Fate of the herbicides glyphosate, glufosinate-ammonium, phenmedipham, ethofumesate and metamitron in two Finnish arable soils

The fate of five herbicides (glyphosate, glufosinate-ammonium, phenmedipham, ethofumesate and metamitron) was studied in two Finnish sugar beet fields for 26 months. Soil types were sandy loam and clay. Two different herbicide-tolerant sugar beet cultivars and three different herbicide application schedules were used. Meteorological data were collected throughout the study and soil properties were thoroughly analysed. An extensive data set of herbicide residue concentrations in soil was collected. Five different soil depths were sampled. The study was carried out using common Finnish agricultural practices and represents typical sugar beet cultivation conditions in Finland. The overall observed order of persistence was ethofumesate > glyphosate > phenmedipham > metamitron > glufosinate-ammonium. Only ethofumesate and glyphosate persisted until the subsequent spring. Seasonal variation in herbicide dissipation was very high and dissipation ceased almost completely during winter. During the 2 year experiment no indication of potential groundwater pollution risk was obtained, but herbicides may cause surface water pollution.     

The effect of root-absorbed PSII inhibitors on Kautsky curve parameters in sugar beet

The effects of the photosystem II inhibitors metamitron and terbuthylazine on the shape of the Kautsky (chlorophyll fluorescence induction) curve were investigated in sugar beet grown in hydroponic culture. The objective of the study was to trace recovery processes following herbicide injury using Kautsky curve parameters. Metamitron is used for selective weed control in sugar beet because it is metabolized in this crop. In contrast, terbuthylazine is toxic to sugar beet. Two hours after treatment, various fluorescence induction curve parameters, such as maximum quantum efficiency (F_V/F_m), the relative changes at the J step (F_vj) and area (the area between the Kautsky curve and maximum fluorescence, F_m), were affected by metamitron at concentration ranges of 70–280 mg active ingredient (a.i.) L^?1 in plants treated at the four‐true‐leaf stage. Shortly after herbicide application, F_v/F_m was more affected by the hydrophilic metamitron [log(K_ow) = 0.83] than by the lipophilic terbuthylazine [log(K_ow) = 3.21], but these differences between compounds were alleviated as metamitron was metabolized and terbuthylazine was not. Terbuthylazine at 1 mg a.i. L^?1 affected sugar beet at the four‐ and six‐true‐leaf stages to the same extent, whereas metamitron at a dose of 140 mg a.i. L^?1 affected much more at four‐ than at the six‐true‐leaf stage. Sugar beet recovered from metamitron injury even at high doses (140 and 280 mg a.i. L^?1). Fluorescence induction curve parameters were similarly affected by terbuthylazine and, although sugar beet recovered from terbuthylazine injury at low doses (<0.2 mg a.i. L^?1), the Kautsky curve was irreversibly affected at higher doses (1–10 mg a.i. L^?1), leading finally to plant death. Older plants were affected later, and recovered sooner, from both herbicides.     

Absorption, translocation and metabolism of metamitron in Chenopodium album

BACKGROUND: In recent years, common lambsquarters (Chenopodium album L.) populations from sugar beet fields in different European countries have responded as resistant to the as‐triazinone metamitron. The populations have been found to have the same D1 point mutation as known for atrazine‐resistant biotypes (Ser₂₆₄ to Gly). However, pot experiments revealed that metamitron resistance is not as clear‐cut as observed with triazine resistance in the past. The objectives of this study were to clarify the absorption, translocation and metabolic fate of metamitron in C. album. RESULTS: Root absorption and foliar absorption experiments showed minor differences in absorption, translocation and metabolism of metamitron between the susceptible and resistant C. album populations. A rapid metabolism in the C. album populations was observed when metamitron was absorbed by the roots. The primary products of metamitron metabolism were identified as deamino‐metamitron and metamitron‐N‐glucoside. PABA, known to inhibit the deamination of metribuzin, did not alter the metabolism of metamitron, and nor did the cytochrome P450 inhibitor PBO. However, inhibition of metamitron metabolism in the presence of the cytochrome P450 inhibitor ABT was demonstrated. CONCLUSION: Metamitron metabolism in C. album may act as a basic tolerance mechanism, which can be important in circumstances favouring this degradation pathway. Copyright © 2011 Society of Chemical Industry     

