Occurrence of Corynespora cassiicola isolates resistant to boscalid on cucumber in Ibaraki Prefecture, Japan

A total of 651 isolates of cucumber corynespora leaf spot fungus ( Corynespora cassiicola ) collected from cucumber in Japan, either with (438 isolates) or without (213 isolates) a prior history of boscalid use, were tested for their sensitivity to boscalid by using a mycelial growth inhibition method on YBA agar medium. Additionally, seven isolates of C. cassiicola obtained from tomato, soybean, eggplant (aubergine) and cowpea in different locations in Japan were tested before boscalid registration. Minimum inhibitory concentration (MIC) and 50% effective concentration (EC₅₀) values for 220 isolates from crops without a prior history of boscalid use ranged from 0·5 to 7·5 μg mL⁻¹ and from 0·04 to 0·59 μg mL⁻¹, respectively. Two hundred and fourteen out of 438 isolates collected from ten cucumber greenhouses in Ibaraki Prefecture, Japan, which received boscalid spray applications showed boscalid resistance, with MIC values higher than 30 μg mL⁻¹. Moreover, resistant isolates were divided into two groups: a moderately resistant (MR) group consisting of 189 isolates with EC₅₀ values ranging from 1·1 to 6·3 μg mL⁻¹, and a very highly resistant (VHR) group consisting of 25 isolates with EC₅₀ values higher than 24·8 μg mL⁻¹. MR isolates were detected from all ten greenhouses, but VHR isolates were detected from only three. As a result of fungus inoculation tests which used potted cucumber plants, control failures of boscalid were observed against resistant isolates. Efficacy of boscalid was remarkably low against VHR isolates in particular. This is the first known report on boscalid resistance in Japan.     

Sensitivity of Sclerotinia homoeocarpa to demethylation-inhibiting fungicides in Ontario, Canada, after a decade of use

In late 2003, nine populations of Sclerotinia homoeocarpa in Ontario Canada (seven of which had been previously sampled in early 1994, prior to the registration of sterol demethylation-inhibiting (DMI) fungicides for turf disease control in Canada) were sampled and tested for sensitivity to propiconazole. Four of the nine populations had not been treated with DMI fungicides during the intervening years, and isolates from these locations were sensitive to propiconazole (geometric mean EC₅₀ values of 0·005–0·012 µ g mL⁻¹, compared with 0·005–0·008 µ g mL⁻¹ for the original 1994 populations). Among the five populations from 2003 that had been exposed to DMI fungicides, mean EC₅₀ values were significantly greater, ranging from 0·020 to 0·048 µ g mL⁻¹. A significant correlation of determination was found between estimated number of fungicide applications and log EC₅₀ ( R ² = 0·832, P  = 0·0001), and the equation predicted that 42·3 applications of propiconazole would be needed to bring a sensitive population (EC₅₀ < 0·01  µ g mL⁻¹) to a resistant level (EC₅₀ > 0·10  µ g mL⁻¹). Fungicide sensitivity vs. duration of fungicide efficacy was also tested, and it was found that isolates with decreased sensitivity were able to more quickly overcome the inhibitory effects of fungicide application, reducing the duration of control from 3 weeks to 2 weeks.     

Development of a semiselective medium for isolating Xanthomonas campestris pv. musacearum from insect vectors, infected plant material and soil

