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).     

Photosynthesis in leaves and siliques of winter oilseed rape (Brassica napus L.)

The rate of photosynthesis and its relation to tissue nitrogen content was studied in leaves and siliques of winter oilseed rape (Brassica napus L.) growing under field conditions including three rates of nitrogen application (0, 100 or 200 kg N ha^-1) and two levels of irrigation (rainfed or irrigated at a deficit of 20 mm). The predominant effect of increasing N application under conditions without water deficiency was enhanced expansion of photosynthetically active leaf and silique surfaces, while the rate of photosynthesis per unit leaf or silique surface area was similar in the different N treatments. Thus, oilseed rape did not increase N investment in leaf area expansion before a decline in photosynthetic rate per unit leaf area due to N deficiency could be avoided. Much less photosynthetically active radiation penetrated into high-N canopies than into low-N canopies. The specific leaf area increased markedly in low light conditions, causing leaves in shade to be less dense than leaves exposed to ample light. In both leaves and siliques the photosynthetic rate per unit surface area responded linearly to increasing N content up to about 2 g m^-2, thus showing a constant rate of net CO₂ assimilation per unit increment in N (constant photosynthetic N use efficiency). At higher tissue N contents, photosynthetic rate responded less to changes in N status. Expressed per unit N, light saturated photosynthetic rate was three times higher in leaves than in silique valves, indicating a more efficient photosynthetic N utilization in leaves than in siliques. Nevertheless, from about two weeks after completion of flowering and onwards total net CO₂ fixation in silique valves exceeded that in leaves because siliques received much higher radiation intensities than leaves and because the leaf area declined rapidly during the reproductive phase of growth. Water deficiency in late vegetative and early reproductive growth stages reduced the photosynthetic rate in leaves and, in particular, siliques of medium- and high-N plants, but not of low-N plants.     

Slow vacuolar channels of non-embryogenic and embryogenic cultures of winter wheat

Slowly activating vacuolar channels (SV), were examined in embryogenic and non-embryogenic cultures of winter wheat using a patch-clamp technique. Four different types of cultures were examined: embryogenic and non-embryogenic calli from embryos, embryogenic and non-embryogenic calli from inflorescences. In a cell-attached mode single SV channel events were recorded. Unitary conductance of single SV channels was between 37 pS and 48 pS and did not significantly depend on the kind of the culture, although it was a tendency that SV channels of embryogenic calli possessed lower unitary conductance than those of non-embryogenic. 2,4-D caused significant lowering of unitary conductance from 48±6 pS in the control culture of embryogenic embryos to 28±6 pS in vacuoles treated. The SV channel density was estimated as 0.34 μm⁻².     

Physiological and yield effects of uniconazole on winter rape (Brassica napus L.)

Uniconazole at various concentrations on rape, at the three-leaf stage, was examined for physiologic and yield effects. Foliar sprays of 10, 25, and 50 mg/liter significantly reduced seedling height, and increased shoot width (stem width before elongation), number of green leaves, and total dry weight at transplanting. Chlorophyll content, superoxide dismutase and catalase activities, root oxidizability (capacity for root oxidation), and ethylene production were also increased. Additionally, the number of branches and pods/plant were increased; and a 7.4, 8.5, and 4.3% increase of seed yield over the controls was observed with treatments at 10, 25, and 50 mg/liter uniconazole, respectively. No significant effects were observed on plant maturity, the seed oil content, or the erucic acid and glucosinolate content. Total oil production significantly increased with 10, 25, and 50 mg/liter by 9.9, 10.6, and 6.8%, respectively, over the controls. These results suggested that uniconazole-induced high productivity was accompanied by increased levels of activities of various antioxidants, including superoxide dismutase and catalase, and by the improvement of root oxidizability and plant vigor.Abbreviations SOD superoxide dismutase - CAT catalase - NBT nitro blue tetrazolium - TTC red tetrazolium - IAA indoleacetic acid     

Effects of light leaf spot (Pyrenopeziza brassicae) on yield of winter oilseed rape (Brassica napus)

