Pea (Pisum sativum L.) is a globally important legume crop cultivated for food or feed. Fusarium wilt, caused by Fusarium oxysporum f. sp. pisi (Fop), is a major constraint on pea production worldwide. Despite China being one of the leading pea producers, knowledge of pea Fusarium wilt and Fop remains relatively limited. In this study, the pea Fusarium wilt samples were collected from 10 provinces between 2015 and 2020 in China, and 79 pathogenic F. oxysporum isolates were obtained. Race identification using a standard differential set revealed that 49 isolates belonged to race 1, 5 to race 2, 16 to race 5, and 9 represented novel virulence types. Screening for 14 Secreted in Xylem (SIX) genes demonstrated that SIX14 was universally present, suggesting a potential role in pathogenicity, whereas other SIX genes occurred in variable combinations that were partially associated with race differentiation. Multi-locus phylogenetic analyses based on five conserved loci resolved the isolates into nine phylogenetic lineages, including five corresponding to known species (F. odoratissimum, F. cugenangense, F. curvatum, F. nirenbergiae, and F. fabacearum) and four potentially novel species. Morphological observations of representative isolates further supported phylogenetic distinctions, although this subtle variation requires additional examination for formal species delimitation. Host range tests revealed that several pea-derived isolates also infected chickpea, lentil, mung bean, faba bean, and common bean, underscoring their broad pathogenic potential within legumes. This study provides the first comprehensive characterization of pea Fusarium wilt pathogens in China, integrating race structure, effector gene distribution, phylogenetic relationships, and host range. The findings highlight the complexity of Fop populations and provide a foundation for improved disease management and resistance breeding.

Soybean (Glycine max) is an important legume crop worldwide. An emerging leaf spot was observed in soybean plants with obvious black necrotic spot symptoms during the disease survey in Changping District, Beijing, China. To confirm the causal agent, the pathogen was isolated from the diseased leaves. Three isolates were obtained and showed a morphology extremely similar to Paramyrothecium vignicola. The isolates were identified by morphological and molecular characteristics. Phylogenetic analyses were performed using multiple gene regions (ITS, cmdA, rpb2, and tub2). The result indicated that the three isolates showed a high similarity (100%) with the known P. vignicola strains. Pathogenicity and host range tests of the isolates were performed on soybean and other legume crops. Three isolates were strongly pathogenic to soybean, hyacinth bean, common bean, faba bean, pea, mung bean, and lentil; moderate pathogenicity on adzuki bean; mild pathogenicity on cowpea and peanut. To screen resistant germplasms for disease control, the screening experiment of inoculum concentration of P. vignicola were performed. The result showed the most suitable concentration of P. vignicola isolate is 1 × 105 spores/mL for evaluation of germplasms resistance. Paramyrothecium species have been frequently identified to cause leaf spot and blight disease on a wide range of vegetables, ornamental plants, and economic crops. To our knowledge, this is the first report of P. vignicolaa inducing leaf spot on soybean worldwide. This study indicates P. vignicola might pose a potential risk to legume crops in the future.

Fusarium wilt (FW), caused by Fusarium oxysporum, poses a significant threat to mungbean (Vigna radiata L.), impacting its yield and quality. In this study, a recombinant inbred line (RIL) population was developed by crossing the highly resistant cultivar Weilv 9002-341 with the highly susceptible line V1128. Assessment of resistance revealed a continuous variation in the average disease index within the resulting population, consistent with the inheritance pattern of quantitative traits. Leveraging an F2:3 segregating population, we conducted linkage mapping analysis and bulked segregant analysis by sequencing, leading to the construction of a genetic linkage map and the identification of a region correlated with resistance. Within this region, 14 novel simple sequence repeat markers were designed to enable refined mapping. A putative resistance locus, spanning 0.17 Mb and encompassing 19 annotated genes, was precisely located. Ultimately, two genes were identified as high-priority candidates conferring resistance. The results of this study lay the foundation for the functional investigation of genes associated with resistance to Fusarium wilt disease in mungbean.

