Fungicidal activity and chemical constitution: XIII.* Active components of commercial formulations containing dinocap

Six isomers, isolated from the mixture of dinitrophenols obtained by the hydrolysis of commercial formulations containing dinocap, were tested in the greenhouse against apple mildew caused by Podosphaera leucotricha. The two most active compounds were 4-(Ié-ethylhexyl)- and 4-(Ié-propylpentyl)-2,6-dinitrophenols, which appear to account for most of the antimildew activity of the mixture. The crotonate of the latter phenol was considerably more effective than dinocap itself, but both crotonates were less effective than the corresponding free phenols. Little evidence of synergism between two ethylhexyldinitrophenols was obtained.     

Degradation of anaerobic reductive dechlorinationproducts of Aroclor 1242 by four aerobic bacteria

We studied the aerobic degradation of eight PCB congeners which comprise from 70 to 85% of the anaerobic dechlorination products from Aroclor 1242, including2-, 4-, 2,4-, 2,6-, 2,2'-, 2,4'-, 2,2',4-, and2,4,4'-chlorobiphenyl (CB), and the biodegradation of their mixtures designed to simulate anaerobic dechlorination profiles M and C. StrainsComamonas testosteroni VP44 and Rhodococcus erythreus NY05 preferentially oxidizeda para-substituted ring, while Rhodococcus sp. RHA1, similar to well known strain Burkholderia sp. LB400, preferably attackedan ortho-chlorinated ring. Strains with ortho-directed attack extensively degraded2,4'- and 2,4,4'-CB into 4-chlorobenzoate, while bacteria with para-directed attack transformed these congeners mostly into potentially problematicmeta-cleavage products. The strains that preferentiallyoxidized an ortho-substituted ring readily degradedseven of the eight congeners supplied individually; only 2,6-CB was poorly degraded. Degradationof 2,2'- and 2,4,4'-CB was reduced when present in mixtures M and C. Higher efficiencies of degradation of the individual congeners and defined PCB mixtures M and C and greater production of chlorobenzoates were observed with bacteria that preferentially attackan ortho-substituted ring. PCB congeners 2,4'-, 2,2',4-, and 2,4,4'-CB canbe used to easily identify bacteria with ortho-directed attack whichare advantageous for use in the aerobic stage of the two-phase (anaerobic/aerobic)PCB bioremediation scheme.     

Screening of new 2,4- and 2,6-dinitroaniline derivates for phytotoxicity and antimitotic activity

Screening of antimitotic activity and phytotoxicity of new 2,4- and 2,6-dinitroaniline derivates has been performed using the Allium-test. All substances that are derivatives of 2,4-dinitroanilines, 2,6-dinitro-(4-fluoromethyl)-aniline, and of (methylsulfonyl) nitrobenzol, proved to change the value of the mitotic index, cause cytogenetic disorders, and have a phytotoxic effect on the roots of Allium cepa seedlings. The results obtained suggest that N-(2,4-dinitrophenyl)-orto-aminobenzoic acid, N, N, -diethyl-2,6-dinitro-4-(trifluoromethyl)-aniline, and 1-methylsulfonyl-3-nitrobenzol are potential herbicides; further studying of their effects is proposed.     

Fungitoxic and phytotoxic effect of some surface-active agents applied for the control of apple powdery mildew

Mixtures of methyl esters of fatty acids (‘Off-Shoot O’) and mixtures of fatty alcohols (‘Off-Shoot T’), applied during the late autumn (November), eradicated apple powdery mildew (caused by Podosphaera leucotricha) from infected buds. Applications in the spring at bud-burst were less effective. The autumn sprays caused little damage to the cultivars Bramley's Seedling, Cox's Orange Pippin and Golden Delicious but on Worcester Pearmain 50% of the buds were killed by a 5% Off-Shoot O spray. The bud-burst sprays were damaging and reduced yield on the Cox and Bramley trees. A 5% a.i. mixture of nonanol/lissapol applied to a range of commercial cultivars was effective in eradicating powdery mildew but was more phytotoxic than the Off-Shoot compounds. Spraying Cox trees later in the winter failed to reduce the damage. Eradication of the overwintering stage of the disease resulted in low spore concentrations during the following spring and early summer, enabling some economies to be made in the summer mildew fungicide programme.     

