On the mode of action of the herbicides cinmethylin and 5-benzyloxymethyl-1, 2-isoxazolines: putative inhibitors of plant tyrosine aminotransferase

BACKGROUND: The mode of action of the grass herbicides cinmethylin and 5‐benzyloxymethyl‐1,2‐isoxazolines substituted with methylthiophene (methiozolin) or pyridine (ISO1, ISO2) was investigated. RESULTS: Physiological profiling using a series of biotests and metabolic profiling in treated duckweed (Lemna paucicostata L.) suggested a common mode of action for the herbicides. Symptoms of growth inhibition and photobleaching of new fronds in Lemna were accompanied with metabolite changes indicating an upregulation of shikimate and tyrosine metabolism, paralleled by decreased plastoquinone and carotenoid synthesis. Supplying Lemna with 10 µM of 4‐hydroxyphenylpyruvate (4‐HPP) reversed phytotoxic effects of cinmethylin and isoxazolines to a great extent, whereas the addition of L ‐tyrosine was ineffective. It was hypothesised that the herbicides block the conversion of tyrosine to 4‐HPP, catalysed by tyrosine aminotransferase (TAT), in the prenylquinone pathway which provides plastoquinone, a cofactor of phytoene desaturase in carotenoid synthesis. Accordingly, enhanced resistance to ISO1 treatment was observed in Arabidopsis thaliana L. mutants, which overexpress the yeast prephenate dehydrogenase in plastids as a TAT bypass. In addition, the herbicides were able to inhibit TAT7 activity in vitro for the recombinant enzyme of A. thaliana. CONCLUSION: The results suggest that TAT7 or another TAT isoenzyme is the putative target of the herbicides. Copyright © 2011 Society of Chemical Industry     

Herbicide pyrazolate causes cessation of carotenoids synthesis in early watergrass by inhibiting 4-hydroxyphenylpyruvate dioxygenase

In aqueous solution, the herbicide pyrazolate [4‐(2,4‐dichlorobenzoyl)‐1,3‐dimethyl‐5‐pyrazolyl p‐toluenesulfonate] is rapidly hydrolyzed to destosyl pyrazolate (DTP), 4‐(2,4‐dichlorobenzoyl)‐1,3‐dimethyl‐5‐hydroxypyrazole, which is an active form of the herbicide. The objective of this study was to examine the effect of pyrazolate and DTP on carotenoids synthesis in susceptible weed, early watergrass (Echinochloa oryzicola Vasing.). Furthermore, their in vitro effect on 4‐hydroxyphenylpyruvate dioxygenase (HPPD) was determined. Roots of the plants at the two‐leaf stage were soaked for 24 h into pyrazolate (5 × 10^–5 mol L^?1) or norflurazon (10^–6 mol L^?1) solution containing 0.5% volume of acetone. At the first sampling time (3 days after treatment: 3 DAT), the chlorophyll content in the third leaves of pyrazolate‐treated plants were not different compared with the untreated control, but it was decreased between 3 and 6 DAT. The declining pattern of β‐carotene in the third leaf of early watergrass was very similar to that of chlorophyll. Both herbicides induced greater accumulation of phytoene in the third leaves of early watergrass 3 DAT, and the levels were kept until 9 DAT. However, feeding of homogentisate reduced the phytoene accumulation only in pyrazolate‐treated plants, suggesting the site of action of the herbicide located in the pathway of plastoquinone synthesis. In a HPPD assay, DTP revealed to inhibit the enzyme with an IC₅₀ value of 13 nmol L^?1 and that of pyrazolate was 52 nmol L^?1. In the pyrazolate solution used in the assay, some of the herbicide possibly has been hydrolyzed to DTP. From the all results obtained, it is strongly suggested that pyrazolate inhibits carotenoids synthesis and causes bleaching on the developing leaves by the similar mechanism with norflurazon, but its action site is not phytoene desaturase and is HPPD.     

