摘要：
Root-knot nematodes (Meloidogyne spp.) are highly destructive pests that cause enormous crop losses worldwide. With the increasing awareness of environmental protection, exploring the potential of biocontrol agents has become crucial for nematode management. Priestia megaterium YB-3 was originally isolated from rice rhizosphere soil. In vitro experiments with the fermentation supernatant of YB-3 exhibited up to 96.0% mortality of M. graminicola second-stage juveniles (J2) and 39.2% egg hatching inhibition rate at 48 h after treatment. In greenhouse and field experiments, the application of YB-3 significantly suppressed the gall index, final nematode and egg populations compared to the untreated control, while it promoted rice (Oryza sativa) growth. YB-3 showed consistently high biocontrol efficacy against M. graminicola. Moreover, YB-3 remarkably enhanced the expression of defence genes involved in salicylic acid (OsNPR1, OsWRKY45, OsPR1a), jasmonic acid (OsJaMYB, OsAOS2) and ethylene (OsACS1) signalling pathways at different levels. In addition, YB-3 steadily colonized rice rhizosphere soil at levels ranging from 3.54 x 103 to 9.08 x 104 cfu/g soil. High-throughput sequencing analysis showed that the application of YB-3 had no significant influence on the rice rhizosphere microbial community structure, and the taxa enriched in the YB-3 treatment were Acidobacteria, Bacteroidetes and Ascomycota. YB-3 effectively suppressed M. graminicola infection, possibly because it colonized the rice rhizosphere, induced plant resistance and increased the relative abundances of beneficial microorganisms. Priestia megaterium YB-3 could be a promising and safe biological component in the integrated management of M. graminicola.
关键词：
Tobacco mosaic virus;alpha diversity;bacterial interaction;beneficial bacteria;sensitivity of bacterial community
摘要：
BACKGROUND: Tobacco mosaic virus (TMV) is one famous plant virus responsible for substantial economic losses worldwide. However, the roles of bacterial communities in response to TMV in the tobacco rhizosphere remain unclear. METHODS: We explored the soil physicochemical properties and bacterial community succession of the healthy (YTH) and diseased (YTD) plants with TMV infection by 16S rRNA gene sequencing and bioinformatics analysis. RESULTS: We found that soil pH in the YTD group was significantly lower than in the YTH group, and the soil available nutrients were substantially higher. The bacterial community analysis found that the diversity and structure significantly differed post-TMV disease onset. With TMV inoculated, the alpha diversity of the bacterial community in the YTD was markedly higher than that in the YTH group at the early stage. However, the alpha diversity in the YTD group subsequently decreased to lower than in the YTH group. The early bacterial structure of healthy plants exhibited higher susceptibility to TMV infection, whereas, in the subsequent stages, there was an enrichment of beneficial bacterial (e.g., Ramlibacter, Sphingomonas, Streptomyces, and Niastella) and enhanced energy metabolism and nucleotide metabolism in bacteria. CONCLUSION: The initial soil bacterial community exhibited susceptibility to TMV infection, which might contribute to strengthening resistance of Tobacco to TMV.
摘要：
Micromelalopha sieversi (Staudinger, 1892) is a significant pest of Poplar trees in China. In this study, we used high-throughput sequencing to sequence the whole mitochondrial genome of M. sieversi. The length of the genome was 15,373 base pairs. The nucleotide composition was 39.8%, 11.5%, 8.0%, and 40.7% for A, C, G, and T, respectively. We used the maximum-likelihood method to construct a molecular phylogenetic tree based on complete mitogenome sequences of 19 Noctuoidea species as ingroups and five Geometroidea species as outgroups. The results indicate that the genus Micromelalopha is closely related to the genus Clostera in family Notodontidae.