Pea (Pisum sativum L.) is one of the most important edible legumes in China, with both planting area and total yield ranking among the highest in the world. Fusarium wilt, caused by Fusarium oxysporum f. sp. pisi (Fop), is a severe factor limiting pea production. The deployment of resistant pea cultivars is the most effective and sustainable strategy for controlling this disease. In the present study, a novel resistance gene PsFwC9, conferring resistance to Fop race 5, was identified in the resistant pure line Chengwan 9-8 (CW9-8), and its candidate gene Psat4g213640 was characterized and functionally validated to be associated with disease resistance. Genetic analysis of the F₂ population derived from the cross between the resistant parent CW9-8 and the susceptible parent Chengwan 9-1 (CW9-1) revealed that PsFwC9 was controlled by a single dominant gene. Based on whole-genome resequencing, bulked segregant analysis sequencing (BSA-seq) and fine mapping, PsFwC9 was localized to an 817.06-kb region on chromosome 4 (i.e. linkage group IV, chr4LG4), flanked by KASP markers A016508 and A016511, and co-segregated with four markers. Haplotype analysis revealed that only the marker A016615 was significantly associated with Fusarium wilt resistance, and this marker was designated as a diagnostic marker for PsFwC9. Marker A016615 was located at 425 699 725 bp on chr4LG4, corresponding to the 277 bp within Psat4g213640, where a 'A/G' single-nucleotide polymorphism caused an amino acid substitution leading to an alteration in protein structure; therefore, Psat4g213640 was identified as the PsFwC9 candidate gene. Quantitative real-time PCR analysis showed no significant difference in the expression levels of Psat4g213640 between CW9-8 and CW9-1. Overexpression of the candidate gene Psat4g213640CW9-8 in the hairy root system significantly enhanced the resistance of CW9-1 to Fusarium wilt, whereas RNA interference-mediated silencing of Psat4g213640CW9-8 reduced the resistance of CW9-8, indicating that Psat4g213640CW9-8 played a crucial role in pea resistance to Fusarium wilt. In addition, subcellular localization showed that the protein encoded by Psat4g213640 was targeted to the endoplasmic reticulum. Collectively, these findings not only enriched the gene resources for disease resistance in pea and provided an important foundation for elucidating the molecular mechanism of PsFwC9-mediated resistance, but also provided important technical support for the practical application of molecular breeding for disease resistance in pea.

Reproductive health is a critical determinant of livestock productivity and economic sustainability. However, it is often compromised by infectious diseases, environmental stressors, and nutritional deficits. Phytogenic extracts—bioactive compounds derived from medicinal plants—have emerged as sustainable alternatives to synthetic antibiotics and hormones, exhibiting antimicrobial, antioxidant, and immunomodulatory properties. These extracts influence key reproductive processes such as follicular development, oocyte maturation, and endometrial health while mitigating the detrimental effects of oxidative stress and pathogenic infections. Recent findings suggest that phytogenic extract can enhance reproductive performance, improve oocyte quality, and support pregnancy outcomes. Despite the growing body of evidence, optimal application strategies and the full breadth of their biological effects remain insufficiently explored. This review focuses on the molecular mechanisms modulated by phytogenic extracts, particularly in the context of hormone regulation, immune modulation, and oxidative stress mitigation. We also identify critical knowledge gaps and propose future research directions to optimize the use of phytogenic extracts as a sustainable approach to enhancing livestock reproductive health.

Mung bean (Vigna radiata L.) is one of the most important legume crops worldwide (Dikr 2023). In August 2023, a severe foliar disease resembling leaf rot was observed on mung bean with ~30% incidence in the field located in Nanning (22°49'N, 108°22'E), Guangxi Province, China. Initially, the leaves develop irregular, water-soaked grayish lesions with a gray-brown center and dark brown edges. Then, the lesions expand and merge into cloud-like patches under high humidity. Lastly, the whole leaf decays. Five infected leaves were collected and cut into 2-3 mm pieces, then were sterilized and cultivated on potato dextrose agar (PDA) and incubated at 25°C under a 12 h photoperiod for 5 days. Three of 10 obtained isolates (LY1, LY2, LY3), were used for further studies. Morphological characteristics were consistent with descriptions of Rhizoctonia solani (Ogoshi 1987). The colony was white initially and then turned to brown with sclerotia, the hyphal were branched at right angles with a septum and a slight constriction at the branch base. DNA was extracted from the three isolates and the three nuclear ribosomal regions (ITS, TEF1, RPB2) were amplified (White et al. 1990; Matheny et al. 2007). The obtained sequences were compared by BLAST analysis and deposited in GenBank (Accession Nos. PV017458 to PV017460 and PV175332 to PV175337). ITS sequences showed 99-100% (688/688, 679/679, 659/661 bp) similarity with R. solani strains (MG387102.1, EF429216.1, OP497979.1). The TEF1 and RPB2 sequences exhibited showed 97 to 99% (1012/1042, 719/724 bp) with R. solani strains (ON042753.1; XM_043331355.1). The three isolates were grouped within the AG-1 IA clade by phylogenetic analyses of ITS sequences (Godoy-Lutz et al. 2008). The pathogenicity of the three isolates was tested on mung bean by placing 0.5 mm-diameter mycelium discs on the leaves of 2-week-old seedlings (n=5), and pure PDA discs were inoculated as controls. All plants were incubated in a moist chamber for 2 days and then transferred to a greenhouse at 25°C. The experiment was repeated twice with three replicates. All isolates were highly pathogenic to the original host, mung bean. Inoculated leaves presented typical leaf rot symptoms consistent with those observed in the field. Based on morphological characteristics and ITS sequences, the identity of the reisolated fungus was confirmed as R. solani, fulfilling Koch's postulates. No pathogens were isolated from controls. Furtherly, the R. solani isolate LY1 was confirmed the host range same as above pathoginicy test, on eight other legume crops (n=5), adzuki bean (Vigna angularis), cowpea (V. unguiculata), pea (Pisum sativum), soybean (Glycine max), lentil (Lens culinaris), hyacinth bean (Lablab Album Semen), common bean (Phaseolus vulgaris), and faba bean (Vicia faba). The experiment was repeated twice. The isolate LY1 exhibited strong virulence on all tested crops, causing symptoms similar to those observed on mung bean and finally leading to plant death. Previously, R. solani was often reported as soil-borne pathogen causing root related diseases (Sneh et al. 1996). To our knowledge, this is the first report of R. solani causing leaf rot on mung bean in China and worldwide. This finding indicates a potential threat to legume crops.

