Viruses infecting winter oilseed rape (Brassica napus ssp. oleifera)

Turnip mosaic virus (TuMV) and cauliflower mosaic virus (CaMV) have been found infecting field crops of winter oilseed rape (Brassica napus ssp. oleifera) in South Warwickshire. Other viruses found include broccoli necrotic yellows virus (BNYV) and a member of the beet western yellows virus group. Systemic leaf symptoms caused by TuMV varied within and between cultivars; the three predominant reaction types were classified as necrotic, mosaic and immune. Some recently introduced cultivars of oilseed rape were more severely affected by TuMV infection than older cultivars. Reactions to CaMV were less varied and immunity was not found. The seed yield from TuMV and CaMV-infected plants was less than that of healthy control plants. This effect was due to infected plants producing either fewer seeds, smaller seeds or both. Germination of seeds from infected plants was unaffected if sown soon after harvest. After storage for one year the germination of seed from a virus infected plant was significantly less than that of seed from a virus-free plant. All commercial cultivars tested were experimentally susceptible to turnip yellow mosaic virus (TYMV) and some American strains of cucumber mosaic virus (CMV).     

A review of sources of resistance to turnip yellows virus (TuYV) in Brassica species

Turnip yellows virus (TuYV; previously known as beet western yellows virus) causes major diseases of Brassica species worldwide resulting in severe yield-losses in arable and vegetable crops. It has also been shown to reduce the quality of vegetables, particularly cabbage where it causes tip burn. Incidences of 100% have been recorded in commercial crops of winter oilseed rape (Brassica napus) and vegetable crops (particularly Brassica oleracea) in Europe. This review summarises the known sources of resistance to TuYV in B. napus (AACC genome), Brassica rapa (AA genome) and B. oleracea (CC genome). It also proposes names for the quantitative trait loci (QTLs) responsible for the resistances, Tu rnip Y ellows virus R esistance (TuYR), that have been mapped to at least the chromosome level in the different Brassica species. There is currently only one known source of resistance deployed commercially (TuYR1). This resistance is said to have originated in B. rapa and was introgressed into the A genome of oilseed rape via hybridisation with B. oleracea to produce allotetraploid (AACC) plants that were then backcrossed into oilseed rape. It has been utilised in the majority of known TuYV-resistant oilseed rape varieties. This has placed significant selection pressure for resistance-breaking mutations arising in TuYV. Further QTLs for resistance to TuYV (TuYR2-TuYR9) have been mapped in the genomes of B. napus, B. rapa and B. oleracea and are described here. QTLs from the latter two species have been introgressed into allotetraploid plants, providing for the first time, combined resistance from both the A and the C genomes for deployment in oilseed rape. Introgression of these new resistances into commercial oilseed rape and vegetable brassicas can be accelerated using the molecular markers that have been developed. The deployment of these resistances should lessen selection pressure for resistance-breaking isolates of TuYV and thereby prolong the effectiveness of each other and extant resistance.     

Performance of cabbage stem flea beetle larvae (Psylliodes chrysocephala) in brassicaceous plants and the effect of glucosinolate profiles

The cabbage stem flea beetle (CSFB), Psylliodes chrysocephala L. (Coleoptera: Chrysomelidae), is one of the most important pests in European winter oilseed rape production. Adult beetles feed on young leaves whereas larvae mine within the petioles and stems. Larval infestation can cause significant crop damage. In this study, the host quality for CSFB of four oilseed rape (Brassica napus L.) cultivars and seven other brassicaceous species with different glucosinolate (GSL) profiles was assessed under controlled conditions. Larval instar weights and mortality were measured after 14 and 21 days of feeding in the petioles of test plants. To study the impact of GSL on the performance of larvae, the GSL contents in petioles from non-infested and infested plants were analysed before, and 21 days after, the start of larval infestation. Larval performance was not significantly different between the four cultivars of oilseed rape, but differed considerably among the other brassicaceous species tested. In comparison to the weight of larvae in the standard B. napus cv. Robust, the larval weight was higher in turnip rape (Brassica rapa L. var. silvestris) and significantly reduced in white mustard (Sinapis alba L.), oil radish (Raphanus sativa L. var. oleiformis), and cabbage (Brassica oleracea L. convar. capitata var. alba). The duration of larval development increased in white mustard and oilseed radish. The GSL profiles of the petioles showed little difference between non-infested and infested plants of oilseed rape whereas the content of aliphatic GSL increased in the infested turnip rape plants. In contrast, the aliphatic and benzenic GSL decreased in infested Indian rape (B. rapa subsp. dichotoma Roxb.). Larval weight was not correlated with the total GSL content of plants, neither before infestation nor 21 days after. Larval weight was positively correlated with progoitrin and 4-hydroxyglucobrassicin. White mustard, which provides inferior host quality for larval development, has the potential to introduce insect resistance into high-yielding oilseed rape cultivars in breeding programmes.     