摘要：
The fall armyworm, known as Spodoptera frugiperda, is a notorious invasive pest wreaking havoc on agricultural crops globally. In the course of this study, a novel variant of the S. litura multiple nucleopolyhedrovirus (SpltMNPV-T0) was isolated from S. litura larvae found on tobacco plants in China. This research delved into the molecular and biological characteristics of SpltMNPV-T0. Electron microscopy revealed that this variant exhibited the characteristics features of a baculovirus. The complete genome of SpltMNPV-T0 was 137,925 bp in length, with a G + C content of 42.88 %, housing a total of 130 open reading frames (ORFs). Phylogenetically, this variant was similar to the published genome of the SpltMNPV-G2 strain, aligning itself with the Alphabaculovirus group II. However, it distinguished itself from the SpltMNPV-II in terms of sequence similarity (76.45 %), shared ORFs (only 63 genes in common), and gene order (exhibiting inversion and reordering). Crucially, SpltMNPV-T0 demonstrated notable effectiveness in controlling third-instar larvae of S. frugiperda and S. litura. Furthermore, the virulence of crude SpltMNPV-T0 matched that of the commercially available virus-based pesticide (SpltMNPV-KY), as evident in terms of mortality rates and speed of kill. These findings yield valu able insights into the molecular biology and functional genomics of this newly discovered variant, characterized by its high virulence. Such knowledge holds promise for advancing bio-control technologies aimed at mitigating the destructive impact of the pest S. frugiperda.
摘要：
Cytochrome P450s play critical roles in the metabolic resistance of insecticides in insects. Previous findings showed that enhanced P450 activity was an important mechanism mediating indoxacarb resistance, and multiple P450 genes were upregulated in indoxacarb resistant strains of Spodoptera litura. However, the functions of these P450 genes in insecticide resistance remain unknown. Here, the P450 inhibitor PBO effectively decreased the resistance of S. litura to indoxacarb. Ten upregulated P450 genes were characterized, all of which were overexpressed in response to indoxacarb induction. Knockdown of nine P450 genes decreased cell viability against indoxacarb, and further silencing of three genes (CYP339A1, CYP340G2, CYP321A19) in larvae enhanced the sensitivity to indoxacarb. Transgenic overexpression of these three genes increased resistance to indoxacarb in Drosophila melanogaster. Moreover, molecular modeling and docking predicted that these three P450 proteins could bind tightly to indoxacarb and N-decarbomethoxylated metabolite (DCJW). Interestingly, these three P450 genes may also mediate cross-resistance to chlorantraniliprole, λ-cyhalothrin and imidacloprid. Additionally, heterologous expression and metabolic assays confirmed that three recombinant P450s could effectively metabolize indoxacarb and DCJW. This study strongly demonstrates that multiple overexpressed mitochondrial and microsomal P450 genes were involved in insecticide resistance in S. litura.
摘要：
Root rot is an important disease of tea plants owing to its unobvious early symptoms and permanent damage (Huu et al. 2016). In 2019, 5% of tea plants displayed symptoms consistent with root rot in a tea plantation (28°09'N, 113°13'E) located in Changsha city, Hunan province of China. The symptoms of the diseased tea plants ranged from wilting leaves to entirely dead. The roots had black lesions and rot typical of this disease. Symptomatic roots were collected, washed with water and disinfected with 75% ethanol, then cut into pieces and sterilized with 0.1% mercuric chloride for 30 s, 75% ethanol for 1 min, and rinsed with sterile water five times. After drying on sterilized filter paper, root tissues were cultured on potato dextrose agar (PDA) medium at 25 oC for 7 days in the dark. Four isolates, CAGF1, CAGF2, CAGF3, and CAGF4 were purified by selecting single spores. All isolates were subjected to a pathogenicity test. A conidial suspension of each strain was collected at a concentration of 2×106 conidia/mL. For the pathogenicity test, two-year-old field grown tea plants were transplanted in plastic pots containing 240 g of the rice grain-bran mixture (inoculated with 