Food legumes play a pivotal role in China’s food security, nutritional health, and green development strategies due to their unique advantages. This paper presents an empirical study on the economic evaluation of scientific research on pest and disease control for food legumes. Using panel data from 31 Chinese provinces from 2008 to 2023, we employ a Double Machine Learning (DML) approach to identify the impact of investment in plant protection research on food legume outputs. The results indicate a steady increase in China’s investment in this field, with an average annual growth rate of 5.19% from 2008 to 2023, and the total investment in 2023 was 2.14 times that of 2008. Investment in plant protection research effectively mitigates output losses and leads to significant production increases. Specifically, a 1% increase in research investment corresponds to a 0.2% increase in food legume output. This effect remains robust across various algorithms, time windows, and control variable settings. Based on these findings, we recommend: (1) increasing financial support and talent acquisition for research on food legume pests and diseases to enhance the stability and sustainability of research investment; (2) strengthening cooperation mechanisms between research institutions and enterprises to leverage their respective strengths and promote the commercialization of research outcomes and regional variety extension; (3) establishing a diversified research investment system that explores a co-construction model guided by the government, involving enterprises, and utilizing public–private partnerships to reconcile the conflict between long research cycles and market demands; (4) fostering a dual-track linkage between regional technological innovation and enterprise product commercialization to improve the efficiency of technology transfer and application; and (5) strengthening R&D in cutting-edge fields like Artificial Intelligence to improve the efficiency and precision of pest and disease control.

This study aimed to investigate the effects of ellagic acid (EA), an antioxidant, on goat sperm quality after freezing and thawing. Goat semen was frozen using Tris-citric acid-glucose (TCG) extender containing 0, 1.25, 2.5, 5, and 10 μM of EA. Egg yolk represented 20 % (v/v) and glycerol represented 5 % (v/v) of the extender's final concentration. Goat sperm post-thaw motility, acrosome integrity, plasma membrane integrity, mitochondrial activity, ATP content, NADH/NAD+ levels, and NADH-CoQ activity were evaluated. Moreover, to elucidate how EA enhanced the goat sperm characteristics, the post-thaw sperm mitochondrial reactive oxygen species (ROS) level, malondialdehyde (MDA) level, oxidative DNA damage, apoptosis, levels of NADH dehydrogenase 1 (MT-ND1) and NADH dehydrogenase 6 (MT-ND6) proteins, and the 4-hydroxynonenal (4-HNE) level were also measured after thawing. The results demonstrated that motility, plasma membrane integrity, and acrosome integrity rates were enhanced in the group treated with 5 μM of EA compared to the other concentrations (0 μM, 1.25 μM, 2.5 μM, 5, and 10 μM). Moreover, mitochondrial activity and ATP content were notably superior in the 5 μM EA group compared to all other treatment groups, along with a considerable decrease in ROS and MDA levels. The 4-HNE level and oxidative DNA damage in sperm were also reduced by EA supplementation. Additionally, it was found that EA (5 μM) significantly (p < 0.05) decreased sperm apoptosis levels. Furthermore, the addition of 5 μM EA maintained the post-thaw sperm MT-ND1 and MT-ND6 levels and reduced the negative impact of ROS on MT-ND1 and MT-ND6, thereby sustaining mitochondrial function for ATP generation. These results suggest that ellagic acid supplementation could maintain goat post-thaw sperm quality by reducing ROS damage and maintaining mitochondrial function for ATP generation. Antioxidant treatments, such as ellagic acid are a useful tool for maintaining frozen-thawed sperm quality.