A semiselective medium was developed for isolating Xanthomonas campestris pv. musacearum ( Xcm ) from infected banana plants, soil and insect vectors. The new medium was named cellobiose-cephalexin agar (CCA) and it contained (L⁻¹): 1 g yeast extract, 1 g glucose, 1 g peptone, 1 g NH₄Cl, 1 g MgSO₄·7H₂O, 3 g K₂HPO₄, 1 g beef extract, 10 g cellobiose, 14 g agar, 40 mg cephalexin, 10 mg 5-fluorouracil and 120 mg cycloheximide. The medium was evaluated for selectivity using 21 bacterial isolates and for plating efficiency using Xcm . The bacterial isolates included a soilborne Xanthomonas species and three pathogenic Xanthomonas strains that infect cassava, cabbage and beans. Although the plating efficiency of Xcm on CCA was lower (59%) than on non-selective yeast extract peptone glucose agar (YPGA), its selectivity was significantly higher, averaging 60 and 82%, when isolating from banana fruits and soil, respectively. CCA was also superior when isolating Xcm from insect vectors, with selectivity of 48–75%, compared with 8–17% on YPGA. Xanthomonas campestris pv. phaseoli did not grow on CCA, while X. campestris pv. campestris and X. axonopodis pv. manihotis grew, but their colonies were smaller than those of Xcm . Twenty-nine out of 33 suspected Xcm strains isolated from plants, soil and insects using CCA were pathogenic when inoculated onto banana plants, indicating that CCA can be a reliable tool in isolating Xcm populations. The medium should prove useful in studies on ecology, epidemiology and management of the banana bacterial wilt pathogen that is currently ravaging bananas in East and Central Africa.     

Reduced sensitivity of Cercospora beticola isolates to sterol-demethylation-inhibiting fungicides

In a survey conducted during October 1995, single-lesion isolates of the sugar beet leaf-spot fungus, Cercospora beticola , were tested for sensitivity to the sterol demethylation inhibiting fungicides (DMIs) flutriafol and bitertanol. The isolates were collected from fields in three different areas of northern Greece. Fields at Serres and Imathia had been sprayed with DMIs for about 15 years to control sugar beet leaf-spot. At the third site, Amyndeon, DMI fungicides had not been used. From each area 150 isolates were tested. ED₅₀ values were calculated for individual isolates by regressing the relative inhibition of colony growth against the natural logarithm of the fungicide concentration. The mean ED₅₀ values for flutriafol for the Serres, Imathia and Amyndeon populations were 1·07, 0·73 and 0·5 µg mL⁻¹, respectively (significantly different at P  = 0·05). For bitertanol the mean ED₅₀ values for the Serres and Imathia populations were 0·72 and 0·81 µg mL⁻¹, respectively, which were not significantly different at P  = 0·05. The mean ED₅₀ value of the Amyndeon population was 0·48 µg mL⁻¹, which was significantly lower than those of the other two populations ( P  < 0·05). A cross-resistance relationship was found to exist between the two triazole fungicides tested when log transformed ED₅₀ values of 60 isolates were subjected to a linear regression analysis ( r  = 0·81).     

Response of interspecific Brassica juncea/Brassica rapa hybrids and their advanced progenies to Albugo candida (white blister)

Transfer of factors for resistance to white blister disease caused by Albugo candida between Brassica species involving two genotypes each of B. juncea and B. rapa was studied in hybrids. More hybrids were obtained by in vivo than in vitro techniques, although an in vitro phase was a prerequisite for the establishment of in vivo hybrids. Hybrids were identified by PCR-based inter-simple sequence repeat (ISSR) markers with both male and female species-specific bands being identified. There was a positive correlation between disease severity and number of days after sowing ( r  > 0·93), the highest being towards pod formation and plant maturity at 110 days after sowing. The plants from F₂ and BC₁ progeny showed higher resistance to A. candida than either of the parents. Plants of B. juncea and B. rapa with high field resistance (disease index < 1·0) were selected from BC₂ and F₂BC₁ generations. The frequency of plants classified as resistant in BC₂ progeny ranged from 4·5 to 39·0% in cross-combinations involving B. juncea genotypes as female parent, compared with 100% in the reciprocal cross involving B. rapa as female parent.     

A semiselective medium for the isolation of Acidovorax valerianellae from soil and plant debris

Bacterial black spot of corn-salad ( Valerianella locusta ) caused by Acidovorax valerianellae appeared in France in the beginning of the 1990s. The disease is now widespread in the main area of corn-salad production and is responsible for high depreciation and significant economic losses every year. A semiselective medium is described for the detection of the bacterium in various environments and to identify primary sources of inoculum. The medium TSAV (tryptic soy broth, 3 g L⁻¹, agar 15 g L⁻¹, 5-fluorouracyl 5 mg L⁻¹, novobiocin 5 mg L⁻¹, propiconazole 5 mg L⁻¹) increased chances of obtaining cultures of A. valerianellae from naturally infested soil and from root-debris. Using TSAV, the pathogen was detected in root-debris up to 39 days after the harvest of a diseased crop. As only a few days usually separates harvest from the next sowing date, soil is likely to be an important source of inoculum for the next crop.     

