O-Carboxymethyl chitosan nanoparticles: A novel approach to enhance water stress tolerance in maize seedlings.

Water stress, a significant abiotic stressor, significantly hampers crop growth and yield, posing threat to food security. Despite the promising potential of nanoparticles (NPs) in enhancing plant stress tolerance, the precise mechanisms underlying the alleviation of water stress using O-Carboxymethyl chitosan nanoparticles (O-CMC-NPs) in maize remain elusive. In this study, we synthesized O-CMC-NPs and delved into their capacity to mitigate water stress (waterlogging and drought) in maize seedlings. Structural characterization revealed spherical O-CMC-NPs with a size of approximately 200 nm. These NPs accumulated near the seed embryo and root tip, resulting in a substantial increase in fresh and dry weights. The application of O-CMC-NPs to water-stressed maize seedlings remarkedly elevated the chlorophyll content and activity of various antioxidant enzymes, including superoxide dismutase (SOD), catalase (CAT), peroxidase (POD), and polyphenol oxidase (PPO). The malondialdehyde (MDA) content was significantly reduced compared to the untreated control. Additionally, the expression of stress-responsive genes, such as ZmSOD, ZmCAT, ZmPOD, ZmTIFY, ZmACO, ZmPYL2, ZmNF-YC12, and ZmEREB180, were significantly upregulated in the O-CMC-NPs treated seedlings. These findings unveil the novel role of O-CMC-NPs in enhancing plant stress tolerance, suggesting their potential application in safeguarding maize seedlings under water stress conditions and facilitating the recovery from oxidative damage.

PxRdl2 dsRNA increased the insecticidal activities of GABAR-targeting compounds against Plutella xylostella.

The utilization of RNA interference (RNAi) for pest management has garnered global interest. The bioassay results suggested the knockout of the PxRdl2 gene significantly increased the insecticidal activities of the γ-aminobutyric acid receptor (GABAR)-targeting compounds (fipronil, two pyrazoloquinazolines, and two isoxazolines), thereby presenting a viable target gene for RNAi-mediated pest control. Consequently, we suggest enhancing the insecticidal activities of GABAR-targeting compounds by knockdown the transcript level of PxRdl2. Furthermore, PxRdl2 dsRNA was expressed in HT115 Escherichia coli to reduce costs and protect dsRNA against degradation. In comparison to in vitro synthesized dsRNA, the recombinant bacteria (ds-B) exhibited superior interference efficiency and greater stability when exposed to UV irradiation. Collectively, our results provide a strategy for insecticide spray that combines synergistically with insecticidal activities by suppressing PxRdl2 using ds-B and may be beneficial for reducing the usage of insecticide and slowing pest resistance.

Distinct roles of two RDL GABA receptors in fipronil action in the diamondback moth (Plutella xylostella).

The phenylpyrazole insecticide fipronil blocks resistance to dieldrin (RDL) γ-aminobutyric acid (GABA) receptors in insects, thereby impairing inhibitory neurotransmission. Some insect species, such as the diamondback moth (Plutella xylostella), possess more than one Rdl gene. The involvement of multiple Rdls in fipronil toxicity and resistance remains largely unknown. In this study, we investigated the roles of two Rdl genes, PxRdl1 and PxRdl2, in P. xylostella fipronil action. In Xenopus oocytes, PxRDL2 receptors were 40 times less sensitive to fipronil than PxRDL1. PxRDL2 receptors were also less sensitive to GABA compared with PxRDL1. Knockout of the fipronil-sensitive PxRdl1 reduced the fipronil potency 10-fold, whereas knockout of the fipronil-resistant PxRdl2 enhanced the fipronil potency 4.4-fold. Furthermore, in two fipronil-resistant diamondback moth field populations, PxRdl2 expression was elevated 3.7- and 4.1-fold compared with a susceptible strain, whereas PxRdl1 expression was comparable among the resistant and susceptible strains. Collectively, our results indicate antagonistic effects of PxRDL1 and PxRDL2 on fipronil action in vivo and suggest that enhanced expression of fipronil-resistant PxRdl2 is potentially a new mechanism of fipronil resistance in insects.

Discovery and Biomimetic Synthesis of a Phloroglucinol-Terpene Adduct Collection from Baeckea frutescens and Its Biogenetic Origin Insight.