Seed germination and viability of herbicide resistant and susceptible Chenopodium album populations after ensiling,digestion by cattle and manure storage

Chenopodium album became a problem weed in sugar beet production, due to resistance to metamitron, a key herbicide in this crop. Dispersal of the seeds from resistant biotypes may occur due to spread by wind, animals, agricultural machinery or manure. This study examined the effect of ensiling, digestion by cattle and storage in slurry and farmyard manure on the germination and viability of the seeds of one susceptible and three resistant C. album populations. After 4 weeks in a maize silo, seed viability of C. album populations was reduced drastically to 0–5%. Incubation for 24 h in the rumen followed by a post‐ruminal digestion in vitro of intact seeds only resulted in a small reduction in viability in one C. album population. Storage in a slurry cellar for 16 weeks reduced the viability of intact seeds of the C. album populations to 25–60%. Only 0–1% of the seeds remained viable after storage in a farmyard manure heap for 4 weeks. An accelerated ageing experiment showed seed persistence to be population specific and less related to seed weight. Keeping a fresh maize silo closed for at least 4 weeks and heaping farmyard manure are excellent preventive measures to limit the spread of resistant C. album seeds between fields.     

Genetic variability among isolates of Fusarium oxysporum from sugar beet

Fusarium yellows, caused by the soil‐borne fungus Fusarium oxysporum f. sp. betae (Fob), can lead to significant yield losses in sugar beet. This fungus is variable in pathogenicity, morphology, host range and symptom production, and is not a well characterized pathogen on sugar beet. From 1998 to 2003, 86 isolates of F. oxysporum and 20 other Fusarium species from sugar beet, along with four F. oxysporum isolates from dry bean and five from spinach, were obtained from diseased plants and characterized for pathogenicity to sugar beet. A group of sugar beet Fusarium isolates from different geographic areas (including nonpathogenic and pathogenic F. oxysporum, F. solani, F. proliferatum and F. avenaceum), F. oxysporum from dry bean and spinach, and Fusarium DNA from Europe were chosen for phylogenetic analysis. Sequence data from β‐ tubulin, EF1α and ITS DNA were used to examine whether Fusarium diversity is related to geographic origin and pathogenicity. Parsimony and Bayesian MCMC analyses of individual and combined datasets revealed no clades based on geographic origin and a single clade consisting exclusively of pathogens. The presence of FOB and nonpathogenic isolates in clades predominately made up of Fusarium species from sugar beet and other hosts indicates that F. oxysporum f. sp. betae is not monophyletic.     

Monitoring the efficacy and metabolism of phenylcarbamates in sugar beet and black nightshade by chlorophyll fluorescence parameters

Desmedipham, phenmedipham and a 50% mixture of the two decreased the maximum quantum efficiency of photosystem II (F(v)/F(m)) and the relative changes at the J step (F(vj)) immediately after spraying in both sugar beet and black nightshade grown in the greenhouse. Sugar beet recovered more rapidly from phenmedipham and the mixture than from desmedipham. Desmedipham and the mixture irreversibly affected F(v)/F(m) and F(vj) in black nightshade at much lower doses than in sugar beet. Black nightshade recovered from phenmedipham injury at the highest dose in the first experiment (120 g AI ha(-1)) but not in the second experiment (500 g AI ha(-1)). The dry matter dose-response relationships and the energy pipeline presentation confirmed the same trend. There was a relatively good correlation between F(vj) taken 1 day after spraying and dry matter taken 2 or 3 weeks after spraying. The differential speed of herbicide metabolism between weed and crop plays an important role in herbicide selectivity and can be studied by using appropriate chlorophyll a fluorescence parameters.     

