay the foundation for the study of improving nutritional value of peanut seed and resistance of peanut plant.

Background: Peanut is one of the most important oil crop species worldwide. NAC transcription factor (TF) genes play important roles in the salt and drought stress responses of plants by activating or repressing target gene expression. However, little is known about NAC genes in peanut., Results: We performed a genome-wide characterization of NAC genes from the diploid wild peanut species Arachis duranensis and Arachis ipaensis, which included analyses of chromosomal locations, gene structures, conserved motifs, expression patterns, and cis-acting elements within their promoter regions. In total, 81 and 79 NAC genes were identified from A. duranensis and A. ipaensis genomes. Phylogenetic analysis of peanut NACs along with their Arabidopsis and rice counterparts categorized these proteins into 18 distinct subgroups. Fifty-one orthologous gene pairs were identified, and 46 orthologues were found to be highly syntenic on the chromosomes of both A. duranensis and A. ipaensis. Comparative RNA sequencing (RNA-seq)-based analysis revealed that the expression of 43 NAC genes was up- or downregulated under salt stress and under drought stress. Among these genes, the expression of 17 genes in cultivated peanut (Arachis hypogaea) was up- or downregulated under both stresses. Moreover, quantitative reverse transcription PCR (RT-qPCR)-based analysis revealed that the expression of most of the randomly selected NAC genes tended to be consistent with the comparative RNA-seq results., Conclusion: Our results facilitated the functional characterization of peanut NAC genes, and the genes involved in salt and drought stress responses identified in this study could be potential genes for peanut improvement.

Long-term monoculture (LTM) decreases the yield and quality of peanut, even resulting in changes in the microbial community. However, the effect of LTM on peanut rhizobial communities has still not been elucidated. In this study, we isolated and characterized peanut rhizobia from 6 sampling plots with different monoculture cropping durations. The community structure and diversity index for each sampling site were analyzed, and the correlations between a peanut rhizobium and soil characteristics were evaluated to clarify the effects on peanut rhizobial communities. The competitive abilities among representative strains were also analyzed. A total of 283 isolates were obtained from 6 sampling plots. Nineteen recA haplotypes were defined and were grouped into 8 genospecies of Bradyrhizobium, with B. liaoningense and B. ottawaense as the dominant groups in each sample. The diversity indexes of the rhizobial community decreased, and the dominant groups of B. liaoningense and B. ottawaense were enriched significantly with extended culture duration. Available potassium (AK), available phosphorus (AP), available nitrogen (AN), total nitrogen (TN) and organic carbon (OC) gradually increased with increasing monoculture duration. OC, TN, AP and AK were the main soil characteristics affecting the distribution of rhizobial genospecies in the samples. A competitive nodulation test indicated that B. liaoningense presented an excellent competitive ability, which was congruent with its high isolation frequency. This study revealed that soil characteristics and the competitive ability of rhizobia shape the symbiotic rhizobial community and provides information on community formation and the biogeographic properties of rhizobia., (Copyright © 2020 Elsevier GmbH. All rights reserved.)

Drought negatively affects the growth and yield of terrestrial crops. Seed priming, pre-exposing seed to a compound, could induce improved tolerance and adaptation to stress in germinated plants. To understand the effects and regulatory mechanism of seed priming with brassinosteroid (BR) on peanut plants, we treated seeds with five BR concentrations and examined dozens of physiological and biochemical features, and transcriptomic changes in leaves under well-watered and drought conditions. We found optimal 0.15 ppm BR priming could reduce inhibitions from drought and increase the yield of peanut, and priming effects are dependent on stage of plant development and duration of drought. BR priming induced fewer differentially expressed genes (DEGs) than no BR priming under well-watered condition. Drought with BR priming reduced the number of DEGs than drought only. These DEGs were enriched in varied gene ontologies and metabolism pathways. Downregulation of DEGs involved in both light perceiving and photosynthesis in leaves is consistent with low parameters of photosynthesis. Optimal BR priming partially rescued the levels of growth promoting auxin and gibberellin which were largely reduced by drought, and increased levels of defense associated abscisic acid and salicylic acid after long-term drought. BR priming induced many DEGs which function as kinase or transcription factor for signal cascade under drought. We proposed BR priming-induced regulatory responses will be memorized and recalled for fast adaptation in later drought stress. These results provide physiological and regulatory bases of effects of seed priming with BR, which can help to guide the framing improvement under drought stress.