Studies on beet western yellows virus in oilseed rape (Brassica napus ssp. oleifera) and sugar beet (Beta vulgaris)

Oilseed rape (Brassica napus L. ssp. oleifera) was studied as a potential overwintering host for the sugar-beet yellowing viruses, beet yellows virus (BYV) and beet mild yellowing virus (BMYV), and their principal vector, Myzus persicae. In spring 1982, plants infected with a virus which reacted positively in enzyme-linked immunosorbent assay (ELISA) with BMYV antibody globulin were found in oilseed-rape crops; none of the plants contained virus which reacted with BYV antibody globulin. This virus was subsequently identified as beet western yellows virus (BWYV). No leaf symptoms could be consistently associated with infection of oilseed rape, but the virus was reliably detected by sampling any leaf on an infected oilseed-rape plant. Some isolates from oilseed rape did infect sugar beet in glasshouse tests, but the proportions of inoculated plants which became infected were low. Apparently there is therefore little danger of much direct transmission of BWYV by M. persicae from oilseed rape to sugar beet in spring. BWYV was introduced to and spread within oilseed-rape crops in autumn by M. persicae, and autumn-sown oilseed rape proved to be a potentially important overwintering host for M. persicae. In a survey of 80 autumn-sown crops of oilseed rape in East Anglia, northern England and Scotland in spring 1983, 78 were shown to be extensively infected with BWYV. Experimental plots of oilseed rape with 100% BWYV-infection yielded approximately 13.4% less oil than plots with 18% virus infection, the result of a decrease in both seed yield and oil content.     

Resistance of transformed and non-transformed oilseed rape cv. HM-81 to the infection with cauliflower mosaic,turnip yellow mosaic and turnip mosaic viruses

Resistance of transformed and non-transformed spring oilseed rape cv. HM-81 to the infection with cauliflower mosaic virus (CaMV), turnip yellow mosaic virus (TYMV) and turnip mosaic virus (TuMV) was studied, to determine the influence of transformation on susceptibility of plants to viruses. For experiments the non-segregating R 1 generation of primary transformant HM-81-JZ and control plants of cv. HM-81 were used. The primary transformant was obtained by inoculation of stems withAgrobacterium rhizogenes 15834. All transformed plants of R 1 generation had typically „transformed“ phenotype. No significant differences were revealed in the resistance of both transformed and non-transformed plants to each virus, as proved by qualitative and quantitative ELISA and visual evaluation of symptoms. Transformed plants infected with turnip yellow mosaic virus showed significantly lower reduction of green mass yield than non-transformed. In the case of CaMV and TuMV infection reduction of yield of transformed and non-transformed plants was almost the same.     

