Molecular Cytogenetic Localization and Characterization of 5S and 25S rDNA Loci for Chromosome Identification in Oilseed Rape (Brassica napus L.)

Chromosome identification in oilseed rape (Brassica napus L.)is extremely difficult using conventional cytogenetic techniquesbecause amphidiploid Brassica species possess numerous verysmall chromosomes with few cytogenetic landmarks. In combinationwith methods for improved chromosome preparations, we used asimplified fluorescencein situ hybridization (FISH) techniqueto localize simultaneously the gene families coding for 5S and25S rDNA in B. napus. The resulting hybridization patterns enabledten of the 19 oilseed rape chromosome pairs to be unequivocallyidentified. Copyright 2000 Annals of Botany Company Brassica napus, oilseed rape, rDNA, molecular cytogenetics, FISH, chromosome identification     

Low Temperature Affects Pattern of Leaf Growth and Structure of Cell Walls in Winter Oilseed Rape (Brassica napus L., var. oleifera L.)

Three-week acclimation of winter oilseed rape (Brassica napusL. var. oleifera L.) plants in the cold (2 °C) resultedin a modified pattern of leaf cell enlargement, indicated bythe increased thickness of young leaf blades and modified dimensionsof mesophyll cells, as compared with non-acclimated tissuesgrown at 20/15 °C (day/night). The thickness of leaf cellwalls also increased markedly during cold acclimation but itdecreased in response to a transient freezing event (5 °Cfor 18 h followed by 6 or 24 h at 2 °C, in the dark). Cellwalls of the upper (adaxial) epidermis were most affected. Theirultrastructure was modified by cold and freezing treatmentsin different ways, as revealed by electron microscopy. Possiblereasons for the cold- and freezing-induced modifications inthe leaf and cell wall morphology and their role in plant acclimationto low temperature conditions are discussed. Copyright 1999Annals of Botany Company Acclimation, Brassica napus var. oleifera, cell wall ultrastructure, cold, freezing, leaf structure, winter oilseed rape.     

Anatomical and Biochemical Changes Associated with the Induction of Oilseed Rape (Brassica napus) Pod Dehiscence by Dasineura brassicae (Winn.)

Dehiscence of pods at an early stage of development is a characteristicof oilseed rape pods damaged by Dasineura brassicae (pod midge).Anatomical examination of pods exhibiting symptoms of infestationrevealed a loss of cohesion between intact cells of the dehiscencezone, a narrow tissue of thin-walled cells present between thevalve margins. Determination of hydrolytic enzyme activity inpericarp tissues of damaged siliquae showed localized enhancementof both polygalacturonase (EC 3.2.1.15 [EC] ) and cellulase (ß1,4 glucanase, EC 3.1.2.4 [EC] ) activity, positionally consistent withthese factors being responsible for the observed cell wall degradation.Mechanistic similarities of midge-induced and maturation-associatedpod dehiscence are discussed. Oilseed rape, Brassica napus L. cv., Bienvenu, pod shatter, pod midge, Dasineura brassicae (Winn.), cellulase, polygalacturonase     

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.     

The Effect of Root Temperature on Growth and Uptake of Ammonium and Nitrate by Brassica napus L. in Flowing Solution Culture: I. GROWTH

Macduff, J. H., Hopper, M. J. and Wild, A. 1987. The effectof root temperature on growth and uptake of ammonium and nitrateby Brassica napus L. in flowing solution culture. I. Growth.—J.exp. Bot. 38: 42–52 Oilseed rape (Brassica napus L. cv. Bien venu) was grown for49 d in flowing nutrient solution at pH 6?0 with root temperaturedecrementally reduced from 20?C to 5?C; and then exposed todifferent root temperatures (3, 5, 7, 9, 11, 13,17 or 25?C)held constant for 14 d. The air temperature was 20/15?C day/nightand nitrogen was supplied automatically to maintain 10 mmolm^–3 NH₄NO₃ in solution. Total dry matter production wasexponential with time and similar at all root temperatures givinga specific growth rate of 0?0784 g g^–1 d^–1. Partitioningof dry matter was influenced by root temperature; shoot: rootratios increased during treatment at 17?C and 25?C but decreasedafter 5 d at 3?C and 5?C. The ratio of shoot specific growthrate: root specific growth rate increased with the ratio ofwater soluble carbohydrates (shoot: root). Concentrations ofwater soluble carbohydrates in shoot and root were inverselyrelated to root temperature; at 3, 5 and 7?C they increasedin stem + petioles throughout treatment, coinciding with a decreasein the weight of tissue water per unit dry matter. These resultssuggest that the accumulation of soluble carbohydrates at lowtemperature is the result of metabolic imbalance and of osmoticadjustment to water stress. Key words: Brassica napus, oilseed rape, root temperature, specific growth rate     

