Mycoviral gene integration converts a plant pathogenic fungus into a biocontrol agent.

Mycovirus-infected fungi can suffer from poor growth, attenuated pigmentation, and virulence. However, the molecular mechanisms of how mycoviruses confer these symptoms remain poorly understood. Here, we report a mycovirus Stemphylium lycopersici alternavirus 1 (SlAV1) isolated from a necrotrophic plant pathogen Stemphylium lycopersici that causes altered colony pigmentation and hypovirulence by specifically interfering host biosynthesis of Altersolanol A, a polyketide phytotoxin. SlAV1 significantly down-regulates a fungal polyketide synthase (PKS1), the core enzyme of Altersolanol A biosynthesis. PKS1 deletion mutants do not accumulate Altersolanol A and lose pathogenicity to tomato and lettuce. Transgenic expression of SlAV1 open-reading frame 3 (ORF3) in S. lycopersici inhibits fungal PKS1 expression and Altersolanol A accumulation, leading to symptoms like SlAV1-infected fungal strains. Multiple plant species sprayed with mycelial suspension of S. lycopersici or S. vesicarium strains integrating and expressing ORF3 display enhanced resistance against virulent strains, converting the pathogenic fungi into biocontrol agents. Hence, our study not only proves inhibiting a key enzyme of host phytotoxin biosynthesis as a molecular mechanism underlying SlAV1-mediated hypovirulence of Stemphylium spp., but also demonstrates the potential of mycovirus-gene integrated fungi as a potential biocontrol agent to protect plants from fungal diseases.

Functional analysis of UDP-glycosyltransferase genes conferring indoxacarb resistance in Spodoptera litura.

UDP-glycosyltransferase (UGT) is the major detoxification enzymes of phase II involved in xenobiotics metabolism, which potentially mediates the formation of insect resistance. Previous transcriptome sequencing studies have found that several UGT genes were upregulated in indoxacarb resistant strains of Spodoptera litura, but whether these UGT genes were involved in indoxacarb resistance and their functions in resistance were unclear. In this study, the UGTs inhibitor, 5-nitrouracil, enhanced the toxicity of indoxacarb against S. litura, preliminarily suggesting that UGTs were participated in indoxacarb resistance. Two UGT genes, UGT33J17 and UGT41D10 were upregulated in the resistant strains and could be induced by indoxacarb. Alignment of UGT protein sequences revealed two conserved donor-binding regions with several key residues that interact with catalytic sites and sugar donors. Further molecular modeling and docking analysis indicated that two UGT proteins were able to stably bind indoxacarb and N-decarbomethoxylated metabolite (DCJW). Furthermore, knockdown of UGT33J17 and UGT41D10 decreased viability of Spli-221 cells and enhanced susceptibility of larvae to indoxacarb. Transgenic overexpression of these genes reduced the toxicity of indoxacarb in Drosophila melanogaster. This work revealed that upregulation of UGT genes significantly contributes to indoxacarb resistance in S. litura, and is of great significance for the development of integrated and sustainable management strategies for resistant pests in the field.

Semi-Supervised Medical Image Segmentation Guided by Bi-Directional Constrained Dual-Task Consistency.

BACKGROUND: Medical image processing tasks represented by multi-object segmentation are of great significance for surgical planning, robot-assisted surgery, and surgical safety. However, the exceptionally low contrast among tissues and limited available annotated data makes developing an automatic segmentation algorithm for pelvic CT challenging. METHODS: A bi-direction constrained dual-task consistency model named PICT is proposed to improve segmentation quality by leveraging free unlabeled data. First, to learn more unmarked data features, it encourages the model prediction of the interpolated image to be consistent with the interpolation of the model prediction at the pixel, model, and data levels. Moreover, to constrain the error prediction of interpolation interference, PICT designs an auxiliary pseudo-supervision task that focuses on the underlying information of non-interpolation data. Finally, an effective loss algorithm for both consistency tasks is designed to ensure the complementary manner and produce more reliable predictions. RESULTS: Quantitative experiments show that the proposed PICT achieves 87.18%, 96.42%, and 79.41% mean DSC score on ACDC, CTPelvic1k, and the individual Multi-tissue Pelvis dataset with gains of around 0.8%, 0.5%, and 1% compared to the state-of-the-art semi-supervised method. Compared to the baseline supervised method, the PICT brings over 3-9% improvements. CONCLUSIONS: The developed PICT model can effectively leverage unlabeled data to improve segmentation quality of low contrast medical images. The segmentation result could improve the precision of surgical path planning and provide input for robot-assisted surgery.