The relationship between development of light leaf spot and yield loss in winter oilseed rape was analysed, initially using data from three experiments at sites near Aberdeen in Scotland in the seasons 1991/92, 1992/93 and 1993/94, respectively. Over the three seasons, single-point models relating yield to light leaf spot incidence (% plants with leaves with light leaf spot) at GS 3.3 (flower buds visible) generally accounted for more of the variance than single-point models at earlier or later growth stages. Only in 1992/93, when a severe light leaf spot epidemic developed on leaves early in the season, did the single-point model for disease severity on leaves at GS 3.5/4.0 account for more of the variance than that for disease incidence at GS 3.3. In 1991/92 and 1992/3, when reasonably severe epidemics developed on stems, the single-point model for light leaf spot incidence (stems) at GS 6.3 accounted for as much of the variance. Two-point (disease severity at GS 3.3 and GS 4.0) and AUDPC models (disease incidence/severity) accounted for more of the variance than the single-point model based on disease incidence at GS 3.3 in 1992/93 but not in the other two seasons. Therefore, a simple model using the light leaf spot incidence at GS 3.3 (x) as the explanatory variable was selected as a predictive model to estimate % yield loss (y_r): y_r= 0.32x– 0.57. This model fitted all three data sets from Scotland, When data sets from Rothamsted, Rosemaund and Thurloxton in England were used to test it, this single-point predictive model generally fitted the data well, except when yield loss was clearly not related to occurrence of light leaf spot. However, the regression lines relating observed yield loss to light leaf spot incidence at GS 3.3 often had smaller slopes than the line produce, by the model based on Scottish data.     

Diagnosis of light leaf spot (Pyrenopeziza brassicae) on winter oilseed rape (Brassica napus) in the UK

Light leaf spot lesions were generally first observed as light green areas on leaves of UK winter oilseed rape crops in January or February and later became brittle and bleached. Elongated lesions, which were brown with indistinct edges, developed on stems in the spring and summer, when lesions were also observed on flower buds, pedicels and pods. Development of diagnostic white pustules (spore masses of Pyrenopeziza brassicae, which erupt through surfaces of infected tissues) for confirmation of light leaf spot infection on symptomless plants or plants with indistinct or ambiguous symptoms in the autumn, winter or spring was enhanced by incubating plants in polyethylene bags. In experiments with artificially inoculated plants, glasshouse-grown plants exposed in infected crops and plants sampled from crops, white pustules developed at all incubation temperatures from 2^oC to 20^oC on infected leaves of different cultivars. The period of incubation required before the appearance of pustules decreased as the time that had already elapsed since the initial infection increased. The longest periods of incubation were required at the lowest temperatures (2^oC or 5^oC) but leaves senesced and abscised from plants most quickly at the highest temperatures (15^oC or 20^oC), suggesting that the optimal incubation temperature was between 10^oC and 15^oC.     

Temperature and daylength control of flower initiation in winter oilseed rape (Brassica napus L.)

The influence of low temperature and daylength on pre-floral growth and flower initiation in winter oilseed rape cv. Mikado was examined under controlled environment conditions at the University of Newcastle upon Tyne during 1985 and 1986.
The vernalisation requirement of Mikado was most effectively fulfilled by temperatures of 6 °C and 9 °C. Plants maintained at both higher and lower temperatures had an extended pre-floral growth phase. The transition from vegetative to reproductive growth in plants maintained at 12 °C was delayed by slow accumulation of the cold requirement, whereas flower initiation appeared to be delayed by limited leaf production, dry matter accumulation and/or assimilate availability in plants grown at 3 °C. The mechanism of floral induction remained unresolved but it was clear that flower initiation was not controlled by low temperature per se . Short days partially substituted for the cold requirement at 12 °C but photoperiodic induction of flower initiation was less important than the influence of low temperature.     

Frequency and distance of pollen dispersal from transgenic oilseed rape (Brassica napus)

The objective of this study was to evaluate pollen dispersal inBrassica napus (oilseed rape). The selectable marker, used to follow pollen movement, was a dominant transgene (bar) conferring resistance to the herbicide glufosinate-ammonium. Transgenic and non-transgenic plants of the cultivar Westar were planted in a 1.1 ha field trial, with the transgenic plants in a 9 m diameter circle at the centre, surrounded by non-transgenic plants to a distance of at least 47 m in all directions. A 1 m circle of non-transgenic plants was sown in the centre of the transgenic area to allow estimation of the level of pollen dispersal when plants were in close contact. Honeybee hives were placed at the trial site to optimize the opportunity for cross-pollination. During the flowering period, regular observations were made of the number of plants flowering and the number and type of insects present in 60 1 m² areas. These areas were located uniformly around the plot at distances of 1, 3, 6, 12, 24, 36 and 47 m from the edge of the 9 m circle of transgenic plants. Seed samples were harvested from each of the 7 distances so that approximately 20% of the circumference of the plot was sampled at each distance. The centre non-transgenic circle was also sampled. Plants were grown from the seed samples and sprayed with glufosinate to estimate the frequency of pollen dispersal at each distance. In order to screen enough samples to detect low frequency cross-pollination events, seed samples were tested in the greenhouse and on a larger scale in the field. Results were confirmed by testing progeny for glufosinate resistance and by Southern blot analysis. The estimated percentage of pollen dispersal in the non-transgenic centre circle was 4.8%. The frequency was estimated to be 1.5% at a distance of 1 m and 0.4% at 3 m. The frequency decreased sharply to 0.02% at 12 m and was only 0.00033% at 47 m. No obvious directional effects were detected that could be ascribed to wind or insect activity.     