Control of Phytophthora root rot of irrigated subterranean clover with potassium phosphonate in Victoria, Australia

Phosphonic acid was more effective in inhibiting the production of sporangia of Phytophthora clandestina in sterile pond water (ED₅₀= 1·4 p.p.m.) than it was in inhibiting the growth of mycelium of the fungus on either corn meal agar (ED₅₀= 13·8p.p.m.) or lima bean agar (ED₅₀= 236 p.p.m.). Experiments under controlled environmental conditions showed that better control of tap root rot of subterranean clover caused by P. clandestina was achieved by application of potassium phosphonate to a pasteurized soil mixture than by a spray on the foliage alone. However, in a soil from a pasture, foliar sprays were more effective than soil treatments. Application of superphosphate at a rate of 250 kg/ha or higher to the soil reduced the effectiveness of sprays. In three experiments in irrigated pastures a spray of potassium phosphonate at 300 or 313 ml/ha, applied to cotyledons of subterranean clover and to soil, reduced severity of Phytophthora root rot and increased the annual production of dry matter of the legume by 1·96 to 5·11 t/ha in comparison with untreated controls.     

Sensitivity of fungi from cereal roots to fluquinconazole and their suppressiveness towards take-all on plants with or without fluquinconazole seed treatment in a controlled environment

The take-all fungus, Gaeumannomyces graminis var. tritici , was highly sensitive to fluquinconazole ( in-vitro EC₅₀ 0·016–0·018 mg L⁻¹), a fungicide developed for use as a seed treatment to control take-all, and to prochloraz (EC₅₀ 0·006 mg L⁻¹). Fungi of other genera that were commonly isolated from cereal roots were sensitive in varying degrees to prochloraz but were relatively insensitive (e.g. Fusarium culmorum , EC₅₀ > 20 mg L⁻¹) or slightly sensitive (e.g. Epicoccum purpurascens , EC₅₀ 0·514 mg L⁻¹) to fluquinconazole. Gaeumannomyces graminis var. graminis and G. cylindrosporus , weak parasites that can protect roots against take-all, and an unnamed Phialophora sp., all closely related to the take-all fungus, were highly or moderately sensitive to fluquinconazole. Alternaria infectoria and E. purpurascens were most consistently effective in suppressing development of take-all on pot-grown wheat plants dual-inoculated with G. graminis var. tritici and the nonpathogen. Take-all was decreased more on dual-inoculated wheat plants grown from seed treated with fluquinconazole or fluquinconazole plus prochloraz than when only an antagonistic fungus ( A. infectoria , E. purpurascens , Fusarium culmorum or Idriella bolleyi ) or a seed treatment was applied. These fungi were less effective in combination with seed treatments on barley. Gaeumannomyces graminis var. graminis and G. cylindrosporus , tested on wheat, suppressed take-all only in the absence of fungicides. It is suggested that the performance of seed treatment containing fluquinconazole against take-all may in some circumstances be enhanced by its partial specificity for the take-all fungus.     