Systematic identification and functional analysis of long noncoding RNAs involved in indoxacarb resistance in Spodoptera litura.

Long noncoding RNAs (lncRNAs) are noncoding transcripts that are more than 200 nucleotides long. They play essential roles in regulating a variety of biological processes in many species, including insects, and some lncRNAs have been found to be associated with insecticide resistance. However, the characteristics and biological functions of lncRNAs involved in indoxacarb resistance are unknown in Spodoptera litura. We performed RNA sequencing in the SS, InRS, and FInRS of S. litura and identified 11 978 lncRNAs, including 3 136 intergenic lncRNAs, 7 393 intronic lncRNAs, and 1 449 anti-sense lncRNAs. Compared with the SS, 51 lncRNAs were upregulated and 134 lncRNAs were downregulated in the two resistant strains, and 908 differentially expressed mRNAs were predicted as the target genes of the 185 differentially expressed lncRNAs. Further analysis showed that 112 of differentially expressed lncRNAs may be associated with indoxacarb resistance by regulating the expression of 14 P450s, seven CCEs, one GST, six UGTs, five ABC transporters, and 24 cuticle protein genes, and 79 of differentially expressed lncRNAs may regulate the expression of 14 detoxification genes and 19 cuticle protein genes to participate in indoxacarb resistance by sponging 10 microRNAs. Interestingly, 47 of differentially expressed lncRNAs may mediate indoxacarb resistance through both lncRNA-mRNA and lncRNA-miRNA-mRNA regulatory pathways. Furthermore, quantitative PCR, RNA interference, and indoxacarb bioassay analyses indicated that overexpressed LNC_004867 and LNC_006576 were involved in indoxacarb resistance. This study provides comprehensive information for lncRNAs of S. litura, and presents evidence that lncRNAs have key roles in conferring insecticide resistance in S. litura.

Transcription factor CncC potentially regulates the expression of multiple detoxification genes that mediate indoxacarb resistance in Spodoptera litura.

The xenobiotic transcription factor cap 'n' collar isoform C (CncC) is considered the central regulator of antioxidant and detoxification genes. Previous research indicated that CncC regulates three-phase enzymes responsible for insecticide resistance. In this study, the SlituCncC gene from Spodoptera litura was obtained and characterized. Quantitative polymerase chain reaction (qPCR) analysis showed that SlituCncC was expressed in all developmental stages and tissues, but was highly expressed in 3rd- and 4th-instar larvae, and in the Malpighian tubule, fat body, and midgut. In addition, SlituCncC was up-regulated and more highly induced with indoxacarb treatment in the indoxacarb-resistant strains compared with the susceptible strain. RNA interference-mediated gene silencing of SlituCncC significantly increased mortality of S. litura when exposed to indoxacarb. Furthermore, comparative transcriptome analysis showed that 842 genes were down-regulated and 127 genes were up-regulated in SlituCncC knockdown S. litura. Further analysis indicated that 18 three-phase enzymes were identified in the down-regulated genes, of which seven were associated with indoxacarb resistance in S. litura. qPCR analysis confirmed that expression of six of these seven genes was consistent with RNA sequencing data. All six detoxification genes were induced by indoxacarb, and the expression patterns were similar to that of SlituCncC. Finally, the CncC-Maf binding site was predicted in all six gene promoters. This study indicates that the transcription factor SlituCncC may regulate multiple detoxification genes that mediate indoxacarb resistance in S. litura.

