A novel spider venom peptide from the predatory mite Neoseiulus barkeri shows lethal effect on phytophagous pests.

The long-term use of pesticides in the field, and the high fertility and adaptability of phytophagous mites have led to resistance problems; consequently, novel safe and efficient active substances are necessary to broaden the tools of pest mite control. Natural enemies of arthropods typically secrete substances with paralytic or lethal effects on their prey, and those substances are a resource for future biopesticides. In this study, two putative venom peptide genes were identified in a parasitic mite Neoseiulus barkeri transcriptome. Recombinant venom NbSP2 peptide injected into Tetranychus cinnabarinus mites was significantly more lethal than recombinant NBSP1. NbSP2 was also lethal to Spodoptera litura when injected but not when fed to third instar larvae. The interaction proteins of NbSP2 in T. cinnabarinus and S. litura were identified by affinity chromatography. Among these proteins, ATP synthase subunit ß (ATP SSß) was deduced as a potential target. Four binding sites were predicted between NBSP2 and ATP SSß of T. cinnabarinus and S. litura. In conclusion, we identified a venom peptide with activity against T. cinnabarinus and S. litura. This study provides a novel component for development of a new biological pesticide.

Amidase, a novel detoxifying enzyme, is involved in cyflumetofen resistance in Tetranychus cinnabarinus (Boisduval).

Amidase is an important hydrolytic enzyme in detoxification metabolism. Amidase hydrolyzes a wide variety of nonpeptide carbon­nitrogen bonds by attacking a cyano group or carbonyl carbon. However, little is known about the relationship between amidase and insecticides. In this study, the amidase activity was significantly higher in cyflumetofen-resistant strain (CyR) than in the susceptible strain (SS) of Tetranychus cinnabarinus, and diethyl-phosphoramidate (an amidase inhibitor) significantly decreased cyflumetofen resistance in T. cinnabarinus. More importantly, an amidase gene, TcAmidase01, was identified in T. cinnabarinus, and the TcAmidase01 overexpression was detected in both two cyflumetofen-resistant strains (CyR and YN-CyR), indicating that it is involved in cyflumetofen resistance in mites. A phylogenetic analysis showed that TcAmidase01 was clustered with deaminated glutathione amidases, which possess hydrolytic activity. The recombinant TcAmidase01 protein showed amidase activity toward succinamate, and the activity could be inhibited by cyflumetofen. High-performance liquid chromatography-mass spectrometry (HPLC-MS) analysis provided evidence that recombinant TcAmidase01 could decompose cyflumetofen by hydrolysis, and the potential metabolites (2-(4-(tert-butyl) phenyl)-2-cyanoacetate and 2-(trifluoromethyl) benzoic acid) were identified. These results show that TcAmidase01 contribute to cyflumetofen-resistance in T. cinnabarinus by hydrolyzing cyflumetofen, and this is the first study to suggest that amidase has a role in insecticides resistance in arthropods.

RNAi targeting ecdysone receptor blocks the larva to adult development of Tetranychus cinnabarinus.

RNA interference (RNAi) is a potentially useful pest control method because of its high specificity. Silencing the expression of important RNAi target genes of pests will block important biological processes and reduce pest damage. Ecdysone is a unique arthropod hormone and the ecdysone receptor (EcR) is a key factor in molting pathway. We investigated the possibility that dsRNA targeting of the EcR of Tetranychus cinnabarinus (TcEcR) could effectively block development from larvae to adults. The mRNA level of TcEcR was highest in the larva stage, and 73.1% of the mites failed to survive the larva stage when TcEcR expression was silenced. Only 11.7% of T. cinnabarinus ingesting dsRNA successfully developed into adults, while 86.7% in the control succeeded in molting across each stage. RNAi significantly increased the developmental intervals of T. cinnabarinus. Under the effects of dsRNA, development times for the larva and first nymph doubled. Phenotype of body size change and death were observed during the development of T. cinnabarinus ingesting dsRNA. These findings suggest that RNAi is a potential means for the control of T. cinnabarinus. Genes in hormone pathways such as EcR are possible RNAi targets.