4 mL of conidial suspension and cultured for 14 days) and 960 g of sterilized soil (Huu et al. 2016). The pots without inoculated mixture served as control group. All the pots were kept in illumination incubators at 25 (o)C and a 12L:12D photoperiod. The pathogenicity test for each strain was repeated three times with three repetitions. Only strain CAGF1 exhibited pathogenicity to tea plants. Symptoms appeared on the third day post inoculation (dpi) and gradually worsened by the 7 dpi. On the 14 dpi, most leaves had died and the roots were black and partially rotten, similar to field symptoms. The reisolated fungus from potted roots was identified as CAGF1 based on ITS region and colony morphology, while isolation was attempted, CAGF1 was not isolated from the control plants, which fulfilled Koch's postulates. On PDA, the colony center of CAGF1 was purple with white margin, while on carnation leaf agar (CLA) medium was white. On CLA medium, macroconidia have 0 to 3 septa, measured 19.1 μm to 41.2 μm × 4.2 μm to 5.4 μm (mean= 31.2 μm × 4.8 μm, n=30). The microconidia were measured as 6.7 μm to 12.8 μm × 2.4 μm to 4.9 μm (mean= 10.1 μm × 3.3 μm, n=30), with 0 to 1 septa. And the chlamydospores were measured as 6.0 to 9.7μm (mean= 7.7μm, n=30). Morphologically, strain CAGF1 was identified as Fusarium oxysporum (Leslie and Summerell 2006). Additionally, the genomic DNA of strain CAGF1 was extracted by cetyltrimethylammonium bromide (CTAB) method, the internal transcribed spacer (ITS), elongation factor 1 alpha (EF-1α) and second largest subunit of RNA polymerase II (RPB2) were amplified using the primers ITS1/ITS4 (White et al. 1990), EF-1/EF-2 (Geiser et al. 2004) and fRPB2-5F/fRPB2-7cR (Liu et al. 1999), respectively. Sequences were deposited in GenBank (ITS, OK178562.1; EF-1α, OK598121.1; RPB2, OP381476.1). BLASTn searches revealed that strain CAGF1 was 100% (ON075522.1 for ITS and JX885464.1 for RPB2) and 99.6% (JQ965440.1 for EF-1α) identical to Fusarium oxysporum species complex (FOSC). Based on phylogenetic analysis, the strain CAGF1 was identified as Fusarium cugenangense, belonging to FOSC. To our knowledge, this is the first report of F. cugenangense causing root rot of tea plants in China. The findings are important for the management of this root rot and the improvement of economic benefits of tea cultivation.
摘要：
Cowpea (Vigna unguiculata L.) is a legume consumed as a high-quality plant protein source in many parts of the world. In August 2023, it was observed that a plant disease affected cowpea growth in Yiyang (28.34°N, 112.55°E), China. The average disease incidence was 10%, resulting in 8.5% economic losses in approximately 3,000 m(2). The symptoms initially appeared as brown lesions near the stem-soil interface and the lesions were colonized by white mycelia. As the disease progressed, the disease symptoms included constriction and brown staining at the base of the stem, covered with a small amount of white mycelia. Eventually, the entire plants withered and collapsed and many sclerotia were scattered on the ground around the diseased stem. Twenty samples (10 sclerotia and 10 diseased tissue fragments) were collected from symptomatic plants for causal agent isolation. Samples were disinfected with 70% ethanol for 30 s, 5% NaClO for 1 min, rinsed three times with sterile water, dried and placed on potato dextrose agar (PDA) plates at 28℃ in the dark. In total, 20 isolates were obtained by the hyphal tip method (Terrones et al. 2022) and showed a consistent phenotype of white cottony mycelia on PDA with an growth rate of 12.9 to 21.3 mm/day (n = 20). Sclerotia formed at five to eight days post inoculation, were initially whitish, turning beige and eventually dark brown. The diameter of mature sclerotia ranged from 0.89 to 2.13 mm (mean = 1.64±0.29 mm; n =50). For pathogen identification, ITS1/ITS4 (White et al. 1990) and EF1-983F/EF1-2218R (Rehner and Buckley 2005) primers were used to amplify the internal transcribed spacer regions (ITS) and translation elongation factor-1 alpha gene (TEF-1α), respectively. The sequences of all 20 isolates showed 99% to 100% similarity withAgroathelia rolfsii sequences from GenBank by BLAST analysis. The sequences of two representative strains, ID1 