The beneficial associations between Arachis hypogaea L. (peanut) and fluorescent Pseudomonas species have been poorly explored despite their predominance in the peanut rhizosphere. The present study explores the mutually beneficial interactions between peanut roots and P. aeruginosa P4 (P4) in terms of their impact on plant growth, defence physiology and the root-rhizobacterial interface. The efficient phosphate solubilizer P4 exhibited biocontrol abilities, including the production of siderophores, pyocyanin, indole-3-acetic acid and hydrogen cyanide. The bacterization of peanut seeds with multi-potential P4 significantly enhanced in vitro seed germination and seedling vigour. Under sand-based gnotobiotic (10 days post-inoculation) and sterile soil-based cultivation systems (30 days post-inoculation), sustained P4 colonization enhanced the peanut root length and dry plant biomass. The subsequent increase in catalase, polyphenol oxidase and phenylalanine ammonia lyase activities with increased phenolic contents in the peanut roots and shoots suggested the systemic priming of defences. Consequently, the altered root exudate composition caused enhanced chemo-attraction towards P4 itself and the symbiotic N 2 -fixing Bradyrhizobium strain. Co-inoculating peanuts with P4 and Bradyrhizobium confirmed the improved total bacterial colonization (∼2 fold) of the root tip, with the successful co-localization of both, as substantiated by scanning electron microscopy. Collectively, the peanut-P4 association could potentially model the beneficial Pseudomonas-driven multi-trophic rhizosphere benefits, emphasizing the plausible role of non-rhizobium PGPR in promoting N 2 fixation., Competing Interests: Declaration of Competing Interest None., (Copyright © 2020. Published by Elsevier GmbH.)

The subspecies fastigiata of cultivated groundnut lost fresh seed dormancy (FSD) during domestication and human-made selection. Groundnut varieties lacking FSD experience precocious seed germination during harvest imposing severe losses. Development of easy-to-use genetic markers enables early-generation selection in different molecular breeding approaches. In this context, one recombinant inbred lines (RIL) population (ICGV 00350 × ICGV 97045) segregating for FSD was used for deploying QTL-seq approach for identification of key genomic regions and candidate genes. Whole-genome sequencing (WGS) data (87.93 Gbp) were generated and analysed for the dormant parent (ICGV 97045) and two DNA pools (dormant and nondormant). After analysis of resequenced data from the pooled samples with dormant parent (reference genome), we calculated delta-SNP index and identified a total of 10,759 genomewide high-confidence SNPs. Two candidate genomic regions spanning 2.4 Mb and 0.74 Mb on the B05 and A09 pseudomolecules, respectively, were identified controlling FSD. Two candidate genes-RING-H2 finger protein and zeaxanthin epoxidase-were identified in these two regions, which significantly express during seed development and control abscisic acid (ABA) accumulation. QTL-seq study presented here laid out development of a marker, GMFSD1, which was validated on a diverse panel and could be used in molecular breeding to improve dormancy in groundnut., (© 2019 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd.)

Background: Stress-induced hormones are essential for plants to modulate their microbiota and dynamically adjust to the environment. Despite the emphasis of the role of the phytohormone ethylene in the plant physiological response to heterospecific neighbour detection, less is known about how this activated signal mediates focal plant rhizosphere microbiota to enhance plant fitness. Here, using 3 years of peanut (Arachis hypogaea L.), a legume, and cyanide-containing cassava (Manihot esculenta Crantz) intercropping and peanut monocropping field, pot and hydroponic experiments in addition to exogenous ethylene application and soil incubation experiments, we found that ethylene, a cyanide-derived signal, is associated with the chemical identification of neighbouring cassava and the microbial re-assemblage in the peanut rhizosphere., Results: Ethylene production in peanut roots can be triggered by cyanide production of neighbouring cassava plants. This gaseous signal alters the microbial composition and re-assembles the microbial co-occurrence network of peanut by shifting the abundance of an actinobacterial species, Catenulispora sp., which becomes a keystone in the intercropped peanut rhizosphere. The re-assembled rhizosphere microbiota provide more available nutrients to peanut roots and support seed production., Conclusions: Our findings suggest that root ethylene acts as a signal with a dual role. It plays a role in perceiving biochemical cues from interspecific neighbours, and also has a regulatory function in mediating the rhizosphere microbial assembly, thereby enhancing focal plant fitness by improving seed production. This discovery provides a promising direction to develop novel intercropping strategies for targeted manipulations of the rhizosphere microbiome through phytohormone signals. Video abstract.