Functional analysis of five trypsin-like protease genes in the oriental fruit fly, Bactrocera dorsalis (Diptera: Tephritidae).

Insect midgut proteases catalyze the release of free amino acids from dietary proteins and are essential for insect normal development. To date, digestive proteases as potential candidates have made great progress in pest control. To clarify the function of trypsin-like protease genes in the digestive system of Bactrocera dorsalis, a serious pest of a wide range of tropical and subtropical fruit and vegetable crops, five trypsin genes (BdTry1, BdTry2, BdTry3, BdTry4 and BdTry5) were identified from transcriptome dataset, and the effects of feeding condition on their expression levels were examined subsequently. RNA interference (RNAi) was applied to further explore their function on the growth of B. dorsalis. The results showed that all the BdTrys in starving midgut expressed at a minimal level but up-regulated upon feeding (except BdTry3). Besides, RNAi by feeding dsRNAs to larvae proved to be an effective method to cause gene silencing and the mixed dsRNAs of the five BdTrys slowed larvae growth of B. dorsalis. The current data suggest that trypsin genes are actively involved in digestion process of B. dorsalis larvae and thereafter play crucial roles in their development.

Insecticide resistance monitoring and metabolic mechanism study of the green peach aphid, Myzus persicae (Sulzer) (Hemiptera: Aphididae), in Chongqing, China.

Myzus persicae (Sulzer) is one of the most important agricultural pests in China, which caused serious losses every year. For resistance monitoring, twelve populations of this pest were collected from tobacco field in Chongqing, China, and their sensitivities to 4 insecticides were tested. Results showed that only WL (RR=6.51) and FJ (RR=6.03) populations have developed minor resistance to imidacloprid, and the others still remained susceptible. One population (NC) has reached a high resistance level to cyhalothrin (RR=41.28), five populations showed medium level (10.36≤RR≤20.45), and the other six remained susceptible (0.39≤RR≤3.53). As regards carbosulfan, three populations have developed medium resistance, four populations showed only minor resistance, and the other five (0.81≤RR≤3.97) were still susceptible. Population SZ developed a medium level (RR=14.83) to phoxim, the other 11 were susceptible (0.29≤RR≤2.41). To analysis the potential resistance mechanism, inhibition effects of synergists and detoxifying enzyme activities were detected. The results indicated that the MFO was the most important detoxifying enzyme conferring imidacloprid resistance, and CarE was most important to cyhalothrin, carbosulfan and phoxim. Our study provided a comprehensive survey of insecticide resistance of M. persicae in Chongqing, and suggested that different counties should take corresponding management to delay the insecticide resistance development and prolong the usefulness of insecticides.

The fenpropathrin resistant Tetranychus cinnabarinus showed increased fecundity with high content of vitellogenin and vitellogenin receptor.

Carmine spider mite, Tetranychus cinnabarinus (Boisduval), an agricultural pest of economically important crops, has developed resistance to a group of pesticides. We have selected a fenpropathrin-resistant strain (FeR) of T. cinnabarinus from the isogenous and susceptible strain (SS), and found that the FeR not only showed resistance but its fecundity also increased. According to the numbers of eggs laid per day of both strains, the FeR was more fertile than SS throughout the life span. To investigate the underlying reason, the protein contents of vitellogenin (vg) and vitellogenin receptor (vgr) were detected, and the results showed both of them were significantly higher in FeR than in SS. Then, the mRNA-expressions of vg and vgr genes were compared between FeR and SS. From the transcriptome data of T. cinnabarinus, we classified two vg genes (designated as Tcvg1 and Tcvg2, respectively) and a vgr gene (designated as Tcvgr). The expressions of Tcvg1, Tcvg2 and Tcvgr were highly associated with the fecundity of the mites that their mRNAs were extremely abundant at the adult stage, but hardly detectable during the developmental period (from egg to deutonymph). In accordance with the protein content, the expression levels of the three genes were all significantly higher in FeR than they were in SS. These results suggested that after resistance selection with fenpropathrin in T. cinnabarinus, the fecundity and the expression of reproduction-related genes (vg and vgr) were significantly higher in fenpropathrin resistant strain than that in susceptible strain.