Composition of the stubble weed flora and its role for Heterodera schachtii in the year preceding sugar beet production

In Germany, sugar beet is often rotated with 2 years of cereal. Extensive fallow periods between cereal harvest and autumn primary tillage allow for a weed flora to develop. Broad‐leaved weeds could potentially be alternate hosts for the common nematode Heterodera schachtii, one of the most important pests of sugar beet. Between 2009 and 2012, annual weeds developing in cereal stubble fields during July to mid‐October in the season prior to sugar beet were surveyed, including known hosts of H. schachtii. Yearly weather patterns and agronomic practices possibly impacted weed species composition and weed population densities. During September, Chenopodium album, Cirsium arvense, Convolvulus arvensis, Mercurialis annua, Polygonum spp., Solanum nigrum and Sonchus spp. occurred at the highest frequencies. Weed hosts of H. schachtii were present, but densities, frequencies and uniformity were limited. In 2010 and 2011, staining for nematodes in roots revealed juvenile penetration of some weeds but few adult stages. No indication of nematode reproduction of H. schachtii was found on these weed hosts. A fairly stable weed flora was detected on stubble fields that could provide some carry over for weed species. An elevated risk for nematode population density build‐up on these weeds was not found and management of these weeds at the observed densities during the stubble period for nematological reasons appeared unnecessary.     

Weed control in glyphosate-tolerant sugarbeet (Beta vulgaris L.)

The effect of glyphosate on weeds was evaluated in greenhouse bioassays with five weed species and compared with a commercial mixture of phenmedipham and ethofumesate. Glyphosate was more active than a mixture of phenmedipham and ethofumesate on the weeds. Solanum nigrum was the most sensitive species to both herbicide treatments. The relative potency of glyphosate between the weeds showed consistency at 50% and 90% control levels for four out of five weed species, which could be ranked independently of control level because of similar response curves. In a field trial on sugarbeet genetically engineered to acquire glyphosate tolerance, a total of 720 g a.i. ha^?1 of glyphosate applied in one, two or three applications gave similar or superior control of weeds to a total of 3.17 kg a.i. ha^?1 mixture of metamitron, phenmedipham and ethofumesate applied in three repeated applications.     

Genetic diversity and pathogenicity of Pseudomonas syringae pv. aptata isolated from sugar beet

Pseudomonas syringae pv. aptata is the causal agent of bacterial leaf spot disease of sugar beet (Beta vulgaris). During 2013, 250 samples were collected from leaf lesions with typical symptoms of bacterial leaf spot in commercial fields of sugar beet in Serbia, and 104 isolates of P. syringae pv. aptata were obtained. Identification and characterization was performed using biochemical, molecular and pathogenicity tests. Identification included LOPAT tests and positive reactions using primers Papt2F and Papt1R specific for P. syringae pv. aptata. Repetitive (rep) sequence‐based PCR typing with ERIC, REP and BOX primers revealed high genetic variability among isolates and distinguished 25 groups of different fingerprinting profiles. Pulse‐field gel electrophoresis (PFGE) and multilocus sequence analysis (MLSA) of representative isolates showed higher genetic variability than in rep‐PCR analysis and distinguished three and four major genetic clusters, respectively. A pathogenicity test performed with 25 representative isolates on four cultivars of sugar beet confirmed the occurrence of leaf spot disease and showed correlation between the most aggressive isolates and the genetic clusters obtained in MLSA. All these findings point to the existence of several lines of P. syringae pv. aptata infection in Serbia that are genetically and pathologically different.     

Joint action of some root-absorbed herbicides in oats (Avena sativa L.)

