Grape gray mold caused by Botrytis cinerea is one of the most important fungal diseases affecting grape production and postharvest quality. The extensive use of fungicides has led to the emergence of resistance in B. cinerea, particularly to quinone outside inhibitors (QoIs) and succinate dehydrogenase inhibitors (SDHI), which compromises the efficacy of disease control. Therefore, detection techniques for fungicide resistance in B. cinerea are needed to provide scientific guidance for disease management. Based on recombinase polymerase amplification (RPA)–CRISPR/Cas12a technology, this study designed specific RPA primer pairs and crRNA targeting resistance-related mutation sites in fungicide target genes cytb and SdhB, incorporated single-stranded fluorescent reporter molecules, and optimized key reaction components. This study achieved rapid fluorescence-based resistance detection within 1 h, with detection sensitivities of 450 and 430 pg/µL for the two fungicide genes.

Phytopathogenic fungi are highly diverse and globally distributed, posing a major threat to agricultural production worldwide. The annual losses caused by plant diseases can reach up to 30

AIMS:To isolate and characterize a rhizosphere-derived Bacillus velezensis strain with biocontrol and plant growth-promoting potential, and to evaluate its efficacy against plant pathogens together with its genomic basis. METHODS AND RESULTS:A bacterial strain (BJ-1141) was isolated from the rhizosphere and identified as B. velezensis based on morphological characteristics and phylogenetic analyses of 16S rRNA and gyrA gene sequences. The strain exhibited broad-spectrum antagonistic activity against 24 plant pathogenic fungi, inhibiting 23 pathogens by more than 50%, with a maximum inhibition rate of 95.57%. BJ-1141 produced hydrolytic enzymes (amylase, protease, cellulase, and pectinase) and plant growth-promoting factors, including siderophores and indole-3-acetic acid (IAA). It also demonstrated phosphorus solubilization, nitrogen fixation, and biofilm formation. Greenhouse experiments showed that root application of BJ-1141 effectively controlled tomato Fusarium wilt caused by Fusarium oxysporum f. sp. lycopersici, achieving control efficacies of 81.13-85.93%, which were higher than those of chemical and commercial microbial treatments. In addition, BJ-1141 significantly increased plant height, biomass, and chlorophyll content in tomato and lettuce seedlings. Whole-genome sequencing revealed a 3 902 965 bp genome containing 13 biosynthetic gene clusters associated with antimicrobial secondary metabolites, including surfactin, fengycin, bacillaene, difficidin, macrolactin H, bacillibactin, and bacilysin. CONCLUSIONS:Bacillus velezensis BJ-1141 is a multifunctional bacterial strain with strong biocontrol efficacy and plant growth-promoting traits, supported by genomic evidence, and represents a promising candidate for sustainable crop disease management.

Strawberry (Fragaria × ananassa) is a commercially important crop in China and is highly susceptible to fungal diseases. A field survey was conducted in commercial strawberry fields in Changping District, Beijing, where symptomatic ’Hongyan’ plants showing fruit rot were collected. Tissue isolation yielded four Penicillium isolates from rotten fruits. Further species delineation was carried out using multi-locus phylogenetic analyses of the Internal Transcribed Spacer (ITS), Calmodulin (CaM), RNA polymerase II second-largest subunit (rpb2), and Beta-tubulin (tub2) genes. Morphological and molecular phylogenetic analyses confirmed that our isolates are Penicillium olsonii. Prior to the pathogenicity essays, a pilot test was conducted to optimize the most suitable surface-sterilization method for strawberry fruit. Results from a pilot test of surface sterilization showed that 75% ethanol for 30 s, followed by 5% NaOCl for 2 minutes, and a final wash with sterile distilled water three times was the most effective treatment for reducing contamination on fruit samples. Pathogenicity assays were conducted on detached, wounded, and non-wounded fruits by inoculating 20 µL of a 1.0 × 106 conidia/mL spore suspension, while sterile distilled water was used as the control, with five replicates. The results demonstrated that P. olsonii caused typical fruit rot symptoms similar to those observed in the field, and the pathogen was reisolated, fulfilling Koch’s postulates. This study identifies P. olsonii causing strawberry fruit rot for the first time worldwide. Our results highlight the phytopathological significance of Penicillium as a field pathogen rather than a typical postharvest pathogens.