Analysis of the relationship between P-glycoprotein and abamectin resistance in Tetranychus cinnabarinus (Boisduval).

Abamectin is an effective acaricide and widely used in the control of Tetranychus cinnabarinus. With the increase of control failures, it is however important to clarify the resistance mechanism to improve the control of this mite. P-glycoprotein (Pgp) is an ATP-dependent drug efflux pump for xenobiotic compounds and is involved in multidrug resistance. In this study, the results showed that verapamil, the specific inhibitor of Pgp, could enhance the lethal effect of abamectin on mites, and this effect is more enhanced in abamectin-resistant strain (AbR, mortality increased 74.51%) than that in susceptible strain (SS, 19.91%). Further analysis showed that the activity of Pgp ATPase in AbR was significantly higher (1.65-fold) than that in SS. After exposure to sublethal concentration of abamectin, the ATPase activity in AbR was significantly increased 1.43-fold to that in control; but there was no significant difference in SS after treatment. Two Pgp gene sequences (TcPgp1 and TcPgp2) from ABCB subfamily were characterized, and their expressions were much more sensitive to abamectin's stimulation in AbR strain than SS. These findings indicate a direct relationship between Pgp and abamectin resistance, and abamectin-induced Pgp expression may be involved in the modulation of abamectin efflux in T. cinnabarinus.

Characteristics of carboxylesterase genes and their expression-level between acaricide-susceptible and resistant Tetranychus cinnabarinus (Boisduval).

Carboxylesterases (CarEs) play important roles in metabolism and detoxification of dietary and environmental xenobiotics in insects and mites. On the basis of the Tetranychuscinnabarinus transcriptome dataset, 23 CarE genes (6 genes are full sequence and 17 genes are partial sequence) were identified. Synergist bioassay showed that CarEs were involved in acaricide detoxification and resistance in fenpropathrin- (FeR) and cyflumetofen-resistant (CyR) strains. In order to further reveal the relationship between CarE gene's expression and acaricide-resistance in T. cinnabarinus, we profiled their expression in susceptible (SS) and resistant strains (FeR, and CyR). There were 8 and 4 over-expressed carboxylesterase genes in FeR and CyR, respectively, from which the over-expressions were detected at mRNA level, but not DNA level. Pesticide induction experiment elucidated that 4 of 8 and 2 of 4 up-regulated genes were inducible with significance in FeR and CyR strains, respectively, but they could not be induced in SS strain, which indicated that these genes became more enhanced and effective to withstand the pesticides' stress in resistant T. cinnabarinus. Most expression-changed and all inducible genes possess the Abhydrolase_3 motif, which is a catalytic domain for hydrolyzing. As a whole, these findings in current study provide clues for further elucidating the function and regulation mechanism of these carboxylesterase genes in T. cinnabarinus' resistance formation.

Expression characteristics of two novel cytochrome P450 genes involved in fenpropathrin resistance in Tetranychus cinnabarinus (Boisduval).