and ID4, were deposited in GenBank. The ITS sequences of ID1 (OR689482) and ID4 (OR689481) were ＞99% similar to A. rolfsii strain QJ7 (593/596 bp; MZ750983) and A. rolfsii strain Kale078 (565/568 bp; MN872304), respectively. Also, TEF-1α sequences of ID1 (OR713735) and ID4 (OR713736) were ＞99% similar to the sequences of A. rolfsii strain HS-Sr (1073/1073 bp; OL416131) and A. rolfsii strain MSB1-2 (1070/1075 bp; MN702790), respectively. Phylogenetic analysis based on ITS and TEF1-α sequences indicated that ID1 and ID4 clustered into the A. rolfsii clade.Based on morphology and sequence analyses, the isolates ID1 and ID4 were identified as A. rolfsii (anamorph Sclerotium rolfsii).Pathogenicity tests were conducted three times on healthy 30-day-old cowpea seedlings. Five plants were inoculated with 6-day-old mycelial discs (6 mm) of ID1 or ID4 at the base of the seedlings (n = 30) while four plants were inoculated with a sterile PDA disc as a control (n = 12). All seedlings were cultivated in a greenhouse with a temperature of 26°C to 28°C and relative humidity 60% to 80% with a 14/10 h light/dark photoperiod. Eight days later, all the fungal inoculated seedlings showed symptoms including brown necrosis and collapse of the stems, and eventual withering of the seedlings. Control plants remained asymptomatic. The causal pathogens were reisolated in PDA plates and identified by ITS sequence analysis, completing Koch's postulates. To our knowledge, this is the first report ofA. rolfsiicausing southern blight oncowpea in China. Early accurate diagnosis will help farmers to adopt suitable practices to control disease outbreaks and reduce losses.
摘要：
The phenylalanine ammonia-lyase (PAL) enzyme catalyses the conversion of l-phenylalanine to trans-cinnamic acid. This conversion is the first step in phenylpropanoid biosynthesis in plants. The phenylpropanoid pathway produces diverse plant metabolites that play essential roles in various processes, including structural support and defence. Previous studies have shown that mutation of the PAL genes enhances disease susceptibility. Here, we investigated the functions of the rice PAL genes using 2-aminoindan-2-phosphonic acid (AIP), a strong competitive inhibitor of PAL enzymes. We show that the application of AIP can significantly reduce the PAL activity of rice crude protein extracts in vitro. However, when AIP was applied to intact rice plants, it reduced infection of the root-knot nematode Meloidogyne graminicola. RNA-seq showed that AIP treatment resulted in a rapid but transient upregulation of defence-related genes in roots. Moreover, targeted metabolomics demonstrated higher levels of jasmonates and antimicrobial flavonoids and diterpenoids accumulating after AIP treatment. Furthermore, chemical inhibition of the jasmonate pathway abolished the effect of AIP on nematode infection. Our results show that disturbance of the phenylpropanoid pathway by the PAL inhibitor AIP induces defence in rice against M. graminicola by activating jasmonate-mediated defence.
摘要：
Tobacco (Nicotiana tabacum L.) is an important economic crop that is widely grown around the world. Its annual production in China is estimated at 2.2 million tons (Berbeć and Matyka 2020). Since 2022, a root rot disease was sporadically observed on tobacco seedlings on cultivar Yunyan 87 in cultivated tobacco fields in the Hunan province of China. A disease incidence of about 10% occurred across 48 ha of tobacco fields. The affected tobacco plants had slow and stunted growth with yellowing leaves. The roots turned grayish brown, decayed, and died. Diseased roots were collected from six fields and cut into small pieces (5 mm ×5 mm) from the edge of the rotted portions, and then sterilized with 70% ethanol for 10 s, 0.1% HgCl2 for 1 min, and washed in sterilized water three times. All the sterilized tissue were placed on potato dextrose agar (PDA) medium and cultured at 26 ℃ in the dark. About 3 days later, colonies with similar morphology were removed and sub-cultured on fresh PDA. A total of six strains were obtained from six tobacco samples. Strains were white and had radial growth on PDA. Hyphae were aseptate and the sporangia were filamentous. The oogonia were subglobose, smooth, 16.04 ± 0.25 µm (n=50) in diameter, and developed on unbranched stalks. The antheridia were barrel shaped and clavate. Oospores were globose, aplerotic or nearly plerotic, measuring 6.62 ± 0.33 µm (n=50). These morphological characteristics were consistent with the description of Pythium spp. (van der Plaats-Niterink 1981). For molecular identification, the internal transcribed spacer (ITS) region of rDNA and cytochrome c oxidase subunit I (Cox I) of a representative isolate, GF-3, were amplified and sequenced (GenBank accession nos. OR228424 for ITS and OR237556 for Cox I) using universal primers ITS1/ITS4 (White et al. 1990) and FM58/FM66, respectively (Villa et al. 2006). BLASTn analysis revealed that the ITS and Cox I sequences were 99.76 % (838/840 bp) and 99.85% (671/672 bp) identical to the corresponding sequences of P. dissotocum strain CBS 166.68 (AY598634.2) and UM982 (MT981147.1), respectively. A neighbor-joining phylogenetic tree based on the Cox I sequence showed that GF-3 grouped in the P. dissotocum branch. Based on morphological and molecular characteristics, GF-3 was identified to be P. dissotocum. For pathogenicity testing, four- to five-leaf-old healthy potted tobacco seedlings of the Yunyan 87 cultivar were inoculated with a zoospore suspension (1 × 105 zoospores/ml), which was induced on V8-juice medium. The zoospore suspension was introduced into the soil around plant roots and 10 mL of inoculum was used for each plant. In the control group, plants were inoculated with sterilized water. All of the treated plants were kept in humid chambers at 26°C under a 12 h/12 h photoperiod. The pathogenicity assays were performed twice, with each treatment having three replicated plants. After 5 days, tobacco seedlings inoculated with P. dissotocum showed symptoms resembling that observed in the field. However, the control plants remained healthy. Pythium dissotocum was re-isolated from the infected plants and identified by morphological and molecular methods, thus confirming Koch's postulates. Pythium dissotocum has been reported causing root rot in other plants, including hydroponic lettuce (McGehee et al. 2018) and spinach (Huo et al. 2020). Also, many Pythium species have recently been recovered from float-bed tobacco transplant production greenhouses (Zhang et al. 2022). However, to our knowledge, this is the first report of root rot on tobacco caused by P. dissotocum in China. Since this disease could greatly affect tobacco seedling establishment in the field, appropriate management strategies need to be developed to reduce further losses in tobacco planting fields.
摘要：
Pre-parasitic second-stage juveniles (pre-J2s) of Meloidogyne graminicola are non-feeding in soil, and their migration, localization and successful invasion of host roots are essential for survival. Chemotaxis is thought to play a central role in host localization, but the mechanisms of chemotaxis are poorly understood. In this study, we cloned and evaluated the molecular characteristics and functions of four chemosensory genes, including Mg-odr-3, Mg-odr-1, Mg-tax-4, and Mg-osm-9. Quantitative real-time PCR (qRT-PCR) analysis revealed that Mg-odr-3 and Mg-osm-9 tended to be expressed in pre-J2s and parasitic J2s (par-J2s), while Mg-odr-1 and Mg-tax-4 were expressed in par-J2s, consistent with their functions during the host-seeking period and establishment of suitable feeding sites in root tips. Root signals from eight plant root extracts, namely, rice, barnyard grass (Echinochloa crus-galli), wheat, soybean, pepper, tomato, eggplant, and marigold, were repulsive to J2s of M. graminicola. Moreover, RNAi silencing of Mg-odr-3, Mg-odr-1, Mg-tax-4, and Mg-osm-9 in J2s attenuated their repulsion to the rice root extract. Furthermore, we found that the response of J2s to root exudates was concentration-dependent, with high concentrations of root exudates repelling J2s. Silenced J2s also showed reduced repulsion in response to high concentrations of rice root exudates. Conversely, M. graminicola J2s were attracted to the root tips of the eight plants, while RNAi-J2s showed defective attraction to root tips. In conclusion, our results showed that chemosensory genes are crucial for the chemotactic host-seeking behaviour of M. graminicola.