Semen preservation involves lengthening sperm’s fertile lifespan without any detrimental effects on its biochemical, functional, and ultrastructural properties. Liquid storage at 4 °C is a ram sperm preservation method. However, this method of storage causes irreversible damage due to cold shocks, osmotic stresses, oxidative stresses, and reductions in sperm metabolism. The present study aims to investigate whether the supplementation of mitochonic acid 5 (MA-5) in a sperm extender could improve chilled ram sperm quality and elucidate its mechanism of action. Ram sperm were diluted with a tris-citrate-glucose extender containing different concentrations of MA-5 (0, 0.1, 1, 10, and 100 nM) and stored at 4 °C for up to 48 h. Sperm motility, membrane integrity, acrosome integrity, mitochondrial membrane potential, reactive oxygen species (ROS) level, ATP content, and the expression of NADPH dehydrogenase subunits 1 (MT-ND1) and NADPH dehydrogenase subunits 6 (MT-ND6) were evaluated. It was observed that compared to the control, the 10 nM MA-5 treatment significantly (p < 0.05) increased total motility (82 ± 3.5% vs. 76 ± 5.9%), progressive motility (67.6 ± 8.2% vs. 51 ± 8.3%), and other parameters (straight-line velocity (VSL), average path velocity (VAP), and curvilinear velocity (VCL)). In addition, 10 nM MA-5 supplementation also improved ram sperm membrane integrity and acrosomal integrity as well increased mitochondrial membrane potential (51.1 ± 0.7% vs. 37.7 ± 1.3%), reduced ROS levels, and elevated adenosine triphosphate (ATP) contents. Furthermore, a Western blot analysis demonstrated that the addition of MA-5 significantly (p < 0.05) increased the expression of MT-ND1 and MT-ND6 proteins in ram sperm, with the 10 nM MA-5 treatment resulting in the highest expression level. These results suggest that MA-5 improves ram sperm quality by maintaining high sperm mitochondrial function during liquid storage at 4 °C.

This investigation aimed to study the effects of varying light exposure durations on ram sperm. A total of 25 rams were randomly divided into five groups. The control group was exposed to light durations of 12 h, while the experimental groups were exposed to light durations of 14, 16, 18, and 20 h. After three months of rearing, semen was collected from each ram four times using the artificial vagina method. The sperm motility parameters, sperm abnormality, sperm concentration, acrosome integrity, membrane integrity, semen volume, and total sperm number were measured. Thereafter, the metabolome, amino acid level, testosterone content, plasma follicle-stimulating hormone (FSH) and luteinizing hormone (LH) levels, and sperm antioxidant capacity were measured. The results showed that the sperm motility, sperm concentration, ejaculation volume, total sperm number, acrosome integrity, and membrane integrity in the 16 h light group were significantly improved compared to the control (p < 0.05), meanwhile the sperm abnormality was decreased. Moreover, we found 345 different metabolites between the control and 16 h light group. Among these, 273 were upregulated and 72 were downregulated. Furthermore, the amino acid content of the seminal plasma in the 16 h light group was significantly increased (p < 0.05) compared to the control. Interestingly, the seminal plasma testosterone content and the levels of FSH and LH in the serum in the 16 h light group were significantly increased (p < 0.05) compared to the control. In terms of the sperm antioxidant capacity, it was observed that the CAT activity was the highest in the group exposed to 16 h of light and decreased at 18 h of light exposure when compared to the control group; however, the CAT activity at 20 h was not different from the control. Additionally, within the 14 to 18 h light exposure range, prolonged light exposure increased the GSH content (p < 0.05), whereas 20 h of light exposure reduced the GSH content. The MDA levels decreased with prolonged light exposure, reaching the lowest point at 16 h (p < 0.05), but increased again at 20 h of light exposure. KEGG analysis indicated that the differential metabolites were mainly involved in metabolic and synthetic activities. Based on the results of this study, we can conclude that the artificial extension of the light duration for 16 h has a positive effect on ram sperm quality.

Powdery mildew caused by Erysiphe pisi DC is a global notorious disease on peas. Deploying resistance pea cultivars is the most efficient and environmentally friendly method for disease control. This study focuses on revealing the resistance genes in three pea germplasms and developing their functional markers for resistance breeding. The identification of resistance genes involved genetic mapping and the sequencing of the pea mildew resistance locus O homolog PsMLO1 gene. To confirm the heredity of three resistant germplasms, they were crossed with susceptible cultivars to generate F1, F2, and F2:3 populations. The F1 generation exhibited susceptibility to E. pisi, whereas the segregation patterns in subsequent generations adhered to the 3:1 (susceptible: resistant) and 1:2:1 (susceptible homozygotes: heterozygotes: resistant homozygotes) ratios, indicating that powdery mildew resistance was governed by a single recessive gene in each germplasm. Analysis of er1-linked markers and genetic mapping suggested that the resistance genes could be er1 alleles in these germplasms. The multiple clone sequencing results of the three homologous PsMLO1 genes showed they were novel er1 alleles, named er1-15, er1-16, and er1-17. The er1-15 and er1-16 were caused by 1-bp deletion at position 335 (A) and 429 (T) in exon 3, respectively, whereas er1-17 was caused by a 1-bp insertion at position 248 in exon 3, causing a frame-shift mutation and premature termination of PsMLO1 protein translation. Their respective functional markers, kompetitive allele-specific PCR (KASP)-er1-15, KASP-er1-16, and KASP-er1-17, were successfully developed and validated in respective mapping populations and pea germplasms. These results provide valuable tools for pea breeding resistance to E. pisi.