Reducing fungicide use is a global priority for agricultural nations. Decision support systems (DSSs) help farmers optimize spraying schedules by assessing crop disease risk. However, a practical DSS specifically designed for grape downy mildew (Plasmopara viticola) has not yet been reported for China. . In this study, we developed a user-friendly DDS to support grape downy mildew management, which serves as a signal for the first spraying application. Furthermore, the tool employs a binary risk classification system (low vs. high) to recommend optimal spraying timings during the secondary infection period. The DSS has been embedded in the WeChat mini-program "Grape Defender" and deployed across 13 provinces in China. Model evaluation was conducted using two datasets: (a) a multi-year (2021-2024) experimental trial conducted across two cities, and (b) climate data collected from 13 provinces. We then compared a model-based fungicide application approach with a regular method in terms of disease incidence and number of spray applications in the two cities, and then we provide prediction services in 13 major grape-growing provinces across China. The DDS reduced fungicide applications by 43.75% in two experimental cities while maintaining comparable disease control, with increase in disease incidence no exceeding 11%relative to the regular management group. Simulations further projected 73 high-risk infection events across 13 provinces in 2025, suggesting that disease management efforts should primarily focus on June to August rather than May. The "Grape Defender" DSS, which will be freely accessible, supports customized strategies tailored to local climates and grape varieties in China.

Tomato (Solanum lycopersicum) is widely grown worldwide, ranking first among vegetable crops. Root diseases of tomatoes can cause serious yield losses. In June 2023 and 2024, tomato root rot symptoms were observed in the greenhouse with 70%-90% incidence approximate number of plants (N=210) in Beizhen City (121°47 ' 30 ''E, 41°35' 45 ''N), Liaoning Province, China. In 2024, the same symptoms were observed in the greenhouse in Shunyi City (116°39 ' 41 ''E, 40°09' 00 ''N), Beijing Province. Initially tomato leaves turned yellow, followed by wilting and withering of the entire plant. Roots and stems of the plants exhibited brown necrosis. Infected root tissues from six individual plants in each of two distinct regions were cut into 24 small fragments (5 × 5 mm) at the interface between healthy and symptomatic root tissue and surface sterilized with 2% NaOCl for 2 min followed by 75% ethanol for 30 s, rinsed five times with sterile water, and placed on potato dextrose agar (PDA). After 3-6 days of incubation at 25℃, hyphal tips from the edges of colonies were transferred to new PDA plates. Nine morphologically similar fungal isolates (JZB3540002-JZB3540010) were obtained from diseased samples. Six days old colonies on PDA were dense, mycelium appressed, white and the reverse was white to light yellow-brown. Conidiophores were straight or flexuous, unbranched, smooth, and had an elliptical apex. Conidia were elliptical, apex rounded, aseptate, and multi-guttulate. Conidia were 6 to 12 × 2 to 5 µm (average 7.70 × 2.95 µm, n=30). Following 14 days of incubation, colonies were hyaline with white mucilaginous masses, and aerial hyphae appeared at the center of the colony (Carlucci et al. 2012; Usami et al. 2012; Yang et al. 2021). For molecular phylogenetic analysis, genomic DNA was extracted from the nine isolates, and the internal transcribed spacer (ITS) region, large subunit 28S rDNA (LSU) and translation elongation factor 1 alpha (TEF-1alpha) gene were amplified using ITS1/ITS4 (White et al. 1990), LROR/LR5 (Vilgalys and Hester 1990), and EF-688F/EF-1251R (Alves et al. 2008) primers respectively. BLAST results indicated that the ITS, LSU, and TEF sequences showed 98 to 100% identity with Plectosphaerella pauciseptata type specimens sequences at NCBI (GenBank KY399823, KY662250, and KY421321). Maximum likelihood (ML) inference phylogenetic tree was constructed to confirm the identity of the nine strains. ML tree reveals that the nine strains clustered with Plectosphaerella pauciseptata with 100% bootstrap support value. Sequence data were deposited in NCBI GenBank with accession numbers PP716848 and PQ621101-PQ621108 (ITS), PP717077 and PQ621369-PQ621376 (LSU), and PQ629447-PQ629455 (TEF). To confirm pathogenicity, roots of 18 three-leaf-stage cultivar 'Ying fen 8' tomato seedlings were wounded and dipped in 50 ml conidial suspension (107 conidia/ml) or sterile water (control) for 30 min. Tomato seedlings were transplanted into new pots and placed in the greenhouse at 22℃(night)/25℃ (day) with natural daylight. After 25 days, inoculated plants had symptoms similar to naturally infected plants, and no symptoms were observed in the control plants. Plectosphaerella pauciseptata was re-isolated from the base of the roots of inoculated tomatoes, and the identification was confirmed based on morphological characteristics and DNA sequence analysis, The experiment was repeated three times with similar results. Plectosphaerella pauciseptata has been reported causing tomato root rot in Italy (Raimondo and Carlucci 2018), however, to our knowledge, this is the first report of P. pauciseptata causing tomato root rot in China. This study provides insight into the root diseases of tomatoes and provides the basis for further investigations to assess and implement effective disease management strategies.