摘要：
Previous research has shown that the pathogenicity and appressorium development of Magnaporthe oryzae can be inhibited by the ATP synthase subunit beta (Atp2) present in the photosynthetic bacterium Rhodopseudomonas palustris. In the present study, transgenic plants overexpressing the ATP2 gene were generated via genetic transformation in the Zhonghua11 (ZH11) genetic background. We compared the blast resistance and immune response of ATP2-overexpressing lines and wild-type plants. The expression of the Atp2 protein and the physiology, biochemistry, and growth traits of the mutant plants were also examined. The results showed that, compared with the wild-type plant ZH11, transgenic rice plants heterologously expressing ATP2 had no significant defects in agronomic traits, but the disease lesions caused by the rice blast fungus were significantly reduced. When infected by the rice blast fungus, the transgenic rice plants exhibited stronger antioxidant enzyme activity and a greater ratio of chlorophyll a to chlorophyll b. Furthermore, the immune response was triggered stronger in transgenic rice, especially the increase in reactive oxygen species (ROS), was more strongly triggered in plants. In summary, the expression of ATP2 as an antifungal protein in rice could improve the ability of rice to resist rice blast.
摘要：
The rice stem borer (RSB, Chilo suppressalis) is a significant agricultural pest that mainly depends on chemical control. However, it has grown to varied degrees of pesticide resistance, which poses a severe threat to rice production and emphasizes the need for safer, more efficient alternative pest management strategies. Here, in vitro and in vivo experiments analyses reveal miR-1579 binds to the critical transcription factor Kruppel ho-mologue 1 (Kr-h1) and negatively regulates its expression. Overexpression of miR-1579 in larvae with signifi-cantly lower levels of Kr-h1 was associated with a decline in larval growth and survival. Furthermore, in female pupae, miR-1579 overexpression led to abnormalities in ovarian development, suggesting that targeting miR-1579 could be a potential management strategy against C. suppressalis. Therefore, we generated transgenic rice expressing miR-1579 and screened three lines that had a single copy of highly abundant mature miR-1579 transcripts. Expectedly, fed with transgenic miR-1579 rice lines were significantly lower survival rates in larvae and high levels of resistance to damage caused by C. suppressalis infestation. These findings suggest that miRNA-mediated RNAi could provide an effective and species-specific strategy for C. suppressalis control.
摘要：
The root-knot nematode Meloidogyne graminicola secretes effectors into rice tissues to modulate host immunity. Here, we characterised MgCRT1, a calreticulin protein of M. graminicola, and identified its target in the plant. In situ hybridisation showed MgCRT1 mRNA accumulating in the subventral oesophageal gland in J2 nematodes. Immunolocalization indicated MgCRT1 localises in the giant cells during parasitism. Host-induced gene silencing of MgCRT1 reduced the infection ability of M. graminicola, while over-expressing MgCRT1 enhanced rice susceptibility to M. graminicola. A yeast two-hybrid approach identified the calmodulin-like protein OsCML31 as an interactor of MgCRT1. OsCML31 interacts with the high mobility group protein OsHMGB1 which is a conserved DNA binding protein. Knockout of OsCML31 or overexpression of OsHMGB1 in rice results in enhanced susceptibility to M. graminicola. In contrast, overexpression of OsCML31 or knockout of OsHMGB1 in rice decreases susceptibility to M. graminicola. The GST-pulldown and luciferase complementation imaging assay showed that MgCRT1 decreases the interaction of OsCML31 and OsHMGB1 in a competitive manner. In conclusion, when M. graminicola infects rice and secretes MgCRT1 into rice, MgCRT1 interacts with OsCML31 and decreases the association of OsCML31 with OsHMGB1, resulting in the release of OsHMGB1 to enhance rice susceptibility.
通讯机构：
[Bai, LY; Bai, LY ; Pan, L ] H;Hunan Agr Univ, Coll Plant Protect, Changsha, Peoples R China.;Hunan Agr Biotechnol Res Inst, Hunan Acad Agr Sci, Changsha, Peoples R China.
