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Tomato (Solanum lycopersicum L.), as one of the most economically important and highly nutritious vegetable crops across the world, is widely cultivated in China, one of the largest tomato-concuming countries in the world (Ye et al., 2020; Wang and Liu, 2021). At present, major bacterial diseases in tomato include bacterial speck disease, tomato bacterial wilt and bacterial canker, all of which affect the tomato production around the world (Rosli et al., 2021; Peritore-Galve et al., 2021; Wang et al., 2022). In April 2022, a new bacterial disease was discovered on leaves, stems and fruits of tomato in a farmer's greenhouse located in Longfeng District in DaQing (125°07`-125°15`E, 46°28`-46°32`N), Heilongjiang Province, China. This field had tomato disease incidences approximately 50%. Apparent brown discolorations were found on fruits, leaves and stems in tomato plants. Symptoms were similar to fungal brown spots caused by Phytophthora infestans of tomato (Zhi et al.,2021; Liu et al.,2021) (Supplementary Figure S1). To isolate and identify the pathogen, the tissues of infected fruits, leaves and stems with typical symptoms were excised from diseased plants separately, and were disinfected with 75% ethanol for 10 s followed by 2% NaClO for 3 min and then washed five to eight times with sterile water (Wang et al., 2017). Afterwards, the samples were plated on nutrient agar (NA) solid medium and incubated. After incubation at 30°C for 2-3 days, bacterial colonies were isolated, then purified on nutrient agar (NA) solid medium at least twice by a streak plate method (Dou et al., 2019; Li et al, 2021; Zhao et al., 2022). White colonies grew on the NA medium after incubating for 2 days, showing round, opaque and smooth, which was similar to characteristics described as Enterobacter cloacae (García-González et al., 2018; Li et al, 2021). To further confirm the speculation on the identity of the isolated bacterium, the fragments of 16S rRNA were amplified and sequenced. The sequence of 16S rRNA was uploaded into GeneBank with accession numbers (OP077195.1). BLAST analysis of the sequence showed 97.68% identity with one corresponding sequence of E. cloacae in GeneBank (namely MK937637.1). Furthermore, a phylogenetic tree based on the sequence of 16S rRNA gene revealed that the isolate was grouped in the same clade as E. cloacae (Supplementary Figure S2). Based on Koch postulates to test pathogenicity of isolated bacteria, bacteria were inoculated on 30 day-old healthy tomato plants with three leaves stages, and the re-isolation of bacteria were carried out after 2 days of inoculation. To confirm pathogenicity, the isolates were cultured on LB medium at 30℃ for 2 days to prepare suspensions and adjusted to an optical density (OD) of 0.2 at A600, with a final concentration of 1ⅹ108 CFU/ml. Eight potted tomato plants were sprayed with bacteria suspensions, and eight control potted plants were sprayed with sterile distilled water. These seedlings were incubated in a chamber at 30°C with a 12 h light/dark photoperiod, with 85% relative humidity. After 2 days, inoculated tomato seedlings showed irregular small spots in leaves and brown necrosis at blade tips, and 8 to 10 days later, the leaves of tomato plants browned and died. The symptoms were the same with those of the initial diseased leaves of tomato plants (Supplementary Figure S1). No symptoms were observed on the control leaves (Supplementary Figure S3). Pathogenicity tests were repeated three biological times with same results. Meanwhile, the bacteria strains were re-isolated from symptomatic inoculated seedlings and confirmed as E. cloacae by culture and sequence methods as above. In China, there are no detailed records about the causal agent of this disease on tomato in a published paper in Chinese and English. To our knowledge, this is the first report of Enterobacter leaf brown necrosis caused by E. cloacae on tomato in China. Those results are of great significance for the production and management of tomato in greenhouse and control of the disease.

The countries taking part in China’s ‘The Belt and Road’ Initiative have achieved greatly increased trade volume and become more firmly interconnected since the initiative was launched in September...

