Screening for resistance to canker (Leptosphaeria maculans) in winter oilseed rape (Brassica napus ssp. oleifera)

A test involving infection of seedlings in a glasshouse was evaluated for potential use in screening breeding material by comparing the results with those of field tests. The two tests correlated well for some resistant selections, particularly those closely related to Jet Neuf, and for very susceptible selections. However, two small groups of selections were resistant in the field but consistently susceptible in the glasshouse. Inoculation of mature plants in the glasshouse resulted in similar discrepancies and therefore gave no advantage over the seedling test. These discrepancies limit the usefulness of glasshouse techniques for routine resistance screening. A very high correlation was found overall between internal and external canker symptoms of the crown in nine field trials. Significant differences were found, however, between selections and cultivars in the relative severity of these two symptoms. Upper stem canker and crown canker were found to be very poorly correlated for individual plants, but highly correlated for eultivar means. The significance of these observations is discussed with respect to scoring canker in the field.     

New genes for resistance to downy mildew (Peronospora parasitica) in oilseed rape (Brassica napus ssp. oleifera)

The response of cotyledons of oilseed rape ( Brassica napus ssp. oleifera ) accessions to infection by isolates of Peronospora parasitica under controlled conditions was assessed on a 0–7 scale (disease reaction). In interactions scored 0–3, 4–5 and 6–7, the host was considered resistant, partially resistant and susceptible, respectively. Accession RES-26, selected from the spring oilseed rape cultivar Janetzkis, was partially resistant to isolate R1 and resistant to isolate P003 of P. parasitica , which distinguishes it from three previously described differential response groups ('A', 'B' and 'C') of accessions in B. napus . The resistance of RES-26 to isolate P003 seemed to be conditioned by a single, partially dominant gene and the resistance of RES-02, which belongs to group 'A' (resistant to R1 and P003), by two independent partially dominant genes. The gene for resistance to P003 in RES-26 is either closely linked, allelic or identical to one of the two genes for resistance in RES-02. Resistance of RES-02 to R1 is conditioned by a single, incompletely dominant gene. The genes for resistance to isolates R1 and P003 in RES-02 are either closely linked, allelic or identical. The cotyledonary leaves of each seedling responded independently when inoculated simultaneously each with a different isolate of the pathogen.     

The identification of a gene for race-specific resistance to Peronospora parasitica (downy mildew) in Brassica napus var. oleifera (oilseed rape)

The oilseed rape cultivar Cresor was resistant to 14 isolates of Peronospora parasitica derived from crops of Brassica napus in the UK. Segregation for resistance to one isolate among F₂ plants and F₃ progeny of crosses between Cresor and the susceptible cultivars Victor and Jet Neuf indicated that resistance was controlled by a single gene. There was evidence that genetic background and environment could influence the phenotypic expression of this resistance. Two sexual progeny isolates derived from a homothallic isolate of P. parasitica avirulent on Cresor were completely virulent on this cultivar. This suggested that the parental isolate was heterozygous at a matching locus or loci for avirulence and demonstrated the race-specific nature of the resistance.     

Identification of quantitative trait loci for resistance against Verticillium longisporum in oilseed rape (Brassica napus)

Verticillium longisporum is one of the major pathogens of oilseed rape (Brassica napus; genome AACC, 2n = 38) in Europe. Current European cultivars possess only a low level of resistance against V. longisporum, meaning that heavy infection can cause major yield losses. The aim of this study was to identify quantitative trait loci (QTL) for resistance against V. longisporum as a starting point for marker-assisted breeding of resistant cultivars. Resistance QTL were localized in a segregating oilseed rape population of 163 doubled haploid (DH) lines derived by microspore culture from the F1 of a cross between two B. napus breeding lines, one of which exhibited V. longisporum resistance derived by pedigree selection from a resynthesized B. napus genotype. A genetic map was constructed comprising 165 restriction fragment length polymorphism, 94 amplified fragment length polymorphism and 45 simple sequence repeats (SSR) markers covering a total of 1,739 cM on 19 linkage groups. Seedlings of the DH lines and parents were inoculated with V. longisporum isolates in four greenhouse experiments performed in Sweden during autumn 1999. In three of the experiments the DH lines were inoculated with a mixture of five isolates, while in the fourth experiment only one of the isolates was used. The intention was to simulate four different environments with variable disease pressure, while still maintaining uniform conditions in each environment to enable reliable disease scoring. The disease index (DI) was calculated by scoring symptoms on a total of 21 inoculated plants per line in comparison to 21 noninoculated plants per line. Using the composite interval mapping procedure a total of four different chromosome regions could be identified that showed significant QTL for resistance in more than one environment. Two major QTL regions were identified on the C-genome linkage groups N14 and N15, respectively; each of these QTL consistently exhibited significant effects on resistance in multiple environments. The presence of flanking markers for the respective QTL was associated with a significant reduction in DI in the inoculated DH lines.     