Crop plants always encounter multiple stresses in the natural environment. Here, the effects of the fungicide zinc thiazole (ZT) on propagation of Ralstonia solanacearum, a bacterial pathogen, were investigated in peanut seedlings under salt stress. Compared with water control, salt stress markedly reduced pathogen resistance in peanut seedlings. However, impaired pathogen resistance was alleviated by treatment with dimethylthiourea, a specific ROS scavenger, or ZT. Subsequently, salt stress or combined salt and pathogen treatment resulted in inhibition of photosynthesis, loss of chlorophyll and accumulation of thiobarbituric acid reactive substances, which could be reversed by ZT. In addition, ZT treatment suppressed the salt stress up-regulated Na+ content and Na+/K+ ratios in peanut roots. Furthermore, salt stress or combined salt and pathogen treatment impaired the activities of antioxidant (e.g. superoxide dismutase/SOD and catalase/CAT), and defense-related (e.g. phenylalanine ammonia lyase /PAL and polyphenol oxidase/PPO) enzymes, which could be rescued by addition of ZT. In contrast, only slight changes of SOD and CAT activities were observed in pathogen-infected seedlings. Similarly, activities of PAL and PPO were slightly modified by salt stress in peanut seedlings. These results suggest that the ZT-enhanced pathogen resistance can be partly attributed to the improvement of photosynthetic capacity and defense enzyme activities, and also the inhibition of Na+/K+ ratios, in this salt-stressed crop plant., Competing Interests: The authors have declared that no competing interests exist.

Water deficit is one of the most serious environmental factors that affect the productivity of crops in the world. Arachis hypogaea is a legume with a high nutritional value and 70% is cultivated in semi-arid regions. This research aimed to study the effect of water deficit on peanut root exudates composition, analyzing the importance of exudates on peanut-PGPR interaction under restrictive water condition. Peanut seedlings were subjected to six treatments: 0 and 15 mM PEG, in combination with non-inoculated, Bradyrhizobium sp. and Bradyrhizobium-Azospirillum brasilense inoculated treatments. We analyzed the 7-day peanut root exudate in response to a water restrictive condition and the presence of bacterial inocula. Molecular analysis was performed by HPLC, UPLC and GC. Bacteria motility, chemotaxis, bacterial adhesion to peanut roots and peanut growth parameters were analyzed. Restrictive water condition modified the pattern of molecules exuded by roots, increasing the exudation of Naringenin, oleic FA, citric and lactic acid, and stimulation the release of terpenes of known antioxidant and antimicrobial activity. The presence of microorganisms modified the composition of root exudates. Water deficit affected the first events of peanut-PGPR interaction and the root exudates favored bacterial mobility, the chemotaxis and attachment of bacteria to peanut roots. Changes in the profile of molecules exuded by roots allowed A. hypogaea-Bradyrhizobium and A.hypogaea-Bradyrhizobium-Azospirillum interaction thus reversing the negative effects of restrictive water condition on peanut growth. These findings have a future potential application to improve plant-PGPR interactions under water deficit by formulating inoculants containing key molecules exuded during stress., (Copyright © 2019 Elsevier Masson SAS. All rights reserved.)