Netilmicin (NET) is an antibiotic widely used in healthcare and agriculture, but it can accumulate in the environment to threat human health. Netilmicin (NET) is an antibiotic used for veterinary purposes, for human therapy and for agricultural purposes. Therefore, there is a need to develop high-sensitive measuring methods to detect NET. Aptamer-based detecting methods are highly sensitive, inexpensive, and portable. In this study, we developed an aptamer-based fluorescence method to detect and quantify NET. NET was first conjugated to magnetic beads by amidation reaction and then NET-coated beads were used as the stationary phase to isolate aptamers by systematic evolution of ligands by exponential enrichment (SELEX) screening method. After ten rounds of SELEX screening, 32 aptamers with NET-binding affinity were obtained and the candidate aptamer APT-21 was finally chosen by comprehensively comparing their secondary structure characters and NET-binding affinity. APT-21 bound to NET with high affinity (Kd = 194.1 nmol/L) and high specificity that it displayed low cross-binding activities on 7 different structural analogs. We also developed a fluorometric assay using SYBR Green I (SG-I) and the APT-21. Key experimental parameters were optimized, including buffer system, SG-I and APT-21 reaction time, SG-I concentration, and aptamer concentration, to improve the detecting sensitivity. Our results suggest that the low limit of detection (LOD) of this method reached a low level of 1.95 nM and it also exhibited a good linear range up to 200 nM. Moreover, we successfully applied our method to detect the NET spiked in tap water and river water with good recoveries in the range from 97% to 111%. In conclusion, our current study isolated a NET-specific aptamer and developed an aptamer-based quantification method, which is promising to apply to detect NET in environmental samples.

Polygalacturonase (PG) gene has been documented as a key candidate for the improvement of fruit firmness, which is a target trait for tomato production because it facilitates transportation and storage. To reduce the expression of the

The non-host specific Pseudomonas syringae phytotoxin Coronatine (COR) causes chlorosis and promotes toxicity by inducing physiological changes in plants. We performed transcriptome analysis to better understand plants' transcriptional and metabolic response to COR. Toward this end, mock-treated and COR-treated soybean plants were analyzed by RNA-Seq. A total of 4,545 genes were differentially expressed between the two treatments, of which 2,170 were up-regulated whereas 2,375 were down-regulated in COR treated samples. Gene annotation and pathway analysis conducted using the Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) databases revealed that the differential genes were involved in photosynthesis, jasmonic acid (JA) synthesis, signal transduction, and phenylpropane metabolism. This study will provide new insights into COR mediated responses and extend our understanding of COR function in plants.

Coronatine (COR) is a non-host specific phytotoxin secreted by Pseudomonas syringae pv. tomato that can induce leaf chlorosis and increase the virulence of pathogens during plant-pathogen interactions. Studies have shown that COR can regulate multiple physiological processes in plants, but its involvement in bacterial pathogenesis and plant growth regulation is not well understood. In this study, transcriptome sequencing was carried out on 4-week-old tomato leaves that were either mock-treated or treated with COR. Transcriptome sequencing led to the identification of 6144 differentially expressed genes (DEGs), of which 4361 genes were downregulated and 1783 genes were upregulated upon COR treatment. To obtain functional information on the DEGs, we annotated these genes using GO and KEGG databases. Functional classification analysis showed that the DEGs were primarily involved in photosynthesis, chlorophyll and carotenoid biosynthesis, jasmonic acid (JA) synthesis and phenylpropane metabolism. A total of 23 genes related to chlorophyll biosynthesis had significant changes, of which 22 genes were downregulated and one gene was upregulated, indicating that chlorophyll biosynthesis was inhibited upon COR treatment. A total of 17 photosystem I related genes and 22 photosystem II related genes involving 20 protein subunits were also downregulated. In the JA synthesis pathway, 25 genes were up regulated, and six genes were downregulated in COR treated samples. COR was also involved in the regulation of multiple secondary metabolites. The identified DEGs will help us better understand the virulence effects and physiological functions of COR and provide a theoretical basis for breeding resistance into economically important crops.

Kidney bean (Phaseolus vulgaris), also being known as common bean, dry bean or french bean, is one of the most precious and highly nutritious legume crop cultivated and consumed worldwide(Blair et ...