Effects of turnip mosaic virus and turnip yellow mosaic virus on the survival, growth and reproduction of wild cabbage (Brassica oleracea)

Wild plants of Brassica oleracea (wild cabbage) are commonly infected with turnip mosaic poty virus (TuMV), turnip yellow mosaic tymovirus (TYMV) and several other viruses. A field experiment in which plants were inoculated either with TuMV or TYMV showed that virus infection significantly reduced survival, growth and reproduction. Relative to water inoculated-controls, plants infected with TYMV had greater mortality, were shorter, had a smaller leaf area and number, showed a greater amount of damage from herbivory and chlorosis, were less likely to flower and produced fewer pods and lower total seed output. Plants infected with TuMV did not appear to be adversely affected at first; however, mortality after 18 months was higher than control plants. Although TuMV infection had no effect on the number of plants flowering, the infected plants did produce fewer pods and a lower total seed output. We conclude that both viruses can significantly affect vegetative and reproductive performance of wild cabbage and hence that introgression of virus resistance (particularly when conferred by a major gene or a transgene) from a crop might increase plant fitness in natural populations of this species. Ecological risk assessments of virus resistance transgenes must do more than survey adult plants in natural populations for the presence of the target virus. Failure to detect the virus could be due to high mortality on infection with the virus.     

The prevalence and spatial distribution of viruses in natural populations of Brassica oleracea

We report a survey of four viruses (beet western yellows luteovirus (BWYV), cauliflower mosaic caulimovirus (CaMV), turnip mosaic potyvirus (TuMV), turnip yellow mosaic tymovirus (TYMV)) in five natural populations of Brassica oleracea in Dorset (UK). All four viruses were common; 43% of plants were infected with BWYV, 60% with CaMV, 43% with TuMV and 18% with TYMV. For each virus there were significant differences in the proportion of infected plants among populations, which were not completely explained by differences in the age of plants. Multiple virus infections were prevalent, with 54% of plants having two or more virus types. There were statistically significant associations between pairs of viruses. The CaMV was positively associated with the other three viruses, and BWYV was also positively associated with TuMV. There was no detectable association between BWYV and TYMV, whereas TuMV and TYMV were negatively associated. We suggest these associations result from BWYV, CaMV and TuMV having aphid vectors in common, as aphids are attracted to plants that already have a virus infection. Infected plants were distributed randomly or were very weakly aggregated within populations. The implications of widespread multiple virus infections in natural plant populations are discussed with respect to the release of transgenic plants expressing virus-derived genes.     

播种期和种植密度对冬油菜籽粒产量和含油率的影响

通过田间试验研究了播种期和种植密度对冬油菜籽粒产量和含油率的影响.结果表明： 播种期主要影响分枝花序籽粒产量,而种植密度不仅影响分枝花序籽粒产量,还对主花序籽粒产量产生一定影响;籽粒含油率不受播种期的影响.主花序籽粒产量占单株籽粒产量的比例随种植密度的增加而升高,主花序籽粒含油率比分枝花序高约1%,因此小区籽粒含油率随种植密度的增加显著升高.研究区冬油菜播种期不能晚于10月中旬,10月下旬播种会显著降低籽粒产量;种植密度在每平方米36～48株可以提高冬油菜籽粒产量和含油率.     

Transformation of microspore-derived embryos of winter oilseed rape (Brassica napus L.) by usingAgrobacterium tumefaciens

Haploid microspore-derived embryos (MDEs) constitute a unique material for the introduction of new traits into winter oilseed rape (Brassica napus). MDEs have been transformed by usingAgrobacterium tumefaciens strains EHA105 and LBA4404, both carrying the binary vector pKGIB containing theuidA gene encoding β-glucuronidase (GUS) and thebar gene as a marker of resistance to phosphinotricin. Transformed embryos expressed GUS and regenerated plants that were resistant to herbicide Basta, as confirmed by a leaf-painting test. Progeny plants of the transformant T-39 were all transgenic, as they inherited T-DNA from their doubled haploid parental plant. Southern-blot analysis confirmed the integration and transmission of T-DNA into T₁ plants. Transformation of MDEs facilitates the obtaining of winter oilseed rape homozygous for the introduced genes.     