Grapevine Botryosphaeria dieback (GBD), caused by Botryosphaeriaceae species, is an important grapevine trunk disease that poses a threat to grape yield and quality in global viticultural regions. Pathogen diagnosis at the species level using morphological methods is difficult and time-consuming. Therefore, this study aimed to develop a rapid and accurate detection method for the pathogens causing GBD. Recombinase polymerase amplification (RPA) with CRISPR/Cas12a cleavage was combined for detecting pathogens associated with GBD, and lateral flow dipsticks were employed to monitor the outcomes. Based on the beta-tubulin sequences of Botryosphaeriaceae and their related species, specific RPA primers and CRISPR/Cas12a CrRNA were designed and subsequently selected for specifically detecting pathogens associated with GBD. Under optimized reaction conditions and systems, the developed RPA/CRISPR-Cas12a detection system specifically detected Botryosphaeriaceae species within 30 min of RPA and 25 min of CRISPR/Cas12a reactions at 37 degrees C. Specificity tests showed that specific fragments were amplified with the RPA primers in the DNA of six Botryosphaeriaceae species found in China, while none of the fragments were amplified in the other 22 nontarget fungal pathogen species of grapevine. The detection sensitivity of this method was 1 pg/mu l, which is equal to that of real-time PCR. In summary, our method is simple to perform, produces visual results, does not rely on expensive equipment, and therefore possesses high practical value, providing an efficient and robust detection platform to accelerate the field detection of pathogens associated with GBD.

Botryosphaeria dieback, caused by the notorious pathogenic agent Lasiodiplodia theobromae, severely threatened the yield and quality of grapes globally. A comprehensive understanding of the fungal pathogenesis and plant defence will be of great importance to the development of efficient control measures. In previous publication, a secreted endopolygalacturonase LtEpg1 from L. theobromae was reported to interact with a SNF1-related protein kinase (SnRK) complex regulatory subunit VvKIN(31 from grapevine. Here, we ectopically expressed LtEpg1 and VvKIN beta 1 genes in Nicotiana benthamiana and profiled the transcriptome of LtEpg1-and VvKIN beta 1-transgenic lines. It was found that, after infection by L. theobromae, a total of 6676 and 4979 differentially expressed genes (DEGs) were identified in LtEpg1-and VvKIN beta 1-transgenic N. benthamiana, respectively, in comparison with wild type. The DEGs were involved in a wide range of physiological processes and immune responses, including photosynthesis, carotenoid biosynthesis, glycolysis, plant pathogen interaction, mitogen-activated protein kinase (MAPK) cascades, phytohormone signal transduction, and phenylpropanoid biosynthesis. Moreover, a total of 4990, 1212 and 4227 DEGs were identified in wild type, LtEpg1-and VvKIN beta 1-transgenic lines infected by L. theobromae, respectively, in comparison with the corresponding line grew under normal condition without inoculation. The DEGs were significantly enriched in photosynthesis and plant hormone signal transduction, suggesting relevant pathways were interrupted during infection. These results provide novel insights for exploring the molecular mechanisms of L. theobromae-grapevine interaction and supply valuable information for the genetic improvement of resistant grapevine in-depth.