Background Callosobruchus chinensis is one of the important postharvest pests in legume growing areas. Bacterial pesticide is a potential alternative method to control storage pests. However, the effect of these pathogen bacteria on storage pests, and the molecular mechanisms of insect response remain to be to investigated. Results Using the next generation sequencing technology, we established a transcriptomic library for C. chinensis larvae in response to Escherichia coli. Total of 355 differential expressed genes (DEGs) were identified, which 178 DEGs were upregulated, and 177 DEGs were downregulated compared to control group. To validate the RNA-seq analysis, 20 DEGs and 14 immune-related genes were selected to perform quantitative polymerase chain reaction (RT-qPCR). These immune-related genes were involved in recognition (peptidoglycan recognition proteins), signal transduction (fibrinogen-related proteins, serine proteinases and NF-κB), and execution effectors (phenoloxidase, defensin, attacin, and antimicrobial peptide). In addition, genes that encode digestive and respiratory enzymes were altered in C. chinensis larvae in response to infection. Some genes that involved in juvenile hormone and insulin pathway appeared to express differentially, suggesting that pathogen infection might lead to developmental arrest. Furthermore, iron homeostasis and chitin metabolism appeared significantly altered after infection. Conclusion In this study, we characterized the immune response of C. chinensis larvae in response to E. coli using RNA-seq, from pathogen recognition, signal transduction, to execution. Some other identified genes were involved in iron homeostasis, respiration, and digestion. A better understanding of molecular response of beetle to pathogen will facilitate us to develop an available strategy to control storage pests.

In recent years, the international crude oil price has become increasingly volatile. It influences the exchange rate changes of relevant countries through economic growth, price level, international balance of payments, and other channels. Such exchange rate fluctuations have caused certain risks for the development of China’s “Belt and Road” Initiative. This article analyzes the impact of oil price changes on the exchange rates of countries. Because the fluctuation of oil prices and exchange rates has shown the characteristics of multiple time scales, this study used the empirical mode decomposition (EMD) method to obtain the long-cycle and short-cycle sequences of oil prices and the exchange rates of various countries, then analyzed the impact of oil price changes on exchange rates under different time scales. The results showed that oil price fluctuations have an impact on the exchange rate changes of countries along the “Belt and Road” under different time scales. However, this effect is asymmetric between oil-producing countries and non-oil-producing countries, and the transmission path of oil prices to exchange rates varies from cycle to cycle.

BACKGROUND Insect tolerance to low oxygen (hypoxia) and high carbon dioxide (hypercapnia) is critical for insect control. On the basis of bioassay, metabolism profiles were built to investigate adaptive mechanisms in bean weevil under hypoxia (2% O2), hypoxia/hypercapnia (2% O2 + 18% CO2) and normoxia (control, 20% O2 + 80% N2) using gas chromatography/time-of-flight mass spectrometry (GC/TOF-MS). RESULTS The growth and development of bean weevils were significantly suppressed by the two hypoxia conditions; hypercapnia enhanced the mortality, but after 24 days of exposure, the surviving insects emerged as adults earlier than those under hypoxia only. Metabolism profiles also showed striking differences in metabolites among the treatment and control groups, both quantitatively and qualitatively. Pairwise comparisons of the three groups showed that 61 metabolites changed significantly, 40 in the hypoxia group and 37 in the hypoxia/hypercapnia group relative to the control group, while only 16 were shared equally by the hypoxia and hypoxia/hypercapnia groups. Increased metabolites were mainly carbohydrates, amino acids and organic acids, while free fatty acids were decreased. Furthermore, the changes were strengthened by the addition of hypercapnia, but excluding free fatty acids. CONCLUSION The findings show that bean weevil has high tolerance to hypoxia or even hypoxia/hypercapnia at biologically achievable levels and provide more direct evidence for stored product insect mechanism regulation under hypoxia stress, especially free fatty acid regulation by hypercapnia but not by hypoxia. © 2016 Society of Chemical Industry

In recent years, botanical insecticides have gradually replaced the traditional synthetic insecticides, with the properties of strong pertinence and environment friendly. Many new botanical insecticides are developed and used, meanwhile insect develop its resistance to these insecticides. Previous studies have proved that Cytochrome P450 monooxygenases (CYPs) play a critical role in insect detoxification. Genomic and proteomic approaches focused on P450 expression and regulation at the transcript and protein levels have made substantial progress. The review covers the recent years’ research of typical botanicals metabolism mediated by CYPs, the relationship between function and structural characters of P450 enzymes, as well as the expression and regulation of related P450 genes in botanicals metabolism.