Oleozon: A novel control strategy against powdery mildew in cucumber

Ozonized sunflower oil (oleozon) is an effective agent for controlling powdery mildew in cucumber. In this study, the mechanisms of oleozon in the control of powdery mildew were determined. The development of Podosphaera xanthii on cucumber leaves treated with oleozon (2%) and water was investigated at different times after inoculation. The germinating conidia, hyphae and conidiophores of the pathogen were severely damaged by oleozon. No visible phytotoxic effect was observed on cucumber after the application of oleozon. This compound had highly preventive effects as well as curative effects against powdery mildew based on in vivo potted seedling assays. The control effects of oleozon were further confirmed in a greenhouse trial. These results may provide a basis for further development of a natural fungicide against cucumber powdery mildew.     

Fungicide programmes for the control of spur blight, powdery mildew and grey mould on raspberry

Five sprays of dichlofluanid (0–1% a. i.) reduced spur blight (Didymella applanata) on canes and powdery mildew (Sphaerotheca macularis) on fruit more than five sprays of captan (0–1% a. i.). Omission of the first three applications resulted in little increase in disease but when the fourth (full bloom) application also was omitted the incidence of disease was comparable with that on untreated plants. The final (fruitlet stage) application did not further reduce disease incidence. Five sprays of dichlofluanid reduced grey mould (Botrytis cinerea) on stored fruit more than five sprays of captan. There was an increase in grey mould with progressive reductions in the number of sprays. Incidence on canes was slight and was unaffected by treatments. A programme of five dichlofluanid sprays applied at 14-day intervals until c. 2 wk before harvest and supplemented by a single early full bloom spray of either dinocap (0–025% a. i.), binapacryl (0–05% a. i.), triforine (0–025% a. i.), benomyl (0–025% a. i.) or replaced by benomyl (0–025% a. i.) all halved both the number of fruits infected by powdery mildew and the number of spur blight lesions on canes, compared with unsprayed controls. Six applications of any of the following fungicides at 7-day intervals from 23 May until 29 June 1977 reduced powdery mildew incidence on fruit, the last fungicide being particularly effective: dinocap (0–0125%), dichlofluanid (0–075%), binapacryl (0–025%), triforine (0–0125%), bupirimate (0–0075%), ditalimfos (0–01875%) and fenarimol (0–0036%). Only triforine and bupirimate reduced the incidence of mildew on shoots. Dichlofluanid, binapacryl, triforine and fenarimol all reduced the amount of spur blight on canes. The largest fruit occurred on plants sprayed with bupirimate, ditalimfos and fenarimol, and late ripening was associated with triforine treatments.     

A new phenol oxidase produced during melanogenesis and encystment stage in the nitrogen-fixing soil bacterium Azotobacter chroococcum

Laccases are copper-containing phenol oxidases that are commonly found in many types of plant, insect, fungi and bacteria. Whilst phenol oxidases have been well characterized in fungal species, laccase-type enzymes originating from bacteria have been much less well defined. Bacteria belonging to the family Azotobacteraceae share many morphological characteristics with strains already known to exhibit polyphenol and phenol oxidase activity; and hence the aim of this work was to identify and characterize a novel laccase from the isolated strain Azotobacter chroococcum SBUG 1484 in an attempt to provide further understanding of the roles such enzymes play in physiological development. Laccase activity was clearly observed through oxidation of 2,6-dimethoxyphenol, other typical substrates including: methoxy-monophenols, ortho- and para-diphenols, 4-hydroxyindole, and the non-phenolic compound para-phenylenediamine. A. chroococcum SBUG 1484 showed production of a cell-associated phenol oxidase when grown under nitrogen-fixing conditions, and was also observed when cells enter the melanogenic and encystment stages of growth. Catechol which is structurally related to melanin compounds was also released from Azotobacter cells into the surrounding culture medium during nitrogen-fixing growth. From our results we propose that a membrane-bound laccase plays an important role in the formation of melanin, which was monitored to correlate with progression of A. chroococcum SBUG 1484 cells into the encystment stage of growth.     

