Aldicarb sulfoxidation by plant root extracts

Soybean root homogenates were found to oxidize aldicarb to aldicarb sulfoxide. Fractionation of soybean root homogenate indicates that the enzyme(s) which oxidize aldicarb to the sulfoxide are largely (63%) in the 25,000g supernatant. In vitro studies of the 25,000g supernatant showed a linear reaction rate at 24 and 34°C for periods of up to 2 and 1 hr, respectively, at the pH optimum of 5.5. Bean and soybean root 25,000g supernatants were the most active on a per milligram of protein basis followed by corn, sorghum, barley, and tomato (8–27 nmol/mg of protein/hr). A 140-fold purification of soybean root aldicarb sulfoxidase was achieved. In vitro studies indicated that methomyl, methiocarb, phorate, Counter, fensulfothion, fenthion, Nemacur, EPTC, vernolate, carboxin, dl-methionine, l-cysteine, l-methionyl-l-serine, thiodiglycolic acid, lipoic acid, thiourea, and thioacetic acid when present at a 10:1 ratio with aldicarb significantly inhibited soybean root aldicarb sulfoxidation.     

Pathogenicity of phylogenetic species in the Fusarium graminearum complex on soybean seedlings in Argentina

Soybean (Glycine max L.) is one of the main crops in Argentina. Most of the studies of pathogenicity in the Fusarium graminearum complex have focused on strains isolated from wheat and maize, and there is little information on strains isolated from soybean. Our objective in the present study was to compare the pathogenicity among soybean isolates of different phylogenetic species within the Fusarium graminearum complex on soybean seedlings under controlled conditions. Six strains representing three different phylogenetic species (F. graminearum, F. meridionale and F. cortaderiae) were identified by partial sequencing of the Translation Elongation Factor -1α gene (TEF-1) and evaluated for pathogenicity. All six strains reduced emergence, mainly by causing pre-emergence damping-off, seedling height and root dry weight and produced abnormal seedlings. The mean disease severity averaged across all isolates was approximately 3.0 in a 0–4 rating scale where 0?=?healthy seedling and 4?=?dead seedling. Significant differences in pathogenicity were observed among F. graminearum, F. meridionale and F. cortaderiae. These results are consistent with the hypothesis that different phylogenetic species within the Fusarium graminearum complex isolated from soybean are pathogenic under controlled conditions to soybean seedlings in Argentina. The present study demonstrates for the first time the pathogenic effect of F. meridionale on soybean in Argentina.     

Identification of <Emphasis Type="Italic">Fusarium</Emphasis> species associated with soybean root rot in Sichuan Province,China

Soybean root rot is a worldwide soil-borne fungal disease threatening soybean production, causing large loss in yield and quality of soybean. Fusarium species are well recognized as the important causal agent of Fusarium root rot, which are often distinct with respect to various factors in different soybean-producing regions around the world. Recently, Fusarium root rot has been frequently reported in Sichuan Province of China, where is unique in its climate and diverse cropping patterns, but it is still unclear about the predominant Fusarium species and their pathogenicity on soybean. In this study, diseased soybean roots were collected from three regions of Sichuan Province during 2014–2015. Based on morphological characteristics and phylogenetic analysis of nucleotide sequences of the ribosomal internal transcribed spacer region and the translation elongation factor 1-α gene, 78 isolates of Fusarium were identified as nine distinct species. Pathogenicity tests showed that seven species of Fusarium were able to infect soybean, but differed in pathogenicity. F. oxysporum, F. equiseti and F. graminearum were the most aggressive species to soybean, whereas F. fujikuroi and F. verticillioides were not pathogenic to soybean. There was a strong positive correlation of the pathogenicity of Fusarium species with seedling emergence and fresh root weight. In addition, the diversity of Fusarium species varied among soybean-growing regions. To our knowledge, this report on population and pathogenicity of Fusarium species, in particular, F. graminearum, associated with soybean root rot in Sichuan Province of southwest China, will be helpful to provide effective control strategies for the disease.     

