Differences in light stability of some carboxylic acid anilide fungicides in relation to their applicability for seed and foliar treatment

Irradiation of carboxin, furcarbanil, pyracarbolid, cyclafuramid, benodanil, mebenil and oxycarboxin by u.v. light (254 nm) for 4 h resulted in 97, 64, 58, 50, 20, 18 and 15% inactivation, respectively. Photoinactivation of the different compounds in aqueous solution or in the solid state was much slower in sunlight than in u.v. light and stability increased in the following order: carboxin < furcarbanil ≤ cyclafuramid ≤ pyracarbolid < mebenil ≤ benodanil < oxycarboxin. The residues of carboxin and furcarbanil on the leaf surface of bean plants were almost completely inactivated after 40 h exposure to sunlight; cyclafuramid lost 85% of its activity. The toxicity of leaf deposits of pyracarbolid, mebenil, benodanil and oxycarboxin decreased by 83, 53, 50 and 41%, respectively after 80 h in sunlight. The compounds with low photostability (e.g. carboxin, furcarbanil and cyclafuramid) are recommended mainly for controlling seed- and soil-borne fungi; pyracarbolid, mebenil, oxycarboxin and benodanil, which proved to be more photostable, appear to be useful fungicides to control rust diseases. Among several photochemical decomposition products of carboxin detected, the sulphoxide and sulphone were identified.     

Photochemical transformation of thiophanate-methyl and thiophanate to alkyl benzimidazol-2-yl carbamates

The systemic fungicides thiophanate-methyl (TPM) and thiophanate (TPE) were transformed in aqueous solutions on glass by irradiation with u.v. and sunlight to the more effective fungitoxic substances methyl benzimidazol-2-yl carbamate (MBC) and ethyl benzimidazol-2-yl carbamate (EBC), respectively. The thiophanate fungicides were not converted when incubated in the dark. The photochemical transformation of TPM and TPE to the corresponding alkyl benzimidazol-2-yl carbamates increased with the exposure time of u.v. irradiation. However, no light catalysed reactions occurred when both fungicides were irradiated with u.v. light in the solid state. The residue of TPM and TPE on leaves of cotton plants following spray application were transformed by the energy of sunlight to the more fungitoxic MBC and EBC.     

Hydrolysis and photolysis of the fungicide triforine

The main decomposition products formed from triforine by hydrolysis in solution and by photolysis were isolated and identified. In aqueous solution at 21°, hydrolysis proceeds rapidly by formation of chloride ions and, through several intermediates, 1-(dihydroxyacetyl)piperazine and piperazine. Photolysis by ultraviolet light in the absence of water leads preferentially to the removal of one side chain, the second side chain being attacked more slowly. In aqueous solution, triforine is rapidly destroyed by ultraviolet light. N-(2,2-Dichlorovinyl)formamide was isolated as an intermediate photodecomposition product.     

Photodecomposition of piperazine in water by ‘sunlight’ ultraviolet radiation

A solution of piperazine in water was photochemically oxidised by the dissolved oxygen when irradiated with ‘sunlight’ ultraviolet radiation obtained with a Hanau Q-300 lamp, or with RUL-300 nm lamps. Sensitisers (riboflavin, acetone and benzophenone) did not increase the rate of the oxidation induced by the RUL-300 nm lamps; however, they (especially riboflavin) accelerated the oxidation induced by the Hanau Q-300 lamp. A large number of products were observed in the sensitised oxidation but they were not analysed further. The irradiation mixture of the non-sensitised photo-oxidation, when 65% of the initial piperazine had been transformed, contained piperazine (35%), glycine (approximately 25%) and three unidentified compounds in equal amounts (approximately 13% each); the same products were obtained with both irradiation systems. The photo-oxidation of piperazine, a metabolite of the fungicide triforine, 1,4-bis(2,2,2-trichloro-l-formamidoethyl)piperaerazine, could thus be a way for the latter's biodegradation at a surface, or in plant tissues, this biodegradation having already been observed.     

Photolysis of fluchloralin in aqueous methanol

The photodegradation of fluchloralin by UV irradiation or sunlight in aqueous methanolic solution has been examined. In the presence of titanium dioxide five photoproducts were obtained, but only four in its absence. One photoproduct, 2, 2'-azoxy-bis(alpha,alpha,alpha-trifluoro-6-nitro-p-toluidine) is reported for the first time as a metabolite of fluchloralin. In natural sunlight the rate of degradation was higher than in UV light and titanium dioxide had almost no effect on the rate of degradation.     