关键词：
Acetolactate synthase (ALS);Mesosulfuron-methyl;gene mutation;homology modelling and docking;Bromus japonicus
摘要：
Introduction Bromus japonicus is one of the most notorious agricultural weeds in China. The long-term use of ALS-inhibiting herbicides has led to rapid evolution of herbicide resistance in B. japonicus. B. japonicus population (BJ-R) surviving mesosulfuron-methyl treatment was collected from wheatland. Here, we aimed to confirm the resistance mechanisms in this putative resistant population. Methods The dose-reponse tests were used to test the resistance level of the B. japonicus to ALS-inhibiting herbicides. Pretreatment with P450 and GST inhibitors and GST activity assays were used to determine whether P450 or GST was involved in the resistance of the BJ-R population. Sanger sequencing was used to analyse the ALS mutation of the BJ-R population. RT-qPCR was used to confirm the the expression levels of the ALS gene in mesosulfuron-methyl -resistant (BJ-R) and-susceptible (BJ-S) B. japonicus. An in vitro ALS activity assay was used to determine the ALS activity of the BJ-R and BJ-S populations. Homology modelling and docking were used to determine the binding energy of the BJ-R and BJ-S populations with ALS-inhibiting herbicides. Results B. japonicus population (BJ-R) was confirmed to be 454- and 2.7-fold resistant to the SU herbicides mesosulfuron-methyl and nicosulfuron, and 7.3-, 2.3-, 1.1- and 10.8-fold resistant to the IMI herbicide imazamox, the TP herbicide penoxsulam, the PTB herbicide pyribenzoxim and the SCT herbicide flucarbazone-sodium, respectively, compared with its susceptible counterpart (BJ-S). Neither a P450 inhibitor nor a GST inhibitor could reverse the level of resistance to mesosulfuron-methyl in BJ-R. In addition, no significant differences in GST activity were found between the BJ-R and BJ-S. ALS gene sequencing revealed a Pro-197-Thr mutation in BJ-R, and the gene expression had no significant differences between the BJ-R and BJ-S. The ALS activity of BJ-R was 106-fold more tolerant to mesosulfuron-methyl than that of BJ-S. Molecular docking showed that the binding energy of the ALS active site and mesosulfuron-methyl was changed from -6.67 to -4.57 kcal mol(-1) due to the mutation at position 197. Discussion These results suggested that the Pro-197-Thr mutation was the main reason for the high resistance level of BJ-R to mesosulfuron-methyl. Unlike previous reports of the cross-resistance pattern conferred by this mutation, we firstly documented that the Pro-197-Thr mutation confers broad cross-resistance spectrums to ALS-inhibiting herbicides in B. japonicus.
通讯机构：
[Youzhi Li] H;Hunan Provincial Engineering & Technology Research Center for Biopesticide and Formulation Processing, Hunan Agricultural University, Changsha 410128, China<&wdkj&>National Research Center of Engineering & Technology for Utilization of Botanical Functional Ingredients, Hunan Agricultural University, Changsha 410128, China<&wdkj&>College of Plant Protection, Hunan Agricultural University, Changsha 410128, China<&wdkj&>Author to whom correspondence should be addressed.
摘要：
Chilo suppressalis is one of the most damaging rice pests in China's rice-growing regions. Chemical pesticides are the primary method for pest control; the excessive use of insecticides has resulted in pesticide resistance. C. suppressalis is highly susceptible to cyproflanilide, a novel pesticide with high efficacy. However, the acute toxicity and detoxification mechanisms remain unclear. We carried out a bioassay experiment with C. suppressalis larvae and found that the LD10, LD30 and LD50 of cyproflanilide for 3rd instar larvae was 1.7 ng/per larvae, 6.62 ng/per larvae and 16.92 ng/per larvae, respectively. Moreover, our field trial results showed that cyproflanilide had a 91.24% control efficiency against C. suppressalis. We investigated the effect of cyproflanilide (LD30) treatment on the transcriptome profiles of C. suppressalis larvae and found that 483 genes were up-regulated and 305 genes were down-regulated in response to cyproflanilide exposure, with significantly higher CYP4G90 and CYP4AU10 expression in the treatment group. The RNA interference knockdown of CYP4G90 and CYP4AU10 increased mortality by 20% and 18%, respectively, compared to the control. Our results indicate that cyproflanilide has effective insecticidal toxicological activity, and that the CYP4G90 and CYP4AU10 genes are involved in detoxification metabolism. These findings provide an insight into the toxicological basis of cyproflanilide and the means to develop efficient resistance management tools for C. suppressalis.