Microsomal sulfoxidation of phorate in the fall armyworm,Spodoptera frugiperda (J. E. Smith)

Microsomal sulfoxidation in fall armyworm (Spodoptera frugiperda) larvae was examined using phorate as substrate. The system required NADPH and was inhibited by CO and by piperonyl butoxide. Sulfoxidase activity was found in the alimentary canal, fat body, and Malpighian tubules, with the midgut being the most active. Microsomal substrates such as aldrin, heptachlor, biphenyl, and methyl parathion significantly inhibited the enzyme activity whereas p-nitroanisole and p-choloro-N-methylaniline had no effect. Enzyme activity increased during larval development, reaching a maximum shortly before pupation. Allelochemicals (monoterpenes, indoles, and flavones), drugs (phenobarbital and 3-methylcholanthrene), and host plants (corn cotton, parsnip, and parsley) significantly increased the enzyme activity. Increased phorate sulfoxidation through enzyme induction was found to decrease oral toxicity of phorate to the larvae. Analyses of internal insecticide revealed that, at various intervals, induced larvae retained less phorate and its oxidative metabolites than the controls. It was concluded that induction of phorate sulfoxidase activity results in an overall increase in the rate of phorate detoxication in this insect.     

Studies on the metabolism of vamidothion and its thioanalog in insecticide-resistant and susceptible house fly strains

The in vivo and in vitro metabolism of vamidothion [O,O-dimethyl S-[2-(1-methylcarbamoyl)-ethylthio] ethylphosphorothiolate] as well as the in vitro metabolism of thiovamidothion [O,O-dimethyl S-[2-(1-methylcarbamoyl)ethylthio] ethylphosphorodithioate] was investigated in insecticide-resistant and susceptible house fly strains. Vamidothion was converted in vivo to the sulfoxide, the principle metabolite, and subsequently to the sulfone at a slower rate. Vamidothion and vamidothion sulfoxide were hydrolyzed at the PS and SC bond. The resulting primary alcohol metabolite was further oxidized to a carboxylic acid followed by decarboxylation. No metabolism of vamidothion or thiovamidothion occurred in vitro without the addition of NADPH. The addition of NADPH resulted in rapid conversion of vamidothion to the sulfoxide, and thiovamidothion was oxidatively metabolized to six metabolic products. No qualitative differences were found between resistant and susceptible strains, but there were signficant quantitative differences. The metabolism was highest in the Rutgers strain followed by Cornell-R, Hirokawa, and then CSMA strain. The route of vamidothion and thiovamidothion metabolism was via the cytochrome P-450-dependent monooxygenase system, and none of the resistant strains showed glutathione S-transferase activity toward vamidothion or thiovamidothion. No further oxidation of vamidothion sulfoxide to the sulfone was observed and also no hydrolysis products were formed, in vitro.     

Monooxygenases from soybean root nodules: Aldrin epoxidase and cinnamic acid 4-hydroxylase

The metabolism of aldrin and trans-cinnamic acid within the root nodules of soybean (Glycine max. cv. Forrest) was investigated. In vitro studies with microsomal preparations revealed the presence of two distinct monooxygenase enzymes requiring NADPH and molecular oxygen. This was shown by different distributions in membrane fractions from sucrose density gradients. In addition, cinnamic acid hydroxylase was sensitive to carbon monoxide (CO), similar to cytochrome P-450-dependent monooxygenases, whereas aldrin epoxidation was not. Cyanide ion, strongly inhibited the epoxidase, without affecting hydroxylase activity. The superoxide radical (O₂⁻) scavengers, superoxide dismutase and norepinephrine, inhibited aldrin epoxidation but allowed greater cinnamic acid 4-hydroxylase activity. These results indicate aldrin epoxidase, in contrast to cinnamic acid hydroxylase, does not involve cytochrome P-450, and may instead utilize a peroxidase-like hemoprotein.     

Subcellular distribution of malathion,phenthoate, and diethylsuccinate carboxylesterases in rat lungs

The subcellular distribution of malathion, phenthoate, and diethylsuccinate carboxylesterases was determined in the lungs of male Sprague-Dawley rats and in rats administered lung toxic doses of bromobenzene and paraquat. In control with the former two substrates, the highest activity was encountered in the 6500g and 12,000g pellets. In addition, significant activity was found in the 100,000g supernatant. These fractions hydrolyzed diethylsuccinate very slowly, and the major diethylsuccinate carboxylesterase activity was recovered in the 100,000g pellet. The bromobenzene treatment had no effect on these carboxylesterases; however, the paraquat treatment significantly decreased the phenthoate and diethylsuccinate carboxylesterases in the 100,000g pellet without altering the activity in the other fractions. The present study suggested that the subcellular distribution of malathion and phenthoate carboxylesterases is different from that in liver. The present study revealed that, in the lungs, the highest total activity for malathion and phenthoate carboxylesterases was found in the 100,000g supernatant, in comparison with liver, where the 100,000g pellet contains a much higher activity of these enzymes. The decrease in specific activities of diethylsuccinate and phenthoate carboxylesterases following the treatment with a pneumotoxicant may serve as an indicator of lung damage.     