Quantitative resistance to symptomless growth of Leptosphaeria maculans (phoma stem canker) in Brassica napus (oilseed rape)

Quantitative resistance to Leptosphaeria maculans in Brassica napus was investigated in field and controlled environments using cultivars Darmor (with quantitative resistance) and Eurol (without quantitative resistance). In field experiments, numbers of phoma leaf spot lesions in autumn/winter and severity of stem canker the following summer were assessed in three growing seasons. There were no differences between Darmor and Eurol in number of leaf lesions in autumn/winter. However, stem cankers were less severe on Darmor than Eurol at harvest the following summer. In controlled-environment experiments, development of leaf lesions at different temperatures (5–25°C) and wetness durations (12–72 h) was investigated using ascospore inoculum; symptomless growth of L. maculans along leaf petioles towards the stem was quantified using quantitative PCR and visualized using GFP-expressing L. maculans ; growth of L. maculans within stem tissues was investigated using GFP-expressing L. maculans . There were more leaf lesions on Darmor than Eurol, although there was no difference between Darmor and Eurol in L. maculans incubation period. There were no differences between Darmor and Eurol in either distance grown by L. maculans along leaf petioles towards the stem or quantity of L. maculans DNA in leaf petioles, but L. maculans colonized stem tissues less extensively on Darmor than Eurol. It was concluded that quantitative resistance to L. maculans operates during colonization of B. napus stems by the pathogen.     

The response of oilseed rape (Brassica napus ssp. oleifera) accessions with different glucosinolate and erucic acid contents to four isolates of Peronospora parasitica (downy mildew) and the identification of new sources of resistance

A total of 101 Brassica napus ssp. oleifera accessions with seed differing in glucosinolate and erucic acid contents were screened for resistance to four isolates of Peronospora parasitica at the cotyledon stage. Two groups of accessions with different resistance factors were identified. Lines that were homogeneous for resistance were selected from seedling populations of accessions that exhibited a heterogeneous reaction to some isolates. The resistance of one group differs from that of cv, Cresor, the only oilseed rape cultivar reported to have an isolate-specific gene for resistance to P. parasitica. The isolate specificity of the second group was identical to that of cv, Cresor, A comparison of the response of host accessions which expressed moderate to full susceptibility at the cotyledon stage, with no clear differential response to any of the four P. parasitica isolates, indicated that those with high glucosinolate and high erucic acid contents (12 accessions) were slightly but significantly less susceptible than those with high glucosinolate and low erucic acid (19 accessions), or low glucosinolate and low erucic acid contents (28 accessions). The mean differences between accessions with low erucic acid but differing in glucosinolate content were inconsistent. The last result was further confirmed by investigating the expression of resistance to three isolates of P. parasitica at three different seedling growth stages among 11 accessions of oilseed rape with seeds low in erucic acid but differing in glucosinolate content.     

Overcoming seed dormancy in oilseed rape (Brassica napus L.) with exogenous compounds

Dormant seeds of oilseed rape (OSR) can persist in the soil and cause OSR volunteers in subsequent crops. Several approaches were tested in the laboratory and in the field to determine whether dormancy induction and seed persistence can be reduced by using dormancy‐breaking exogenous compounds. In a laboratory experiment, OSR seeds were coated with KNO₃, micronutrients, or gibberellic acid (GA) prior to a secondary dormancy test. In a field experiment, seeds were coated in a manner analogous to the laboratory experiment, and then buried 10 cm deep in the soil for 2.5 months. In a practical demonstration, OSR plants were sprayed with either urea ammonium nitrate (UAN) or a commercial product containing GA prior to seed maturity. Seed coating (laboratory and field experiments) reduced secondary dormancy and seed persistence in the field by up to 99%. The efficiency of the treatments for mitigating secondary dormancy (laboratory and field experiments) in decreasing order was GA > micronutrients > KNO₃ > control. With pre‐maturity spraying (practical demonstration), UAN reduced primary dormancy by up to 77% and the development of secondary dormancy by up to 38%; GA had no effect. Dormancy and seed persistence of OSR seeds may be reduced by a pre‐maturity UAN treatment of OSR mother plants, or by applying appropriate exogenous compounds to OSR seeds.     