A phloroglucinol-terpene adduct (PTA) collection consisting of twenty-four molecules featuring three skeletons was discovered from Baeckea frutescens. Inspired by its biosynthetic hypothesis, we synthesized this PTA collection by reductive activation of stable phloroglucinol precursors into highly reactive ortho-quinone methide (o-QM) intermediates and subsequently Diels-Alder cycloaddition. We also demonstrated, for the first time, the generation process of the active o-QM by performing dynamic NMR and HPLC-MS monitoring experiments. Moreover, the PTA collection showed significant antifeedant effect toward the Plutella xylostella larvae.

Synthesis and antiphytoviral activity of α-aminophosphonates containing 3, 5-diphenyl-2-isoxazoline as potential papaya ringspot virus inhibitors.

α-Aminophosphonates compounds containing 3,5-diphenyl-2-isoxazoline were synthesized and evaluated for their bioactivity. Seventeen of them showed good bioactivity (protection effect > 50%) in vivo against papaya ringspot virus, while two of them (V29 and V45) exhibited excellent antiviral activity (both 77.8%). In the latter case, the antiviral activity was close to that of antiphytovirucides ningnanmycin and dufulin (both 83.3%) at 500 mg/L. The preliminary structure-activity relationships indicated that the bioactivity was strongly influenced by the substituents.

DMSO-mediated palladium-catalyzed cyclization of two isothiocyanates via C-H sulfurization: a new route to 2-aminobenzothiazoles.

DMSO was found to activate arylisothiocyanates for self-nucleophilic addition. A subsequent intramolecular C-H sulfurization catalyzed by PdBr2 enables access to a wide range of 2-aminobenzothiazole derivatives in moderate to good yields. This is the first example of a DMSO-mediated Pd-catalyzed synthesis of 2-aminobenzothiazoles through cyclization/C-H sulfurization of two isothiocyanates.

HFIP-Promoted Bischler Indole Synthesis under Microwave Irradiation.

1,1,1,3,3,3-Hexafluoropropan-2-ol (HFIP) was found to be effective for the Bischler indole synthesis under microwave irradiation in the absence of a metal catalyst. Under the catalysis of HFIP, a wide range of α-amino arylacetones were successfully transformed into indole derivatives with moderate to good yields.

AtLHT1 Transporter Can Facilitate the Uptake and Translocation of a Glycinergic-Chlorantraniliprole Conjugate in Arabidopsis thaliana.

Understanding of the transporters involved in the uptake and translocation of agrochemicals in plants could provide an opportunity to guide pesticide to the site of insect feeding. The product of Arabidopsis thaliana gene AtLHT1 makes a major contribution to the uptake into the roots of free amino acids and some of their derivatives. Here, a chlorantraniliprole-glycine conjugate (CAP-Gly-1) was tested for its affinity to AtLHT1 both in planta and in vitro. Seedlings deficient in AtLHT1 exhibited a reduction with respect to both the uptake and root-to-shoot transfer of CAP-Gly-1; plants in which AtLHT1 was constitutively expressed were more effective than wild type in term of their root uptake of CAP-Gly-1. Protoplast patch clamping showed that the presence in the external medium of CAP-Gly-1 was able to induce AtLHT1 genotype-dependent inward currents. An electrophysiology-based experiment carried out in Xenopus laevis oocytes expressing AtLHT1 showed that AtLHT1 had a high in vitro affinity for CAP-Gly-1. The observations supported the possibility of exploiting AtLHT1 as a critical component of a novel delivery system for amino acid-based pesticide conjugates.

Family of Ricinus communis Monosaccharide Transporters and RcSTP1 in Promoting the Uptake of a Glucose-Fipronil Conjugate.