Development of Oilseed Rape Buds, Flowers, and Pods In Vitro

A technique for growing buds, flowers, and pods of oilseed rape(Brassica napus L. cv. Haplona) on stem explants in vitro hasbeen developed. Open flowers and young pods underwent normaldevelopment on a basal medium of minerals, vitamins, and sucrosebut the development of buds was less successful. Young buds(3 mm long) did not develop and only limited development ofthe older buds (5 mm long) took place. Some 3 mm-long buds wereinduced to develop to open flowers by adding naphthyl aceticacid or gibberellic acid. Pod and seed set in open flowers werenot affected by adding plant growth substances to the medium,but pod elongation and pod dry weight were promoted by gibberellicacid, 105 M, and benzyl amino purine, 10⁷ M, respectively. Reducingthe supply of sucrose or minerals to open flowers reduced seedset, pod elongation and pod weight but did not affect pod set.The physiological significance of the results is discussed. Key words: In vitro cultures, oilseed rape, pod development, flower development     

Nodulation of Oilseed Rape (Brassica napus) by Rhizobia

Nodules were induced on the non-legume oilseed rape, followingenzyme treatment of seedling roots and inoculation with Rhizobiumlegnminosarum, Bradyrhizobtum 32H1 or a mixture of R. lott withBradyrhizobium 32H1 in the presence of PEG. A Nod– strainof R leguminosarum also induced nodules, but a Nod– strainfailed to elicit this response Nodules induced on oilseed rapewere morphologically similar, when examined by light microscopyand cryo-scanning electron microscopy, to those induced on rootsof white clover by R trifolu. Transmission electron microscopyshowed rhizobia within cells of the nodules These observationsare discussed with respect to the extension of Rhizobium symbiosisto non-legumes. Key words: Brassica napus, Bradyrhizobium, Rhizobium, non-legumes, nodulation, transmission electron microscopy, cryo-scanning electron microscopy, acetylene reduction, cell wall degrading enzymes     

Effect of sulphate deficiency on the plasma membrane polypeptide composition of Brassica napus

Highly purified oilseed rape (Brassica napus) root plasma membranefractions were prepared and their polypeptide patterns analysedby two-dimensional gel electrophoresis. Sulphur starvation enhancesthe sulphate uptake capacity of B. napus roots. The relativeabundance of several polypeptides increased significantly andspecifically after sulphur starvation. Several of them (37,38, 60, and 65 kDa), found in sulphur-starved plants, were moreabundant in a phase-partitioned membrane fraction treated withTriton X-100/KBr, indicating that they are intrinsic polypeptides.One polypeptide (47 kDa) was identified in the in vitro translationproducts of the roots mRNAs as specific for S-starved plants.It was also present among the intrinsic polypeptides specificfor —S plants. These plasma membrane polypeptides mightbe involved in sulphate uptake. Key words: Sulphate, sulphur-starvation, plasma membrane, polypeptides, root, transport     

Transformation of Brassica napus L. with a 'Disarmed' Octopine Plasmid of Agrobacterium tumefaciens: Molecular Analysis and Inheritance of the Transformed Phenotype

Phenotypically normal and fertile transgenic Brassica napuscv. Westar plants were obtained following co-cultivation ofstem epidermal explants with an Agrobacterium tumefaciens straincontaining a disarmed octopine tumour-inducing plasmid pTiB6S3-SE.The A. tumefaciens cells also contained pMON316, a cointegratevector carrying genes for kanamycin resistance and a scorablemarker nopaline synthase. Transformants were selected by theirability to grow in the presence of 100 µg cm^-3 kanamycin.Transformation was confirmed by the activities of neomycin phosphotransferaseII and nopaline synthase enzymes and by Southern blots. Thekanamycin resistance trait was transferred to the progeny ofthe self-fertilized plants. Key words: Transformation, octopine Ti-plasmid, oilseed rape     

Seed transmission of turnip yellow mosaic virus in winter turnip and winter oilseed rapes

This is the first record of seed transmission of turnip yellow mosaic virus (TYMV) in oilseed and turnip rapes. The seed transmission of TYMV in a naturally infected winter turnip rape (Brassica napus var. silvestris) cultivar Perko PVH was investigated. By ELISA 1.6%, 3.2% and 8.3% seed transmission of the virus was found in seed of plants from three localities. The proportion of infected seeds produced by artificially infected plants of winter oilseed rape (Brassica napus ssp. oleifera) and winter turnip rape cultivars was determined. The virus transmission rate, expressed as the proportion of virus-infected plants which germinated from the seed was for the oilseed rape cvs Jet Neuf 0.1%, Solida 0.4%, Silesia 0.8%, Darmor 1.2%, SL-507 0.2%, SL-509 0.0% and for the winter turnip rape cv. Perko 1.5%. ELISA cannot be used in direct tests on bulk seed lots to estimate proportion of infected seed, but must be used on germinated seedlings.     