Low probability of chloroplast movement from oilseed rape (Brassica napus) into wild Brassica rapa

Pollen-mediated movement of transgenes from transplastomic oilseed rape (Brassica napus) into wild relatives will be avoided if chloroplasts are maternally transmitted. We assess the probability of chloroplast exchange between conventional oilseed rape and wild Brassica rapa to model the future behavior of transplastomic cultivars. Primers specific to cpDNA were used to demonstrate maternal inheritance of chloroplasts in 47 natural hybrids between cultivated B. napus and wild B. rapa. We conclude that there will be no or negligible pollen-mediated chloroplast dispersal from oilseed rape. Transgene introgression could still occur in mixed populations, however, if B. napus acted as the recurrent female parent. Rate of transfer would then depend on the abundance of mixed populations, their persistence as mixtures, and hybridization frequency within stands. A low incidence of sympatry (0.6-0.7%) between wild B. rapa and cultivated B. napus along the river Thames, UK, in 1997 and 1998, suggests mixed stands will form only rarely. Eighteen feral populations of B. napus also showed a strong tendency toward rapid decline in plant number, seed return, and ultimately, extinction within 3 years. Conversely, hybrid production is significant in mixed stands, and the absence of control practices means that oilseed rape will have slightly greater persistence. We infer that some introgression from transplastomic B. napus into B. rapa is inevitable in mixed populations even though such populations will occur infrequently and will tend to lose B. napus plants relatively quickly. Chloroplast exchange will be extremely rare and scattered.     

Mapping a high oleic acid mutation in winter oilseed rape (Brassica napus L.)

Two winter oilseed rape mutant lines, 7488 and 19661, with a high oleic (HO) acid content in the seed oil were characterized phenotypically. In both mutant lines the HO trait was monogenically inherited. Segregation analysis in an F₂ population derived from a cross between 7488 and 19661 showed the two mutations to be allelic. From a comparison of seed, leaf and root fatty acid composition it was concluded that fad2, the endoplasmic oleic acid desaturase, is affected by the mutation. In a bulked segregant analysis three AFLP markers linked to this mutation were detected and localized on the genetic map of Brassica napus. The markers mapped near the locus of one copy of the fad2 gene in the rapeseed genome. Received: 16 February 2000 / Accepted: 28 March 2000     

Regrowth of spring canola (Brassica napus) after defoliation

Aims

Regrowth of dual-purpose canola after grazing is important for commercial success and the aim of this research was to investigate the effects of defoliation on the development, growth, photosynthesis and allocation of carbohydrates.

Methods

We conducted two pot experiments in which defoliation was conducted at multiple intensities with scissors. Experiment 1 determined changes in flowering date due to defoliation while Experiment 2 investigated the effects of defoliation on growth, photosynthesis and allocation of carbohydrates in canola.

Results

Time to the appearance of the first flower was delayed by up to 9 days after the removal of all leaves at the start of stem elongation (GS30), and up to 19 days if the elongating bud was also removed. Stem growth rate decreased by 56–86 % due to defoliation and tap roots did not increase in mass when plants were completely defoliated. Leaf area continued to expand at the same rate as in un-defoliated plants. The new leaf area established per gram of regrowth biomass over 20 days was 158 cm².g^-1 for the complete defoliation treatments compared with 27 cm².g^?1 for the half-defoliated treatment and 13 cm².g^?1 for the un-defoliated treatment. Despite a reduction in total biomass of up to 60 %, the proportion of dry matter partitioned to the leaves was 18 % for all treatments within 20 days after defoliation. Total non-structural carbohydrate levels were reduced rapidly in the stem by day two (predominately sucrose) and the tap root by day four (predominately starch) after defoliation and did not recover to match un-defoliated plant levels within 20 days. Residual leaves on defoliated plants maintained photosynthetic rate compared with the same leaf cohorts on un-defoliated plants in which photosynthetic rate decreased to 39 % by day 12.