Detection of Xanthomonas oryzae pv. oryzae in seeds using a specific TaqMan probe.

Xanthomonas oryzae pv. oryzae is the pathogen that causes bacterial leaf blight in rice. Bacterial leaf blight is the main cause for severe rice underproduction in many countries. However, with conventional methods it is difficult to quickly and reliably distinguish this pathogen from other closely related pathogenic bacteria, especially X. oryzae pv. oryzicola, the causal organism of bacterial leaf streak in rice. We have developed a novel and highly sensitive real-time method for the identification of this specific bacteria based on a TaqMan probe. This probe is designed to recognize the sequence of a putative siderophore receptor gene cds specific to X. oryzae pv. oryzae, and can be identified from either a bacterial culture or naturally infected rice seeds and leaves in only 2 h. The sensitivity of the method is 100 times higher than that of the current polymerase chain reaction (PCR) gel electrophoresis method for diagnosis.

Electroantennogram and behavioral responses of the imported fire ant, Solenopsis invicta Buren, to an alarm pheromone component and its analogues.

A characteristic behavior in ants is to move rapidly to emission sources of alarm pheromones. The addition of ant alarm pheromones to bait is expected to enhance its attractiveness. To search for candidate compounds for bait enhancement in fire ant control, 13 related alkylpyrazine analogues in addition to synthetic alarm pheromone component were evaluated for electroantennogram (EAG) and behavioral activities in Solenopsis invicta. Most compounds elicited dose-dependent EAG and behavioral responses. There exists a correlation between the EAG and behavioral responses. Among the 14 tested alkylpyrazines, three compounds, 2-ethyl-3,6(5)-dimethyl pyrazine (1), 2,3,5-trimethylpyrazine (7), and 2,3-diethyl-5-methylpyrazine (12), elicited significant alarm responses at a dose range of 0.1-1000 ng. Further bait discovery bioassay with the three most active alkylpyrazines demonstrated that food bait accompanied by sample-treated filter paper disk attracted significantly more fire ant workers in the first 15 min period. EAG and behavioral bioassays with pure pheromone isomers accumulated by semi-preparative high-performance liquid chromatography demonstrated that 2-ethyl-3,6-dimethylpyrazine was significantly more active than 2-ethyl-3,5-dimethylpyrazine.

[Distribution and risk assessment of polycyclic aromatic hydrocarbons in urban soils: a review].

With rapid urbanization, the contamination of polycyclic aromatic hydrocarbons (PAHs) has become one of the major factors threatening urban soil quality and human health. This paper summarized the current studies on the PAHs in urban soils, with the focus on the anthropogenic and natural factors affecting the distribution of PAHs in urban soils, analytical methods on the pollution sources of PAHs, risk assessment of urban soil contamination by PAHs, and especially, applications of geo-statistics in the spatial analysis and risk assessment of PAHs. The potential issues and research trends about the PAHs contamination of urban soils were also discussed.

[Ecological distributions of airborne fungi in outdoor environments in Beijing, China].

An investigation on fungal types, concentrations, and their dynamic variation in outdoor environments was carried out in three different functional areas around one year in Beijing. Results show that the fungal concentrations varied widely and the average was (1164.8 +/- 73.2) CFU x m(-3), ranging from 23.6CFU x m(-3) to 13 959.5 CFU x m(-3). The most common culturable airborne fungi in all seasons and all functional areas were Cladosporium , nonsporulating fungi, Alternaria, Penicillium and Aspergillus. The most dominant fungus was Cladosporium, which contributed to more than 1/3 of the total. The fungal levels in culture and education region (CER) and garden green region (GGR) were highest in the fall and summer, and lowest in the winter and spring, while the seasonal variation in main traffic line (MTL) was not significant. The fungal concentrations in CER and GGR were significantly higher than in MTL (p <0.05). No statistically significant difference exists between CER and GGR.