Host–parasite relationships of Meloidogyne incognita on spinach

Host–parasite relationships in root-knot disease of spinach caused by Meloidogyne incognita race 1 were studied under glasshouse conditions. Nematode-induced mature galls were large and usually contained one or more females and egg masses with eggs. Feeding sites were characterized by the development of giant cells containing granular cytoplasm and many hypertrophied nuclei. The cytoplasm in these giant cells was aggregated alongside the thickened cell walls. Stelar tissues within galls appeared disorganized. The relationship between initial nematode population density ( P _i) in a series from 0–128 eggs and second-stage juveniles per cm³ soil and growth of spinach cv. Symphony F₁ seedlings was tested under glasshouse conditions. A Seinhorst model [ y = m  + (1 −  m ) z ^P–T ] was fitted to fresh top- and total plant-weight data for inoculated and control plants. Tolerance limits ( T ) of spinach cv. Symphony F₁ to M. incognita race 1 for fresh top and total plant weights were 0·25 and 0·5 eggs and second-stage juveniles per cm³ soil, respectively. The minimum relative values for fresh top and total plant weights were zero in both cases at P _i ≥ 32 eggs and second-stage juveniles per cm³ soil. Root galling was least at low initial population densities and greatest at 16 eggs and second-stage juveniles per cm³ soil. Maximum nematode reproduction rate was 33·1-fold at the lowest P _i.     

Effects of nitrogen, phosphorus, potassium and calcium nutrition on strawberry anthracnose

The effects of a range of concentrations of four nutrients – nitrogen, phosphorus, potassium and calcium – in fertilizer solutions on the severity of anthracnose on strawberry cv. Nyoho cultivated under a noncirculation hydroponics system were determined after inoculation with Colletotrichum gloeosporioides . Crop growth and tissue nitrogen, phosphorus, potassium and calcium contents of the entire above-ground parts of the plant were also investigated. Elevated nitrogen and potassium concentrations in the fertilizer solution increased disease severity in contrast to phosphorus and calcium. Treatment with either NH₄ or NO₃ nitrogen was not significantly different. The dry weight of the strawberry plants increased significantly with elevated concentrations of nitrogen ( R ² = 0·9078) and phosphorus ( R ² = 0·8842), but was not influenced by the elevated amounts of potassium ( R ² = 0·8587) and calcium ( R ² = 0·6526) concentrations.     

Effect of water potential on mycelial growth and perithecial production of Monosporascus cannonballus in vitro

The effects of osmotic water potential (Ψ_s) on mycelial growth and perithecial production of Monosporascus cannonballus , the cause of root rot and vine decline of melons, were examined at 25°C on potato dextrose agar (PDA) amended with KCl, NaCl or sucrose. Patterns of the growth responses of four isolates to decreasing Ψ_s were similar for each of the osmotica. Compared with growth on nonamended PDA (−0·3 MPa), growth of all isolates increased as Ψ_s was reduced to −0·8 MPa. Maximum growth occurred at Ψ_s values of −0·6 to −0·8 MPa. Growth was not reduced below that on nonamended PDA until Ψ_s was reduced to −1·8 MPa, and a 50% reduction in growth did not occur until Ψ_s was reduced to < −2·5 MPa. Reproduction was much more sensitive to reduced Ψ_s than was mycelial growth, and perithecia were produced only at Ψ_s ≥ −0·7 or −0·8 MPa on PDA amended with KCl or NaCl, respectively. Three isolates produced perithecia on PDA amended with sucrose only at Ψ_s ≥ −0·6 MPa, but the fourth isolate produced perithecia at ≥ −1·9 MPa. Colonization of the xylem early in disease development may provide an essential source of water for subsequent reproduction in the root cortex during plant senescence. Postharvest cultivation to expose and desiccate roots may prevent reproduction even when temperatures lethal to hyphae are not attained.     

土壤棉黄萎病菌选择性培养基——棉选一号

 棉花黄萎病的发生发展与土壤中微菌核的数量有着密切关系.本文介绍一种用于检验土壤内棉黄萎病菌的选择性培养基——棉选一号.其成份为:NaNO₃ 2g、MgSO₄·7H₂O 0.5g、K₂HPO₄ 1g、Fe₂（SO₄）₃·XH₂O 0.01g.KCl 0.5g、蔗糖5g、琼脂15g、蒸馏水1000ml、氯霉素300mg、井岗霉素2.5ppm、五氯硝基苯350ppm、克菌丹0.5ppm.这种培养基能有效地分离自然土壤中黄萎病菌的微菌核,而极少受其他微生物的干扰,由于培养基上菌落大、黑色、微菌核呈放射状,故易用肉跟计数微菌核的数量.     