Characterisation of bound residues of [3H]triforine in the straw of barley grown in the field

Barley was grown in an experimental field and, at the growth stage J (during the stem extension stage when the second node of the stem was formed and the next-to-last leaf just visible), the aerial part of the plants was sprayed with an aqueous emulsion of a mixture of triforine and [³H]triforine (uniformly labelled in the piperazine ring) at the recommended rate of 250 g triforine ha^?1. Barley was harvested when ripe, and the straw was analysed separately. Extraction of the straw with methanol left methanol-insoluble solids containing an amount of radioactivity (the bound residue) which represented 88 % of the tritium (as with all the subsequently quoted percentages, relative to the total tritium incorporated into the straw). Methanol acidified with hydrochloric acid extracted a further 8% of the triforine-derived bound residues as radioactive iminodiacetic acid (0.4%). glycine (3.2%), serine (2.0%), ethanolamine (0.2%) and unidentified compounds (2.2%); a neutral detergent solution extracted a further 3% of the radioactive triforine-derived bound residues as unidentified compounds; these radioactive compounds, which were dissolved by the acidified methanol and by the neutral detergent solution, were thus complexed in the straw to straw constituents. An acid detergent solution extracted a further 58% of the total tritium, which was incorporated by means of σ chemical bonds into the hemicelluloses fraction. Acid hydrolysis of the hemicelluloses yielded a mixture of mono-saccharides; these were converted into a mixture of osazones (50%), recrystallised several times, and had a constant specific radioactivity, indicating that the mono-saccharides were tritiated. The plant solids (which contained 19% of the total tritium), left by the acid-detergent extraction, were processed and separated into the tritium-containing cellulose (13%) and lignin (6%) fractions. No piperazine was observed in the bound residue of straw. Biochemical pathways are suggested for the radioactive metabolites of [³H]triforine.     

小麦赤霉菌分生孢子产生方法的初步研究

 1.一般赤霉菌在不同培养基上产生的分生孢子较少,但经阳光照射后,其在马铃薯琼脂培养基上产生极多。
2.阳光照射能促进小麦赤霉菌产生大量的分生孢子(每毫升2080万),开培养皿盖照射较关盖照射产生分生孢子的数量稍多,照射时间也缩短一倍以上。赤霉菌经阳光连续照射后,可以不断产生较多的分生孢子。
3.不同光线对赤霉菌产生分生孢子的效用各异。紫外线、日光灯能使其产生分子孢子,红外线和可见光无作用,其中紫外线照射距离17公分,关培养皿盖照射4小时,产生分生孢子最多。
4.赤霉菌经紫外线照射后,温度、湿度、氧气对其产生分生孢子的形态和数量有一定的影响,在处理后3小时就可以形成新的分生孢子。     

紫外线对Bt伴孢晶体的损伤和腐植酸的保护作用

用SDS-PAGE电泳分析和生物测定等方法研究了紫外线对苏云金芽孢杆菌（Bacillus thuringiensis,简称Bt）伴孢晶体的损伤及腐植酸对Bt伴孢晶体的保护作用。结果表明，紫外线对Bt伴孢晶体有明显的损伤，伴孢晶体紫外线辐射3h后溶解性能及生物活性基本丧失。腐植酸对紫外线有较强的吸收作用，能有效地减轻紫外线对Bt伴孢晶体的损伤。     

Characterisation of bound residues of [3H]triforine in barley grain grown in the field