Towards the primary target of chloroacetamides –new findings pave the way

This review reports on research of the last ten years to find the primary target enzyme for chloroacetamides. As could be shown first with the green alga Scenedesmus, the formation of very‐long‐chain fatty acids (VLCFAs) is severely impaired. Subsequently, in short‐term experiments, labelled malonate or stearate could be incorporated into leaf discs of cucumber, barley or leek seedlings. While the formation of ‘normal’ long‐chain fatty acids (up to C18) was not influenced, phytotoxic chloroacetamides strongly inhibited the synthesis of VLCFAs of C20, 22 and 24, with I₅₀ values of 10–100 nM . Inhibition depends on the amide structure and on stereospecificity. Also cafenstrole or recently developed tetrazolinones and phosphosulfonates were found active to inhibit fatty‐acid elongation. Subsequently, a cell‐free elongase assay was developed using a microsomal preparation from leek seedlings (Allium porrum L), [¹⁴C]malonyl‐CoA and C18, 20, or C22 acyl‐CoA primer substrates. All elongation steps were strongly affected by those phytotoxic herbicides which were also active in vivo. The inhibitors form a tight‐binding complex with the condensing elongase enzyme system which develops with time and lowers the I₅₀ values markedly. Apparently, a nucleophilic attack of the inhibitor takes place at the specific target enzyme. Acyl‐CoA elongation inhibition is correlated with growth inhibition of the intact cell. Due to the low I₅₀ values and the specific inhibition, we assume that impaired VLCFA‐formation is the primary phytotoxic impact of chloroacetamides and functionally related structures. © 2000 Society of Chemical Industry     

Inhibition of carotenogenesis by substituted diphenyl ethers of the m-phenoxybenzamide type

Diphenyl ethers exhibit different modes of action according to their chemical constitution. Diphenyl ethers of the m-phenoxybenzamide type, which were found to be effective on carotenogenesis resulting in an accumulation of colorless carotenoid precursors, mostly phytoene, indicative of inhibition of desaturation, are discussed. As seen with other carotenoid biosynthesis inhibitors, a concurrent loss of chlorophyll was observed as a secondary effect caused by the absence of protective carotenoids. In contrast to peroxidative p-nitrodiphenyl ethers like oxyfluorfen (2-chloro-4-trifluoromethylphenyl-3′-ethoxy-4′-nitrophenyl ether), the m-phenoxybenzamides assayed showed the same phytotoxic mode of action in the dark as observed when using heterotrophic Scenedesmus cultures. As expected, chlorophylls were not affected. The decrease of carotenoids was not due to their degradation but to inhibited carotenogenesis. Examination of carotenoid fractions show that the m-phenoxybenzamides, e.g., 3-(2,5-dimethylphenoxy)-N-ethylbenzamide, used here act similarly to 2-phenylpyridazinones like norflurazon [4-chloro-5-methylamino-2-(2-trifluoromethylphenyl)-pyridazin-3(2H)one]. All these inhibitors strongly decrease the α- and β-carotene content, while xanthophyll content is not lowered as much.     

类胡萝卜素生物合成抑制剂研究进展

概述了类胡萝卜素生物合成抑制剂类除草剂的作用机理以及八氢番茄红素去饱和酶(phytoene desaturase, PD酶)抑制剂的结构-活性关系。简要介绍了进入商品化开发应用的类胡萝卜素生物合成抑制剂类除草剂品种以及它们的除草活性。     

Mode of action of new diethylamines in lycopene cyclase inhibition and in photosystem II turnover

The new bleaching herbicidal compound N,N‐diethyl‐N‐(2‐undecynyl)amine (NDUA) is identified here as an inhibitor of lycopene cyclase and is compared with the known cyclase inhibitors N,N‐diethyl‐N‐[2‐(4‐chlorophenylthio)ethyl]amine (CPTA) and N,N‐diethyl‐N‐[2‐(4‐methylphenoxy)ethyl]amine (MTPA). HPLC separation of chloroplast pigments shows lycopene accumulation in NDUA treated tissue. Variation in chain length of the undecynylamine moeity of NDUA from 7 to 21 C atoms reveals an optimum of 11 to 14 C atoms for herbicidal activity. A series of seven further analogues of NDUA and CPTA reveals the structural elements necessary for inhibition of lycopene cyclase. The effect of NDUA derivatives on photosynthesis has been studied in Chlamydomonas reinhardtii. Photosynthesis is highly sensitive, particularly towards the C14 and longer chain length analogues at nanomolar concentrations. It is shown that the breakdown of photosynthesis by NDUA is due to interference with the turnover of the D1 protein of the photosystem II reaction centre that requires the continous biosynthesis of the two reaction‐centre β‐carotene moieties in the reassembly phase. The D1 protein disappearance is most marked under strong light conditions. The depletion of photosystem II occurs before total pigment bleaching. This newly recognized mechanism in herbicidal activity is also the basis for the mode of action of other lycopene cyclase inhibitors as well as phytoene desaturase inhibitors. © 2001 Society of Chemical Industry     