The role of the apple spray programme in the protection of fruit buds from powdery mildew

Comparisons of programmes, which differed in the dates on which high-volume sprays of dinocap (0.019 %) were omitted, indicated that applications in the period from green cluster to early fruitlet protected the bourse buds of apple cv. Cox's Orange Pippin from infection, and so controlled the number of mildewed blossom trusses in the following year. This role of the sprays in preventing primary mildew declined after the early fruitlet stage. The existence of two phases of the disease was confirmed, and although dinocap at late blossom and early fruitlet contributed to the control of preliminary infections of secondary mildew on vegetative shoots, the latter phase did not become severe until after the peak invasion of bourse buds. Fewer than 20% of the fruit buds which produced primary-mildewed blossom in 1968 were succeeded by healthy fruit buds in the next year; this poor recovery was not improved by dinocap applied during the flowering period in 1968.
Fruit-set and crop of James Grieve and Cox's Orange Pippin were reduced in 1968 by dinocap applied at green cluster and pink bud, or at full blossom and petal-fall, but no deleterious effects were produced by equivalent sprays in 1969. The need for mildew control during flowering is discussed in relation to phytotoxic sensitivity.     

Aerobic Mineralization of 2,6-Dichlorophenol by Ralstonia sp. Strain RK1

A new aerobic bacterium was isolated from the sediment of a freshwater pond close to a contaminated site at Amponville (France). It was enriched in a fixed-bed reactor fed with 2,6-dichlorophenol (2,6-DCP) as the sole carbon and energy source at pH 7.5 and room temperature. The degradation of 2,6-DCP followed Monod kinetics at low initial concentrations. At concentrations above 300 μM (50 mg · liter⁻¹), 2,6-DCP increasingly inhibited its own degradation. The base sequence of the 16S ribosomal DNA allowed us to assign the bacterium to the genus Ralstonia (formerly Alcaligenes). The substrate spectrum of the bacterium includes toluene, benzene, chlorobenzene, phenol, and all four ortho- and para-substituted mono- and dichlorophenol isomers. Substituents other than chlorine prevented degradation. The capacity to degrade 2,6-DCP was examined in two fixed-bed reactors. The microbial population grew on and completely mineralized 2,6-DCP at 2,6-DCP concentrations up to 740 μM in continuous reactor culture supplied with H₂O₂ as an oxygen source. Lack of peroxide completely stopped further degradation of 2,6-DCP. Lowering the acid-neutralizing capacity of the medium to 1/10th the original capacity led to a decrease in the pH of the effluent from 7 to 6 and to a significant reduction in the degradation activity. A second fixed-bed reactor successfully removed low chlorophenol concentrations (20 to 26 μM) with hydraulic residence times of 8 to 30 min.     

Effect of Substituted β-Phenoxyacrylates on Shoot Elongation of Barnyard-grass

Twenty-seven substituted ethyl β-phenoxyacrylates were prepared to clarify the relationship between chemical structure and inhibition of the shoot elongation of barnyard-grass. ortho-Halo-substitution greatly enhanced the inhibitory activity, the order of which was Cl>F≧Br>I.

The cis/trans ratio of each preparation was approximately 1:4 on the basis of the NMR spectrum, and the activity of trans isomers was about 100 times higher than that of cis isomers in the case of the 2-chloro- and 2,4-dichloro-derivatives. Among the compounds tested, ethyl trans- β-(2,4-dichlorophenoxy) acrylate was found to be the most effective in inhibiting shoot elongation.     

Regiospecificity of Chlorophenol Reductive Dechlorination by Vitamin B12s

Vitamin B₁₂, reduced by titanium (III) citrate to vitamin B_12s, catalyzes the reductive dechlorination of chlorophenols. Reductive dechlorination of pentachlorophenol and of all tetrachlorophenol and trichlorophenol isomers was observed. Reaction of various chlorophenols with vitamin B₁₂ favored reductive dechlorination at positions adjacent to another chlorinated carbon, but chlorines ortho to the hydroxyl group of a phenol were particularly resistant to reductive dechlorination, even if they were also ortho to a chlorine. This resulted in a reductive dechlorination pattern favoring removal of para and meta chlorines, which differs substantially from the pattern exhibited by anaerobic microbial consortia.     

The effectivity of Tilletiopsis albescens in biocontrol of powdery mildew

Tilletiopsis albescens grows well on powdery mildew fungi inoculated on barley or cucumber leaves and causes collapse of the colonies. Application of ballistospores or cut mycelium was equally effective for biocontrol, and the effectiveness tended to increase exponentially with the concentration of germinating units (conidia and cut mycelium) applied. Seventy percent relative humidity or more is required for effective biocontrol. Two applications of T. albescens in the period from 3 days before to 3 days after inoculation with powdery mildew were more effective than one. Applications before inoculation or 7 days after inoculation with powdery mildew had little effect. T. albescens followed the powdery mildew as it was disseminated to uninoculated leaves, but this did not result in an effective biocontrol. The potential for using T. albescens for biocontrol of powdery mildews is discussed.     