Endophytic fungi play important roles in the modification of ecosystem productivity; however, the underlying mechanisms are only partly understood. A 2-year field plot experiment verified that the endophytic fungus Phomopsis liquidambaris increased peanut ( Arachis hypogaea L.) yields and significantly increased nodulation and N 2 fixation regardless of whether N fertilizers were added. Root exudates collected from P. liquidambaris -colonized plants significantly improved nodulation and N 2 fixation. Rhizosphere stimulation experiments further showed that colonized root exudates had significantly decreased soil nitrate (NO 3 - ) concentrations, with decreased abundance and diversity of ammonia oxidizing archaea (AOA). In contrast, the abundance and diversity of diazotrophs significantly increased, and most diazotrophs identified were peanut nodulation-related strains ( Bradyrhizobium sp.). P. liquidambaris symbiosis increased the expression of phenolic and flavonoid synthesis-related genes, and the derived phenolics and flavonoids could effectively increase the chemotaxis, biofilm formation, and nodC gene expression (nodulation-related biological processes) of the Bradyrhizobium strain. Metabolic pattern analysis showed that phenolics and flavonoids are more likely to accumulate to higher levels in the rhizosphere soil of peanuts colonized with P. liquidambaris Finally, a synthetic root exudate experiment further confirmed the underlying mechanisms for the P. liquidambaris -induced improvement in nodulation and N 2 fixation, i.e., that the specific root exudates derived from P. liquidambaris colonization decrease nitrate concentration and increase the population and biological activities of peanut nodulation-related Bradyrhizobium species, which beneficially enhance peanut- Bradyrhizobium interactions. Therefore, this study is the first to provide new insight into a positive relationship between an exotic endophytic fungus, crop nodulation, and N 2 fixation increase. IMPORTANCE Endophytic fungi play an important role in balancing the ecosystem and boosting host growth; however, the underpinning mechanisms remain poorly understood. Here, we found that endophytic fungal colonization with P. liquidambaris significantly increased the productivity, nodulation, and N 2 fixation of peanuts through the secretion of specific root exudates. We provide a reasonable mechanism explaining how P. liquidambaris promotes peanut nodulation and N 2 fixation, whereby the specific root exudates produced by P. liquidambaris colonization decrease rhizosphere soil nitrate (NO3 - ) and increase the population and biological activities of peanut-nodulating-related Bradyrhizobium strains, which is beneficial to enhancing the peanut- Bradyrhizobium symbiotic interaction. Our study provides reliable empirical evidence to show the mechanism of how an exotic endophytic fungus drives an increase in nodulation and N 2 fixation, which will be helpful in erecting a resource-efficient and sustainable agricultural system., (Copyright © 2019 Xie et al.)

This study was conducted to investigate the changes in leaf physiological parameters to abiotic stress induced by different levels of cement dust. On day 15, Arachis hypogaea L. plants (sowing day was considered as day 0) were divided into six groups, and cement was sprinkled over plants with the help of hand pump, twice a week at T 1 (5 g pot -1 ), T 2 (8 g pot -1 ), T 3 (10 g pot -1 ), T 4 (15 g pot -1 ), T 5 (20 g pot -1 ), and T 0 /control (0 g pot -1 ), until fruit maturity. Morphometric parameters such as root and shoot length, leaf area, and seed weight were significantly higher in T 0 , while the minimum was recorded in T 5 . Physiological analyses of leaves and roots revealed a remarkable reduction (p < 0.05) in sugar, amino acid, and protein contents, while the concentration of enzymatic antioxidants was increased in cement-treated plants. The concentration of abscisic acid in leaves was significantly higher in treatment groups as compared with control, while gibberellic acid concentration was low. Strikingly, cement dust decreases the level of leaf photosynthetic pigments, reduces stomatal conductance, and adversely affects photosynthesis. Leaf histological analysis revealed confirmatory evidence of stomatal closure, cell damage, reduced cell area, and abridged leaf thickness. Salient features of the present study provide useful evidence to estimate cement dust as a critical abiotic stress factor, which has adverse effects on photosynthesis, leaf anatomical features, stomatal functioning, and productivity. Our work opens new avenues for a deep portfolio of cement-based stress mediating pathophysiology in Arachis hypogaea.