Conclusions

The rapid recovery of leaf area in defoliated canola was facilitated by the sustained high photosynthetic rate in remaining leaves, rapid mobilisation of stored sugars (stem) and starch (root), and a cessation of root and stem growth.     

The role of phosphorus in aluminium-induced citrate and malate exudation from rape (Brassica napus)

Exudation of organic anions is believed to be a common tolerance mechanism for both aluminium toxicity and phosphorus deficiency. Nevertheless, which of these stresses that actually elicit the exudation of organic anions from rape ( Brassica napus L) remains unknown, and the combined effects of Al toxicity and P deficiency on rape have not been reported before. Therefore, in the current study, Brassica napus var. Natane nourin plants grown with or without 0.25 m M P were exposed to 0 or 50 µ M AlCl₃ and several parameters related to the exudation of organic anions from the roots were investigated. Eight days of P deficiency resulted in a significant growth reduction, but P deficiency alone did not induce exudation of organic anions. In contrast, Al strongly induced organic acid exudation, while simultaneously inhibiting root growth. Increased in-vitro activity of citrate synthase (CS, EC 4.1.3.7), malate dehydrogenase (MDH, EC 1.1.1.37) and phosphoenolpyruvate carboxylase (PEPC, EC 4.1.1.31), together with reduced root respiration, indicated that the Al-induced accumulation and subsequent exudation of citrate and malate were associated with both increased biosynthesis and reduced metabolism of citric and malic acid. Phosphorus-sufficient plants showed more pronounced aluminium-induced accumulation and exudation of organic anions than P-deficient plants. A divided root chamber experiment showed the necessity of direct contact between Al and roots to elicit exudation of organic anions. Prolonged exposure (10 days) to Al resulted in a decrease in the net exudation of citrate and malate, and the rate of decrease was much more rapid in P-deficient plants than in P-sufficient plants. It is concluded that P nutrition affects the level of Al-induced synthesis and exudation of organic anions. However, the mechanism needs further investigation.     

Methods for assessment of light leaf spot (Pyrenopeziza brassicae) on winter oilseed rape (Brassica napus) in the UK

Light leaf spot (Pyrenopeziza brassicae) was assessed as % plants with light leaf spot, % leaves with light leaf spot or % leaf area with light leaf spot in winter oilseed rape field experiments done at different sites (Rothamsted, Hertfordshire; Boxworth, Cambridgeshire; near Aberdeen, Scotland), with different cultivars (e.g. Bristol and Capitol), different fungicide treatments, on plants sampled at different dates. Regression analyses on data from these experiments showed that there were consistently good relationships between % leaves with light leaf spot and % plants with light leaf spot for plants sampled during the autumn and winter, until the % plants with light leaf spot approached 100%. The slopes and positions of regression lines were sometimes affected by cultivar, fungicide treatment or sampling date, but not by site. The relationship between % leaf area with light leaf spot (square root-transformed) and % leaves with light leaf spot was less consistent than that between % leaves with light leaf spot and % plants with light leaf spot and was sometimes affected by cultivar, fungicide treatment or sampling date but not by site. The relationship between % leaf area with light leaf spot (square root-transformed) and % plants with light leaf spot was also inconsistent and was sometimes affected by cultivar, fungicide treatment, sampling date and site.     

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.     

Application of a probabilistic model for analysing the abortion of seeds and pods in winter oilseed rape (Brassica napus)

In winter oilseed rape (WOSR), only a subset of ovules can develop into seeds in the majority of pods. Any difficulty during the process of seed production may result in seed or pod abortion. This study aimed to reproduce the process of seed development in WOSR based on a limited number of parameters. As a result of the complexity of the developmental patterns of WOSR, it is challenging to identify the roles of various factors that influence seed production using an experimental approach. Here, we present a stochastic probabilistic model of seed development. The generalised least squares method was implemented to estimate the model parameters using the experimental data. Experiments were done in Grignon (France) in 2008 and 2009. The variations in the parameters were analysed according to the following four factors: year, pod rank, inflorescence position and ramification‐clipping treatment. The year had no effect on the number of ovules per ovary (μ) and the probability of seed viability (p). The proportion of effective pollen grains (k) significantly decreased with pod rank at the end of the main stem. Inflorescence position influenced the number of ovules per ovary (μ: 30.8–33.8 from top to bottom) and the parameter k. The mean number of seeds per pod on the main stem and the bottom ramification along the stem was larger than the other ramifications within one plant. Ramification‐clipping treatment increased the number of ovules per ovary (μ: 31 for control plants and 32 for clipped plants), the parameter k and the number of seeds per pod (p). This effect could be due to the competition for assimilates between the pods and seeds. Furthermore, the distribution parameters of the pollen number per stigma (m) remained stable, and the probability of pod survival (Bo) varied with different factors, including the year, pod rank and inflorescence position. Our results indicate that pollen germination is a factor that determines final seed number. This model can identify the impact of each of the factors that lead to the abortion of seeds and pods in WOSR, which include the position effect, assimilate competition and pollination limitation. However, further studies on the pollination process in WOSR should help to refine this model.     