Type III effectors AvrRpm1 and AvrRpt2 promote bacterial growth dependent on a COI1-mediated pathway in the absence of the RPM1 and RPS2 resistance proteins. The type III effectors, AvrRpm1 and AvrRpt2, promote bacterial virulence by suppressing host defense responses. The defense suppressing activities of AvrRpm1 and AvrRpt2 are best studied in the absence of the resistance proteins RPM1 and RPS2, which induce defense responses to them. We tested whether the type III effectors could modulate a CORONATINE INSENSITIVE1 (COI1)-mediated hormone signaling pathway to promote virulence. COI1 has been demonstrated to contribute in the induction of chlorosis during Pseudomonas syringae infection. By comparing the activity of inducibly expressed AvrRpm1-HA or AvrRpt2-HA in rpm1rps2 and rpm1rps2coi1 backgrounds, we demonstrate that both effectors promote bacterial growth dependent on a COI1-mediated pathway and additively with the action of coronatine (COR) and that AvrRpt2-HA induces COI1-dependent chlorosis. Further, PATHOGENESIS RELATED1 (PR-1) expression resulting from inducible expression of AvrRpm1-HA or AvrRpt2-HA is elevated in coi1 plants consistent with the effectors activating JA-signaling to antagonize SA-signaling. In addition, we found that AvrRpm1-HA or AvrRpt2-HA requires COI1 to promote bacterial growth through suppression of both SA-dependent and SA-independent defense responses. Collectively, these results indicate that type III effectors AvrRpm1 and AvrRpt2 promote bacterial virulence by targeting a COI1-dependent signaling pathway.

Oxygen is essential for aerobic life, and hypoxia has very severe consequences. Organisms need to overcome low oxygen levels to maintain biological functions during normal development and in disease states. The mechanism underlying the hypoxic response has been widely investigated in model animals such as Drosophila melanogaster and Caenorhabditis elegans. Hypoxia-inducible factor (HIF), a key gene product in the response to oxygen deprivation, is primarily regulated by prolyl hydroxylase domain enzymes (PHDs). However, recent findings have uncovered novel HIF-independent functions of PHDs. This review provides an overview of how invertebrates are able to sustain hypoxic damages, and highlights some recent discoveries in the regulation of cellular signalling by PHDs. Given that some core genes and major pathways are evolutionarily conserved, these research findings could provide insight into oxygen-sensitive signalling in mammals, and have biomedical implications for human diseases.

Plant pathogens deploy an array of virulence factors to suppress host defense and promote pathogenicity. Numerous strains of Pseudomonas syringae produce the phytotoxin coronatine (COR). A major aspect of COR function is its ability to mimic a bioactive jasmonic acid (JA) conjugate and thus target the JA-receptor COR-insensitive 1 (COI1). Biological activities of COR include stimulation of JA-signaling and consequent suppression of SA-dependent defense through antagonistic crosstalk, antagonism of stomatal closure to allow bacterial entry into the interior of plant leaves, contribution to chlorotic symptoms in infected plants, and suppression of plant cell wall defense through perturbation of secondary metabolism. Here, we review the virulence function of COR, including updates on these established activities as well as more recent findings revealing COI1-independent activity of COR and shedding light on cooperative or redundant defense suppression between COR and type III effector proteins.

Insect tolerance to low oxygen (hypoxia) and high carbon dioxide (hypercapnia) is critical for insect control. On the basis of bioassay, metabolism profiles were built to investigate adaptive mechanisms in bean weevil under hypoxia (2% OThe growth and development of bean weevils were significantly suppressed by the two hypoxia conditions; hypercapnia enhanced the mortality, but after 24 days of exposure, the surviving insects emerged as adults earlier than those under hypoxia only. Metabolism profiles also showed striking differences in metabolites among the treatment and control groups, both quantitatively and qualitatively. Pairwise comparisons of the three groups showed that 61 metabolites changed significantly, 40 in the hypoxia group and 37 in the hypoxia/hypercapnia group relative to the control group, while only 16 were shared equally by the hypoxia and hypoxia/hypercapnia groups. Increased metabolites were mainly carbohydrates, amino acids and organic acids, while free fatty acids were decreased. Furthermore, the changes were strengthened by the addition of hypercapnia, but excluding free fatty acids.The findings show that bean weevil has high tolerance to hypoxia or even hypoxia/hypercapnia at biologically achievable levels and provide more direct evidence for stored product insect mechanism regulation under hypoxia stress, especially free fatty acid regulation by hypercapnia but not by hypoxia. © 2016 Society of Chemical Industry.