In vitro somatic growth and reproduction of phenylamide-resistant and -sensitive isolates of Phytophthora erythroseptica from infected potato tubers in Idaho

Pink rot of potato, most commonly caused by Phytophthora erythroseptica , is a major field and post-harvest problem in southern Idaho, USA, particularly since 1998 when isolates resistant to the phenylamide fungicide metalaxyl-M (mefenoxam) were detected. Isolates of P. erythroseptica were collected from infected tubers in 2001 and 2002 from six Idaho counties and tested for resistance to metalaxyl-M on amended agar. Metalaxyl-M resistant (MR) and metalaxyl-M-sensitive (MS) isolates were identified in six counties; 160 isolates were highly resistant, seven moderately resistant and 57 sensitive to metalaxyl-M with mean EC₅₀ values of 182, 23 and 0·5 mg L⁻¹ ai metalaxyl-M, respectively. Mycelial growth rates and oospore production in agar were assessed for 20 MS and 20 MR isolates at 10, 15, 20, 25 and 30°C. Growth rates of MR isolates were between 2·5 and 3·1 times greater ( P  < 0·05) than those of MS isolates at 10, 15, 20 and 25°C, and oospore production was between 6·8 and 20·5 times greater ( P  < 0·0001) for MR than for MS isolates at the same temperatures. Colony growth in V8 broth at 18°C was greater for MR than MS isolates ( P  < 0·0032). However, zoospore production at 18°C was greater for MS than for MR isolates ( P  < 0·0109), and zoospore production m m ⁻¹ of colony circumference was also greater for MS than for MR isolates, 14 191 and 9959, respectively ( P  = 0·0109). Sexual reproduction of MR isolates in nature may be greater than MS isolates, but MS isolates may be more asexually fit based on the fitness parameters studied.     

Sensitivity of Phytophthora infestans to flumorph: in vitro determination of baseline sensitivity and the risk of resistance

The sensitivity of 127 Phytophthora infestans isolates to flumorph was determined in 2003 and 2004. The isolates originated from two geographical regions and showed similar levels of sensitivity in both years. Baseline sensitivities were distributed as a unimodal curve with EC₅₀ values for growth of mycelia ranging from 0·1016 to 0·3228  µ g mL⁻¹, with a mean of 0·1813 (± 0·0405) µ g mL⁻¹. There was no cross-resistance between flumorph and metalaxyl. Laboratory studies were conducted to evaluate the risk of P. infestans developing resistance to flumorph. Mutants resistant to metalaxyl or flumorph were obtained by treating mycelium of wild-type isolates with ultraviolet radiation. Metalaxyl-resistant mutants were obtained with a high frequency and exhibited resistance factor values (EC₅₀ resistant/EC₅₀ sensitive phenotypes) of more than 100, while flumorph-resistant mutants were obtained at much lower frequencies and had very small resistance factors (1·5–3·2). There was cross-resistance between flumorph and dimethomorph, but not with azoxystrobin or cymoxanil. Most flumorph-resistant mutants showed decreases in hyphal growth in vitro and in sporulation both in vitro and on detached leaf tissues. These studies suggested that the risk of resistance developing was much lower for flumorph than metalaxyl. However, as P. infestans is a high-risk pathogen, appropriate precautions against resistance development should be taken.     