Edge distributions of Ceutorhynchus assimilis and its parasitoid Trichomalus perfectus in a crop of winter oilseed rape (Brassica napus)

The edge distributions of Ceutorhynchusassimilis and its parasitoid Trichomalusperfectus within a crop of winter oilseed rape were investigated between May and July using yellow water-traps arranged on a grid system. A randomisation test, which compared the observed arrangement of trap counts with permuted arrangements, was used to quantify the observed tendency of C. assimilisto congregate at the edges as it migrated into the crop; this tendency declined during the season. Trichomalus perfectus were distributed at the edge of the crop only during the early phase of its immigration in May, which occurred later than immigration of its host. It is considered that this simple means of quantifying insect spatial distribution could prove a useful tool in precision targeting of insecticides. This revised version was published online in July 2006 with corrections to the Cover Date.     

Factors affecting development of phoma canker (Leptosphaeria maculans) on stems of winter oilseed rape (Brassica napus) in southern England

In winter oilseed rape experiments at Rothamsted in 1997/98 (cvs Lipton and Capitol), 1998/99 (cv. Apex) and 1999/2000 (cvs Apex, Lipton and Capitol), development of crown canker and phoma stem lesions in spring was related to development of phoma leaf spot in the previous autumn/winter. There were differences in thermal time (degree‐days) from the first appearance of phoma leaf spot (autumn) to the first appearance of crown canker (spring) between cultivars (cvs Lipton and Capitol, 1220–1240; cv. Apex, 1120–1140 degree‐days) but not between growing seasons. In 1998/99 and 1999/2000, fungicide (November) treatment delayed the start of crown canker development in the spring but did not affect the rate of increase in severity. In 1997/98, fungicide treatments did not delay the appearance of crown canker but decreased the rate of increase in crown canker severity. In all three seasons, fungicide treatments generally decreased the proportions of plants at harvest with crown canker severity scores 3 or 4 and increased the proportions with scores 0 or 1. There were differences between seasons in the distributions of crown canker severity scores at harvest. The severity of both crown canker and phoma stem lesions increased linearly with accumulated degree‐days in plots with or without fungicide treatment in 1997/98 (cv. Lipton), 1998/99 (cv. Apex) and 1999/2000 (cv. Apex). Regressions showed that severity of crown canker at harvest in July was related to severity in the spring in 1997/98 (early June, cv. Lipton), 1998/99 and 1999/2000 (April, cv. Apex).     

Persistence of seeds from crops of conventional and herbicide tolerant oilseed rape (Brassica napus)

A series of rotation experiments at five sites over four years has explored the environmental and agronomic implications of growing herbicide tolerant oilseed rape and sugar beet. This paper reports on the population dynamics of volunteer rape (Brassica napus). The experiments compared four winter oilseed rape (WOSR) cultivars: a conventional cultivar (Apex) and three developmental cultivars either genetically modified (GM) to be tolerant to glyphosate or glufosinate, or conventionally bred to be tolerant to herbicides of the imidazolinone group. Seed losses at harvest averaged 3575 seeds m(-2) but ranged from less than 2000 up to more than 10000 seeds m(-2). There was a rapid decline in seed numbers during the first few months after harvest, resulting in a mean loss of seeds of 60%. In subsequent seasons, the seedbank declined much more slowly at four of the five sites (ca 20% per year) and the models predicted 95% seed loss after approximately 9 years. Seed decline was much faster at the fifth site. There were no clear differences between the four cultivars in either the numbers of seeds shed at harvest or in their subsequent persistence. The importance of the persistence of GM rape seeds, in the context of the coexistence of GM and non-GM crops and the role of good management practices that minimize seed persistence, are discussed.