Insecticide-resistant pollen beetles (Meligethes aeneus F) found in Danish oilseed rape (Brassica napus L) fields

The pollen beetle is the most important pest in Danish oilseed rape fields. In 2001, we screened a broad range of pollen beetle populations for pyrethroid and dimethoate resistance. A standard dip-test was used to test insecticide resistance in 18 populations collected from oilseed winter and spring rape fields. The beetles were treated with four different insecticides: the pyrethroids tau-fluvalinate, lambda-cyhalothrin and esfenvalerate, and the organophosphate dimethoate. The results show that up to 99% of the pollen beetles survived Danish standard doses of pyrethroids and up to 36% of the beetles survived standard doses of dimethoate.     

Competitiveness and yield impact of volunteer oilseed rape (Brassica napus) in winter and spring wheat (Triticum aestivum)

Journal of Plant Diseases and Protection - Volunteer oilseed rape is a relevant weed in many crop rotations due to seed persistence and dormancy. With the intro-duction of imidazolinone-tolerant...     

Distribution and metabolism of D/L-, L- and D-glufosinate in transgenic, glufosinate-tolerant crops of maize (Zea mays L ssp mays) and oilseed rape (Brassica napus L var napus)

The aim of the present study was to determine whether post-emergence application of glufosinate to transgenic crops could lead to an increase in residues or to the formation of new, hitherto unknown metabolites. Transgenic oilseed rape and maize plants were treated separately with L-glufosinate, D-glufosinate or the racemic mixture. Whereas about 90% of the applied D-glufosinate was washed off by rain and only 5-6% was metabolised, 13-35% of the applied L-glufosinate remained in the form of metabolites and unchanged herbicide in both transgenic maize and oilseed rape. The main metabolite was N-acetyl-L-glufosinate with total residues of 91% in oilseed rape and 67% in maize, together with small amounts, of 5% in oilseed rape and 28% in maize, of different methylphosphinyl fatty acids. These metabolites were probably formed from L-glufosinate by deamination and subsequent decarboxylation. The residues were distributed in all fractions of the plants, with the highest contents in treated leaves and the lowest in the grains (0.07-0.3% in maize and 0.4-0.6% in oilseed rape). There was no indication of an accumulation of total residues or of residue levels above the official tolerances for glufosinate.     

Selection of bacterial antagonists for the biological control of Rhizoctonia solani in oilseed rape (Brassica napus)

Spore-forming bacteria isolated from oilseed rape (OSR) roots were tested for biological control activity against Rhizoctonia solani damping-off of OSR, using a simple slurry coat formulation. From a total of 239 strains which were tested, Bacillus subtilis 205 was selected as the most promising biocontrol agent (BCA). A known isolate of B. subtilis (NCIMB 12376), which had previously shown activity against R. solani in cotton, was used as a standard comparison throughout. In an in vivo screening, pre-emergence damping-off disease control by B. subtilis 205 was excellent, with plant stands being almost as high as non-diseased controls. However, post-emergence survival was less good, although the surviving plants exhibited significantly reduced hypocotyl rot. Growth-room trials which assessed the effect of ambient temperature on the biological control efficacy of B. subtilis strains 12376 and 205 showed that isolate 205 was more effective at lower temperatures. Disease control by isolate 205 at 15°C/10°C (day/night) suggests that this bacterium could operate under autumn sowing field conditions. Further investigations also indicated that the addition of iron (III) to the BCA formulation may enhance disease control by both bacilli. The potential of bacterial BCAs for the control of seedling diseases is discussed.     

Visual and PCR assessment of light leaf spot (Pyrenopeziza brassicae) on winter oilseed rape (Brassica napus) cultivars

Methods to assess light leaf spot ( Pyrenopeziza brassicae ) on winter oilseed rape cultivars were compared in laboratory, controlled-environment and field experiments. In controlled-environment experiments with seedling leaves inoculated at GS 1,4, the greatest differences in percentage area affected by P. brassicae sporulation were observed with inoculum concentrations of 4 × 10³ or 4 × 10⁴ spores mL⁻¹, rather than 4 × 10² or 4 × 10⁵ spores mL⁻¹, but older leaves had begun to senesce before assessment, particularly where they were severely affected by P. brassicae . In winter oilseed rape field experiments, a severe light leaf spot epidemic developed in 2002/03 (inoculated, September/October rainfall 127·2 mm) but not in 2003/04 (uninoculated, September/October rainfall 40·7 mm). In-plot assessments discriminated between cultivars best in February/March in 2003 and June in 2004, but sometimes failed to detect plots with many infected plants (e.g. March/April 2004). Ranking of cultivar resistance differed between seedling experiments done under controlled-environment conditions and field experiments. The sensitivity of detection of P. brassicae DNA extracted from culture was greater using the PCR primer pair PbITSF/PbITSR than using primers Pb1/Pb2. P. brassicae was detected by PCR (PbITS primers) in leaves from controlled-environment experiments immediately and up to 14 days after inoculation, and in leaves sampled from field experiments 2 months before detection by visual assessment.     