Pathogenicity of the root-knot nematode Meloidogyne javanica on potato

Host–parasite relationships and pathogenicity of Meloidogyne javanica on potatoes (newly recorded from Malta) were studied under glasshouse and natural conditions. Potato cvs Cara and Spunta showed a typical susceptible reaction to M. javanica under natural and artificial infections, respectively. In potato tubers, M. javanica induced feeding sites that consisted of three to four hypertrophied giant cells per adult female. Infection of feeder roots by the nematode resulted in mature large galls which usually contained at least one mature female and egg mass. In both tubers and roots, feeding sites were characterized by giant cells containing granular cytoplasm and many hypertrophied nuclei. Cytoplasm in giant cells was aggregated alongside the thickened cell walls. Stelar tissues within galls appeared disorganized. The relationship between initial nematode population density ( P ) [0–64 eggs + second-stage juveniles (J2s) per cm³ soil] and growth of cv. Spunta potato seedlings was tested under glasshouse conditions. A Seinhorst model [ y = m  + (1 −  m ) z ^{( P − T )}] was fitted to fresh shoot weight and shoot height data of nematode-inoculated and control plants. Tolerance limits ( T ) for fresh shoot weight and shoot height of cv. Spunta plants infected with M. javanica were 0·50 and 0·64 eggs + J2s per cm³ soil, respectively. The m parameter in that model (i.e. the minimum possible y -values) for fresh shoot weight and shoot height were 0·60 and 0·20, respectively, at P  = 64 eggs + J2s per cm³ soil. Root galling was proportional to the initial nematode population density. Maximum nematode reproduction rate was 51·2 at a moderate initial population density ( P  = 4 eggs + J2s per cm³ soil).     

Fungicide sensitivity in Swedish isolates of Phaeosphaeria nodorum

This is the first report of variability in sensitivity of Phaeosphaeria nodorum to the fungicide azoxystrobin, and also reports on sensitivity to propiconazole, prothioconazole and cyprodinil. An in vitro sensitivity test of 42 isolates from five Swedish winter wheat fields, collected in 2003–2005, was performed on malt extract agar amended with six concentrations of each fungicide. Four isolates collected during the early 1990s, before azoxystrobin had been commercially used in agriculture, were used as references. Fragments of DNA from 231 isolates, including the reference isolates, were sequenced for the genes of cytochrome b and CYP51 in search for amino acid substitutions known to cause loss of sensitivity to strobilurins and triazoles, respectively. The majority of the P. nodorum isolates possessed the amino acid substitution G143A, associated with loss of sensitivity in fungi to strobilurins, except in one field where only half of the isolates had the substitution. The EC₅₀ values varied between 0·66 mg L⁻¹ to estimations far above 1000 mg L⁻¹, with an estimated median value of 366 mg L⁻¹. The EC₅₀ values of the reference isolates ranged from 0·02 to 80·72 mg L⁻¹. The P. nodorum population is still sensitive to propiconazole, prothioconazole and cyprodinil, even though some isolates varied in sensitivity to triazoles. Part of the CYP51 gene, a possible target for triazole sensitivity, was sequenced but no nonsynonymous substitutions were found.     

Comparison of extraction procedures and determination of the detection threshold for Clavibacter michiganensis ssp. michiganensis in tomato seeds

Several seed extraction procedures, used for detection of Clavibacter michiganensis ssp. michiganensis ( Cmm ) in naturally infected and artificially infested tomato seed lots were evaluated. Extraction methods that included grinding the seeds were significantly better at detecting the pathogen in three different seed lots than methods that used only soaking. The detection threshold of Cmm in relation to seed sample size was determined by adding naturally infected seeds into samples of three different sizes. Cmm was detected by agar plating assay, on three media (CNS, mSCM, D₂ANX), and by direct PCR from seeds and Bio-PCR (bacteria cultured on agar media prior to PCR). In samples of 10 000 seeds containing one infected seed, Cmm could be detected only by Bio-PCR and in only one replicate out of five. In samples containing five or 10 infected seeds per 10 000 seeds, three of five and five of five replicates, respectively, were detected by the three detection methods. In samples of 5000 seeds, one infected seed could be detected in all five replicates only after adding a concentration step. A high correlation ( R ² = 0·9448) between artificially infested seeds and the disease incidence was found. Seed lots infested with less than 58 colony-forming units (CFU) per g did not cause disease under glasshouse conditions, whereas lots with about 1000 CFU g⁻¹ caused disease in 78 plants out of 2000.     