ABA is an important messenger that acts as a signaling mediator for regulating the adaptive response of plants to drought stress. Two production pathways, de novo biosynthesis and hydrolysis of glucose-conjugated ABA by β-glucosidase (BG), increase cellular ABA levels in plants. ABA catabolism via hydroxylation by 8'-hydroxylase (CYP707A), or conjugation by uridine diphosphate glucosyltransferase (UGT), decreases cellular ABA levels. The transport of ABA through ATP-binding cassette (ABC)-containing transporter proteins, members of ABC transporter G family (ABCG), across plasma membrane (PM) is another important pathway to regulate cellular ABA levels. In this study, based on our previously constructed transcriptome of peanut leaves in response to drought stress, fourteen candidate genes involved in ABA production (including AhZEP, AhNCED1 and AhNCED3, AhABA2, AhAAO1 and AhAAO2, AhABA3, AhBG11 and AhBG24), catabolism (including AhCYP707A3, AhUGT71K1 and AhUGT73B4) and transport (including AhABCG22-1 and AhABCG22-2), were identified homologously and phylogenetically, and further analyzed at the transcriptional level by real-time RT-PCR, simultaneously determining ABA levels in peanut leaves in response to drought. The high sequence identity and very similar subcellular localization of the proteins deduced from 14 identified genes involved in ABA production, catabolism and transport with the reported corresponding enzymes in databases suggest their similar roles in regulating cellular ABA levels. The expression analysis showed that the transcripts of AhZEP, AhNCED1, AhAAO2 and AhABA3 instead of AhABA2, AhNCED3 and AhAAO1 in peanut leaves increased significantly in response to drought stress; and that the AhBG11 and AhBG24 mRNA levels were rapidly and significantly up-regulated, with a 4.83- and 4.58-fold increase, respectively at 2-h of drought stress. The genes involved in ABA catabolism AhCYP707A3, AhUGT71K1 instead of AhUGT73B4 were significantly induced in response to drought stress. The expression of two closely related peanut ABCG genes, AhABCG22.1 and AhABCG22.2, was significantly up-regulated in response to drought stress. The ABA levels rapidly began to accumulate within 2 h (a 56.6-fold increase) from the start of drought stress, and peaked at 10 h of the stress. The highly and rapidly stress up-regulated expressions of genes involved in ABA production and transport, particularly AhNCED1, AhBG11 and AhBG24, and AhABCG22.1 and AhABCG22.2, might contribute to the rapid ABA accumulation in peanut leaves in response to drought. In response to drought stress, ABA accumulation levels in peanut leaves agree well with the up-regulated expressions of ABA-producing genes (AhZEP, AhNCED1, AhAAO2, AhABA3, AhBG11 and AhBG24) and PM-localized ABA importer genes (AhABCG22-1 and AhABCG22-2), in spite of the simultaneously induced ABA catabolic genes (AhCYP707A3 and AhUGT71K1), although the induction of catabolic genes was much lower than that of biosynthetic gene (AhNCED1). This difference in induction kinetics of gene expression may define the significant accumulation of drought-induced ABA levels. These results suggest that ABA homeostasis in peanut leaves in response to drought maintained through a balance between the production, catabolism and transport, rather than simply by the biosynthesis., Competing Interests: The authors have declared that no competing interests exist.

The peanut red spider mite, Tetranychus ogmophallos Ferreira and Flechtmann (Acari: Tetranychidae), is an important pest of peanut in Brazil and is considered a quarantine pest in other countries. This study investigates the development, reproduction, survival and life table parameters of T. ogmophallos on five peanut cultivars-three with high levels of oleic acid [high oleic] and recently released: Granoleico, IAC OL 3 and IAC 503; and two regular and previously used by farmers: Runner IAC 886, IAC Tatu ST 3-and two breeding lines (L. 8008 and L. 322) in the growth chamber. There were differences between the developmental times of all immature stages, the oviposition period, fecundity and adult longevity of T. ogmophallos reared on the peanut cultivars and breeding lines. The longest duration of the immature stage and lowest fecundity occurred on cultivars Granoleico and Runner IAC 886, and breeding line L. 322, which also displayed the lowest intrinsic rate of increase (r), finite rate of increase (λ) and net reproductive rate (R 0 ) and the shortest mean generation time (T). The highest fitness occurred on the IAC Tatu ST, IAC OL 3, IAC 503 and L. 8008. Our study shows that peanut cultivars belonging to the group with high oleic trait and recently released (IAC OL 3 and IAC 503) are susceptible to T. ogmophallos, except Granoleico, which is resistant. The regular and previously used cultivar Runner IAC 886 and breeding line L. 322 are resistant to the peanut red spider mite.