Enhancing the systemic distribution of a bioactive compound by exploiting the vascular transport system of a plant presents a means of reducing both the volume and frequency of pesticide/fungicide application. The foliar uptake of the glucose-fipronil conjugate N-[3-cyano-1-[2,6-dichloro-4-(trifluoromethyl)phenyl]-4-[(trifluoromethyl)sulfinyl]-1H-pyrazol-5-yl]-1-(ß-d-glucopyranosyl)-1H-1,2,3-triazole-4-methanamine (GTF) achieved in castor bean (Ricinus communis) and its transport via the phloem are known to be mediated by monosaccharide transporter(s) [MST(s)], although neither the identity of the key MST(s) involved nor the mechanistic basis of its movement have yet to be described. On the basis of homology with Arabidopsis thaliana sugar transporters, the castor bean genome was concluded to harbor 53 genes encoding a sugar transporter, falling into the eight previously defined subfamilies INT, PMT, VGT, STP, ERD6, pGlucT, TMT, and SUT. Transcriptional profiling identified the product of RcSTP1 as a candidate for mediating GTF uptake, because this gene was induced by exposure of the plant to GTF. When RcSTP1 was transiently expressed in onion epidermis cells, the site of RcSTP1 deposition was shown to be the plasma membrane. A functional analysis based on RcSTP1 expression in Xenopus laevis oocytes demonstrated that its product has a high affinity for GTF. The long-distance root-to-shoot transport of GTF was enhanced in a transgenic soybean chimera constitutively expressing RcSTP1.

Preparation, Characterization and Intracellular Imaging of 2,4-Dichlorophenoxyacetic Acid Conjugated Gold Nanorods.

Visualizing the biodistribution of pesticides inside living cells is great importance for enhancing targeting of pesticides. Here we reported for the first time that gold nanorods (Au NRs) with size of 39.4 nm x 11.3 nm could be used as a fluorescent tracer to examine the distribution of a typical herbicide, 2,4-dichlorophenoxyacetic acid (2,4-D), in tobacco bright yellow 2 (BY-2) cells. The nanostructures of hybrid materials were analyzed by using Raman spectra and X-ray photoelectron spectroscopy (XPS), including spectra assignments and electronic property. These data revealed 2,4-D has successfully conjugated MP-Au NRs according to Raman and XPS. The biodistribution of the conjugates inside BY-2 cells was directly examined at 12 and 24 h by the two-photon microscopy. The intensity of two-photon luminescence (TPL) inside cells demonstrated that the conjugates could be localized and excluded by BY-2 cells. Thus, this labeling approach opens up new avenues to the facile and efficient labeling of pesticides.

Glycinergic-Fipronil Uptake Is Mediated by an Amino Acid Carrier System and Induces the Expression of Amino Acid Transporter Genes in Ricinus communis Seedlings.

Phloem-mobile insecticides are efficient for piercing and sucking insect control. Introduction of sugar or amino acid groups to the parent compound can improve the phloem mobility of insecticides, so a glycinergic-fipronil conjugate (GlyF), 2-(3-(3-cyano-1-(2,6-dichloro-4-(trifluoromethyl)phenyl)-4-((trifluoromethyl)sulfinyl)-1H-pyrazole-5-yl)ureido) acetic acid, was designed and synthesized. Although the "Kleier model" predicted that this conjugate is not phloem mobile, GlyF can be continually detected during a 5 h collection of Ricinus communis phloem sap. Furthermore, an R. communis seedling cotyledon disk uptake experiment demonstrates that the uptake of GlyF is sensitive to pH, carbonyl cyanide m-chlorophenylhydrazone (CCCP), temperature, and p-chloromercuribenzenesulfonic acid (pCMBS) and is likely mediated by amino acid carrier system. To explore the roles of amino acid transporters (AATs) in GlyF uptake, a total of 62 AAT genes were identified from the R. communis genome in silico. Phylogenetic analysis revealed that AATs in R. communis were organized into the ATF (amino acid transporter) and APC (amino acid, polyaminem and choline transporter) superfamilies, with five subfamilies in ATF and two in APC. Furthermore, the expression profiles of 20 abundantly expressed AATs (cycle threshold (Ct) values <27) were analyzed at 1, 3, and 6 h after GlyF treatment by RT-qPCR. The results demonstrated that expression levels of four AAT genes, RcLHT6, RcANT15, RcProT2, and RcCAT2, were induced by the GlyF treatment in R. communis seedlings. On the basis of the observation that the expression profile of the four candidate genes is similar to the time course observation for GlyF foliar disk uptake, it is suggested that those four genes are possible candidates involved in the uptake of GlyF. These results contribute to a better understanding of the mechanism of GlyF uptake as well as phloem loading from a molecular biology perspective and facilitate functional characterization of candidate AAT genes in future studies.

Flavonoids from Pronephrium megacuspe.