In growth chambers the phytotoxicity of binary mixtures of four herbicides was compared using an Additive Dose Model as reference. Of the four herbicides used, lenacil was the most potent whereas ethofumesate was the least potent herbicide; chloridazon and metamitron were equally potent. The Additive Dose Model implicitly presupposes that at any one response level the herbicides of a mixture can replace each other in proportion to their relative potency when applied separately. Metamitron and chloridazon mixtures appeared to follow the reference model. The efficacy of mixtures of lenacil and chloridazon or lenacil and metamitron, however, were increased in comparison with the herbicides applied separately. On the other hand, lenacil and ethofumesate or metamitron and ethofumesate were less phytotoxic than expected from the Additive Dose Model. The results are discussed in relation to the mode of action of the compounds. Action combinée de certains herbicides absorbés par les racines en culture d'avoine Avena sativa L. La phytotoxicité de mélanges binaires de quatre herbicides a fait l'objet d'une comparaison en phytotron, utilisant comme témoin un Modèle Dose Additive. Parmi les quatre herbicides utilisés, le lénacile s'est révélé le plus et l'éthofumésate le moins actif; le chloridazone et le métamitrone ont fait preuve d'une activitéégale. Le Modèle Dose Additive présuppose qu'à tout niveau d'activité chaque herbicide faisant partie d'un mélange puisse se substituer à l'autre en fonction de son efficacité relative en traitement simple. Les mélanges métamitrone–chloridazone semblent s'accorder au modèle de référence. Cependant, l'efficacité des mélanges chloridazone–lénacile et lénacile–métamitrone a augmenté par rapport à l'application de chaque élément séparément. D'autre part, les mélanges lénacile–éthofumesate et métamitrone–éthofumesate ont fait preuve d'une phytotoxicité moindre que celle qu'annonçait le modèle. Ces résultats sont discutés par rapport au mode d'action des composés. Kombinierte Wirkung verschiedener wurzelwirksamer Herbizide auf Avena sativa L. In Klimakammern wurde die Phytotoxizität von Zweier-Mischungen von vier Herbiziden untersucht und mit einem Modell additiver Dosierungen verglichen. Von den vier untersuchten Herbiziden war Lenacil das wirksamste und Ethofumesat das schwächste; Chloridazon und Metamitron wirkten gleich stark. Das Modell additiver Dosierungen setzt stillschweigend voraus, dass auf jedem Wirkungsniveau die Herbizide einer Mischung einander im Verhältnis ihrer relativen Wirksamkeiten, wenn allein appliziert, ersetzen können. Metamitron + Chloridazon Mischungen scheinen dem Vergleichsmodell zu folgen. Die Wirkungen der Mischungen von Lenacil + Chloridazon oder Lenacil + Metamitron, hingegen, waren im Vergleich zu den Effekten der einzeln applizierten Komponenten gesteigert. Andrerseits waren Lenacil + Ethofumesat oder Metamitron + Ethofumesat weniger phytotoxisch, als das Modell hätte erwarten lassen. Die Resultate werden mit Bezug auf die Wirkungsmechanismen der Komponenten diskutiert.     

Phylogenetic relationships and virulence assays of Fusarium secorum from sugar beet suggest a new look at species designations

Fusarium spp. are responsible for significant yield losses in sugar beet (Beta vulgaris) with Fusarium oxysporum f. sp. betae most often reported as the primary causal agent. Recently, a new species, F. secorum, was reported to cause disease in sugar beet but little is known on the range of virulence within F. secorum or how this compares to the virulence and phylogenetic relationships previously reported for Fusarium pathogens of sugar beet. To initiate this study, partial translation elongation factor 1-α (TEF1) sequences from seven isolates of F. secorum were obtained and the data were added to a previously published phylogenetic tree that includes F. oxysporum f. sp. betae. Unexpectedly, the F. secorum strains nested into a distinct group that included isolates previously reported as F. oxysporum f. sp. betae. These results prompted an expanded phylogenetic analysis of TEF1 sequences from genomes of publicly available Fusarium spp., resulting in the additional discovery that some isolates previously reported as F. oxysporum f. sp. betae are F. commune, a species that is not known to be a sugar beet pathogen. Inoculation of sugar beet with differing genetic backgrounds demonstrated that all Fusarium strains have a significant range in virulence depending on cultivar. Taken together, the data suggest that F. secorum is more widespread than previously thought. Consequently, future screening for disease resistance should rely on isolates representing the full diversity of the Fusarium population that impacts sugar beet.     