Although citrate transporters are involved in iron (Fe) translocation and aluminum (Al) tolerance in plants, to date none of them have been shown to confer both biological functions in plant species that utilize Fe-absorption Strategy I. In this study, we demonstrated that AhFRDL1, a citrate transporter gene from peanut (Arachis hypogaea) that is induced by both Fe-deficiency and Al-stress, participates in both root-to-shoot Fe translocation and Al tolerance. Expression of AhFRDL1 induced by Fe deficiency was located in the root stele, but under Al-stress expression was observed across the entire root-tip cross-section. Overexpression of AhFRDL1 restored efficient Fe translocation in Atfrd3 mutants and Al resistance in AtMATE-knockout mutants. Knocking down AhFRDL1 in the roots resulted in reduced xylem citrate and reduced concentrations of active Fe in young leaves. Furthermore, AhFRDL1-knockdown lines had reduced root citrate exudation and were more sensitive to Al toxicity. Compared to an Al-sensitive variety, enhanced AhFRDL1 expression in an Fe-efficient variety contributed to higher levels of Al tolerance and Fe translocation by promoting citrate secretion. These results indicate that AhFRDL1 plays a significant role in Fe translocation and Al tolerance in Fe-efficient peanut varieties under different soil-stress conditions. Given its dual biological functions, AhFRDL1 may serve as a useful genetic marker for breeding for high Fe efficiency and Al tolerance., (© The Author(s) 2019. Published by Oxford University Press on behalf of the Society for Experimental Biology. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

High oil and protein content make tetraploid peanut a leading oil and food legume. Here we report a high-quality peanut genome sequence, comprising 2.54 Gb with 20 pseudomolecules and 83,709 protein-coding gene models. We characterize gene functional groups implicated in seed size evolution, seed oil content, disease resistance and symbiotic nitrogen fixation. The peanut B subgenome has more genes and general expression dominance, temporally associated with long-terminal-repeat expansion in the A subgenome that also raises questions about the A-genome progenitor. The polyploid genome provided insights into the evolution of Arachis hypogaea and other legume chromosomes. Resequencing of 52 accessions suggests that independent domestications formed peanut ecotypes. Whereas 0.42-0.47 million years ago (Ma) polyploidy constrained genetic variation, the peanut genome sequence aids mapping and candidate-gene discovery for traits such as seed size and color, foliar disease resistance and others, also providing a cornerstone for functional genomics and peanut improvement.

Microorganisms associated with roots are thought to be part of the so-called extended plant phenotypes with roles in the acquisition of nutrients, production of growth hormones, and defense against diseases. Since the crops selectively enrich most rhizosphere microbes out of the bulk soil, we hypothesized that changes in the composition of bulk soil communities caused by agricultural management affect the extended plant phenotype. In the current study, we performed shotgun metagenome sequencing of the rhizosphere microbiome of the peanut (Arachis hypogaea) and metatranscriptome analysis of the roots of peanut plants grown in the soil with different management histories, peanut monocropping and crop rotation. We found that the past planting record had a significant effect on the assembly of the microbial community in the peanut rhizosphere, indicating a soil memory effect. Monocropping resulted in a reduction of the rhizosphere microbial diversity, an enrichment of several rare species, and a reduced representation of traits related to plant performance, such as nutrients metabolism and phytohormone biosynthesis. Furthermore, peanut plants in monocropped soil exhibited a significant reduction in growth coinciding with a down-regulation of genes related to hormone production, mainly auxin and cytokinin, and up-regulation of genes related to the abscisic acid, salicylic acid, jasmonic acid, and ethylene pathways. These findings suggest that land use history affects crop rhizosphere microbiomes and plant physiology.