A chemical investigation of the whole plant of Pronephrium megacuspe (Bak.) Holtt. led to the isolation of four flavonoids, pronephrones A-D (1-4), which were firstly reported. The chemical structures of four compounds were established using spectroscopic methods. These isolates were further evaluated for cytotoxicity to ovarian cells of Spodoptera litura Fabricius.

Biosynthesis of Gold Nanoparticles Using Novel Bamboo (Bambusa chungii) Leaf Extracts.

Gold nanoparticles (Au NPs) have drawn significant interest because of their antisotropic physical properties in biomedical applications. In this paper, we reported the application of bamboo (Bambusa chung) leaf extracts, previously not exploited, in the synthesis of Au NPs at ambient temperature. The average dimension of quasi-spherical Au NPs was 28.8±4.5 nm by transmission electron microscopy (TEM). The UV-vis spectroscopy gave an optimal reaction time of 180 min in the process of bioreduction. The organic shell of Au NPs was characterized by Fourier transform infrared (FTIR) spectra and thermogravimetric analysis (TGA), suggesting that the main compositions of the organic shell were hydroxyflavones. The X-ray diffraction (XRD) studies indicated the Au NPs were (111) oriented. This eco-friendly method for the synthesis of Au NPs was simple, amenable for large scale commercial production and biological applications to future in vivo imaging and cancer therapy.

ß-Glucosidase involvement in the bioactivation of glycosyl conjugates in plants: synthesis and metabolism of four glycosidic bond conjugates in vitro and in vivo.

Mobile glucose-pesticide conjugates in the phloem are often restricted by decreases in biological activity. However, plants can bioactivate endogenous glucosides, which are assumed as able to bioactivate exogenous conjugates. In this study, four glycosidic bonds (O-, S-, N-, and C-glycosidic bonds) of glucose-pesticide conjugates were designed and synthesized, and then metabolism assays were carried out in vitro and in vivo. Results showed that ß-glucosidases played a role in the hydrolysis of O-glycosidic bond conjugates in Ricinus communis L. The liberated aglycons possessed insecticidal activities against Plutella xylostella L. and Spodoptera litura F. These results could help establish methods of circumventing the mutual exclusivity of phloem mobility and biological activity by hydrolyzing endogenous ß-glucosidases.

Synthesis of rotenone-O-monosaccharide derivatives and their phloem mobility.

Six monosaccharide derivatives of rotenone were designed and synthesized to assess whether rotenone could become phloem mobile by the addition of a monosaccharide group. Phloem mobility experiments showed that only D-glucose conjugates exhibit phloem transport properties in castor bean (Ricinus communis L.) seedlings. Two D-glucose conjugates, 2-O-ß-D-glucopyranosyldemethylrotenone and 6'-O-ß-D-glucopyranosyldalpanol, had significantly obtained systemicity compared with that of rotenone, and 6'-O-ß-D-glucopyranosyldalpanol was more mobile than 2-O-ß-D-glucopyranosyldemethylrotenone. Coupling with a monosaccharide core is a reasonable method for conferring phloem mobility on insecticides, but phloem mobility is also affected by the parent molecule and the position of the monosaccharide.

2,4-Dichlorophenoxyacetic acid functionalized gold nanoparticles: synthesis, characterization and biological effects.

Understanding and visualizing the biodistribution of agricultural chemicals inside cells and living plants is very important for enhancing targeting and changing the application approaches of chemicals. Here, a novel material was synthesized through 2,4-dichlorophenoxyacetic acid functionalized small gold nanoparticles (2,4-D-MP-Au NPs). The successful modification of Au NPs (4.46 ± 0.70 nm) was ascertained by UV-vis, TEM, FTIR and XPS. TGA data revealed about 1197 molecules of 2,4-D were coupled to the surface of one Au nanoparticle, which was sufficient for bioapplications. The optical imaging of 2,4-D-MP-Au NPs inside BY-2 cells was directly examined, revealing that 2,4-D-MP-Au NPs could be internalized in BY-2 cells by the two-photo microscopy and endocytosis, as the internalization mechanism was energy dependent for 2,4-D-MP-Au NPs. Furthermore, the biodistribution of 2,4-D-MP-Au NPs in Ricinus cotyledons was measured, revealing that 2,4-D-MP-Au NPs could enter into mesophyll cells of Ricinus cotyledons; the cell recognition was enhanced after 2,4-D conjugated Au NPs. These results indicate that the conjugates have great potential for applications on bioimaging and biolabeling for agricultural chemicals in plant physiology.