The phytotoxin coronatine (COR) promotes various aspects of Pseudomonas syringae virulence, including invasion through stomata, growth in the apoplast, and induction of disease symptoms. COR is a structural mimic of active jasmonic acid (JA) conjugates. Known activities of COR are mediated through its binding to the F-box-containing JA coreceptor CORONATINE INSENSITIVE1. By analyzing the interaction of P. syringae mutants with Arabidopsis thaliana mutants, we demonstrate that, in the apoplastic space of Arabidopsis, COR is a multifunctional defense suppressor. COR and the critical P. syringae type III effector HopM1 target distinct signaling steps to suppress callose deposition. In addition to its well-documented ability to suppress salicylic acid (SA) signaling, COR suppresses an SA-independent pathway contributing to callose deposition by reducing accumulation of an indole glucosinolate upstream of the activity of the PEN2 myrosinase. COR also suppresses callose deposition and promotes bacterial growth in coi1 mutant plants, indicating that COR may have multiple targets inside plant cells.

Advances in horticulture plant biotechnologies provide new opportunities for researchers to study the field of vegetative propagation and genetic engineering. Developments of clonal propagation methods, especially somatic embryogenesis (SE), have numerous potential applications. This paper reviewed progress of research on SE in horticultural plants in last decade; analyzed plant regeneration having both direct and indirect SE from the characteristics of occurrence means, but mainly in an indirect way; and discussed the impact factors of SE, as well as reviewed the research in the practical applications of horticulture plants SE in the practice.

Regulating the air in low-oxygen environments protects hermetically stored grains from storage pests damage. However, pests that can tolerate hypoxic stress pose a huge challenge in terms of grain storage. We used various biological approaches to determine the fundamental mechanisms of Tribolium castaneum to cope with hypoxia. Our results indicated that limiting the available oxygen to T. castaneum increased glycolysis and inhibited the Krebs cycle, and that accumulated pyruvic acid was preferentially converted to lactic acid via anaerobic metabolism. Mitochondrial aerobic respiration was markedly suppressed for beetles under hypoxia, which also might have led to mitochondrial autophagy. The enzymatic activity of citrate synthase decreased in insects under hypoxia but recovered within 12 h, which suggested that the beetles recovered from the hypoxia. Moreover, hypoxia-reperfusion resulted in severe oxidative damage to insects, and antioxidant levels increased to defend against the high level of reactive oxygen species. In conclusion, our findings show that mitochondria were the main target in T. castaneum in response to low oxygen. The beetles under hypoxia inhibited mitochondrial respiration and increased antioxidant activity after reoxygenation. Our research advances the field of pest control and makes it possible to develop more efficient strategies for hermetic storage.

Plant disease resistance (R) proteins recognize potential pathogens expressing corresponding avirulence (Avr) proteins through 'gene-for-gene' interactions. RPM1 is an Arabidopsis R-protein that triggers a robust defense response upon recognizing the Pseudomonas syringae effector AvrRpm1. Avr-proteins of phytopathogenic bacteria include type III effector proteins that are often capable of enhancing virulence when not recognized by an R-protein. In rpm1 plants, AvrRpm1 suppresses basal defenses induced by microbe-associated molecular patterns. Here, we show that expression of AvrRpm1 in rpm1 plants induced PR-1, a classical defense marker, and symptoms including chlorosis and necrosis. PR-1 expression and symptoms were reduced in plants with mutations in defense signaling genes (pad4, sid2, npr1, rar1, and ndr1) and were strongly reduced in rpm1 rps2 plants, indicating that AvrRpm1 elicits defense signaling through the Arabidopsis R-protein, RPS2. Bacteria expressing AvrRpm1 grew more on rpm1 rps2 than on rpm1 plants. Thus, independent of its classical 'gene-for-gene' activation of RPM1, AvrRpm1 also induces functionally relevant defenses that are dependent on RPS2. Finally, AvrRpm1 suppressed host defenses and promoted the growth of type III secretion mutant bacteria equally well in rps2 and RPS2 plants, indicating that virulence activity of over-expressed AvrRpm1 predominates over defenses induced by weak activation of RPS2.

Construction of transgenic plants is central to modern plant molecular genetics. Inducible systems permit spatial and temporal control of transgene expression. One commonly used inducible system relies on the use of dexamethasone to activate an endogenously expressed hybrid transcription factor, which positively regulates the expression of the gene of interest (Aoyama and Chua, Plant J 11:605-612, 1997). We have developed Arabidopsis plants using this inducible system to drive expression of a bacterial type III effector protein. The effector, AvrRpm1, elicits either strong cell death or weak cell death and chlorosis depending on the genetic background of the plant. Using these reagents, we examine several properties of the inducible system in Arabidopsis, including the timing of induction, the ability to tune the level of transgene expression by altering the concentration of applied dexamethasone, and the movement of dexamethasone within the plant.