Lasiodiplodia theobromae (L. theobromae) is the causative agent of grapevine canker disease, which is a serious threat to global grape production. Currently, no effective fungicides are available to manage this disease. In this study, LtmilR2 was identified as an Argonaute1 (AGO1)-dependent small RNA produced by L. theobromae. Functional analysis revealed that both LtAGO1 and LtmilR2 negatively regulated the pathogenicity of L. theobromae. Through degradome sequencing, we identified LtRASGEF-a guanine nucleotide exchange factor involved in RAS signalling-as the target gene of LtmilR2. LtmilR2 suppressed the expression of LtRASGEF. Pathogenicity assays demonstrated that LtRASGEF was essential for fungal virulence. Furthermore, following infection, the expression of LtmilR2 rapidly decreased, while LtRASGEF expression quickly increased and positively regulated infection. Finally, we observed that the growth of L. theobromae was inhibited when an RNA duplex targeting LtmilR2 was delivered via the star polycation (SPc) nanocarrier, suggesting that LtmilR2 may serve as an RNA-based fungicide target for grapevine canker disease management.

Grapevine trunk diseases (GTD), particularly Botryosphaeria dieback caused by Lasiodiplodia theobromae, pose a significant threat to global viticulture. Current study aimed to develop a biocontrol solution using the locally isolated BJ-3 bacterial strain. We assessed the genomic biocontrol potential of BJ-3 through genome mining, evaluated its efficacy against 19 fungal pathogenic strains in vitro, and validated its efficiency against L. theobromae through spore inhibition assay, grapevine shoot inoculation and field experiments. Based on morphological characteristics and phylogenomic analysis, BJ-3 was identified as Bacillus halotolerans. Genome mining revealed eleven antimicrobial biosynthesis gene clusters, encoding secondary metabolites including bacillibactin, bacilysin, bacillaene, fengycin, subtilosin, aurantinin, thailanstatin, and laterocidine; many of which are associated with biocontrol activities. In vitro assays demonstrated BJ-3′s broad-spectrum antagonistic capacity, with inhibition rates ranging from 55.56 % to 91.62 % against economically important GTD pathogens, including Botryosphaeria dothidea (76.18 %), Neofusicoccum parvum (70.59 %), Diaporthe sojae (91.62 %), D. eres (81.31 %), and L. theobromae (69.89 %). In the spore germination assay, BJ-3 achieved 98.47 % inhibition at 4 × 107 CFU/mL after 10–12 h of incubation. Preventive treatments with BJ-3 at a concentration of 1.08 × 108 CFU/mL resulted in 74.39 % control efficiency against L. theobromae. In the field experiment, BJ-3 effectively controlled Botryosphaeria dieback disease, with an inhibition rate of 71.94 %. These findings highlight the potential of the B. halotolerans BJ-3 strain as a biofungicide for managing Botryosphaeria dieback disease in grapevine, providing a foundation for developing B. halotolerans BJ-3-based antimicrobial secondary metabolites in future studies.

Peach (Prunus persica L.) is one of the most important and oldest stone fruits grown in China. Even though P. persica is one of the most commonly grown stone fruits in China, little is known about the biodiversity of microfungi associated with peach branch diseases. In the present study, samples were collected from a wide range of peach growing areas in China, and fungal pathogens associated with peach branch diseases were isolated. In total, 85 isolates were obtained and further classified into nine genera and 10 species. Most of the isolates belonged to Botryosphaeriaceae (46), including Botryosphaeria, Diplodia, Neofusicoccum, Phaeobotryon, and Lasiodiplodia species; Ascochyta, Didymella, and Nothophoma species representing Didymellaceae were also identified. Herein, we introduce Ascochyta prunus and Lasiodiplodia pruni as novel species. In addition, we report the first records of Nothophoma pruni, Neofusicoccum occulatum, and Phaeobotryon rhois on peach worldwide, and Didymella glomerata, Nothophoma quercina, and Phaeoacremonium scolyti are the first records from China. This research is the first comprehensive investigation to explore the microfungi associated with peach branch disease in China. Future studies are necessary to understand the pathogenicity and disease epidemiology of these identified species.