The mode of action of RH‐1965: a new phenylpyrimidinone bleaching herbicide

RH‐1965 is a new bleaching herbicide which causes newly developing leaf tissue to emerge devoid of photosynthetic pigments. Mode‐of‐action studies revealed that RH‐1965 inhibited the accumulation of both total chlorophyll and β‐carotene. Concomitantly, it induced the accumulation of the β‐carotene precursors, phytoene, phytofluene and, in particular, ξ‐carotene. Inhibition of chlorophyll accumulation by RH‐1965 is attributed to the photo‐oxidative destruction of chlorophyll in the absence of β‐carotene because RH‐1965 blocked chlorophyll accumulation to a greater extent under high light (50–330 µE m⁻² s⁻¹) than under low light (0.8 µE m⁻² s⁻¹) conditions. Radish (Raphanus sativus L) and barnyardgrass (Echinochloa crus‐galli (L) Beauv) were very senstive to RH‐1965. Under high light (330 µE m⁻² s⁻¹), the I₅₀ values for inhibition of chlorophyll accumulation were 0.10 and 0.15 µM , respectively. Wheat (Triticum aestivus L), on the other hand, was much less sensitive to RH‐1965 (I₅₀ = 1.4 µM ). It is concluded that the mode of action of RH‐1965 involves the inhibition of ξ‐carotene desaturation. © 2000 Society of Chemical Industry     

Carotenogenic inhibition by norflurazon in wheat

Wheat (Triticum aestivum L. cv Holley) seedlings were exposed to [N-¹⁴CH₃]norflurazon in nutrient solution studies. The ¹⁴CH₃ group was incorporated into a compound eluting on GLC at a relative retention temperature R_f equivalent to n-C₂₁ H₃₆ and mass spectrometry validated a 295 MW. The concentration of [N-¹⁴CH₃]norflurazon and/or Rl[¹⁴C]norflurazon which resulted in carotenogenesis inhibition was 0.07 μM in the water contained in the leaves. The concentration of norflurazon required for phytoene accumulation as a mode-of-action was ca. 140 × the concentration of norflurazon required for geranylgeraniol accumulation. Geranylgeraniol accumulated at 1 ppbw (3.2 nM) norflurazon and phytofluene accumulated throughout the norflurazon concentration series (1 to 1000 ppbw). Carotene content was increased by 1 to 16 ppbw norflurazon but was decreased by 64 ppbw norflurazon. Thus, two modes-of-activity for norflurazon are documented that depend upon concentration of the toxicant in the tissue. Norflurazon demethylation in prephytoenepyrophosphate synthesis resulted in a C₂₁ conjugate and increased concentrations of GGPP and phytoene in the tissue. At approximately 31 ppbw norflurazon, an inhibition of phytoene dehydrogenation occurred and phytoene accumulated. At 62 ppbw norflurazon, phytofluene hydrogenation inhibition occurred and phytofluene accumulated while β-carotene synthesis was inhibited. These inhibitions may possibly be reversible when substrate concentrations are in excess.     

Increased activity and reduced sensitivity of glutamine synthetase in glufosinate‐resistant vetiver (Vetiveria zizanioides Nash) cells