The carmine spider mite, Tetranychus cinnabarinus, which is also considered as the red form of Tetranychus urticae, is one of the most serious mite pests on crops. It is capable of rapidly developing resistance to acaricides, and has caused difficulty in controlling. However, the resistance mechanism of this mite remains unclear at molecular level. As a member of main detoxification enzymes, cytochrome P450 monooxygenases (CYPs or P450s) play important roles in the development of acaricide resistance in arthropods. In this study, two novel P450 genes (CYP389B1 and CYP392A26) were identified and characterized from T. cinnabarinus. The opening reading frames (ORFs) of CYP389B1 and CYP392A26 contained 1545 and 1488 nucleotides, which encode 514 and 495 amino acids, respectively. Phylogenetic analysis showed that CYP389B1 and CYP392A26 were most closely related to CYP389B1 and CYP392A4 from T. urticae, respectively. When treated with piperonyl butoxide (PBO), an inhibitor of P450s, the resistance ratio of fenpropathrin-resistant (FeR) strain decreased from 101- to 75-fold, which suggested a correlation between P450 and fenpropathrin-resistance in T. cinnabarinus. Furthermore, constitutive over-expressions of CYP389B1 and CYP392A26 were detected in FeR strain. Meanwhile, the expressions of CYP389B1 and CYP392A26 were inducible in FeR strain after treatment in 6, 12 and 24 h with LC30 of fenpropathrin; especially, the expression of CYP392A26 increased to a markedly high level (20.88-fold higher than in the control) after treatment in 6 h. However, there was no significant difference between treatment and control in susceptible strain. Furthermore, stage specific expression profiles of these two genes did not show significant difference among developing stages, except for eggs, in which the mRNA levels were quite low. The results indicate that CYP389B1 and CYP392A26 were involved in the fenpropathrin-resistance in T. cinnabarinus, and the expression of CYP392A26 was more sensitive to fenpropathrin stress. These findings provide clues for further elucidating the function and regulation mechanism of these two cytochrome P450 genes in T. cinnabarinus.

Reference Gene Validation for Quantitative PCR Under Various Biotic and Abiotic Stress Conditions in Toxoptera citricida (Hemiptera, Aphidiae).

The regulation of mRNA expression level is critical for gene expression studies. Currently, quantitative reverse transcription polymerase chain reaction (qRT-PCR) is commonly used to investigate mRNA expression level of genes under various experimental conditions. An important factor that determines the optimal quantification of qRT-PCR data is the choice of the reference gene for normalization. To advance gene expression studies in Toxoptera citricida (Kirkaldy), an important citrus pest and a main vector of the Citrus tristeza virus, we used five tools (GeNorm, NormFinder, BestKeeper, ΔCt methods, and RefFinder) to evaluate seven candidate reference genes (elongation factor-1 alpha [EF1α], beta tubulin [ß-TUB], 18S ribosomal RNA [18S], RNA polymerase II large subunit (RNAP II), beta actin (ß-ACT), alpha tubulin, and glyceraldhyde-3-phosphate dehydrogenase) under different biotic (developmental stages and wing dimorphism) and abiotic stress (thermal, starvation, and UV irradiation) conditions. The results showed that EF1α and 18S were the most stable genes under various biotic states, ß-ACT and ß-TUB during thermal stress, EF1α and RNAP II under starvation stress, and RNAP II, ß-ACT, and EF1α under UV irradiation stress conditions. This study provides useful resources for the transcriptional profiling of genes in T. citricida and closely related aphid species.

The Essential Role of Vitellogenin Receptor in Ovary Development and Vitellogenin Uptake in Bactrocera dorsalis (Hendel).

The vitellogenin receptor (VgR) functions as an essential component in uptaking and transporting vitellogenin (Vg) in female adults, which is involved in ovary development and oviposition. This study aimed to clarify the molecular characteristics and function of VgR in the oriental fruit fly Bactrocera dorsalis (Hendel). Here, we identified the full-length of BdVgR (GenBank Accession No. JX469118), encoding a 1925 residue (aa) protein with a 214.72 kDa molecular mass and several typical motifs of low-density lipoprotein receptor superfamily (LDLR). Phylogenic analysis suggested that BdVgR was evolutionary conserved with other Dipteran VgRs. The expression of BdVgR was exclusively detected in the ovaries rather than head, thorax or other tissues. The developmental expression patterns showed that the signal of BdVgR was detectable in very beginning of adult stage, and positively correlated with the growth rate of ovaries and the expression levels of its ligands. In addition, we also demonstrated that the expression level of BdVgR, and ovary development were significantly suppressed after being injected with BdVgR-targeted dsRNA. Together, all of these results indicated that BdVgR was critical for yolk protein absorption and ovary maturation in B. dorsalis, playing a vital role in female reproduction.