Gray mold caused by Botrytis cinerea is a destructive disease in grapes. Although the preharvest use of pesticides can control it, fungicide resistance in B. cinerea is now common. We used an Effect and Less Spraying Control (ELSC) method for applying fungicides effective against B. cinerea on grapes. The spraying schedule was determined by exploring the key stages of B. cinerea invasion using field and in vitro inoculation tests. The results indicated that the stage most vulnerable to pathogen invasion is the full-bloom stage. The disease incidence/severity in this stage is highest compared with the pre-bloom, 10-days-after-full-bloom, bunch-closure and veraison stages. Given the inoculation results and the threat of residual infected petals, the ELSC method established an optimum spray schedule at full bloom and 10 days after full bloom. To evaluate the ELSC method, four kinds of fungicides were used in an experimental trial in Beijing in 2015 and 2017; Shanghai in 2016; and Hebei in 2019 and 2021. Fludioxonil was the most effective fungicide, followed by Pythium oligandrum (bio-fungicide), difenoconazole + azoxystrobin and pyraclostrobin. ELSC was more effective against B. cinerea than the traditional control schedule, when comparing the disease severity (i.e., 0.07 ± 0.10% in ELSC and 0.49 ± 0.014% in the traditional practice when using fludioxonil). The average yield per hectare in ELSC confirmed that spraying during flowering does not have a deleterious effect on grape yield. It produced a 1224.37 00 kg/ha greater yield than the control group when fludioxonil was applied.

采用菌丝生长速率法和孢子萌发法测定了 7种杀菌剂对可可毛色二孢CSS-01s的室内生物活性,筛选出3种抑制效果较好的杀菌剂;通过对离体葡萄绿枝条及盆栽葡萄幼苗新梢进行人工接种病原菌,进一步评价杀菌剂的防治效果.结果表明,7种杀菌剂对可可毛色二孢菌丝生长和孢子萌发表现出不同的抑制活性,其中戊唑醇、氯氟醚菌唑、啶酰菌胺和氟啶胺对菌丝生长的抑制作用较强,EC500分别为0.116、0.137、0.109 μg/mL和0.119 μg/mL;戊唑醇、氯氟醚菌唑、氟啶胺对孢子萌发的抑制作用较强,EC50分别为0.420、0.595 μg/mL和1.885 μg/mL.CSS-01s接种离体葡萄绿枝条试验中,戊唑醇100、200 mg/L,氯氟醚菌唑100、300 mg/L和氟啶胺100、200 mg/L的防治效果无显著差异,在57.81％～65.31％;CSS-01s接种葡萄幼苗新梢试验中,氟啶胺200 mg/L的防治效果最好,为77.56％,与氟啶胺100 mg/L、氯氟醚菌唑100、300 mg/L和戊唑醇200 mg/L之间无显著差异,但显著高于戊唑醇100mg/L.研究结果显示,氯氟醚菌唑、戊唑醇和氟啶胺可以作为推荐杀菌剂,经进一步的田间药效评价用于葡萄溃疡病的防治.

葡萄顶枯病(Eutypa dieback)是世界各地许多葡萄生产地区发生最具破坏性的病害之一,目前广泛分布在美国、澳大利亚、加拿大、中国等28个国家.葡萄顶枯病由多种病原菌侵染引起,防治难度较大.目前,文献主要报道了该病害的发生规律,以及针对致病性最强、也是最常见的致病菌Eutypa lata的防治研究.通过综述该病害的症状、病原、发生规律和防治措施,为国内该病害的防治提供新思路.