Barley was grown in an experimental field and at growth stage J (during the stem extension stage when the second node of the stem was formed and the next-to-last leaf was just visible), the aerial part of the plants was sprayed with an aqueous emulsion of a mixture of triforine and [³H]triforine (uniformly labelled in the piperazine ring) at the normal rate of 250 g triforine ha^?1. The barley was harvested when ripe, and the grain was analysed separately. Extraction of the grain with methanol left methanol-insoluble solids containing an amount of radioactivity (the bound residue) which represented 75% of the total radioactivity incorporated into the grain. Methanol acidified with hydrochloric acid extracted a further 7% of the triforine-derived bound residues in the form of radioactive iminodiacetic acid (1.1%), glycine (3.3 %), serine (0.9%), ethanolamine (0.2%) and unidentified compounds (1.5 %); in the grain, these compounds or their precursors had thus been complexed to grain constituents. Aqueous 0.03M sodium hydroxide extracted a further 27% of the total tritium which had been incorporated by means of α chemical bonds into the protein fraction; acid hydrolysis of the proteins yielded radioactive glycine (9.2%), serine (3.9%) and unidentified compounds (13.9%) which could have been a mixture of a large number of other amino-acids. The plant solids (which contained 41% of the total tritium) left after the alkaline aqueous extractions, were processed and separated into tritiated cellulose (4%) and starch (37%) fractions. The starch was hydrolysed and the resulting glucose was converted into the osazone (34%). After being recrystallised several times, the osazone contained a constant specific radioactivity, indicating that [³H]glucose was present. This glucose may be considered as having a carbon skeleton mainly originating biochemically from some metabolites of [³H]triforine; this tritiated glucose may also be considered as indirect evidence of the biochemical oxidation, by transamination, of the metabolites of [³H]triforine; in barley grain, the tritiated glucose (or at least a part of it) was anabolised into proteins. No piperazine was observed in the bound residues in the grain.     

Resistance to triforine: A nonexistent problem?

Permanent resistance to triforine in Cladosporium cucumerinum was obtained after UV irradiation of spore suspensions and selection of resistant strains in the presence of triarimol or triforine. About 50 strains were examined for mycelial growth on malt agar, viability upon routine subculturing, growth and inhibition by triforine in thin-layer chromatographic bioassays, and virulence towards cucumber seedlings. In addition, sporulation and spore germination, as well as effects of ergosterol were investigated in a restricted number of strains. Although all strains differed considerably from each other in these characteristics, they were quite similar in a reduced overall-fitness. These triforine-resistant mutants of C. cucumerinum might, therefore, have a greatly reduced chance of survival in competition with the wild-type strain, as was actually demonstrated for two strains in greenhouse experiments with mixed-inoculations of cucumber seedlings. Such a reduced chance of survival might also explain why under practical conditions development of resistance to triforine has not been observed yet.The triforine-resistant C. cucumerinum strains showed cross-resistance to a number of systemic fungicides known or supposed to inhibit ergosterol biosynthesis, viz. Denmert, fenarimol, imazalil, triadimefon and triarimol. The practical implications and potential hazards of such cross-resistance for the use of already established fungicides are discussed.     

Antifungal mode of action of triforine

Rapidly growing mycelia of Aspergillus fumigatus treated with 10 μg/ml triforine (N,N′-bis-(1-formamido-2,2,2-trichloroethyl)-piperazine) showed little or no inhibition in dry weight increase prior to 2 h. By 2.5–3 h, triforine inhibited dry weight increase by 85%. The effects of triforine on protein, DNA, and RNA syntheses corresponded to the effect on dry weight increase both in time of onset and magnitude. Neither glucose nor acetate oxidation were inhibited by triforine.Ergosterol synthesis was almost completely inhibited by triforine even in the first hour after treatment. Inhibition of ergosterol synthesis was accompanied by an accumulation of the ergosterol precursors 24-methylenedihydrolanosterol, obtusifoliol, and 14α-methyl-Δ^{8, 24 (28)}-ergostadienol. Mycelia treated with 5 μg/ml of triarimol (α-(2,4-dichlorophenyl)-α-phenyl-5-pyrimidinemethanol) also accumulated the same sterols as well as a fourth sterol believed to be Δ^{5, 7}-ergostadienol.Identification of 4,4-dimethyl-Δ^{8, 24 (28)}-ergostadienol in untreated mycelia indicates that the C-14 methyl group is the first methyl group removed in the biosynthesis of ergosterol by A. fumigatus. The lack of detectable quantities of 4,4-dimethyl-Δ^{8, 24 (28)}-ergostadienol in triforine or triarimol-treated mycelia and the accumulation of C-14 methylated sterols in treated mycelia suggests that both fungicides inhibit sterol C-14 demethylation. The accumulation of Δ^{5, 7}-ergostadienol in triarimol-treated mycelia further implies that triarimol also inhibits the introduction of the sterol C-22(23) double bond.Two strains of Cladosporium cucumerinum tolerant to triforine and triarimol were also tolerant to the fungicide S-1358 (N-3-pyridyl-S-n-butyl-S′-p-t-butylbenzyl imidodithiocarbonate).     