Exploitation of host plant preferences in pest management strategies for oilseed rape (Brassica napus)

New control strategies for insect pests of arable agriculture are needed to reduce current dependence on synthetic insecticides, the use of which is unsustainable. We investigated the potential of a simple control strategy to protect spring‐sown oilseed rape, Brassica napus L. (Brassicaceae), from two major inflorescence pests: the pollen beetle, Meligethes aeneus (Fabricius) (Coleoptera: Nitidulidae), and the seed weevil, Ceutorhynchus assimilis (Paykull) (Coleoptera: Curculionidae), through exploitation of their host plant preferences. The strategy comprised, for the main crop, Starlight [an oilseed rape cultivar with relatively low proportions of alkenyl glucosinolates in the leaves (thereby releasing lower levels of attractive isothiocyanates than conventional cultivars)] and turnip rape, Brassica rapa (L.) (Brassicaceae), as a trap crop. We tested the system in laboratory, polytunnel semifield arena, and field experiments. The odours of Starlight were less attractive in olfactometer tests to both pests than those from a conventional cultivar, Canyon, and the plants were less heavily colonized in both polytunnel and field experiments. Turnip rape showed good potential as a trap crop for oilseed rape pests, particularly the pollen beetle as its odour was more attractive to both pests than that of oilseed rape. Polytunnel and field experiments showed the importance of relative growth stage in the system. As turnip rape flowers earlier than oilseed rape, beetles would be maintained on turnip rape past the damage‐susceptible growth stage of oilseed rape. The development of a pest control regime based on this strategy is discussed.     

Injury to oilseed rape caused by mirid bugs (Lygus) (Heteroptera: Miridae) and its effect on seed production

Injury by Lygus spp. to oilseed rape, Brassica napus L. and Brassica campestris L., was assessed based on laboratory and field studies in Alberta, Canada. The visible injuries consisted of lesions on the surfaces of stems, buds, flowers and pods similar to those described for other crops. They caused buds and flowers to abscise and seeds to collapse, and reduced the weight of healthy seeds produced per pod. The plants compensated for bud loss so that no net reduction in the number of pods occurred. However, in some situations the damage to buds resulted in a reduced seed yield that increased with the amount of injury. The plants also compensated for flower loss so that no net reduction in the number of pods occurred, but seed yield declined as injury increased. Plants did not compensate for seeds that collapsed as a result of lygus feeding. The feeding activity of lygus bugs reduced seed yield in oilseed rape.     

Feeding preferences of woodpigeons and flea-beetles for oilseed rape and turnip rape

Winter turnip rape Brassica rapa ssp. oleifera was shown to have reduced palatability to woodpigeons, enhanced susceptibility to adult flea beetle feeding and reduced susceptibility to larval flea-beetle infection when compared with winter oilseed rape (B. napus) cultivars in field trials. Levels of leaf waxes were negatively correlated with feeding preferences of adult flea beetles. Analysis of volatiles from damaged leaves showed that while all cultivars produce a similar range of nitriles, cyanoepithioalkanes and isothiocyanates, derived from 3-butenyl, 4-pentenyl and phenylethyl glucosinolates, B. rapa leaves produced relatively high levels of 1-methylpropyl isothiocyanate, although there was significant plant-to-plant variation. The possible involvement of this mustard oil glycoside and variation in epicuticular waxes in plant-herbivore interactions are discussed.     

Effect of a turnip rape <Emphasis Type="Italic">(Brassica rapa)</Emphasis> trap crop on stem-mining pests and their parasitoids in winter oilseed rape <Emphasis Type="Italic">(Brassica napus)</Emphasis>

Concerns about the negative effects of chemical control of oilseed rape (Brassica napus L.) pests on non-target species, human safety, and development of insecticide resistance, require alternative control strategies such as the use of trap crops and biocontrol to be developed. Psylliodes chrysocephala(L.) (Coleoptera: Chrysomelidae) (cabbage stem flea beetle) and Ceutorhynchus pallidactylus (Marsh.) (Coleoptera: Curculionidae) (cabbage stem weevil) are two major stem-mining pests of oilseed rape. This study investigated the phenology of these pests and their main parasitoids in the UK, the potential use of turnip rape (Brassica rapa L.) as a trap crop to reduce oilseed rape infestation, and the effects of insecticide treatment on pest incidence and larval parasitism. Water trap samples, plant dissections and pest larval dissections were done to determine: the incidence of adult pests and their parasitoids, the level of plant infestation by the pests and percentage larval parasitism, respectively. The turnip rape trap crop borders reduced P. chrysocephalabut not C. pallidactylus infestation of oilseed rape plots. Treatment of the trap crop with insecticide had little effect on either pest or parasitoid incidence in the oilseed rape. TersilochusmicrogasterSzép. andT. obscurator Aub. (Hymenoptera: Ichneumonidae) were the main larval parasitoids of P. chrysocephalaand C. pallidactylus, respectively. Tersilochus microgasteris reported for the first time in the UK. The implications for integrated pest management are discussed.     