Botrytis cinerea, the causal agent of gray mold, is one of the most destructive pathogens of cherry tomatoes, causing fruit decay and economic loss. Fludioxonil is an effective fungicide widely used for crop protection and is effective against tomato gray mold. The emergence of fungicide-resistant strains has made the control of B. cinerea more difficult. While the genome of B. cinerea is available, there are few reports regarding the large-scale functional annotation of the genome using expressed genes derived from transcriptomes, and the mechanism(s) underlying such fludioxonil resistance remain unclear. The present study prepared RNA-sequencing (RNA-seq) libraries for three B. cinerea strains (two highly resistant (LR and FR) versus one highly sensitive (S) to fludioxonil), with and without fludioxonil treatment, to identify fludioxonil responsive genes that associated to fungicide resistance. Functional enrichment analysis identified nine resistance related DEGs in the fludioxonil-induced LR and FR transcriptome that were simultaneously up-regulated, and seven resistance related DEGs down-regulated. These included adenosine triphosphate (ATP)-binding cassette (ABC) transporter-encoding genes, major facilitator superfamily (MFS) transporter-encoding genes, and the high-osmolarity glycerol (HOG) pathway homologues or related genes. The expression patterns of twelve out of the sixteen fludioxonil-responsive genes, obtained from the RNA-sequence data sets, were validated using quantitative real-time PCR (qRT-PCR). Based on RNA-sequence analysis, it was found that hybrid histidine kinase, fungal HHKs, such as BOS1, BcHHK2, and BcHHK17, probably involved in the fludioxonil resistance of B. cinerea, in addition, a number of ABC and MFS transporter genes that were not reported before, such as BcATRO, BMR1, BMR3, BcNMT1, BcAMF1, BcTOP1, BcVBA2, and BcYHK8, were differentially expressed in the fludioxonil-resistant strains, indicating that overexpression of these efflux transporters located in the plasma membranes may associate with the fludioxonil resistance mechanism of B. cinerea. All together, these lines of evidence allowed us to draw a general portrait of the anti-fludioxonil mechanisms for B. cinerea, and the assembled and annotated transcriptome data provide valuable genomic resources for further study of the molecular mechanisms of B. cinerea resistance to fludioxonil.

One of the most destructive diseases, Gibberella stalk rot (GSR), caused by Fusarium graminearum, reduces maize yields significantly. An induced resistance response is a potent and cost-effective plant defense against pathogen attack. The functional counterpart of JAs, coronatine (COR), has attracted a lot of interest recently due to its ability to control plant growth and stimulate secondary metabolism. Although several studies have focused on COR as a plant immune elicitor to improve plant resistance to pathogens, the effectiveness and underlying mechanisms of the suppressive ability against COR to F. graminearum in maize have been limited. We investigated the potential physiological and molecular mechanisms of COR in modulating maize resistance to F. graminearum. COR treatment strongly enhanced disease resistance and promoted stomatal closure with H2O2 accumulation, and 10 μg/mL was confirmed as the best concentration. COR treatment increased defense-related enzyme activity and decreased the malondialdehyde content with enhanced antioxidant enzyme activity. To identify candidate resistance genes and gain insight into the molecular mechanism of GSR resistance associated with COR, we integrated transcriptomic and metabolomic data to systemically explore the defense mechanisms of COR, and multiple hub genes were pinpointed using weighted gene correlation network analysis (WGCNA). We discovered 6 significant modules containing 10 candidate genes: WRKY transcription factor (LOC100279570), calcium-binding protein (LOC100382070), NBR1-like protein (LOC100275089), amino acid permease (LOC100382244), glutathione S-transferase (LOC541830), HXXXD-type acyl-transferase (LOC100191608), prolin-rich extensin-like receptor protein kinase (LOC100501564), AP2-like ethylene-responsive transcription factor (LOC100384380), basic leucine zipper (LOC100275351), and glycosyltransferase (LOC606486), which are highly correlated with the jasmonic acid–ethylene signaling pathway and antioxidants. In addition, a core set of metabolites, including alpha-linolenic acid metabolism and flavonoids biosynthesis linked to the hub genes, were identified. Taken together, our research revealed differentially expressed key genes and metabolites, as well as co-expression networks, associated with COR treatment of maize stems after F. graminearum infection. In addition, COR-treated maize had higher JA (JA-Ile and Me-JA) levels. We postulated that COR plays a positive role in maize resistance to F. graminearum by regulating antioxidant levels and the JA signaling pathway, and the flavonoid biosynthesis pathway is also involved in the resistance response against GSR.