The use of digital image analysis and real‐time PCR fine‐tunes bioassays for quantification of Cercospora leaf spot disease in sugar beet breeding

Cercospora leaf spot, caused by the fungus Cercospora beticola, is a major fungal sugar beet disease worldwide and the cause of significant yield losses. The disease is most successfully countered by the introduction of genetic tolerance into elite sugar beet hybrids. To this end, breeding programmes require high quality biological assays allowing discrimination of minor differences between plants within a segregating population. This study describes the successful implementation of image analysis software in the bioassays for quantification of necrotic lesions at different stages of C. beticola infection, allowing selection on minor phenotypic differences during the sugar beet breeding process for C. beticola resistance. In addition, a real‐time PCR assay was developed for the quantification of C. beticola pathogen biomass in infected beet canopy. The use of both techniques, even in an early stage of infection, fine‐tunes current bioassays, allowing more accurate and efficient selection of resistant breeding material.     

Determination of sugarbeet herbicides in soil samples by hplc

Determination of sugarbeet herbicides such as chloridazon, metamitron and phenmedipham in soil samples is described. After extraction with acetone, pesticides were determined by HPLC on an RP-18 column using methanol/water as mobile phase. Average recoveries were 82% for chloridazon, 93% for metamitron and 77% for phenmedipham. Quantification limits were 3·5 μg kg^?1 for chloridazon, 6·3 μg kg^?1 for metamitron and 3·6 μg kg^?1 for phenmedipham.     

Evaluation of virulence and pathogenicity of Alternaria patula on French marigold (Tagetes patula)

Alternaria patula, the cause of French marigold leaf black spot and flower blight, was first isolated from seeds of French marigold cv. Queen Sophia. It is described as a new species of Alternaria and has a considerable morphological variation with a preferential pathogenicity to Asteraceae, Solanaceae and Cucurbitaceae plants. Alternaria patula produces an array of pectin depolymerases that can break 1,4-α-glycosidic bonds either by hydrolysis of polygalacturonases (PG, E.C. 3.2.1.15) or via trans-elimination of pectate lyases (PL, E.C. 4.2.2.2) and pectin lyases (PNL, E.C. 4.2.2.10). This study is the first to emphasize the variability and significance to pathogenesis of the pectinolytic enzymes of A. patula that target various pectic polymers structures during host tissue invasion. Alternaria patula also produced zinniol derivatives as non-host-specific toxins (nHSTs), albeit without phytotoxic symptoms in French marigold. The management of Alternaria blight caused by A. patula via the application of pyrifenox and antagonistic Bacillus amyloliquefaciens effectively reduced disease severity, without adverse effects on French marigold in both in vitro and in vivo bioassays.     

New insights into virus yellows distribution in Europe and effects of beet yellows virus,beet mild yellowing virus,and beet chlorosis virus on sugar beet yield following field inoculation

Beet yellows virus (BYV), beet mild yellowing virus (BMYV), beet chlorosis virus (BChV), and beet mosaic virus (BtMV) cause virus yellows (VY) disease in sugar beet. The main virus vector is the aphid Myzus persicae. Due to efficient vector control by neonicotinoid seed treatment over the last decades, there is no current knowledge regarding virus species distribution. Therefore, Europe-wide virus monitoring was carried out from 2017 to 2019, where neonicotinoids were banned in 2019. The monitoring showed that closterovirus BYV is currently widely spread in northern Europe. The poleroviruses BMYV and BChV were most frequently detected in the northern and western regions. The potyvirus BtMV was only sporadically detected. To study virus infestation and influence on yield, viruses were transmitted to sugar beet plants using viruliferous M. persicae in quadruplicate field plots with 10% inoculation density simulating natural infection. A plant-to-plant virus spread was observed within 4 weeks. A nearly complete infection of all plants was observed in all treatments at harvest. In accordance with these findings, a significant yield reduction was caused by BMYV and BChV (−23% and −24%) and only a moderate reduction in yield was observed for BYV (−10%). This study showed that inoculation at low densities mimics natural infection, and quick spreading induced representative yield effects. Within the background of a post-neonicotinoid era, this provides the basis to screen sugar beet genotypes for the selection of virus tolerance/resistance and to test the effectiveness of insecticides for the control of M. persicae with a manageable workload.     