Inheritance of oilseed rape (Brassica napus) RAPD markers in a backcross progeny with Brassica campestris

Different cultivars/transgenic lines of oilseed rape (Brassica napus) were crossed (as females) with different cultivars/populations of Brassica campestris. All cross combinations produced seed, with an average seed set per pollination of 9.8. Backcrossing of selected interspecific hybrids (as females) to B. campestris resulted in a much lower seed set, average 0.7 seed per pollination. In the single backcross progeny where a large enough population (92 plants) was obtained for analysis, 33 B. napus specific RAPD markers were investigated to determine the extent of transfer of oilseed rape genetic material into this population. Markers were transferred to the backcross generation with frequencies ranging from 26% to 91%. Almost all of the markers (30/33) were transferred in a frequency not significantly different from 50%. Analysis of the pairwise segregation of markers revealed that 23 markers could be assigned to six linkage groups, most probably reflecting six B. napus C-chromosomes. The presence of backcross plants with recombinant genotypes suggests that complex genetic processes can take place during interspecific hybridisation and backcrossing in these Brassica species. The implications of our results for the possible choice of integration sites of transgenes in oilseed rape are discussed.     

The germination and emergence of seeds of winter oilseed rape stored and sown in admixture with pelleted methiocarb

The possibility that pellets of the molluscicide Draza (a.i. methiocarb) might reduce field emergence of seeds of winter oilseed rape (Brassica napus, cv. Jet Neuf) when sown admixed, laboratory germination when stored admixed, or field emergence when both stored and sown admixed was the subject of three separate investigations. Admixture at sowing did not influence the emergence of five seed lots (P > 0·05). Neither did admixture influence loss in germination or change in moisture content during 2 months' storage of one lot in four different regimes (P > 0·05). Field emergence following 2 months' storage in paper bags was not influenced by admixture (P > 0·05), whereas a 12% reduction in field emergence resulting from storage in admixture within polyethylene bags was detected (P < 0·05). Thus short-term storage within paper sacks of winter oilseed rape seeds in admixture with slug pellets containing methiocarb did not damage the seeds. The emergence of five seed lots differed considerably within and between field sowings. Tentative advice on the calculation of sowing rates for different seed lots of winter oilseed rape is provided to account for this variation.     

Endophytic Bacteria Induce Growth Promotion and Wilt Disease Suppression in Oilseed Rape and Tomato

To determine whether bacteria isolated from within plant tissue can have plant growth-promotion potential and provide biological control against soilborne diseases, seeds and young plants of oilseed rape (Brassica napus L. cv. Casino) and tomato (Lycopersicon lycopersicum L. cv. Dansk export) were inoculated with individual bacterial isolates or mixtures of bacteria that originated from symptomless oilseed rape, wild and cultivated. They were isolated after surface sterilization of living roots and stems. The effects of these isolates on plant growth and soilborne diseases for oilseed rape and tomato were evaluated in greenhouse experiments. We found isolates that not only significantly improved seed germination, seedling length, and plant growth of oilseed rape and tomato but also, when used for seed treatment, significantly reduced disease symptoms caused by their vascular wilt pathogens Verticillium dahliae Kleb and Fusarium oxysporum f. sp. lycopersici (Sacc.), respectively.     