The herbicide flamprop-M-methyl has a new antimicrotubule mechanism of action

BACKGROUND: The herbicidal mode of action of flamprop‐M‐methyl [methyl N‐benzoyl‐N‐(3‐chloro‐4‐fluorophenyl)‐D ‐alaninate] was investigated. RESULTS: For initial characterization, a series of bioassays was used, which indicated a mode of action similar to that of mitotic disrupter herbicides. Cytochemical fluorescence studies, which included monoclonal antibodies against polymerized tubulin, were applied to elucidate effects on mitosis and microtubule assembly in maize roots. When seedlings were root treated with 50 µM of flamprop‐M‐methyl, cell division activity in meristematic root tip cells ceased within 4 h. The compound severely disturbed the orientation of spindle and phragmoblast microtubules, leading to defective spindle and phragmoblast structures. Cortical microtubules were only slightly affected. In late anaphase and early telophase cells, phragmoblast microtubules were disorganized in multiple arrays that hampered regular cell plate deposition in cytokinesis. Microtubules of the spindle apparatus were found attached to chromosomal kinetochores, but did not show regular organization associated with a zone of microtubule‐organizing centres at the opposite ends of the cell. On account of this loss of spindle organization, chromosomes remained in a condensed state of prometaphase or metaphase. Unlike known microtubule disrupter herbicides, flamprop‐M‐methyl and its biologically active metabolite flamprop did not inhibit soybean tubulin polymerization to microtubules in vitro at 50 µM . In contrast, soybean plants responded sensitively to the compounds. CONCLUSION: The results indicate that flamprop‐M‐methyl is a mitotic disrupter herbicide with a new antimicrotubule mechanism of action that affects orientation of spindle and phragmoblast microtubules, possibly by minus‐end microtubule disassembly. Copyright © 2008 Society of Chemical Industry     

Reduction of Striga hermonthica parasitism on maize using soybean rotation

Striga hermonthica is a serious parasite of cereals in most of the semi-arid savanna zone of West Africa, causing substantial yield loss. It has been observed that some soybean cultivars are capable of stimulating germination of S. hermonthica seed, which would reduce the seed bank in the soil. This study was undertaken to quantify the effect of a soybean crop (compared with a sorghum control) on S. hermonthica emergence in subsequent maize in three farmers' fields in northern Nigeria. Soybean cultivar TGx 1740-7F, previously identified as efficacious for S. hermonthica seed germination, was grown at four densities without P fertilizer to test the effect of increasing plant density of soybean on subsequent S. hermonthica parasitism. The effect of P as single super phosphate was tested on the two highest soybean densities. S. hermonthica parasitism on maize was significantly lower after unfertilized soybean than after the sorghum control treatment at two of three trial sites. Soybean rotation increased maize yield by approximately 90% for the three sites combined. Increasing soybean plant density did not result in lower emerged S. hermonthica. Application of P to soybean at the higher soybean densities resulted in higher root length density, lower emerged S. hermonthica on maize (P<0.15), and significantly higher maize yield. The results suggest that an efficacious cultivar of soybean reduces S. hermonthica parasitism on a succeeding maize crop and that the effect is increased by application of P to the soybean.     

Some factors affecting the susceptibility of two nematode species to phorate

The uptake and metabolism of phorate has been compared in two species of free-living, soil nematodes, Aphelenchus avenae and Panagrellus redivivus, which differ greatly in their sensitivity to this pesticide. The dynamics of uptake and metabolism are analyzed using a simple catenary model. Estimated rate constants indicated a much greater rate of phorate metabolism in P. redivivus than in A. avenae, and qualitative differences in the toxic metabolites of phorate produced by the two species were also found. The toxicological importance of these factors is discussed.     