Metabolism of [3H]triforine in barley grown in the field: The unbound radioactive residue

Barley was sown and grown normally in an experimental field. At the growth stage J,

1 Growth stage J: during the stem extension stage; second node of stem formed and next-to-last leaf just visible.

the aerial part of the plants was sprayed with an aqueous emulsion of a mixture of triforine and [³H]triforine (uniformly labelled in the piperazine ring) using the recommended dose rate of about 240g triforine ha^?1. The barley was harvested when ripe and the straw and grain were analysed separately. The total radioactivity concentration was 20 times higher in straw than in grain. In straw and grain, 12 and 25% respectively of the total incorporated radioactivity in each of these tissues was methanol soluble. The composition of the methanol-soluble radioactive residue was investigated and was shown to contain triforine and its metabolites which were free and unbound in barley straw and grain. No radioactive piperazine was observed, in spite of the high detection sensitivity for radioactivity. The concentrations of triforine and its identified metabolites in straw and grain respectively (mg kg^?1, relative to the fresh weight of tissue) were: triforine, 0.034 and 0.0018; N-[(2,2,2-trichloro-1-(piperazin-1-yl)ethyl]formamide), 0.009 and 0.0006; iminodiacetic acid, 0.021 and 0.001; glycine, 0.043 and 0.0033. Other radioactive water-soluble and very polar, unidentified compounds were observed, corresponding to advanced metabolic products of triforine.     

Photolysis of pirimicarb in water under natural and simulated sunlight conditions

The photostability of pirimicarb, (2-dimethylamino-5,6-dimethylpyrimidin-4-yl dimethylcarbamate) in aqueous media, under natural and artificial sunlight irradiation conditions, is reported. The pH of the irradiated solutions remained constant during degradation time. The photodegradation mechanism seemed to be similar under both conditions, but the half-life was found to be about three times longer under natural than under artificial conditions. Four main photo-products ( 1 , 2 , 3 and 4 ) were isolated and identified by spectroscopic methods. It is proposed that a common photodegradation pathway exists for pirimicarb under both irradiation conditions. The results obtained from the photodegradation of pirimicarb and of its four main products isolated on solid phase permit the construction of a possible photodegradation pathway. Photoproduct 4 (2-methyl-formylamino-5,6-dimethyl-4-hydroxypyrimidine) is here described for the first time.     

Metabolism of [2,5-14c]piperazine in barley plants

The shoots of barley plants root-treated with [2,5-¹⁴C]piperazine were analysed 30 days after treatment. Methanol extraction left a solid residue which contained 31.9% of ¹⁴C (all percentages refer to the total of ¹⁴C incorporated into the shoots); further extraction with acidified methanol and dimethyl sulphoxide dissolved respectively 3.2% and 5.8% of ¹⁴C. The initial methanol extract contained radioactive piperazine (16.8%), iminodiacetic acid (8.6%), glycine (15.4%), oxalic acid (7.2%), and un-identified compounds (20.1%). In barley, piperazine is the product of the most advanced metabolism so far identified of the fungicide triforine, 1,4-bis(2,2,2-trichloro-1-formamidoethyl)piperazine; the results obtained here show that piperazine is certainly not the end-product of the metabolism of triforine in barley.     

光对入侵性植物黄顶菊种子萌发及植株生长的影响

入侵性杂草黄顶菊原产南美,2003年在我国河北省衡水湖首次报道。采用室内控制试验,对光与黄顶菊种子发芽及植株生长的关系进行了研究。结果表明：黄顶菊种子属光敏型,种子需要光刺激才能发芽。但其萌发对光强要求不严,1 000 lx光照强度30 ℃培养6、12 h和24 h转入暗培养,5 d后黄顶菊种子发芽率分别为67.0%、88.0%和95.8%。黄顶菊种子出苗与光照关系密切,播种在土壤表面、0.5 cm和1 cm深土层的种子出苗率分别为96.0%、8.0%和0。随光照强度减弱,植株的生物量及繁殖力显著降低。黄顶菊在35%自然光强下生长时其生物量、结实数比自然光强下分别降低55.0%和55.6%。上述结果为预测黄顶菊的适生区域及制订有效的防除策略提供了科学依据。     