Background: Plant basic leucine zipper (bZIP) transcription factors play crucial roles in plant growth, development, and abiotic stress responses. However, systematic investigation and analyses of the bZIP gene family in peanut are lacking in spite of the availability of the peanut genome sequence., Results: In this study, we identified 50 and 45 bZIP genes from Arachis duranensis and A. ipaensis genomes, respectively. Phylogenetic analysis showed that Arachis bZIP genes were classified into nine groups, and these clusters were supported by several group-specific features, including exon/intron structure, intron phases, MEME motifs, and predicted binding site structure. We also identified possible variations in DNA-binding-site specificity and dimerization properties among different Arachis bZIPs by inspecting the amino acid residues at some key sites. Our analysis of the evolutionary history analysis indicated that segmental duplication, rather than tandem duplication, contributed greatly to the expansion of this gene family, and that most Arachis bZIPs underwent strong purifying selection. Through RNA-seq and quantitative real-time PCR (qRT-PCR) analyses, the co-expressed, differentially expressed and several well-studied homologous bZIPs were identified during seed development stages in peanut. We also used qRT-PCR to explore changes in bZIP gene expression in response to salt-treatment, and many candidate bZIPs in groups A, B, and S were proven to be associated with the salt-stress response., Conclusions: This study have conducted a genome-wide identification, characterization and expression analysis of bZIP genes in Arachis genomes. Our results provide insights into the evolutionary history of the bZIP gene family in peanut and the funcntion of Arachis bZIP genes during seed development and in response to salt stress.

Drought is known to limit carbon assimilation in plants. However, it has been debated whether photosynthesis is primarily inhibited by stomatal or non-stomatal factors. This research assessed the underlying limitations to photosynthesis in peanuts (Arachis hypogaea L.) grown under progressive drought. Specifically, field-grown peanut plants were exposed to either well-watered or drought-stressed conditions during flowering. Measurements included survey measurements of gas exchange, chlorophyll fluorescence, PSII thermotolerance, pigment content, and rapid A-C i response (RACiR) assessments. Drought significantly decreased stomatal conductance with consequent declines in photosynthesis (A N ), actual quantum yield of PSII, and electron transport rate (ETR). Pigment contents were variable and depended on stress severity. Stomatal closure on stressed plants resulted in higher leaf temperatures, but F v /F m and PSII thermotolerance were only slightly affected by drought. A strong, hyperbolic relationship was observed between stomatal conductance, A N , and ETR. However, when RACiR analysis was conducted, drought significantly decreased A N at C i values comparable to drought-stressed plants, indicating non-stomatal limitations to A N . The maximum rate of carboxylation and maximum electron transport rate were severely limited by drought, and chloroplast CO 2 concentration (C C ) declined substantially under drought along with a comparable increase in partitioning of electron flow to photorespiration. Thus, while stomatal conductance may be a viable reference indicator of water deficit stress in peanut, we conclude that declines in A N were largely due to non-stomatal (diffusional and metabolic) limitations. Additionally, this is the first study to apply the rapid A-C i response method to peanut, with comparable results to traditional A-C i methods., (Published by Elsevier GmbH.)

The distribution of rhcRST and rhcJ-C1 fragments located in different loci of the type III secretion system (T3SS) gene cluster in the peanut-nodulating bradyrhizobia isolated from Guangdong Province, China was investigated by PCR-based sequencing. T3SS was detected in approximately one-third of the peanut bradyrhizobial strains and the T3SS-harboring strains belonging to different Bradyrhizobium genomic species. Diverse T3SS groups corresponding to different symbiotic gene types were defined among the 23 T3SS-harboring strains. The same or similar T3SS genes were detected in different genospecies, indicating that interspecies horizontal transfer of symbiotic genes had occurred in the Bradyrhizobium genus., (Copyright © 2018. Published by Elsevier GmbH.)