Chickpea (Cicer arietinum L.) is an important legume crop worldwide. An emerging disease, basal stem rot with obvious wilt symptoms, was observed in the upper part of chickpea plants during the disease survey in Qiubei County of Yunnan Province. Three fungal isolates (ZD36-1, ZD36-2, and ZD36-3) were obtained from the diseased tissue of chickpea plants collected from the field. Those isolates were morphologically found to be similar to Diaporthe aspalathi. Molecular sequence analyses of multiple gene regions (ITS, tef1, tub2, cal, and his3) indicated that the three isolates showed a high identity with D. aspalathi. Pathogenicity and host range tests of the isolates were performed on the original host chickpea and eight other legume crops. The isolates were strongly pathogenic to chickpea and appeared highly pathogenic to soybean, cowpea, and mung bean; moderated or mild pathogenic to adzuki bean and common bean; however, the isolates did not cause symptoms on grass pea (Lathyrus sativus). Diaporthe aspalathi was previously reported as a main pathogen causing the southern stem canker in soybean. To our knowledge, this is the first report of D. aspalathi inducing basal stem rot on chickpea worldwide.

Several fungal pathogens cause root rot of common bean, among which Fusarium spp. are the most common pathogens causing Fusarium root rot (FRR) worldwide. FRR has been becoming an increasingly severe disease of common bean in China, but the species of Fusarium spp. have remained unclear. Thus, this study was performed to identify the pathogen causing common bean root rot in Liangcheng County, Inner Mongolia, China. Nineteen Fusarium-like isolates were obtained after pathogen isolation and purification. The pathogenicity test indicated that eight isolates caused severe disease symptoms on common bean, while 11 other isolates were not pathogenic. The eight pathogenic isolates, FCL1–FCL8, were identified as Fusarium cuneirostrum by morphological characterization and phylogenetic analysis using partial sequences of EF-1α, ITS, 28S, and IGS regions. Host range test showed that the representative F. cuneirostrum isolate FCL3 was also pathogenic to mung bean, while not pathogenic to adzuki bean, chickpea, cowpea, faba bean, pea, and soybean. Moreover, 50 common bean and 50 mung bean cultivars were screened for resistance to FRR, and seven highly resistant or resistant cultivars of common bean were identified, while no resistant cultivars of mung bean were screened. This study revealed that F. cuneirostrum was one of common bean FRR pathogens in Inner Mongolia and it could induce mung bean root rot as well. To our knowledge, this is the first report of F. cuneirostrum causing FRR of common bean in China.

Objective: The present study aimed to investigate the effect of β-nicotinamide mononucleotide (NMN) supplementation on ram sperm quality during storage at 4°C in vitro.Methods: Tris-citric acid-glucose solution containing different doses of NMN (0, 30, 60, 90, and 120 μM) was used to dilute semen collected from rams and it was stored at 4°C. Sperm motility, plasma membrane integrity as well as acrosome integrity were evaluated at 0, 24, and 48 h time points after storage at 4°C. In addition, sperm mitochondrial activity, lipid peroxidation (LPO), malondialdehyde (MDA) content, reactive oxygen species (ROS) content, glutathione (GSH) content, superoxide dismutase (SOD) activity, and apoptosis were measured at 48 h time point after storage at 4°C.Results: Results demonstrate that the values obtained for sperm motility, acrosome integrity, and plasma membrane integrity in the NMN treatments were significantly higher than control (p<0.05). The addition of 60 μM NMN significantly improved ram sperm mitochondrial activity and reduced LPO, MDA content, and ROS content compared to control (p<0.05). Interestingly, sperm GSH content and SOD activity for the 60 μM NMN treatment were much higher than those observed for control. NMN treatment also decreased the level of Cleaved-Caspase 3, Cleaved-Caspase 9, and Bax while increasing Bcl-2 level in sperm at 48 h time point after storage at 4°C.Conclusion: Ram sperm quality can be maintained during storage at 4°C with the addition of NMN at 60 μM to the semen extender. NMN also reduces oxidative stress and apoptosis. Overall, these findings suggest that NMN is efficient in improving the viability of ram sperm during storage at 4°C in vitro.