Infection of brassica leaf protoplasts by turnip yellow mosaic virus

Summary Conditions optimal for the infection of Brassica leaf protoplasts by turnip yellow mosaic virus (TYMV) were found to be: pH 5.4–5.8 with citrate buffer; concentration of poly-L-ornithine, 0.8–1.0 g/ml; concentration of TYMV, 0.2–1.0 g/ml. More than 90% of Brassica rapa and B. sinensis protoplasts were infected under these conditions. TYMV replication in Brassica protoplasts was followed by quantitating the virus extracted from protoplasts and separated by sucrose density gradient centrifugation. Brassica protoplasts produced 1 to 2x10⁶ TYMV particles per cell within 48 hrs. Infection by TYMV induced the formation of polyplasts (aggregates of chloroplasts) in Brassica protoplasts. Polyplast formation paralleled the appearance of TYMV-specific immunofluorescence and could thus be used conveniently to determine the number of infected protoplasts. TYMV replication in Brassica protoplasts was resistant to actinomycin D and chloramphenicol, but was completely inhibited by cycloheximide.     

The competitive effects of volunteer barley (Hordeum vulgare) on the growth of oilseed rape (Brassica napus)

The effects of competition from volunteer barley (Hordeum vulgare) on the growth and yield of oilseed rape (Brassica napus) were investigated in four experiments over three seasons. The growth of rape in the autumn was reduced by 50 - 91 % by competition from 400 barley plants m^-2. A lower barley density of 200 plants m^-2 had less effect but still reduced growth of rape by 65 - 81% in two of the experiments and 25 - 40% in the other two. During winter and spring the barley decreased in vigour and in the spring the rape started to recover, especially on the early drilled (23 - 30 August) plots. The rape sown in mid-September recovered less quickly. In Experiment 3, herbicides applied in November to control barley did not result in increased growth of rape in winter but led to greater recovery in spring. The barley died during the winter in Experiments 2 and 4, even in the absence of herbicides. Despite the marked effects of barley on the growth of rape in the autumn, yields on plots that had previously contained 200 barley plants m^-2 were reduced by a maximum of only 16% in three of the experiments. In Experiment 3, where the barley was most competitive, this density and 400 plants m^-2 lowered yields by 39% and 78%, respectively. Where a herbicide was used in November to control the barley these yield losses were reduced to 5%. In many rape crops the cost of herbicide treatment would be greater than the financial returns from the expected increase in yield resulting from the control of weeds. Possible reasons for the small loss in yield of rape from barley densities that had substantial effects on the growth of rape in the autumn are discussed.     

A strain of radish mosaic virus occurring in turnip in Yugoslavia

Fields of turnip (Brassica rapa L. var. rapa) in Yugoslavia often contained plants infected with radish mosaic virus (RaMV). Yugoslav isolates resemble the type strain in host range, symptoms, physical properties, particle morphology and flea-beetle transmission and are serologically closely related to the type strain, but did not infect radish and are designated as the European strain. Light microscopy of infected turnip and some other cruciferous species revealed the characteristic inclusion bodies in the cytoplasm.     

Trace elements bioavailability to winter wheat (Triticum aestivum L.) grown subsequent to high biomass plants in a greenhouse study

Multielement-contaminated agricultural land requires the adaptation of agronomic practices to meet legal requirements for safe biomass production. The incorporation of bioenergy plants with, at least, moderate phytoextraction capacity into crop rotations with cereals can affect trace elements (TE) phytoavailability and, simultaneously, constitute economic revenues for farmers outside the food or forage sector. Hence, in a crop rotation pot study sunflower (Helianthus annuus L.), modified for high biomass and TE accumulation by chemical mutagenesis, was compared to winter oilseed rape (Brassica napus L.) as pre-crop. On two agricultural soils with different TE loads, the crops´ potential for phytoextraction and for impacts on TE uptake by subsequent winter wheat (Triticum aestivum L.) was studied. The results showed that rape tolerated high-level mixed contamination with metals (Cd, Pb and Zn) and As more than sunflower. In both soils, labile metals concentration increased and soil acidity remained high following sunflower. Furthermore, enhanced grain As accumulation in subsequent wheat was observed. By contrast, soil acidity and Cd or Zn accumulation of subsequent wheat decreased following rape. In the short term, moderate phytoextraction was superimposed by nutrient use or rhizosphere effects of pre-crops, which should be carefully monitored when designing crop rotations for contaminated land.