Vetiver (Vetiveria zizanioides Nash) cells derived from an inflorescence were cultured in a modified N6 liquid medium supplemented with 10 µm 2,4‐D and 10 mm proline. Exponentially growing cell suspensions were subcultured with a selection medium containing glufosinate (ammonium dl ‐homoalanin‐4‐yl(methyl)phosphinate). The glufosinate‐resistant cells which can grow in a medium containing 5 × 10^?5 M glufosinate was selected by a stepwise selection, and its I₅₀ value was determined to be 4.2 × 10^?5 M. The growth of susceptible cells was inhibited by lower concentrations of glufosinate and its I₅₀ value was 2.5 × 10^?7 M. This indicated that the selected cells were 170‐fold resistant compared with the susceptible cells. Glutamine synthetase (GS) activity of the resistant cells was twice as high as that of the susceptible cells. The I₅₀ values of glufosinate were 3.2 × 10^?5 M and 9.0 × 10^?7 M for GS from the resistant and susceptible cells, respectively. The accumulation of ammonia caused by GS inhibition was higher in the susceptible cells. Absorption of [3,4–¹⁴C]glufosinate was not significantly different between the resistant and susceptible cells. Both cell types did not metabolize glufosinate. These results suggest that the resistance of the selected vetiver cell suspension to glufosinate is mainly due to increased GS activity and its decreased sensitivity to the herbicide.     

Physiological and biochemical modes of action of the diphenylether aclonifen

Aclonifen belongs to the diphenylether (DPE) chemical family among which potent herbicides with a photodependent mode of action can be found. For years aclonifen has been used in several types of cultures. However its biochemical mode of action remains unclear although it was listed as a carotenoid synthesis inhibitor by the Herbicide Resistance Action Committee (HRAC). As a matter of fact, corn seedling leaves treated with 10⁻⁴ M aclonifen and maintained in the dark no longer contained carotenoids but showed an accumulation of a compound having all the characteristics of phytoene. That demonstrated aclonifen ability to inhibit the desaturases catalyzing the transformation of phytoene into carotenoids. Moreover, aclonifen (5 × 10⁻⁵ M) was responsible for a photodependent cell destruction (necrosis) of cucumber cotyledons typically due to protoporphyrin IX accumulation. The same phenomenon was demonstrated in aclonifen-treated etiolated corn seedlings (10⁻⁴ M) that showed an accumulation of protoporphyrin IX, reaching 62 ng/g of leaf fresh mass and reactive oxygen species production under light. On these two cases (cucumber cotyledons and etiolated corn seedlings), aclonifen was acting as a typical DPE, as demonstrated by the accumulation of protoporphyrin IX. As a whole, aclonifen was shown here, to act on two different biochemical pathways including carotenoid synthesis on the one hand, as well as protoporphyrinogen oxidase, in the chlorophyll synthesis pathway, on the other for the same range of concentrations.     

Biocontrol of sclerotinia stem rot (Sclerotinia sclerotiorum) of soybean using novel Bacillus subtilis strain SB24 under control conditions

Sclerotinia stem rot (SSR), caused by Sclerotinia sclerotiorum, is a major disease of soybean in Canada. Laboratory and greenhouse experiments were conducted to evaluate potential effectiveness of cell suspensions, cell‐free culture filtrates and broth cultures of Bacillus subtilis strain SB24 for suppression of SSR. The SB24 cell suspensions and cell‐free culture filtrates significantly reduced mycelial growth of S. sclerotiorum by 50 to 75% and suppressed sclerotial formation by > 90%. The severity on soybean was negatively correlated (r < ?0·84, P < 0·01) to the concentrations of cell suspension, cell‐free culture filtrate and broth culture applied. The cell suspension and broth culture preparations significantly (P < 0·01) reduced SSR severity by 45 to 90% at concentrations ranging from 5 × 10⁶ to 10⁹ CFU mL^?1. The most effective concentration was 5 × 10⁸ CFU mL^?1 for all three preparations, reducing the severity by 60 to 90%. The B. subtilis SB24 was most effective in reducing disease severity when applied ≤ 24 h before plant inoculation with S. sclerotiorum and a significant effectiveness was observed up to 15 days after plant inoculation. The population density of B. subtilis on soybean leaves decreased by 1·5 to 2·5 log units over 15 days under field conditions, and by 0·8 log units over 5 weeks under control conditions. The decrease in population density was significantly correlated with rainfall in the field (r < ?0·93, P < 0·01), suggesting that the biocontrol bacteria may be washed away by rain.     

Mechanism of herbicidal activity of a new cyclohexane-1,3-dione, tepraloxydim to Poa annua L.