Inducible Expression of Mu-Class Glutathione S-Transferases Is Associated with Fenpropathrin Resistance in Tetranychus cinnabarinus.

The carmine spider mite, Tetranychus cinnabarinus (Boisduval), is a serious pest on a variety of economically important crops widely distributed in China, and its resistance to acaricides has quickly developed. In this study, we fully sequenced 13 GST genes of T. cinnabarinus (TcGSTs). The phylogenetic tree showed that five of them belonged to the delta class and the other eight belonged to the mu class. The alignment of gene sequences and comparison of gene expressions between a fenpropathrin-resistant strain (FR) and a susceptible strain (SS) showed that neither point mutation nor overexpression was detected in TcGSTs. However, when challenged by a sublethal dose of fenpropathrin, the mRNA levels of three GSTs from the mu class (TCGSTM2, TCGSTM3, and TCGSTM8) highly increased in FR, while in SS, the expression of these genes was still at the same level under the treatment. In conclusion, specific TcGSTs were identified that were inducible to stimulation by fenpropathrin, and proved that TcGSTs in FR were not constantly expressed at a high level, but could react much more quickly under the stress of fenpropathrin than SS.

Characterization of Bactrocera dorsalis serine proteases and evidence for their indirect role in insecticide tolerance.

The oriental fruit fly Bactrocera dorsalis (Hendel) causes devastating losses to agricultural crops world-wide and is considered to be an economically important pest. Little is known about the digestive enzymes such as serine proteases (SPs) in B. dorsalis, which are important both for energy supply and mitigation of fitness cost associated with insecticide tolerance. In this study, we identified five SP genes in the midgut of B. dorsalis, and the alignments of their deduced amino acid sequences revealed the presence of motifs conserved in the SP superfamily. Phylogenetic analyses with known SPs from other insect species suggested that three of them were trypsin-like proteases. Analyses of the expression profiles among the different developmental stages showed that all five genes were most abundant in larvae than in other stages. When larvae were continuously fed on diet containing 0.33 µg/g ß-Cypermethrin, expression of all five genes were upregulated in the midgut but the larval development was delayed. Biochemical assays were consistent with the increased protease activity exhibited by SPs in the midgut after treatment with ß-Cypermethrin. Taken together, these findings provide evidence for the hypothesis that enhanced SP activity may play an indirect role in relieving the toxicity stress of insecticide in B. dorsalis.

Functional analysis of SDR112C1 associated with fenpropathrin tolerance in Tetranychus cinnabarinus (Boisduval).

Short-chain dehydrogenases/reductases (SDRs) are ubiquitously distributed across diverse organisms and play pivotal roles in the growth, as well as endogenous and exogenous metabolism of various substances, including drugs. The expression levels of SDR genes are reportedly upregulated in the fenpropathrin (FEN)-resistant (FeR) strain of Tetranychus cinnabarinus. However, the functions of these SDR genes in acaricide tolerance remain elusive. In this study, the activity of SDRs was found to be significantly higher (2.26-fold) in the FeR strain compared to the susceptible strain (SS) of T. cinnabarinus. A specific upregulated SDR gene, named SDR112C1, exhibited significant overexpression (3.13-fold) in the FeR population compared with that in the SS population. Furthermore, the expression of SDR112C1 showed a significant increase in the response to FEN induction. Additionally, knockdown of the SDR112C1 gene resulted in decreased SDR activity and reduced mite viability against FEN. Importantly, heterologous expression and in vitro incubation assays confirmed that recombinant SDR112C1 could effectively deplete FEN. Moreover, the overexpression of the SDR112C1 gene in Drosophila melanogaster significantly decreased the toxicity of FEN to transgenic fruit flies. These findings suggest that the overexpression of SDR SDR112C1 is a crucial factor contributing to FEN tolerance in T. cinnabarinus. This discovery not only enhances our understanding of SDR-mediated acaricide tolerance but also introduces a new family of detoxification enzymes to consider in practice, beyond cytochrome P450s, carboxyl/choline esterases and glutathione S-transferases.