A gene encoding a serine-rich DnaJIII protein called AdDjSKI that has a 4Fe-4S cluster domain was found to be differentially upregulated in the wild peanut, Arachis diogoi in its resistance responses against the late leaf spot causing fungal pathogen Phaeoisariopsis personata when compared with the cultivated peanut, Arachis hypogaea. AdDjSKI is induced in multiple stress conditions in A. diogoi. Recombinant E. coli cells expressing AdDjSKI showed better growth kinetics when compared with vector control cells under salinity, osmotic, acidic and alkaline stress conditions. Overexpression of this type three J-protein potentiates not only abiotic stress tolerance in Nicotiana tabacum var. Samsun, but also enhances its disease resistance against the phytopathogenic fungi Phytophthora parasitica pv nicotianae and Sclerotinia sclerotiorum. In the present study we show transcriptional upregulation of APX, Mn-SOD and HSP70 under heat stress and increased transcripts of PR genes in response to fungal infection. This transmembrane-domain-containing J protein displays punctate localization in chloroplasts. AdDjSKI appears to ensure proper folding of proteins associated with the photosynthetic machinery under stress., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Drought is one of the environmental factors that most affects peanut cultivation in semi-arid regions, resulting in economic losses to growers. However, growth promoting bacteria are able to reduce water deficit damage in some plant species. In this context, this study aimed to evaluate the interaction of Bradyrhizobium strains reducing water stress effects on peanut genotypes by antioxidant enzymes activities, leaf gas exchanges and vegetative growth, as well as to determine the taxonomic positioning of strain ESA 123. The 16S rRNA gene of ESA 123 was amplified by PCR and sequenced by dideoxy Sanger sequencing method. An experiment was performed in greenhouse with three peanut genotypes (BRS Havana, CNPA 76 AM and 2012-4), two Bradyrhizobium strains (SEMIA 6144 and ESA 123), a mineral source of N and an absolute control (without N) under two water regimes (with and without irrigation). Seeds of peanut were sown and the plants were grown until 30 days after emergence. On the 20th day, the water deficit plants group had their irrigation suspended for 10 days. At in silico analyzes, ESA 123 presented 98.97% similarity with the type strain of B. kavangense. Leaf gas exchange was affected by water deficit; as well as alteration of antioxidant activities and reduction of vegetative growth variables. However, some plants inoculated with SEMIA 6144 and ESA 123 strains presented lower reductions and increment of some evaluated variables, mainly the ones inoculated with the ESA 123 strain, Bradyrhizobium sp. from the semi-arid region of Northeast Brazil. This data suggests beneficial effects of the peanut-Bradyrhizobium interaction in a water stress condition, specially with the ESA 123 strain.

Wild peanut relatives (Arachis spp.) are genetically diverse and were selected throughout evolution to a range of environments constituting, therefore, an important source of allelic diversity for abiotic stress tolerance. In particular, A. duranensis and A. stenosperma, the parents of the reference Arachis A-genome genetic map, show contrasting transpiration behavior under limited water conditions. This study aimed to build a comprehensive gene expression profile of these two wild species under dehydration stress caused by the withdrawal of hydroponic nutrient solution. For this purpose, roots of both genotypes were collected at seven time-points during the early stages of dehydration and used to construct cDNA paired-end libraries. Physiological analyses indicated initial differences in gas exchange parameters between the drought-tolerant genotype of A. duranensis and the drought-sensitive genotype of A. stenosperma. High-quality Illumina reads were mapped against the A. duranensis reference genome and resulted in the identification of 1,235 and 799 Differentially Expressed Genes (DEGs) that responded to the stress treatment in roots of A. duranensis and A. stenosperma, respectively. Further analysis, including functional annotation and identification of biological pathways represented by these DEGs confirmed the distinct gene expression behavior of the two contrasting Arachis species genotypes under dehydration stress. Some species-exclusive and common DEGs were then selected for qRT-PCR analysis, which corroborated the in silico expression profiling. These included genes coding for regulators and effectors involved in drought tolerance responses, such as activation of osmosensing molecular cascades, control of hormone and osmolyte content, and protection of macromolecules. This dataset of transcripts induced during the dehydration process in two wild Arachis genotypes constitute new tools for the understanding of the distinct gene regulation processes in these closely related species but with contrasting drought responsiveness. In addition, our findings provide insights into the nature of drought tolerance in wild germoplasm, which might be explored as novel sources of diversity and useful wild alleles to develop climate-resilient crop varieties., Competing Interests: The authors have declared that no competing interests exist.