Tepraloxydim [(EZ)‐(RS)‐2‐{1‐[(2E)‐3‐chloroallyloxyimino]propyl}‐3‐hydroxy‐5‐perhydropyran‐4‐ylcyclohex‐2‐en‐1‐one] showed high activity against annual bluegrass (Poa annua L.), which is relatively tolerant to sethoxydim [(±)‐2‐(1‐ethoxyiminobutyl)‐5‐[2‐(ethylthio)propyl]‐3‐hydroxycyclohex‐2‐en‐1‐one]. Absorption and translocation rates of tepraloxydim and sethoxydim were higher in P. annua than in Setaria faberi, but the absorption and translocation patterns of tepraloxydim in the two plants were similar to those of sethoxydim. Metabolic rates of tepraloxydim and sethoxydim in P. annua and S. faberi were found to be similar. The concentration for 50% inhibition (I₅₀) of acetyl‐coenzyme A carboxylase (ACCase) with tepraloxydim was approximately 3 × 10^?6 mol L^?1 for P. annua and 7 × 10^?7 mol L^?1 for S. faberi. For sethoxydim, the I₅₀ was found to be 2 × 10^?6 mol L^?1 with the enzyme of S. faberi, while sethoxydim showed a slight effect on ACCase from P. annua activity, even at 10^?4 mol L^?1. The strong inhibition of ACCase with tepraloxydim is considered to be the major factor contributing to the high herbicidal activity against P. annua. Measuring the whole plant growth response, the ratio of the tepraloxydim I₅₀ dose of P. annua to that of S. faberi (P/S) was found to be 2.4, while the P/S ratio of sethoxydim and a tepraloxydim analog with a propyl chain at R₂ were 56.3 and 73.3, respectively. The herbicidal activity against P. annua was remarkably influenced by the length of the R₂ alkyl chain, while the effect on S. faberi was not affected. Acetyl‐coenzyme A carboxylase from P. annua also exhibited a higher resistance to the tepraloxydim analog with a propyl chain than to tepraloxydim. These results suggest that a binding site structure of cyclohexane‐1,3‐diones in the ACCase differs between P. annua and S. faberi.     

Reversal of norflurazon carotenogenesis inhibition by isomers

Norflurazon (0, 0.1, 0.2, 0.4, or 0.8 μM) was applied concomitantly with desmethyl norflurazon (DMN), dichloropyridazinone (DCP), or the wrong isomer (WI) of norflurazon (0, 0.1, 0.2, 0.4, 0.8, 1.6, or 3.3 μM) to wheat (Triticum aestivum L. cv. Holley) grown in sand. After 14 days, carotenogenesis was inhibited by norflurazon and the inhibition was partially reversed by DMN, DCP, and WI. These reversals were observed at norflurazon concentrations ≤250 ≤ ∼0.823 μM in the potting medium. Carotene contents in norflurazon (0.4 μM) + no isomer, DMN, WI, or DCP (3.3 μM) were 5.4, 40.7, 28.6, and 22.2%, respectively, of that present in the untreated control. Therefore, these materials might function as antidotes to soil residues of norflurazon. Partitioning of norflurazon and DMN among triolein (TG), phosphatidylcholine (PC), and water was attained via isopycnic centrifugation. Norflurazon was highly soluble in PC and accumulated in PC. DMN was not soluble in TG and was soluble in water and PC at a ratio of 0.5 Change in water solubility when norflurazon is demethylated to DMN may be the basis for lack of bleaching influence of DMN. DMN, DCP, and WI partially reversed norflurazon carotenogenesis inhibition in the concentration range of norflurazon associated with phytoene synthesis and the low range of norflurazon concentrations associated with phytoene desaturase.     