Enantioselective biocatalytic reactions on (±)-aryl alkyl ketones with native and modified porcine pancreatic lipase

Abstract

Porcine pancreatic lipase (PPL), pre-incubated with acetophenone in tetrahydrofuran, fails to recognize ortho- and para-acyloxy functions with respect to the nuclear carbonyl group in (±)-2,4-diacyloxyphenyl alkyl ketones and produces novel aryl alkyl ketones in moderate-to-highly optically active forms; this result supports the hypothesis on the mechanism of action of PPL in deacylation reactions on peracylated polyphenolics.     

Co-metabolism of di- and trichlorobenzoates in a 2-chlorobenzoate-degrading bacterial culture: Effect of the position and number of halo-substituents

The degradative properties towards chlorobenzoates by a 2-chlorobenzoate-degrading mixed culture (2MC) were investigated. Although 2MC did not grow on 2,3,5-; 2,3,6- and 2,4,6-trichlorobenzoates, it was able to completely oxidise 2,3,5-trichlorobenzoate and 2,5-dichlorobenzoate. This was degraded through the intermediate formation of 4-chlorocatechol and 3-cis,cis-chloromuconate, suggesting a dioxygenation in 1,2 position. 4-Chlorobenzoate; 3,4- and 2,6-dichlorobenzoate; 2,4,6- and 2,3,6-trichlorobenzoate did not undergo any co-metabolic transformation, the only 2,4-dichlorobenzoate was scarcely transformed into a dichlorinated phenol as end-product. 2MC was constituted of three prominent species: Stenotrophomonas maltophilia, Cupriavidus necator and Flavobacterium sp. Ring-hydroxylating dioxygenase genes (BenA) were retrieved in S. maltophilia and C. necator, indicating the presence of a “modified ortho pathway” in the culture. On the basis of its degradative properties, 2MC could be a good candidate for use in the removal of ortho- and/or meta-chlorobenzoates that are the major end-products of PCB co-metabolism.     

Molecular phylogenetic analyses reveal a close relationship between powdery mildew fungi on some tropical trees and Erysiphe alphitoides, an oak powdery mildew

To investigate the phylogenetic relationships among the powdery mildew fungi of some economically important tropical trees belonging to Oidium subgenus Pseudoidium, we conducted molecular phylogenetic analyses using 30 DNA sequences of the rDNA internal transcribed spacer (ITS) regions and 26 sequences of the domains D1 and D2 of the 28S rDNA obtained from the powdery mildews on Hevea brasiliensis (para rubber tree), Anacardium occidentale (cashew), Bixa orellana, Citrus spp., Mangifera indica (mango), and Acacia spp. The results indicate that the powdery mildew fungi isolated from these tropical trees are closely related to one another. These powdery mildews are also closely related to E. alphitoides (including Erysiphe sp. on Quercus phillyraeoides). Because of the obligate biotrophic nature of the powdery mildew fungi, the relationship between powdery mildews and their host plants is conservative. However, the present study suggests that a particular powdery mildew species has expanded its host ranges on a wide range of the tropical trees. This article also suggests that a powdery mildew fungus distributed in temperate regions of the Northern Hemisphere expanded its host ranges onto tropical plants and may be a good example of how geographical and host range expansion has occurred in the Erysiphales.     

Temporal isolation explains host-related genetic differentiation in a group of widespread mycoparasitic fungi

Understanding the mechanisms responsible for divergence and specialization of pathogens on different hosts is of fundamental importance, especially in the context of the emergence of new diseases via host shifts. Temporal isolation has been reported in a few plants and parasites, but is probably one of the least studied speciation processes. We studied whether temporal isolation could be responsible for the maintenance of genetic differentiation among sympatric populations of Ampelomyces, widespread intracellular mycoparasites of powdery mildew fungi, themselves plant pathogens. The timing of transmission of Ampelomyces depends on the life cycles of the powdery mildew species they parasitize. Internal transcribed spacer sequences and microsatellite markers showed that Ampelomyces populations found in apple powdery mildew (Podosphaera leucotricha) were genetically highly differentiated from other Ampelomyces populations sampled from several other powdery mildew species across Europe, infecting plant hosts other than apple. While P. leucotricha starts its life cycle early in spring, and the main apple powdery mildew epidemics occur before summer, the fungal hosts of the other Ampelomyces cause epidemics mainly in summer and autumn. When two powdery mildew species were experimentally exposed to Ampelomyces strains naturally occurring in P. leucotricha in spring, and to strains naturally present in other mycohost species in autumn, cross‐infections always occurred. Thus, the host‐related genetic differentiation in Ampelomyces cannot be explained by narrow physiological specialization, because Ampelomyces were able to infect powdery mildew species they were unlikely to have encountered in nature, but instead appears to result from temporal isolation.     