Sperm motility is an important factor in the migration of sperm from the uterus to the oviduct. During sperm preservation in vitro, sperm generates excessive ROS that damages its function. This study aims to investigate whether the addition of pyrroloquinoline quinone (PQQ) to the diluted medium could improve chilled ram sperm quality, and then elucidates the mechanism. Ram semen was diluted with Tris-citric acid-glucose (TCG) medium containing different doses of PQQ (0 nM, 10 nM, 100 nM, 1000 nM, 10,000 nM), and stored at 4 °C. Sperm motility patterns, plasma membrane integrity, acrosome integrity, mitochondrial membrane potential, reactive oxygen species (ROS) levels, malondialdehyde (MDA) levels, superoxide dismutase (SOD) activity, and ATP levels were measured after preservation. Furthermore, the expressions of NADH dehydrogenase 1 (MT-ND1) and NADH dehydrogenase 6 (MT-ND6) in sperm were also detected by western blotting. In addition, sperm capacitation and the ability of sperm to bind to the zona pellucina were also evaluated. It was observed that the addition of PQQ significantly (p < 0.05) improved ram sperm motility, membrane integrity, and acrosome integrity during preservation. The percentage of sperm with high mitochondrial membrane potential in the PQQ treatment group was much higher than that in the control. In addition, supplementation of PQQ also decreased the sperm MDA and ROS levels, while increasing ATP levels. Interestingly, the levels of MT-ND1 and MT-ND6 protein in sperm treated with PQQ were also higher than that of the control. Furthermore, the addition of 100 nM PQQ to the medium decreased ROS damage in MT-ND1 and MT-ND6 proteins. The addition of 100 nM PQQ significantly (p < 0.05) increased protein tyrosine phosphorylation in ram sperm after induced capacitation. Furthermore, the value of the sperm–zona pellucida binding capacity in the 100 nM PQQ treatment group was also much higher than that of the control. Overall, during chilled ram- sperm preservation, PQQ protected ram sperm quality by quenching the ROS levels to reduce ROS damage and maintain sperm mitochondrial function, and preserved the sperm’s high ability of fertilization.

Glucagon-like peptide-1 (GLP-1) is a peptide hormone involved in regulating insulin secretion and energy metabolism. It typically needs to bind to Glucagon-like peptide-1 Receptor (GLP-1R) in the body to exert its regulatory effects. Exenatide-4 (EX-4) is a synthetic GLP-1 analogue that is widely used as a weight loss and blood sugar-lowering medication due to its high stability and slow degradation rate. Since there is no current research on whether GLP-1 affects energy metabolism and sperm motility, it is necessary thatthe effects of GLP-1 analogues on sperm motility and energy metabolism will be investigated by treating sheep sperm with EX-4. Our results showed that GLP-1R was present in sheep sperm, and expressed in the head of the sperm. After concentration screening, it was found that 300 pM EX-4 was most effective for improving sheep sperm motility. Incubating sperm with EX-4 resulted in a significant increase in LDH, G6PDH, lipase activity, and ATP content (P < 0.05), while triglyceride content significantly decreased (P < 0.05). Additionally, EX-4 significantly promoted insulin secretion in sheep sperm (P < 0.05). When EX-4 was used in combination with GLP-1R inhibitor (GLP-1R AB), the levels of LDH, G6PDH, lipase activity, ATP content, and insulin concentration significantly decreased (P < 0.05), while triglyceride content significantly increased (P < 0.05). The further results showed that EX-4 effectively promoted cholesterol efflux in sheep sperm (P < 0.05), which is beneficial for sperm energy acquisition and maturation. Both insulin receptor inhibitors (IR AB) and GLP-1R AB reduced the promoting effect of EX-4 on cholesterol efflux in sheep sperm (P < 0.05). Our other results revealed that EX-4 regulates sperm metabolism through the GLP-1R/PI3K/Akt pathway, enhancing energy levels in sheep sperm. Generally, sperm motility is closely related to metabolic levels, and it is believed that EX-4 enhances the activity of some metabolic enzymes in sheep sperm by activating this signaling pathway, thereby promoting energy acquisition, maturation, and significantly improving sperm motility.

A Fusarium wilt resistance gene FwS1 on pea chromosome 6 was identified and mapped to a 91.4 kb region by a comprehensive genomic-based approach, and the gene Psat6g003960 harboring NB-ARC domain was identified as the putative candidate gene. Pea Fusarium wilt, incited by Fusarium oxysporum f. sp. pisi (Fop), has always been a devastating disease that causes severe yield losses and economic damage in pea-growing regions worldwide. The utilization of pea cultivars carrying resistance gene is the most efficient approach for managing this disease. In order to finely map resistance gene, F2 populations were established through the cross between Shijiadacaiwan 1 (resistant) and Y4 (susceptible). The resistance genetic analysis indicated that the Fop resistance in Shijiadacaiwan 1 was governed by a single dominant gene, named FwS1. Based on the bulked segregant analysis sequencing analyses, the gene FwS1 was initially detected on chromosome 6 (i.e., linking group II, chr6LG2), and subsequent linkage mapping with 589 F2 individuals fine-mapped the gene FwS1 into a 91.4 kb region. The further functional annotation and haplotype analysis confirmed that the gene Psat6g003960, characterized by a NB-ARC (nucleotide-binding adaptor shared by APAF-1, R proteins, and CED-4) domain, was considered as the most promising candidate gene. The encoding amino acids were altered by a “T/C” single-nucleotide polymorphism (SNP) in the first exon of the Psat6g003960, and based on this SNP locus, the molecular marker A016180 was determined to be a diagnostic marker for FwS1 by validating its specificity in both pea accessions and genetic populations with different genetic backgrounds. The FwS1 with diagnostic KASP marker A016180 could facilitate marker-assisted selection in resistance pea breeding in pea. In addition, a comparison of the candidate gene Psat6g003960 in 74SN3B and SJ1 revealed the same sequences. This finding indicated that 74SN3B carried the candidate gene for FwS1, suggesting that FwS1 and Fwf may be closely linked or an identical resistant gene against Fusarium wilt.