THE PHOTOLYTIC DEGRADATION OF PICLORAM

Summary. The effects of sunlight and ultraviolet light (253.7 nm) on 4-amino-3,5,6-trichloropicolinic acid (picloram) in a basic aqueous solution were investigated. Electron-capture gas chromatography determinations showed that approximately 20% of a 2 × 10⁻² M concentration of picloram was degraded for each 48-hr exposure to ultraviolet light of an intensity of 200 μW/cm². Decomposition by sunlight was slower and more variable.
Precipitation of the degradation products with AgNO₃ indicated that two chloride ions were liberated for each molecule of picloram that was degraded. Titrations indicated that acids were formed in the degradation process, and ultraviolet spectra showed that the pyridine nucleus was destroyed. Chromatograms of the methylated decomposition products showed that at least five products were formed.
The effect of the addition of a free radical scavenger, hydroquinone, on the rate of decomposition was investigated but no definite conclusions could be drawn.
La dégradation photolytique du piclorame     

Metabolism of the fungicide triforine in barley plants

The metabolic fate of [³H]-triforine in barley plants has been studied. 15 and 30 days after treatment, unchanged triforine amounted to 57.5 and 43.2% respectively, and of three soluble metabolites detected, piperazine (0.3 and 4.0%) and N-[2, 2,2-trichloro-1-(piperazin-1-yl)ethyl] formamide (12.9 and 8.4%), the latter for the first time as a metabolite in plants, were identified using t.1.c. with four solvent systems. The solid residue contained 23.1 and 38.2% of bound label after 15 and 30 days respectively.     

Effects of fungicides on powdery mildew, tree growth and cropping of apple

In a 6-year orchard experiment, seven fungicide programmes were assessed for control of powdery mildew (Podosphaera leucotricha) and for long-term effects on growth and cropping of the apple cultivar Cox's Orange Pippin. Programmes based on binapacryl, bupirimate, fenarimol, nitrothal-isopropyl and triadimefon gave better control than dinocap or triforine. The heaviest cumulative yields of marketable fruit were obtained with the non-systemic fungicides binapacryl and nitrothal-isopropyl, the former significantly outyielding triadimefon and triforine over 6 years. Tree vigour, as assessed by extension shoot length and internode length, was found to be better after 4-5 years of programmes based on binapacryl than on triadimefon or fenarimol. A relation was demonstrated between cumulative marketable yield and mean annual mid-season incidence of secondary mildew on extension shoot leaves.     

Metabolism of some sterol inhibitors in fungi and higher plants,with special reference to the selectivity of fungicidal action

The metabolism of triforine, chloraniformethan, the α-(pyrimidin-5-yl)benzhydryl alcohols (fenarimol, nuarimol and triarimol), the trityl-azoles (fluotrimazole and clotrimazole), and the morpholine types of sterol inhibitors is reviewed; the metabolism of the azolyl-alkane derivatives (mainly triadimefon and triadimenol) is discussed in detail. Redox and hydrolytic reactions are of primary importance. Enzymic inactivation may be one factor influencing fungicide selectivity. Metabolism is the dominant factor of selectivity if it represents the activation process, as with triadimefon. Transformations in higher plants do not differ significantly from those occurring in fungi, except that factors such as the formation of conjugates with natural compounds of plant tissues also play a role, as with triforine and triadimenol. The selectivity of fungitoxic action may be influenced by metabolism both in the host plant and the pathogen.     

Antifeedant activity of solvent extracts of neem seed kernel against Spodoptera litura F. and their persistency against sunlight through encapsulation

Abstract

Different solvent extracts of neem seed kernels were evaluated against Spodoptera litura F. on cauliflower, Brassica oleracea var. botrytis leaves. Based on this evaluation, aqueous extract was dissolved in ethanol, and methanol extract in ethanol, and dissolved parts were designated as fraction I and fraction II, respectively. The extracts having more antifeedant activity were encapsulated to achieve stability against the sunlight. Among the solvent extracts tested at 1.0% concentration, methanol extract provided maximum protection (100%) of the leaves followed by ethanol (98.39%) and aqueous (93.01%) extracts. Fraction I and fraction 11 were equally effective at 0.1 % concentration against S. litura larvae and checked more than 70% of leaf damage. However, such extracts were found to be unstable when exposed to sunlight. The pre‐gelatinized starch‐encapsulated products, viz. ρre‐gel I and pre‐gel II were quite stable and afforded more than 68.0% protection to the cauliflower leaves even after 6 days of exposure to sunlight.