Development of herbicide resistance in weeds in a crop rotation with acetolactate synthase‐tolerant sugar beets under varying selection pressure

The development of acetolactate synthase (ALS) tolerant sugar beet provides new opportunities for weed control in sugar beet cultivation. The system consists of an ALS?inhibiting herbicide (foramsulfuron + thiencarbazone‐methyl) and a herbicide‐tolerant sugar beet variety. Previously, the use of ALS‐inhibitors in sugar beet was limited due to the susceptibility of the crop to active ingredients from this mode of action. The postulated benefits of cultivation of the ALS‐tolerant sugar beet are associated with potential risks. Up to now, with no relevant proportion of herbicide‐tolerant crops in Germany, ALS‐inhibitors are used in many different crops. An additional use in sugar beet cultivation could increase the selection pressure for ALS‐resistant weeds. To evaluate the impact of varying intensity of ALS‐inhibitor use on two weed species (Alopecurus myosuroides and Tripleurospermum perforatum) in a crop rotation, field trials were conducted in Germany in two locations from 2014 to 2017. Weed densities, genetic resistance background and crop yields were annually assessed. The results indicate that it is possible to control ALS‐resistant weeds with an adapted herbicide strategy in a crop rotation including herbicide‐tolerant sugar beet. According to the weed density and species, the herbicide strategy must be extended to graminicide treatment in sugar beet, and a residual herbicide must be used in winter wheat. The spread of resistant biotypes in our experiments could not be attributed to the integration of herbicide‐tolerant cultivars, although the application of ALS‐inhibitors promoted the development of resistant weed populations. Annual use of ALS‐inhibitors resulted in significant high weed densities and caused seriously yield losses. Genetic analysis of surviving weed plants confirmed the selection of ALS‐resistant biotypes.     

Identifying key components of weed beet management using sensitivity analyses of the GeneSys-Beet model in GM sugar beet

Genetically-modified (GM) sugar beet varieties tolerant to non-selective herbicides would be useful for managing weed beet, an annual form of Beta vulgaris impossible to eliminate with herbicides in sugar beet. However, it is highly probable that the herbicide-tolerance transgene would be transmitted to the weed through pollen flow. It is therefore essential to study how weed beet. particularly Herbicide-Tolerant (HT) populations, develop in cropping systems and how to optimise crop succession and management for controlling these weeds. As multiple interactions and long-term effects make field experiments impractical, we carried out a simulation study with a deterministic and mechanistic model, G ene S ys- B eet , which quantifies weed beet dynamics and gene flow in cropping systems with interactions with climate, soil structure and hydro-thermal conditions. The sensitivity analysis consisted of 250 000 random combinations of input variables to rank cropping system components according to their effect on both total and GM weed beet infestations. Frequency of sugar beet crops, crop succession, manual and mechanical weeding and tillage were identified as the most important variables. Several cultivation techniques must be combined to efficiently control weed beet. Our recommendations are complex, but a delayed return of sugar beet in the rotation. Harvest should be followed as soon as possible by a shallow tilling; tillage should always be as shallow and as early as possible, except before sugar beet where mouldboard ploughing is advisable. If possible, sowing dates should be delayed. Sugar beet should be weeded mechanically and/or manually, aiming at late and efficient, rather than early or frequent operations. Herbicides should be applied whenever possible and target all weed beet stages and genotypes. Set-aside must be cut as frequently and as late as possible.     