Population dynamics and molecular adaption of Tetranychus cinnabarinus to long-term thermal stress.

BACKGROUND: Global warming is a general trend in the current era. Temperature is one of the most important nonbiological factors that affects the development, life cycle and distribution of arthropods, which are a major component of agriculture pests. This study focused on life-table parameters and the molecular adaption of Tetranychus cinnabarinus under long-term thermal stress. RESULTS: The life tables of T. cinnabarinus were constructed at room temperature (26 °C) and high temperature (34 °C). Results showed that although the lifespan of the mites was shortened, the developmental periods of egg, larva and nymph stages were accelerated, and the peak egg-laying period came earlier at high temperature, which resulted in faster expansion of pest mite population. RNA-seq was used to reveal the thermal adaption mechanism according to differentially expressed genes. Combined with transcriptome data and quantitative polymerase chain reaction (qPCR) verification, MAPK, CAT, HSP20 and HSP70 were found highly expressed at 34 °C, which were associated with thermal adaption of T. cinnabarinus. RNAi analysis proved that expression of HSP20 was closely related to the survival of mites at high temperature. CONCLUSION: These results indicated that long-term high temperature treatment was beneficial to the expansion of the T. cinnabarinus population. The genes involved in heat tolerance of T. cinnabarinus such as MAPK-HSP pathway provides ideas for subsequent control measures. © 2023 Society of Chemical Industry.

Uridine Diphosphate Glycosyltransferases (UGTs) Involved in the Carotenoid-Based Body Color Difference between Tetranychus cinnabarinus (Red) and Tetranychus urticae (Green).

It has long been disputed whether Tetranychus cinnabarinus and Tetranychus urticae belong to the same genus, with T. cinnabarinus regarded as a red form of T. urticae. However, it is unclear why T. urticae and T. cinnabarinus have different body colors. Since carotenoids are responsible for the color of many organisms, the carotenoid profiles of T. cinnabarinus and T. urticae were compared by HPLC. There was no difference in carotenoid type, but T. cinnabarinus contained significantly more neoxanthin, astaxanthin, α-carotene, ß-carotene, and γ-carotene, which may contribute to the deep red color. The transcriptome sequencing of both species identified 4079 differentially expressed genes (DEGs), of which 12 were related to carotenoid metabolism. RNA interference (RNAi) experiments demonstrated that silencing seven of these DEGs resulted in the different accumulation of carotenoid compounds in T. cinnabarinus and T. urticae. In addition, the body of T. urticae turned yellow after two days of feeding with UGT double-stranded RNAs and ß-UGT small interfering RNAs. In conclusion, differences in the carotenoid profiles of T. urticae and T. cinnabarinus may be responsible for the different body colors.

A pH- and enzymatic-responsive nanopesticide to control pea aphids and reduce toxicity for earthworms.

Thiacloprid is a new chlorinated nicotinoid insecticide against stinging-oral pests, such as aphids. It is less toxic to bees but more toxic to earthworms. In this study, a pH- and amylase-responsive MOF (ZIF-8) was constructed for site-specific delivery of thiacloprid to control pea aphids and more safety for earthworms. Thiacloprid from α-cyclodextrin@Thiacloprid@ZIF-8 (α-CD@T@ZIF-8) could be released quickly in pea aphids, which was ascribed to disintegration of ZIF-8 at low pH values in pea aphid intestines and degradation of α-CD under the action of α-amylase. The release results showed a significant pH dependence of α-CD@T@ZIF-8, with an approximately 65 % release amount at pH = 7 and a 95 % release amount at pH = 5 for 7 d. The results of the pot experiment and biosafety showed that for α-CD@T@ZIF-8, 88 % pea aphids could be killed compared with 32 % aphids for commercially available formulation on the 7th day after application. Meanwhile the LC50 of thiacloprid OD was 0.034 µg/cm2 and the LC50 of α-CD@T@ZIF-8 was 0.564 µg/cm2 on earthworms, and it was more safety for pea and lower acute toxicity and enrichment for the earthworms. α-CD@T@ZIF-8 could be used for intelligently controlled release of other insecticides against aphids.