Sensitivity of Botrytis cinerea from vegetable greenhouses to boscalid

Between 2004 and 2006, 228 isolates of Botrytis cinerea from two regions in China were characterized for baseline sensitivity to boscalid, a new active ingredient that interferes with succinate ubiquinone reductase in the electron transport chain. The isolates showed similar sensitivity in different years and regions. Baseline sensitivities were distributed as unimodal curves with mean EC₅₀ values of 1·07 (± 0·11) and 0·42 (± 0·05) mg L⁻¹ for inhibition of mycelial growth and conidial germination, respectively. Laboratory studies were conducted to evaluate the risk of development of resistance to boscalid. Boscalid-resistant mutants were obtained by UV-treatment at lower frequencies and with smaller resistance factors than pyrimethanil-resistant mutants. All boscalid-resistant mutants were also significantly more sensitive to Qo inhibitors than their wild-type parents and showed reduced sporulation in vitro and pathogenicity on aubergine leaves. The results suggested that the risk of resistance developing for boscalid was lower than for pyrimethanil. However, as B. cinerea is a high-risk pathogen, appropriate precautions against resistance development should be taken. Synergism between the activity of boscalid and that of kresoxim-methyl was observed.     

Nematicidal activity of essential oils and organic amendments from Asteraceae against root-knot nematodes

The essential oil of Chrysanthemum coronarium flowerheads showed strong nematicidal activity in vitro and in growth-chamber experiments. Essential oil concentrations of 2, 4, 8 and 16  µ L mL⁻¹, significantly reduced hatch, J ₂ survival (determined by final value and area under curves of cumulative percentage hatch or mortality) and reproduction rate of Meloidogyne artiellia in vitro , with the lowest values occurring at 16  µ L mL⁻¹. In pot trials with chickpea cv. PV 61, essential oil concentrations of 10–40  µ L per 500 cm³ soil, applied on sterile cotton pellets, also significantly reduced the nematode's reproduction rate. The biological processes of mortality and hatching/reproduction were adequately described by the monomolecular and expanded negative exponential models, respectively. Effectiveness of soil amendment with either flowers, leaves, roots or seeds of C. coronarium , and flowers from several species of Asteraceae ( Chrysanthemum segetum , Calendula maritima , Calendula officinalis and Calendula suffruticosa ) at 5 g per 500 cm³ soil was tested for suppression of M. artiellia and growth of chickpea cv. PV 61 under growth-chamber conditions. In these tests, flowers of all five Asteraceae species and various parts of C. coronarium significantly reduced reproduction rates of M. artiellia , by 83·0–95·9%, with the minimum rates occurring in infected chickpea plants amended with flowers of C. officinalis and C. suffruticosa . The in vitro and in planta results suggest that the essential oil of C. coronarium and organic amendments from Asteraceae species may serve as nematicides.     

A qualitative quick-test for detection of herbicide resistance to tribenuron-methyl in Papaver rhoeas

A qualitative seed-based method useful for the detection of resistance to the herbicide tribenuron-methyl in Papaver rhoeas L. is described. Seeds were germinated on 35 mL of a 1.3% agar medium containing 2 g KNO₃ L^–1 in 8.5 cm Petri dishes in a growth chamber under 20 μmol s^–1 m^–2 of fluorescent light. When 0.24 μM tribenuron-methyl or more was added, growth in susceptible plants stopped after the cotyledon stage and they turned chlorotic. The resistant plants continued developing new leaves. The same effect was achieved when 0.2 g gibberellin (GA₃) L^–1 and 7.68 μM tribenuron-methyl or 0.5 g GA₃ L^–1 and 61.44 μM tribenuron-methyl were added. Germination percentage rose with gibberellin in the presence or absence of the herbicide. Plants developed rapidly, with only about 14 d needed to finish the test but sometimes root growth was reduced because of the addition of gibberellin. In the absence of gibberellin but in the presence of the herbicide, plants grew more slowly and developed smaller leaves with a 17-d evaluation period requirement. The test was validated with pot experiments in a greenhouse and also with field trials. The best combination was found to be 0.2 g GA₃ L^–1 and 7.68 μM tribenuron-methyl, assuring homogenous germination and testing of dormant seeds but avoiding root inhibition associated with too much gibberellin.