Effects of Root Temperature and Form of Nitrogen Nutrition on Nitrate Reductase Activity in Oilseed Rape (Brassica napus L.)

The effect of root temperature and form of inorganic nitrogensupply on in vitro nitrate reductase activity (NRA) was studiedin oilseed rape (Brassica napus L. cv. bien venu). Plants weregrown initially in flowing nutrient solution containing 10 µMNH₄NO₃ and then supplied with either nitrate or ammonium for15 d at root temperatures of 3, 7, 11 or 17 °C. Shoot temperatureregime was similar for all plants; 20/15 °C, day/night.Root NRA was highest when roots were grown at 3 and 7 °C.In laminae and petioles NRA was highest when roots were 11 or17 °C. The plants supplied with ammonium had much lowerlevels of NRA in roots after 5 d than the plants supplied onlywith nitrate. NRA in the laminae of plants supplied with ammoniumwas low relative to that in plants supplied with nitrate onlywhen root temperature was 11 or 17 °C. Values of the apparent activation energy (E_a) of NR, calculatedfrom the Arrhenius equation, in laminae and petioles were differentfrom roots suggesting difference in enzyme conformation. Evidencethat the temperature at which roots were growing affected E_awas equivocal. Oilseed rape, Brassica napus L., activation energy, ammonium, Arrhenius equation, nitrate, root temperature, nitrate reductase     

Improving of oilseed rape lateral root formation by physiological analogues of auxin

The aim of the present work was to study the effect of auxin physiological analogue TA-12 [1-(2-chloroethoksicarbonylmethyl)-4-naphthalenesulfonic acid calcium salt] on the formation of oilseed rape lateral root and on the mitotic activity of apical meristem cells. Spring oilseed rape (Brassica napus L. ssp. oleifera annua Metzg.) cultivar ‘Mascot’ was chosen as a test object. Anatomical, cytological and histological studies on root development suggest that compound TA-12 induces the activity of parent root pericycle cells, stimulates the formation of lateral roots and enhances the division of apical meristem cells. The auxin transport inhibitor 2,3,5-triiodobenzoic acid suppresses the division of apical meristem cells, while this process is restored by the auxin physiological analogue TA-12 and naphthaleneacetic acid. The compound TA-12, by stimulating primary root growth and lateral root induction, optimised the formation of the oilseed rape root system.     

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.     

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.     

Gene dispersal from transgenic crops

The risk of release of genetically modified oilseed rape (Brassica napus) was investigated in relation to interspecific gene flow with hoary mustard (Hirschfeldia incana). Microscopic studies showed polymorphism within the population of hoary mustard for pollen germination on oilseed rape flowers. The transgenic herbicide-resistant and a commercial cultivar of oilseed rape were not different for pollen behaviour and ovule fertilization. Pollen tube growth was slow and erratic in interspecific crosses. Fertilization efficiency of oilseed rape and hoary mustard pollen in interspecific crosses was 15% and 1.3%, respectively, of that in intraspecific crosses. This unequal efficiency in reciprocal crosses was confirmed by hybrid seed set in pods. There was no post-zygotic barrier to the development of hybrid embryos in hoary mustard pods. Up to 26 spontaneous hybrids per male sterile oilseed rape plant, and one per hoary mustard plant, were obtained in field experiments. Hybrids were identified by isozyme electrophoresis, morphology and cytology. All hybrids were triploid with 26 chromosomes, and had low fertility. They produced 0.5 seeds per plant after spontaneous backcrossing with hoary mustard. Some of these descendants were produced from unreduced gametes. Our results suggest that gene flow is likely to occur, but its actual frequency under crop growing conditions remains to be estimated.     