Molecular Basis for the Selectivity of the Succinate Dehydrogenase Inhibitor Cyflumetofen between Pest and Predatory Mites.

Acaricides that act as inhibitors of the mitochondrial succinate dehydrogenase (SDHIs) provide excellent control of phytophagous mites but display limited toxicity to predatory mites and other beneficial organisms. However, the molecular mechanism of selectivity is not fully understood. Here, we first confirm that SDHI acaricides are over 10,000-fold more toxic to spider mites than predatory mites. Next, we show that differential penetration, pro-acaricide activation, or metabolism are most likely not the main reason for this selectivity. In contrast, the inhibition of AB-1 on the SDH target is approximately 200-fold more potent in spider mites compared to predatory mites, revealing strong target-site selectivity. Strikingly, a key motif associated with differential binding was identified and validated by gene editing in Drosophila. Our findings contribute to understanding the selectivity of SDHIs, which can be used for the rational design of selective acaricides in support of an integrated pest management.

The mechanism of coumarin inhibits germination of ryegrass (Lolium perenne) and its application as coumarin-carbon dots nanocomposites.

BACKGROUND: As an important plant allelochemical, coumarin can effectively inhibit the germination of various seeds. However, little is known about the inhibition mechanism of coumarin on weed seed germination. Moreover, the herbicidal activity of coumarin is needed to be improved as a natural pesticide. RESULTS: Coumarin had the highest inhibition effect on the ryegrass (Lolium perenne) seed, where coumarin disturbed the hormone pathway by decreasing the content of gibberellic acid 3, resulting in the reduction of amylase activity and consumption of starch during the germination process of ryegrass seed. Moreover, coumarin induced decreased activity of catalase and subsequently led to the accumulation of hydrogen peroxide and malondialdehyde, causing oxidative stress during the germination process of ryegrass seed. Furthermore, to enhance the herbicidal activity of coumarin, carbon dots (CDs) modified with polyetherimide were prepared, characterized, and then combined with coumarin to form coumarin-carbon dots (Cm-CDs) nanocomposites. Compared with coumarin, Cm-CDs nanocomposites significantly increased the herbicidal activity of coumarin on ryegrass, which implies that Cm-CDs nanocomposites could be used as a potential formulation to improve the herbicidal activity of coumarin. CONCLUSION: This study not only reveals the mechanism of coumarin on ryegrass germination, but also develop Cm-CDs nanocomposites to enhance the herbicidal activity of coumarin. Our findings will stimulate the application of Cm-CDs nanomaterials as an effective and environmentally friendly formulation in agriculture. © 2023 Society of Chemical Industry.

Retinoid X receptor 1 is a specific lethal RNAi target disturbing chitin metabolism during hatching of Tetranychus cinnabarinus.

RNA interference (RNAi) can be developed as an alternative method of chemical pesticides for pest control. In this study, we noticed a specifically expressed gene (retinoid X receptor 1, TcRXR1) in the egg stage of T. cinnabarinus. RNAi was applied to investigate the function of TcRXR1. Results showed that with continuous feeding of dsTcRXR1, the larvae of T. cinnabarinus could still successfully develop to adult, which was in accordance with the low expression of TcRXR1 out of egg stage. High mortality of eggs was observed after eggs were treated with dsTcRXR1. To investigate the downstream genes of TcRXR1, the RNA samples after successful RNAi of TcRXR1 were analyzed by transcriptome analysis. According to function annotation of differentially expressed genes, 6 genes were selected for their potential function with the phenotype of dsTcRXR1, and among them, a chitinase gene (TcCHT-E) attained a high expression level in the late stage of egg, peaking just after the expression peak of TcRXR1. Mortality of eggs was observed under the effect of dsTcCHT-E as well as dsTcRXR1. In conclusion, TcRXR1 is a specific RNAi target for control of T. cinnabarinus, and its lethal mechanism might be disturbing chitin metabolism hatching of egg.