Design and synthesis of N-alkyl-N'-substituted 2,4-dioxo-3,4-dihydropyrimidin-1-diacylhydrazine derivatives as ecdysone receptor agonist.

Based on the discovery of thymine as an ecdysteroid agonist, a series of 1,4-disubstituted diacylhydrazine derivatives with a thymine moiety were designed and synthesized. The activities of these compounds against Spodoptera litura (Fabricius) were evaluated by the insect immersion method. Results showed that compound 2h with an N-cyclohexylmethyl substituent exhibits the most potent agonist activity with a median lethal concentration of 23.21 µg/mL. This compound also caused malformation of molting larvae and adults. Compound 2 h was further demonstrated as an ecdysteroid agonist by reporter gene assay on the Spodoptera frugiperda cell line (Sf9 cells). A molecular docking study indicated that hydrophobic interactions and the formation of hydrogen bonds between the compounds and the ecdysone receptor play critical roles in promoting the binding affinity of the compound. The structure of compound 2h may serve as a favorable template for the development of new ecdysteroid agonists with a pyrimidinedione moiety.

Synthesis of a series of monosaccharide-fipronil conjugates and their phloem mobility.

To test the effect of adding different monosaccharide groups to a non-phloem-mobile insecticide on the phloem mobility of the insecticide, a series of conjugates of different monosaccharides and fipronil were synthesized using the trichloroacetimidate method. Phloem mobility tests in castor bean ( Ricinus communis L.) seedlings indicated that the phloem mobility of these conjugates varied markedly. L-Rhamnose-fipronil and D-fucose-fipronil displayed the highest phloem mobility among all of the tested conjugates. Conjugating hexose, pentose, or deoxysugar to fipronil through an O-glycosidic linkage can confer phloem mobility to fipronil in R. communis L. effectively, while the -OH orientation of the monosaccharide substantially affected the phloem mobility of the conjugates.

A proteomic approach provides new insights into the control of soil-borne plant pathogens by Bacillus species.

Beneficial microorganisms (also known as biopesticides) are considered to be one of the most promising methods for more rational and safe crop management practices. We used Bacillus strains EU07, QST713 and FZB24, and investigated their inhibitory effect on Fusarium. Bacterial cell cultures, cell-free supernatants and volatiles displayed varying degrees of suppressive effect. Proteomic analysis of secreted proteins from EU07 and FZB24 revealed the presence of lytic enzymes, cellulases, proteases, 1,4-ß-glucanase and hydrolases, all of which contribute to degradation of the pathogen cell wall. Further proteomic investigations showed that proteins involved in metabolism, protein folding, protein degradation, translation, recognition and signal transduction cascade play an important role in the control of Fusarium oxysporum. Our findings provide new knowledge on the mechanism of action of Bacillus species and insight into biocontrol mechanisms.

High quality gold nanorods and nanospheres for surface-enhanced Raman scattering detection of 2,4-dichlorophenoxyacetic acid.

Nearly monodisperse Au nanorods (NRs) with different aspect ratios were separated from home-synthesized polydisperse samples using a gradient centrifugation method. The morphology, size and its distribution, and photo-absorption property were analyzed by transmission electron microscopy, atomic force microscopy and UV-visible spectroscopy. Subsequently, using colloidal Au NRs (36.2 nm ×10.7 nm) with 97.4% yield after centrifugation and Au nanospheres (NSs) (22.9 ± 1.0 nm in diameter) with 97.6% yield as Au substrates, surface-enhanced Raman scattering (SERS) spectra of 2,4-dichlorophenoxyacetic acid (2,4-D) were recorded using laser excitation at 632.8 nm. Results show that surface enhancement factors (EF) for Au NRs and NSs are 6.2 × 10(5) and 5.7 × 10(4) using 1.0 × 10(-6) M 2,4-D, respectively, illustrating that EF value is a factor of ~10 greater for Au NRs substrates than for Au NSs substrates. As a result, large EF are a mainly result of chemical enhancement mechanisms. Thus, it is expected that Au NPs can find a comprehensive SERS application in the trace detection of pesticide residues.