Herbicide-resistant crops: yield penalties and weed thresholds for oilseed rape (Brassica napus L.)

An expérimental procedure was designed to provide a simple model for types of analyses necessary to determine weed density thresholds for advantageous use of crop plants engineered for herbicide resistance. Oilseed rape (Brassica napus L., cv. Tower) biotypes resistant (RES) and susceptible (SUS) to atrazine were used as model crop plants, and wild oat (Avena fatua L.) was used as the model weed. Along a wild oat density gradient equivalent to 0–128 plants m^?2, RES plants consistently experienced biomass and yield reductions of approximately 10–20% compared to SUS plants. When atrazine was applied at 1.5 kg ha^?1 to control wild oats competing with RES plants, RES biomasses and yields were stabilized at the same level as that where 25–30 wild oats m^?2 reduce yields of SUS plants. This implies that with wild oat densities of 25–30 plants m^?2, it becomes agronomically advantageous to crop with RES plants plus atrazine rather than to crop with higher-yielding SUS plants.     

Effects of cultivar resistance and fungicide application on stem canker of oilseed rape (Brassica napus) and potential interseasonal transmission of Leptosphaeria spp. inoculum

Phoma stem canker is a damaging disease of oilseed rape (Brassica napus) that causes annual yield losses to UK oilseed rape growers worth approximately £100 million, despite the use of fungicides. In the UK, oilseed rape is sown in August/September and harvested in the following July. The disease epidemics are initiated by ascospores released from Leptosphaeria spp. pseudothecia (ascocarps) on stem stubble in the autumn/winter. Control of this disease is reliant on the use of cultivars with “field resistance” and azole fungicides. This study investigated the effects of cultivar resistance and application of the fungicide prothioconazole on the severity of stem canker before harvest and the subsequent production of pseudothecia on the infected stubble under natural conditions in the 2017/2018, 2018/2019, and 2019/2020 cropping seasons. The application of prothioconazole and cultivar resistance decreased the severity of phoma stem canker before harvest, and the subsequent production of Leptosphaeria spp. pseudothecia on stubble in terms of pseudothecial density. Results showed that stems with less severe stem cankers produced fewer mature pseudothecia of Leptosphaeria spp. on the infected stubble. This investigation suggests that the most sustainable and effective integrated control strategy for phoma stem canker in seasons with low quantities of inoculum is to use cultivars with medium or good field resistance and apply only one spray of prothioconazole when required.     

Major gene resistance in Brassica napus (oilseed rape) is overcome by changes in virulence of populations of Leptosphaeria maculans in France and Australia

Resistance of Brassica napus (oilseed rape, canola) conferred by three different major resistance genes has been overcome by changes in virulence of Leptosphaeria maculans populations in France and Australia. In South Australia where B. napus cultivars with major gene resistance derived from Brassica rapa ssp. sylvestris were grown extensively, resistance was rendered ineffective within 3 years of commercial release of the cultivar. Disease severity was higher on cultivars with sylvestris-derived resistance than cultivars with polygenic resistance. This Australian situation is compared to that in France, where resistance conferred by the Rlm1 gene was overcome nation-wide in 5 years under commercial cropping practices, and also where a source of resistance introgressed into B. napus from B. juncea was rendered inefficient in 3 years in experimental field plots near Rennes.     

Early autumn soil disturbance decreases persistence of volunteer summer‐annual oilseed rape (Brassica napus)

Volunteer summer‐annual oilseed rape (sOSR; Brassica napus) is an ongoing concern in Canadian crop production. Large harvest seed losses and secondary dormancy in this species generate a persistent volunteer seedbank. Yield loss in subsequent crops, potential sOSR oil profile contamination and herbicide‐resistance trait introgression create a need for effective sOSR seedbank management. This field study evaluated the effects of timing and type of implement of post‐harvest soil disturbance and seeding a winter cereal on volunteer sOSR population persistence and demographic life‐stage transition rates at five locations in Manitoba, Canada. Following sOSR harvest and supplemental seed rain, seedbank densities ranged from 6770 to 15360 and 50 to 2610 seeds m^?2 among sites in autumn and spring respectively. In contrast to European research on winter‐annual oilseed rape, early autumn soil disturbance, shortly after sOSR harvest, was the best strategy to decrease volunteer sOSR persistence (3% population persistence from autumn to spring, compared with 6% in zero tillage). Substantial autumn seedling recruitment (38% of the autumn seedbank) and subsequent winterkill contributed to lower population persistence. Soil disturbance in spring stimulated spring seedling recruitment compared with other disturbance timings (11% and 3% of the spring seedbank, respectively). The implement used for soil disturbance and seeding winter wheat (Triticum aestivum) had minimal effect on population persistence. This research showed that timing of post‐harvest soil disturbance should be utilised as an effective tactic to decrease population persistence of volunteer sOSR via stimulation of autumn seedling recruitment and concomitant winterkill.     