HomePlant DiseaseVol. 107, No. 8First Report of Pythium myriotylum Causing Root Rot on Snap Bean in China PreviousNext DISEASE NOTE OPENOpen Access licenseFirst Report of Pythium myriotylum Causing Root Rot on Snap Bean in ChinaDong Deng, Juechen Long, Suli Sun, Fengjing Song, Wenqi Wu, Canxing Duan, and Zhendong ZhuDong DengInstitute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081, China, Juechen Longhttps://orcid.org/0000-0003-4990-3043Chongqing Academy of Agricultural Sciences, Chongqing 402160, China, Suli Sunhttps://orcid.org/0000-0002-0741-018XInstitute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081, China, Fengjing SongQingdao Academy of Agricultural Sciences, Shandong 266100, China, Wenqi WuInstitute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081, China, Canxing Duanhttps://orcid.org/0000-0002-6534-1426Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081, China, and Zhendong Zhu†Corresponding author: Z. D. Zhu; E-mail Address: [email protected]https://orcid.org/0000-0002-6867-0591Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081, ChinaAffiliationsAuthors and Affiliations Dong Deng1 Juechen Long2 Suli Sun1 Fengjing Song3 Wenqi Wu1 Canxing Duan1 Zhendong Zhu1 † 1Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081, China 2Chongqing Academy of Agricultural Sciences, Chongqing 402160, China 3Qingdao Academy of Agricultural Sciences, Shandong 266100, China Published Online:28 Jul 2023https://doi.org/10.1094/PDIS-11-22-2659-PDNAboutSectionsView articlePDFSupplemental ToolsAdd to favoritesDownload CitationsTrack Citations ShareShare onFacebookTwitterLinked InRedditEmailWechat View articleIn April 2021, severe root rot of snap bean lines at the seedling stage occurred at sunlight greenhouses of Qingdao Academy of Agricultural Sciences, Shandong Province, China, which caused 10 to 30% plant wilt and death. To determine the causal agent of the disease, three mixed soil samples (about 2 kg each) were taken from infested plots of three greenhouses. A bioassay was used to detect the root rot pathogen in the soil samples. The sieved soil samples were loaded into 500-ml paper cups with holes at the bottom and filled to about three-quarters of the cup. Each sample was repeated with three cups, and each cup was sown with five seeds of bean cultivar Longyundou 21 and watered to saturation. The planted cups were kept on racks in a glasshouse at 25°C. Two weeks after planting, seedlings growing in infected soils showed symptoms such as stunt, necrotic streaks extending upward from the base of the stem, root browning and rot, and wilt. Many oospores were observed in diseased root and stem epidermal tissues under a microscope. Isolates such as Pythium sp. were consistently isolated from stems and roots of diseased plants on potato dextrose agar (PDA). Five single-hyphal tip isolates (BPM1 to BPM5) were used for identification of morphological and molecular characteristics and pathogenicity testing. The hyphae were hyaline and nonseptate; sporangia were filamentous inflated; oogonia were terminal or intercalary and 27.0 to 37.0 μm (average 32.3 μm) in diameter; antheridia were one to five per oogonium and diclinous, occasionally monoclinous, with a breadth of 4.0 to 12.1 μm; and oospores were mostly aplerotic and 22.2 to 30.8 μm (average 27.4 μm) in diameter, with 0.9- to 2.0-μm-thick walls. Total genomic DNA of five isolates was extracted from mycelia by using the Fungi Genomic DNA Extraction Kit (Solarbio, Beijing, China). The internal transcribed spacer (ITS) gene and cytochrome oxidase subunit II (Cox II) gene were amplified and sequenced using the primer pairs ITS1/ITS4 and FM58/FM66 as described by White et al. (1990) and Martin and Tooley (2003), respectively. The resulting sequences of the five isolates were identical within their respective ITS or Cox II gene. ITS and Cox II sequences of the isolate BMP4 were uploaded to GenBank with accession numbers OP477340 and OP490301, respectively. A BLAST search of the obtained sequences at GenBank revealed that the ITS sequence of the isolate BMP4 had 100% similarity with that of Pythium myriotylum isolates (MT471374, MN629356, KY019278, KJ162354, and KF761218), and the Cox II sequence shared 100% identity with the P. myriotylum strain MAFF 235183 (AB095057) as well. The corresponding Pythium spp. ITS and Cox II sequences were obtained from GenBank to construct phylogenetic trees. The five isolates were clustered in the same clade with other P. myriotylum isolates in the two corresponding phylogenetic trees. The inoculum layer method was used to test pathogenicity (Walker and Schmitthenner 1984), with bean cultivar Longyundou 21. The five isolates cultured on PDA for 10 days were transferred to the planting paper cups and placed 3 cm below the seed when planted in fresh vermiculite, and sterile PDA was used as a control. After sowing, the cups were kept in a glasshouse at 22 to 25°C and watered one to two times a day. Two weeks after inoculation, all inoculated plants showed symptoms of severe stunt and root rot, whereas the control plants remained healthy. P. myriotylum was reisolated from inoculated plants. To our knowledge, this is the first report of P. myriotylum causing root rot on snap bean in China. Since the pathogen has a wide host range (van der Plaats-Niterink 1981), the disease management should be further investigated to avoid major economic losses.The author(s) declare no conflict of interest.References:Martin, F. N., and Tooley, P. W. 2003. Mycologia 95:269. https://doi.org/10.2307/3762038 Crossref, ISI, Google Scholarvan der Plaats-Niterink, J. 1981. Stud. Mycol. 21:1. https://www.studiesinmycology.org/index.php/issue/23-studies-in-mycology-no-21 Google ScholarWalker, A. K., and Schmitthenner, A. F. 1984. Crop Sci. 24:487. https://doi.org/10.2135/cropsci1984.0011183x002400030013x Crossref, ISI, Google ScholarWhite, T. J., et al. 1990. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, CA. Crossref, Google ScholarFunding: Funding was provided by the China Agriculture Research System of MOF and MARA (CARS-08) and the Scientific Innovation Program of the Chinese Academy of Agricultural Sciences.The author(s) declare no conflict of interest.DetailsFiguresLiterature CitedRelated Vol. 107, No. 8 August 2023SubscribeISSN:0191-2917e-ISSN:1943-7692 Download Metrics Article History Issue Date: 29 Aug 2023Published: 28 Jul 2023Accepted: 15 Mar 2023 Page: 2559 Information© 2023 The American Phytopathological SocietyFundingChina Agriculture Research System of MOF and MARAGrant/Award Number: CARS-08Chinese Academy of Agricultural SciencesKeywordsoomycete diseasepathogen identificationPhaseolus vulgarissunlight greenhouseThe author(s) declare no conflict of interest.PDF download