Water Stress-Induced Changes in the Morphology of the Powdery Mildew, Leveillula taurica (Lèv.) Arn.

Water stress in Capsiucm annuum L., induced by means of polyethylene glycol influenced the length and branching frequency of conidiophores and conidial dimensions of the powdery mildew, Leveillula taurica (Lèv.) Arn. Conidiophore branching frequency and length as well as conidial length and width decreased with decreasing relative water content. The availability of water to host plants seems to have a direct effect on the growth vigour and development of the powdery mildew possibly is suggested that water stress may be imposed on host plants by the dry harmattan season resulting in reduced growth vigour of the powdery mildew. Morphology of powdery mildew developing under low water potential may be altered.     

Simultaneous modification of three homoeologs of TaEDR1 by genome editing enhances powdery mildew resistance in wheat

Wheat (Triticum aestivum L.) incurs significant yield losses from powdery mildew, a major fungal disease caused by Blumeria graminis f. sp. tritici (Bgt). enhanced disease resistance1 (EDR1) plays a negative role in the defense response against powdery mildew in Arabidopsis thaliana; however, the edr1 mutant does not show constitutively activated defense responses. This makes EDR1 an ideal target for approaches using new genome‐editing tools to improve resistance to powdery mildew. We cloned TaEDR1 from hexaploid wheat and found high similarity among the three homoeologs of EDR1. Knock‐down of TaEDR1 by virus‐induced gene silencing or RNA interference enhanced resistance to powdery mildew, indicating that TaEDR1 negatively regulates powdery mildew resistance in wheat. We used CRISPR/Cas9 technology to generate Taedr1 wheat plants by simultaneous modification of the three homoeologs of wheat EDR1. No off‐target mutations were detected in the Taedr1 mutant plants. The Taedr1 plants were resistant to powdery mildew and did not show mildew‐induced cell death. Our study represents the successful generation of a potentially valuable trait using genome‐editing technology in wheat and provides germplasm for disease resistance breeding.     

Effect of Molecular Characteristics on Cellular Uptake,Subcellular Localization,and Phototoxicity of Zn(II) N-Alkylpyridylporphyrins

Tetra-cationic Zn(II) meso-tetrakis(N-alkylpyridinium-2 (or -3 or -4)-yl)porphyrins (ZnPs) with progressively increased lipophilicity were synthesized to investigate how the tri-dimensional shape and lipophilicity of the photosensitizer (PS) affect cellular uptake, subcellular distribution, and photodynamic efficacy. The effect of the tri-dimensional shape of the molecule was studied by shifting the N-alkyl substituent attached to the pyridyl nitrogen from ortho to meta and para positions. Progressive increase of lipophilicity from shorter hydrophilic (methyl) to longer amphiphilic (hexyl) alkyl chains increased the phototoxicity of the ZnP PSs. PS efficacy was also increased for all derivatives when the alkyl substituents were shifted from ortho to meta, and from meta to para positions. Both cellular uptake and subcellular distribution of the PSs were affected by the lipophilicity and the position of the alkyl chains on the periphery of the porphyrin ring. Whereas the hydrophilic ZnPs demonstrated mostly lysosomal distribution, the amphiphilic hexyl derivatives were associated with mitochondria, endoplasmic reticulum, and plasma membrane. A comparison of hexyl isomers revealed that cellular uptake and partition into membranes followed the order para > meta > ortho. Varying the position and length of the alkyl substituents affects (i) the exposure of cationic charges for electrostatic interactions with anionic biomolecules and (ii) the lipophilicity of the molecule. The charge, lipophilicity, and the tri-dimensional shape of the PS are the major factors that determine cellular uptake, subcellular distribution, and as a consequence, the phototoxicity of the PSs.