In vitro inhibition of α-amylase and protease by nematocides in Cicer arietinum L

The in vitro effects of four systemic nematocides, i.e., aldicarb, carbofuran, oxamyl, and phorate, on the α-amylase and protease activities in Cicer arietinum has been revealed. All four nematocides markedly inhibited the activities of both the enzymes, with a general tendency of increased inhibition with corresponding increase in the concentrations of the nematocides. There was complete inhibition of α-amylase activity by the highest concentration (500 μM) of aldicarb and carbofuran, while oxamyl at the same concentration showed the same effects on protease activity. The lowest concentration (10 μM) was almost ineffective.     

吲哚乙酸引导下三唑醇在大豆植株中的传导与积累研究初报

将吲哚乙酸与三唑醇发生酯化反应后生成的吲哚乙酸三唑醇酯 ,在0.5 mmol/L浓度下水培和喷雾处理6~8叶期大豆植株,同时以相同浓度的三唑醇及其与吲哚乙酸的混合物为对照,色谱法测定不同时间植株不同部位吲哚乙酸三唑醇酯和三唑醇的含量。结果表明,与对照相比,吲哚乙酸三唑醇酯在大豆中具有明显的双向传导和向根部积累的特点。喷雾处理后,在12~60 h之间内吸量和根部积累量均出现最大值。     

The effect of fentin hydroxide (triphenyltin hydroxide) on soybean [Glycine max (L.) Merr.] and rice (Oryza sativa L.) photosynthesis,respiration, and leaf ultrastructure

Soybean [Glycine max (L.) Merr., cv. Swift] plants at the second trifoliate leaf stage and rice (Oryza sativa L. cv. Starbonnet) plants 25 cm tall were treated with 0, 0.56, and 2.24 kg/ha of fentin hydroxide (triphenyltin hydroxide) to determine the effect of this fungicide on photosynthesis, respiration, and leaf ultrastructure. Photosynthesis and respiration were measured with an infrared CO₂ analyzer in an open flow system prior to fentin hydroxide application and at 4, 24, 48, and 96 hr after treatment with fentin hydroxide. No significant detrimental effects on photosynthesis or respiration were evident in either soybean or rice through 96 hr after treatment. Tissue samples from soybean and rice plants, 9 days after fentin hydroxide application, examined for ultrastructure changes with the transmission electron microscope showed no effect due to the fungicide treatment.     

Root exudation of prometryn and its metabolites from Chinese celery (Apium graveolens)

Root exudates from Chinese celery (Apium graveolens) and Chinese cabbage (pak choi, Brassica chinensis) plants treated by prometryn, an herbicide, were qualitatively and quantitatively investigated and compared under hydroponic cultivation. Prometryn and its metabolites released into the nutrient solution were analyzed by ultra-performance liquid chromatograph coupled with orbitrap mass spectrometer to investigate whether this xylem-mobile herbicide is exuded from the roots. The results showed that celery and pak choi had different root exudation profiles. Celery metabolized prometryn to prometryn sulfoxide and released both compounds from the roots. In contrast, pak choi barely metabolized or actively released prometryn from the roots. The concentration of prometryn sulfoxide released from celery after 96 hr was 21 µg/L, which was nearly one-third that of released prometryn. Our results indicate that the root exudation and translocation of xylem-mobile herbicides could be significant in plants and are highly species dependent compared with phloem-mobile herbicides.     

Seed treatments associated with resistance inducers for management of <Emphasis Type="Italic">Pratylenchus brachyurus</Emphasis> in soybean

Pratylenchus brachyurus is one of the main limiting factors of soybean yield in Brazil, particularly because of inefficiency of the control methods when used individually. The present study aimed to assess the effect of associated methods, using seed treatment with nematicides (ST) and resistance inducers (RI), on nematode control in soybean plant in both field and greenhouse conditions. A field assay was conducted in an infested field and nematode population was assessed at sowing, 45, 75, and 100 days after sowing and the yield measured at end of crop cycle. The experiment was repeated in greenhouse. In another experiment, that was conducted in two different periods in a greenhouse, seed treatments and resistance inducers, alone or combined, were assessed under two initial populations of P. brachyurus (low IP = 500 specimens and high IP = 2000 specimens). The treatments did not reduce the number of nematodes g^?1 of root in field assay, but all seed treatments effectively controlled nematode population in the greenhouse assay. Most treatments reduced the number of nematodes g^?1 of root when nematode initial population was low (IP = 500) but when initial nematode population was high (IP = 2000) combinations of treatments which includes abamectin inhibited P. brachyurus reproduction. Chemical products did not affect yield but acibenzolar-S-methyl, alone or associated with other products, generally inhibit plant growth.     