通讯机构：
[Guo-Hua Huang] C;College of Plant Protection, Hunan Agricultural University, Changsha 410128, China<&wdkj&>Hunan Provincial Key Laboratory for Biology and Control of Plant Diseases and Insect Pests, Hunan Agricultural University, Nongda Road, Furong District, Changsha 410128, China<&wdkj&>Author to whom correspondence should be addressed.
通讯机构：
[Huan Yu; Gong Chen; Huan Yu Huan Yu Huan Yu; Gong Chen Gong Chen Gong Chen] H;Hunan Provincial Key Laboratory for Biology and Control of Plant Diseases and Insect Pests, Hunan Agricultural University, Changsha, Hunan, People's Republic of China<&wdkj&>College of Plant Protection, Hunan Agricultural University, Changsha, Hunan, People's Republic of China
摘要：
In recent years, carbon-based materials catalyzing peroxymonosulfate (PMS) for green degradation of persistent or-ganic pollutants have attracted increasing attention. However, PMS activation by hydrochar composite (e.g. hydrochar-montomorillonite) has rarely been investigated. Herein, a simple preparation, low-cost and eco-friendly catalyst of hydrochar-montmorillonite composite (HC-Mt) was prepared to firstly catalyze PMS for the degradation of dicamba (DIC). The as-prepared HC-Mt showed a remarkably better catalyzing performance for PMS than pure hydrochar (HC) due to its good physicochemical characteristics and abundant oxygen-containing groups. Further-more, the electron spin resonance (ESR) and quenching tests revealed that active species such as SO4 center dot-, center dot OH and O2 center dot- all participated in the degradation process. DIC sites on C6, Cl 10, and O15 exhibited higher reactivity according to the density functional theory (DFT) calculation, which were easily attacked by active species. The DIC degradation mainly occurred via hydroxyl substitution, decarboxylation, oxidation and ring-cleavage and finally most of the inter-mediates were mineralized into CO2 and H2O. Finally, the phytotoxicity assessment was measured by the germination growth situation of tobacco and mung beans in the presence of DIC (with or without treatment by HC-Mt/PMS). The result showed that HC-Mt/PMS could significantly reduce the phytotoxicity of DIC to crops, suggesting that catalyzing PMS using HC-Mt was environmentally friendly. Therefore, this work did not only provide a novel catalyzing PMS strategy using hydrochar composite for wastewater treatment, but also give a new idea for herbicide phytotoxicity management.
通讯机构：
[Lianyang Bai; Lang Pan] A;Authors to whom correspondence should be addressed.<&wdkj&>College of Plant Protection, Hunan Agricultural University, No. 1, Nongda Road, Furong District, Changsha 410128, China
摘要：
Asia minor bluegrass (Polypogon fugax) is a common and problematic weed throughout China. P. fugax that is often controlled by acetyl-CoA carboxylase (ACCase) inhibitors in canola fields. Herein, we confirmed a P. fugax population (R) showing resistance to all ACCase inhibitors tested with resistance indexes ranging from 5.4–18.4. We further investigated the resistance mechanisms of this R population. Molecular analyses revealed that an amino acid mutation (Asp-2078-Gly) was present in the R population by comparing ACCase gene sequences of the sensitive population (S). In addition, differences in susceptibility between the R and S population were unlikely to be related to herbicide metabolism. Furthermore, a new derived cleaved amplified polymorphic sequence (dCAPS) method was developed for detecting the Asp-2078-Gly mutation in P. fugax efficiently. We found that 93.75% of plants in the R population carried the Asp-2078-Gly mutation, and all the herbicide-resistant phenotype of this R population is inseparable from this mutation. This is the first report of cross resistance to ACCase inhibitors conferred by the Asp-2078-Gly target-site mutation in P. fugax. The research suggested the urgent need to improve the diversity of weed management practices to prevent the widespread evolution of herbicide resistance in P. fugax in China.