Effects of different cultivation systems (soil management,nitrogen fertilization) on the epidemics of fungal diseases in oilseed rape (Brassica napus L. var. napus)

Journal of Plant Diseases and Protection - The effects of different cultivation systems on disease incidence and severity in oilseed rape were investigated on two sites in Schleswig-Holstein in the...     

Impact of alpha-cypermethrin on honey bees foraging on spring oilseed rape (Brassica napus) flowers in field conditions

BACKGROUND: Cruciferous oil-bearing crops have gained in importance worldwide. The expansion of the growing area of these crops has caused a proliferation of pests. Exposure to organophosphate, carbamate and pyrethroid insecticides has been associated with bee poisoning in food crops. This study examines the repellent effect of alpha-cypermethrin on the number of foraging honey bees, Apis mellifera L., on fields of spring oilseed rape, Brassica napus L. var. oleifera. RESULTS: The first experiment was conducted on differently sprayed 10 m(2) experimental plots where alpha-cypermethrin was applied at different times. Another experiment was conducted on a 4 ha seed production field divided into two parts: one part was treated with alpha-cypermethrin and the other was not treated with this insecticide. The results show that there was no difference in the number of honey bees between alpha-cypermethrin-treated and untreated patches. The result persisted through three observation years, regardless of varying flower and honey bee densities. CONCLUSION: No repellent effect of the insecticide on honey bees was found even 24 h after spraying. The density of oilseed rape flowers most likely played a major role in choosing the foraging area.     

The control of sclerotinia stem rot on oilseed rape (Brassica napus): current practices and future opportunities

Sclerotinia stem rot (SSR) caused by the phytopathogenic fungus Sclerotinia sclerotiorum is a major disease of oilseed rape (Brassica napus). During infection, large, white/grey lesions form on the stems of the host plant, perturbing seed development and decreasing yield. Due to its ability to produce long‐term storage structures called sclerotia, S. sclerotiorum inoculum can persist for long periods in the soil. Current SSR control relies heavily on cultural practices and fungicide treatments. Cultural control practices aim to reduce the number of sclerotia in the soil or create conditions that are unfavourable for disease development. These methods of control are under increased pressure in some regions, as rotations tighten and inoculum levels increase. Despite their ability to efficiently kill S. sclerotiorum, preventative fungicides remain an expensive gamble for SSR control, as their effectiveness is highly dependent on the ability to predict the establishment of microscopic infections in the crop. Failure to correctly time fungicide applications can result in a substantial cost to the grower. This review describes the scientific literature pertaining to current SSR control practices. Furthermore, it details recent advances in alternative SSR control methods including the generation of resistant varieties through genetic modification and traditional breeding, and biocontrol. The review concludes with a future directive for SSR control on oilseed rape.     

Overexpression of an nsLTPs-like antimicrobial protein gene (LJAMP2) from motherwort (Leonurus japonicus) enhances resistance to Sclerotinia sclerotiorum in oilseed rape (Brassica napus)

Sclerotinia stem rot caused by Sclerotinia sclerotiorum is one of the most important diseases of oilseed rape worldwide and leads to considerable yield losses. In this study, a non-specific lipid transfer protein-like antimicrobial protein gene (LJAMP2) from motherwort (Leonurus japonicus) was introduced into oilseed rape (Zhongyou 821) by Agrobacterium-mediated transformation. In vitro experiments revealed that the mycelial growth of S. sclerotiorum was significantly inhibited when supplied with crude leaf extracts from transgenic oilseed rape plants overexpressing LJAMP2. Furthermore, in vivo studies showed that transgenic LJAMP2 plants had enhanced resistance to S. sclerotiorum. Semi-quantitative RT-PCR analysis showed that the LJAMP2 gene was transcribed in all transformed plants. In addition, we also found that overexpression of LJAMP2 in transgenic plants caused constitutive activation of the defense-related gene PR-1 and an increase of H₂O₂ production, but did not enhance PDF1.2 expression. Our results suggest that constitutive expression of the LJAMP2 gene from motherwort seeds might be exploited to improve the resistance of oilseed rape against S. sclerotiorum.