Quantitative reproductive potential of Heterodera schachtii on weeds typical for late summer fallow in sugar beet rotations

In Europe, sugar beet is often produced in a 3‐year rotation with cereals, leaving stubble fields fallow from cereal harvest until primary tillage in autumn in the year prior to sugar beet production. The weed flora on such fields could include host plants of Heterodera schachtii that is one of the most important pests of sugar beet. Crop sequences with non‐hosts and cover cropping with resistant cruciferous hosts during this period have been crucial for its management. Availability of resistant and tolerant sugar beet cultivars could entice growers to forego cover cropping, exacerbating weed problems during the fallow period. The objective of this study was to determine the reproductive potential of H. schachtii on weeds that develop during this period. Under glasshouse conditions, reproduction on 39 plant species was compared with that on oilseed radish and sugar beet of differing nematode host status. In 2 years in field microplots, 18 previously tested species were grown in H. schachtii‐infested soil during the typical fallow period at 60 plants m^?2, and nine of these species were also grown at 180 plants m^?2. There were variable results between years after 8 weeks of growth, but most weeds allowed lower reproduction (<10%) than the susceptible sugar beet; only Stellaria media at 180 plants m^?2 and Thlaspi arvense at both plant densities increased nematodes. Such weed densities may seldom occur under commercial conditions; thus, weed management for nematological considerations during the stubble period may have limited importance.     

Identification of sugar beet germplasm EL51 as a source of resistance to post-emergence Rhizoctonia damping-off

Resistance of sugar beet seedlings to Rhizoctonia damping-off caused by Rhizoctonia solani has not been described. A series of preliminary characterisations using a single susceptible host and four different R. solani isolates suggested the disease progression pattern was predictable. Two AG-4 isolates and a less virulent AG-2-2 isolate (W22) showed a comparable pattern of disease progression in the growth chamber where disease index values increased for the first 5–6 days, were relatively constant for the next 7–8 days, and declined thereafter. Seedlings inoculated with a highly virulent AG-2-2 isolate (R-1) under the same conditions showed similar patterns for the first 4 days post-inoculation; however disease index values continued to increase until seedling death at 13–14 days. Similar results were observed in the greenhouse, and a small expanded set of other germplasm lines were screened. One tested germplasm accession, EL51, survived seedling inoculation with R. solani AG-2-2 R-1, and its disease progress pattern was characterised. In a field seedling disease nursery artificially inoculated with R. solani AG-2-2 R-1, seedling persistence was high with EL51, but not with a susceptible hybrid. Identification of EL51 as a source of resistance to Rhizoctonia damping-off may allow investigations into the Beta vulgaris–Rhizoctonia solani pathosystem and add value in sugar beet breeding.     

A new molecular method for the rapid detection of a metamitron‐resistant target site in Chenopodium album

BACKGROUND: Resistance to photosystem II inhibitors—triazines (atrazine) and triazinones (metamitron, metribuzin)—in Chenopodium album L. is caused by the serine 264 to glycine mutation in the D1 protein. This mutation has been detected in C. album collections from Belgium with unsatisfactory metamitron efficacy in the field and was confirmed in greenhouse resistance bioassays. Incomplete herbicide efficacy in practice can also be caused by reduced uptake due to environmental conditions. Hence, for reliable differentiation and resistance identification, a rapid method for mutation detection in the target gene psbA is required. RESULTS: Dose–response curves obtained in herbicide greenhouse assays with metamitron‐resistant and ‐susceptible reference biotypes showed that a dose of 2 L ha^?1 metamitron was suitable for discrimination. A psbA PCR‐RFLP was developed, based on the presence of a FspBI restriction enzyme recognition site, covering D1 codon 264 in susceptible genotypes. A paper‐based DNA extraction allowed direct processing of leaf samples already in the field. In order to detect the mutation even in mixed seed samples, a nested PCR‐RFLP was also developed. CONCLUSION: The method allows exhaustive surveys screening C. album leaf or seed samples for the occurrence of the D1 Ser264Gly mutation to confirm or disprove metamitron resistance in the case of unsatisfactory control. Copyright © 2010 Society of Chemical Industry