Mitochondrial impacts of insecticidal formate esters in insecticide‐resistant and insecticide‐susceptible Drosophila melanogaster

BACKGROUND: Previous research on insecticidal formate esters in flies and mosquitoes has documented toxicity profiles, metabolism characteristics and neurological impacts. The research presented here investigated mitochondrial impacts of insecticidal formate esters and their hydrolyzed metabolite formic acid in the model dipteran insect Drosophila melanogaster Meig. These studies compared two Drosophila strains: an insecticide‐susceptible strain (Canton‐S) and a strain resistant by cytochrome P450 overexpression (Hikone‐R). RESULTS: In initial studies investigating inhibition of mitochondrial cytochrome c oxidase, two proven insecticidal materials (hydramethylnon and sodium cyanide) caused significant inhibition. However, for insecticidal formate esters and formic acid, no significant inhibition was identified in either fly strain. Mitochondrial impacts of formate esters were then investigated further by tracking toxicant‐induced cytochrome c release from mitochondria into the cytoplasm, a biomarker of apoptosis and neurological dysfunction. Formic acid and three positive control treatments (rotenone, antimycin A and sodium cyanide) induced cytochrome c release, verifying that formic acid is capable of causing mitochondrial disruption. However, when comparing formate ester hydrolysis and cytochrome c release between Drosophila strains, formic acid liberation was only weakly correlated with cytochrome c release in the susceptible Canton‐S strain (r² = 0.70). The resistant Hikone‐R strain showed no correlation (r² < 0.0001) between formate ester hydrolysis and cytochrome c release. CONCLUSION: The findings of this study provide confirmation of mitochondrial impacts by insecticidal formate esters and suggest links between mitochondrial disruption, respiratory inhibition, apoptosis and formate‐ester‐induced neurotoxicity. Copyright © 2009 Society of Chemical Industry     

<Emphasis Type="Italic">White clover mosaic virus</Emphasis>-induced gene silencing in pea

Double-stranded RNAs formed in secondary structures and replicative intermediates of viral genomes are thought to strongly elicit RNA silencing. This phenomenon is known as virus-induced gene silencing (VIGS). VIGS is a powerful tool for modifying gene expression in host plants. We constructed a virus vector based on White clover mosaic virus (WClMV) and demonstrated VIGS of phytoene desaturase (PDS) in pea. Photobleaching of tissues, caused by VIGS of PDS, was observed in restricted areas of upper leaves and stems. We confirmed that the PDS mRNA and subgenomic RNAs of WClMV were reduced in the photobleached tissues.     

Metabolism of fungicidal cyanooximes, cymoxanil and analogues in various strains of Botrytis cinerea

BACKGROUND: The metabolism of cymoxanil [1‐(2‐cyano‐2‐methoxyiminoacetyl)‐3‐ethylurea] and fungicidal cyanooxime analogues was monitored on three phenotypes of Botrytis cinerea Pers. ex Fr. differing in their sensitivity towards cymoxanil. For this purpose, labelled [2‐¹⁴C]cymoxanil was added either to the culture medium of these strains or to its cell‐free extract. RESULTS: In the culture medium of the most sensitive strain, four main metabolites were detected. Three were isolated and identified. Cymoxanil was quickly metabolised by at least three concurrent enzymatic pathways: (i) cyclisation leading, after hydrolysis, to ethylparabanic acid, (ii) reduction giving demethoxylated cymoxanil, (iii) hydrolysis followed by reduction and then acetylation leading to N‐acetylcyanoglycine. In the cell‐free extract of the same strain, only the first and the second of these enzymatic reactions occurred. By comparing the metabolic profile of the most sensitive strain with that of the less sensitive ones, it was shown that the decrease in sensitivity to cymoxanil correlates with a reduced acetylcyanoglycine formation. Among all metabolites, only N‐acetylcyanoglycine is active against the most sensitive strain. Moreover, in a culture of this strain, two other fungicidal cyanooximes were also metabolised into this metabolite. CONCLUSION: The formation of N‐acetylcyanoglycine may play an important role in the fungitoxicity of cymoxanil and cyanooxime derivatives. Copyright © 2008 Society of Chemical Industry     

Studies on the interaction between the biocontrol agent,Serratia plymuthica A30, and blackleg‐causing Dickeya sp. (biovar 3) in potato (Solanum tuberosum)