Studies on the mechanisms of accumulation of phorate by the free-living nematode Panagrellus redivivus

Accumulation and metabolism of the organophosphate pesticide, phorate by the free-living nematode Panagrellus redivivus was studied under anaerobic conditions and in the presence of carbon monoxide or SKF 525A, and compared with results obtained under normal, aerobic conditions and using heat-killed nematodes. Both phorate hydrolysis and side-chain oxidation were inhibited under anaerobic conditions and with heat-killed nematodes, but only hydrolysis was affected in the presence of carbon monoxide. The much greater phorate accumulation in the nematodes under anaerobic conditions and with carbon monoxide could be explained by this metabolic inhibition. SKF 525A had no significant effect on phorate metabolism or accumulation. The possible mechanisms involved in phorate breakdown by P. redivivus are discussed.     

Real-time quantitative PCR assays for evaluation of soybean varieties for resistance to the stem and root rot pathogen Phytophthora sojae

Phytophthora root and stem rot caused by Phytophthora sojae is one of the most destructive disease of soybeans in the world. Effective management of the disease depends on selection and use of soybean varieties resistant to the disease. Fast and reliable procedures are vital to screen soybean varieties against the pathogen. Novel real-time quantitative (qPCR) assays were developed for both absolute and relative quantification of P. sojae in infected root tissues. QPCR assays were based on the detection of the internal transcribed spacer (ITS) gene of the pathogen and 18S ribosomal gene of the host plant. Absolute qPCR allowed the detection of as low as 10 femtograms (fg) of P. sojae DNA in soybean roots. Relative qPCR, employing the comparative threshold cycle (Ct) method, was effective and reliable for quantification of P. sojae DNA normalized to plant DNA in infected soybean root tissues. P. sojae DNA quantities detected in both qPCR assays had high correlations with disease severity index (DSI) ratings of soybean varieties. QPCR assays developed in this study were useful for determination of the levels of P. sojae DNA in different varieties of soybean and for evaluation of them for relative resistance to the pathogen.     

Circadian changes in the activity of some esterases in house crickets and their effect on the inhibitory action of phorate

A culture of house crickets Acheta domisticus (L) was maintained under light-dark, 12:12 photoperiod. The activities of cholinesterase (ChE) and carboxylesterase (CarE) of rhythmic adults were assayed at different times of the day. Both enzymes showed a daily rhythmic pattern of activity. ChE had its highest activity level at 1200 hr, while CarE showed two peaks of activity, at 1200 and 2400 hr. The lowest level of activity for both enzymes was recorded shortly after the onset of darkness. The effect of these rhythmic fluctuations on the inhibitory action of phorate was studied. Insects were topically treated with a sublethal dose of phorate. Phorate application took place at different intervals to coincide with different magnitudes of enzymes activity. ChE inhibition did not differ significantly in relation to the time of phorate application, however, CarE inhibition was much higher when phorate was applied at the beginning of the dark phase. Inhibited ChE showed a rapid recovery only in insects treated in the light phase.     

Pathogenicity of Fusarium graminearum and F. meridionale on soybean pod blight and trichothecene accumulation

Soybean (Glycine max) is the most important crop in Argentina. At present Fusarium graminearum is recognized as a primary pathogen of soybean in several countries in the Americas, mainly causing seed and root rot and pre‐ and post‐emergence damping off. However, no information about infections at later growth stages of soybean development and pathogenicity of F. graminearum species complex is available. Therefore, the objectives of this study were to compare the pathogenicity of F. graminearum and F. meridionale isolates towards soybean under field conditions and to evaluate the degree of pathogenicity and trichothecene production of these two phylogenetic species that express different chemotypes. Six isolates of F. graminearum and F. meridionale were evaluated during 2012/13 and 2013/14 soybean growing seasons for pod blight severity, percentage of seed infected in pods and kernel weight reduction. The results showed a higher aggressiveness of both F. graminearum and F. meridionale species during the 2013/14 season. However, the differences in pathogenicity observed between the seasons were not reflected in a distinct trichothecene concentration in soybean seeds at maturity. Fusarium meridionale isolates showed similar pathogenicity to F. graminearum isolates but they were not able to produce this toxin in planta during the two field trials.