本试验旨在探究饲粮中添加不同水平壳寡糖(COS)对肉兔生长性能、养分表观消化率、血清生化指标及机体免疫功能的影响.试验选取日龄[(35±2)日龄]相近、健康状况良好、体重相近的肉兔300只(公母各占1/2),随机分为6组,每组5个重复,每个重复10只兔.试验采用单因素完全随机设计,对照组包括空白对照组(基础饲粮)、抗生素对照组(基础饲粮+20 mg/kg多黏菌素E)和4个COS组(基础饲粮+50、100、150和200 mg/kg COS).预试期7 d,正试期35 d.结果表明:1)与空白对照组相比,饲粮添加50、100、150 mg/kg COS能有效提高肉兔平均日增重(ADG)(P<0.05);150 mg/kg COS组料重比(F/G)效果最好;50 mg/kg COS组腹泻率最低.2)添加COS对屠宰性能有较好的改善作用,150 mg/kg COS组肉兔全净膛重显著高于空白对照组(P<0.05);与空白对照组相比,各COS组肉兔全净膛率均显著提高(P<0.05).3)与空白对照组相比,添加COS显著提高肉兔对干物质(DM)、总能(GE)、粗蛋白质(CP)表观消化率(P<0.05),50 mg/kg COS组效果最好.4)与抗生素对照组相比,添加50 mg/kg COS可显著降低肉兔血清中葡萄糖(GLU)、甘油三酯(TG)和总胆固醇(TC)含量(P<0.05).5)与空白对照组、抗生素对照组相比,50和150 mg/kg COS组肉兔胸腺重均显著提高(P<0.05);100 mg/kg COS组肉兔血清中免疫球蛋白G(IgG)、免疫球蛋白M(IgM)及补体4(C4)的含量最高;200 mg/kg COS组血清中免疫球蛋白A(IgA)含量最高.综上所述,饲粮中添加COS能够通过提高养分表观消化率及其在体内的代谢水平,从而改善肉兔的生长性能,同时可通过提高免疫器官重和血清免疫球蛋白含量来提高机体免疫功能.本试验条件下,饲粮中COS适宜添加水平为50～100 mg/kg.

The NmrA-like proteins have been reported to be important nitrogen metabolism regulators and virulence factors in herbaceous plant pathogens. However, their role in the woody plant pathogen Lasiodiplodia theobromae is less clear. In the current study, we identified a putative NmrA-like protein, Lws1, in L. theobromae and investigated its pathogenic role via gene silencing and overexpression experiments. We also evaluated the effects of external carbon and nitrogen sources on Lws1 gene expression via qRT-PCR assays. Moreover, we analyzed the molecular interaction between Lws1 and its target protein via the yeast two-hybrid system. The results show that Lws1 contained a canonical glycine-rich motif shared by the short-chain dehydrogenase/reductase (SDR) superfamily proteins and functioned as a negative regulator during disease development. Transcription profiling revealed that the transcription of Lws1 was affected by external nitrogen and carbon sources. Interaction analyses demonstrated that Lws1 interacted with a putative GATA family transcription factor, LtAreA. In conclusion, these results suggest that Lws1 serves as a critical regulator in nutrition metabolism and disease development during infection.

根结线虫是具有毁灭性危害的植物内生寄生虫,臭氧水漫灌土壤是较为理想的替代化学药剂的防治方法,但还未见在茄子上使用的报道.为了探究高浓度臭氧水漫灌处理土壤对茄子根结线虫的田间防治效果及对茄子生长的影响,本研究在茄子定植前对土壤进行连续2次臭氧水(9 mg/L)漫灌处理,并对漫灌处理前后和茄子整个生长期的根结线虫病害发生情况及茄子生长情况进行跟踪调查.结果表明,与对照相比,结果期和生长后期根结线虫数量分别下降91.67％和95.97％;幼苗期、结果期和生长后期的防治效果分别为100％、55.36％和65.52％;处理样本的株高显著提高.因此,高浓度臭氧水漫灌处理土壤对茄子根结线虫的防治效果较好且可减少农药使用量,对茄子生长安全,值得推广应用.

本研究首先克隆鹅FOXO3基因编码区(CDS)序列,构建FOXO3基因的真核表达载体,然后将其转染至鹅卵泡颗粒细胞中,利用荧光定量PCR方法检测FOXO3基因对细胞凋亡和自噬相关基因mRNA表达水平的影响.结果显示,本研究成功克隆了鹅FOXO3基因CDS序列,并由此构建了真核表达载体pcDNA3.1-FOXO3.鹅卵泡颗粒细胞FOXO3基因过表达实验发现,过表达组凋亡相关基因Bax、Bcl-2、Caspase3、Fasl的mRNA表达水平较对照组均升高(P<0.05),而自噬相关基因LC3和Beclin1的mRNA表达水平较对照组均降低(P<0.05).本研究结果表明,FOXO3基因可能影响鹅卵泡颗粒细胞中凋亡及自噬相关基因的表达,进而调控颗粒细胞的凋亡和自噬.