Interactions between Serratia plymuthica A30 and a blackleg‐causing biovar 3 Dickeya sp. were examined. In a potato slice assay, S. plymuthica A30 inhibited tissue maceration caused by Dickeya sp. IPO2222 when co‐inoculated at a density at least 10 times greater than that of the pathogen. In glasshouse experiments, population dynamics of the antagonist and of the pathogen in planta were studied by dilution plating and confocal laser scanning microscopy (CLSM) using fluorescent protein‐tagged strains. Pathogen‐free minitubers were vacuum‐infiltrated with DsRed‐tagged Dickeya sp. IPO2222 and superficially treated during planting with a water suspension containing GFP‐tagged S. plymuthica A30. A30 reduced the blackleg incidence from 55% to 0%. Both the pathogen and the antagonist colonized the seed potato tubers internally within 1 day post‐inoculation (dpi). Between 1 and 7 dpi, the population of A30 in tubers increased from 10¹ to c. 10³ CFU g^?1 and subsequently remained stable until the end of the experiment (28 dpi). Populations of A30 in stems and roots increased from c. 10² to c. 10⁴ CFU g^?1 between 7 and 28 dpi. Dilution plating and CLSM studies showed that A30 decreased the density of Dickeya sp. populations in plants. Dilution plating combined with microscopy allowed the enumeration of strain A30 and its visualization in the vascular tissues of stem and roots and in the pith of roots, as well as its adherence to and colonization of the root surface. The implications of these finding for the use of S. plymuthica A30 as a biocontrol agent are discussed.     

Mesotrione control and pigment concentration of large crabgrass (Digitaria sanguinalis) under varying environmental conditions

BACKGROUND: Mesotrione is a carotenoid biosynthesis‐inhibiting herbicide currently labeled for crabgrass (Digitaria spp.) control. Mesotrione control of large crabgrass has been reported to vary with temperature and relative humidity; however, the effect of irradiance on mesotrione efficacy has not previously been reported. Likewise, little is known about pigment concentrations of Digitaria spp. The present research investigated the effects of mesotrione on large crabgrass, Digitaria sanguinalis (L.) Scop., control and pigment concentrations under varying irradiance at three temperatures. RESULTS: Mesotrione (0.28 kg ha^?1) control of large crabgrass did not differ between temperature levels (18, 26 and 32 °C). Control was similar at tested irradiance levels (600, 1100 and 1600 µmol m^?2 s^?1). Mesotrione reduced large crabgrass chlorophyll a, chlorophyll b and total carotenoid concentrations, as well as chlorophyll a to b ratios. Treated plant bleaching was highest 7 days after treatment (DAT) but decreased by 21 DAT. Treated plants were less than 10% necrotic 3 and 7 DAT but nearly 35% necrotic 21 DAT. Treated large crabgrass bleaching was highest and photochemical efficiency was lowest 7 DAT. These results indicate that some plant recovery occurs prior to 21 DAT. CONCLUSION: Although mesotrione efficacy has previously been reported to vary according to environmental factors, mesotrione control of large crabgrass did not vary with measured temperature and irradiance levels in this study. On account of crabgrass convalescence, secondary applications of mesotrione may control large crabgrass more effectively when applied prior to 21 DAT. Copyright © 2009 Society of Chemical Industry     

The inhibitory mode of action of the pyridazinone herbicide norflurazon on a cell-free carotenogenic enzyme system

The inhibition site of the phenylpyridazinone herbicide, norflurazon [SAN 9789, 4-chloro-5-(methylamino)-2-(3-trifluoromethylphenyl)-pyridazin-3(2H)one] was determined in a cell-free carotenogenic enzyme system from a mutant strain of Phycomyces blakesleeanus (Mucoraceae). The presence of norflurazon resulted in a reduced flow of radioactivity from [2-¹⁴C]mevalonic acid to phytoene (7,8,11,12,7′,8′,11′,12′-octahydro-ψ,ψ-carotene) and β-carotene (β,β-carotene), whereas an increased incorporation occurred in the C₃₀ terpenoids, squalene, and ergosterol. Furthermore, radioactivity accumulated in geranylgeranyl pyrophosphate. Since no radioactivity was found in prephytoene pyrophosphate and the radioactivity in phytoene decreased upon addition of norflurazon, this herbicide exerts its primary inhibitory action on the reaction catalyzed by phytoene synthetase. The nonbleaching phenylpyridazinone BAS 13761 [4-chloro-5-methoxy-2-phenyl-pyridazin-3(2H)-one] did not show this effect. Other inhibitory sites of norflurazon, either on prenyl pyrophosphate synthetase or on the desaturation of phytoene, were excluded.