Stalk rot caused by Fusarium verticillioides (Fv) is one of the most destructive diseases in maize production. The defence response of root system to Fv invasion is important for plant growth and development. Dissection of root cell type-specific response to Fv infection and its underlying transcription regulatory networks will aid in understanding the defence mechanism of maize roots to Fv invasion. Here, we reported the transcriptomes of 29 217 single cells derived from root tips of two maize inbred lines inoculated with Fv and mock condition, and identified seven major cell types with 21 transcriptionally distinct cell clusters. Through the weighted gene co-expression network analysis, we identified 12 Fv-responsive regulatory modules from 4049 differentially expressed genes (DEGs) that were activated or repressed by Fv infection in these seven cell types. Using a machining-learning approach, we constructed six cell type-specific immune regulatory networks by integrating Fv-induced DEGs from the cell type-specific transcriptomes, 16 known maize disease-resistant genes, five experimentally validated genes (ZmWOX5b, ZmPIN1a, ZmPAL6, ZmCCoAOMT2, and ZmCOMT), and 42 QTL or QTN predicted genes that are associated with Fv resistance. Taken together, this study provides not only a global view of maize cell fate determination during root development but also insights into the immune regulatory networks in major cell types of maize root tips at single-cell resolution, thus laying the foundation for dissecting molecular mechanisms underlying disease resistance in maize.

Cryopreservation generates a substantial quantity of ROS in semen, leading to a decline in sperm quality and fertilization capacity. The objective of this study was to investigate the effects of resveratrol and its optimal concentration on ram sperm quality after cryopreservation. Ram semen was diluted with a freezing medium containing different concentrations of resveratrol (0, 25, 50, 75, and 100 μM). After thawing, various sperm parameters such as total motility, progressive motility, acrosome integrity, plasma membrane integrity, mitochondrial membrane potential, glutathione (GSH) content, glutathione synthase (GPx) activity, superoxide dismutase (SOD) activity, catalase (CAT) activity, lipid peroxidation (LPO) content, malondialdehyde (MDA) content, ROS level, SIRT1 level, DNA oxidative damage, and AMPK phosphorylation level were assessed. In addition, post-thaw sperm apoptosis was evaluated. Comparatively, the addition of resveratrol up to 75 μM significantly improved the sperm motility and sperm parameters of cryopreserved ram sperm. Specifically, 50 μM resveratrol demonstrated a notable enhancement in acrosome and plasma membrane integrity, antioxidant capacity, mitochondrial membrane potential, adenosine triphosphate (ATP) content, SIRT1 level, and AMPK phosphorylation levels compared to the control group (p < 0.05). It also significantly (p < 0.05) reduced the oxidative damage to sperm DNA. However, detrimental effects of resveratrol were observed at a concentration of 100 μM resveratrol. In conclusion, the addition of 50 μM resveratrol to the cryopreservation solution is optimal for enhancing the quality of cryopreserved ram sperm.