Determination of a Critical Nitrogen Dilution Curve for Winter Oilseed Rape

Several controlled environmental and field experiments werecarried out to define the critical nitrogen dilution curve forwinter oilseed rape, cultivar Goeland. This curve is describedby the following power equation:N=4.48 W^-0.25,whereNis the totalnitrogen concentration in the shoot biomass andWthe shoot biomass.This curve has been validated over the range of shoot dry matterof 0.88 to 6.3 t ha^-1. For lower shoot biomasses this equationoverestimated the critical nitrogen concentration; we proposea constant value of 4.63 (Nis expressed in reduced N, whichis a more stable N fraction in the shoot at these stages ofdevelopment). These results have been validated in several pedoclimaticconditions in France on a single variety in 1994 and 1995. Thehigher position of this curve relative to the C₃species referencecurve (Greenwoodet al.,Annals of Botany67: 181–190, 1990)can be explained by the experimental conditions obtained byGreenwoodet al. (1990); therefore, all their rape data are ratherclose to the critical curve that we propose. The differencesfound between wheat and winter oilseed rape critical N dilutioncurves correspond to their respective leaf:stem dry matter ratioand the specific leaf loss phenomenon occuring in rape. Winteroilseed rape has a higher capacity of N accumulation in itsshoot than wheat for the same aerial dry matter. The proportionof nitrate in shoots rises with the nitrogen nutrition index(N.N.I.) and is more important for rapeseed than for wheat forthe same N.N.I. This difference is especially high at the beginningof flowering when the shade provided by the canopy of rapeseedflowers decreases nitrate reductase activity.Copyright 1998Annals of Botany Company Winter oilseed rape;Brassica napusL.; plant N concentration; nitrate; reduced N; shoot biomass; critical nitrogen concentration; dilution curve; N productivity.     

Attributes of Individual Flowers of Brassica napus L. are Affected by Defoliation but not by Intraspecific Competition

To investigate the effect of intraspecific competition on floweringin Brassica napus‘Westar’, a series of 30 pots wasestablished that spanned a range of one–96 plants perpot. In each pot, the following attributes of individual flowerswere quantified: petal length; petal width; stamen length; pistillength; pollen production; diameter of pollen grains; and nectarproduction. Certain plants contained a transgenic marker thatenabled the vigour of their pollen to be quantified by screeningthe progeny from post-pollination trials against conventionalmale competitors. Plant size was strongly affected by plantdensity; plants in the lowest density pots comprised ten-timesmore dry biomass than individuals in the highest density pots.However, none of the attributes of individual flowers variedwith density. In contrast, the number of flowers per plant declinedrapidly with density. In the face of resource scarcity, theplants apparently conserved flower size rather than flower number.There was no difference in the response to density between transgenicand conventional plants, but transgenic plants obtained morefertilizations than expected in post-pollination competitionagainst conventional competitors. A separate experiment demonstratedthat B. napus is, nevertheless, capable of plasticity in certainflower attributes (petal size, stamen length) in response todefoliation. Explanations for the stability of flower size relativeto flower number are discussed. Copyright 2001 Annals of BotanyCompany Brassica napus, defoliation, density effects, flowers, flower size, intraspecific competition, nectar production, oilseed rape, pollination, transgenic plants     

Spontaneous hybridization between oilseed rape (Brassica napus) and weedy B. campestris (Brassicaceae): a risk of growing genetically modified oilseed rape

Frequencies of spontaneous hybridization between oilseed rape (Brassica napus L.) and weedy Brassica campesíris ssp. campestris L. were measured in agricultural fields. Hybrids were identified by enzyme electrophoresis, random amplified polymorphic DNA (RAPD) analysis, chromosome counting, morphology, and pollen fertility. When the two species were mixed 1:1, B. campestris produced 13% hybrid seeds and oilseed rape 9%. In two experiments with single plants of the self-incompatible B. campestris widely spaced within fields of oilseed rape, 56% and 93% hybrid seeds were produced. Analysis of a weedy population of B. campestris in oilseed rape revealed 60% hybrid seeds. Backcrossing of the hybrids to the weedy species seems to occur as supported by the finding in a natural population of two B. campestris-like plants with a marker specific to oilseed rape. The results suggest that transgenes could be dispersed from oilseed rape to B. campestris.     

Role of Sucrose in Microspore Embryo Production in Brassica napus ssp. oleifera

Dun well, J. M. and Thurling, N. 1985. Role of sucrose in microsporeembryo production in Brassica napus ssp. oleifera.—J.exp. Bot. 36: 1478–1491. One cultivar of winter oil seed rape (Brassica napus ssp. oleifera)and three cultivars of spring rape were used in a study of theeffects of sucrose on microspore survival and embryo inductionin cultured anthers. A preliminary study on the winter cultivar(Fiona) revealed that the osmotic pressure of the supernatantof anther homogenates was equivalent to a solution of 17% sucrose.A study of microspore survival and embryo induction in thiscultivar on media containing either 8 %, 12%, 16% or 20% sucroserevealed the highest survival (after 16 d) and the greatestnumber of anthers with induced embryos (after 42 d) occurredon the highest sucrose concentration. A subsequent study on three spring cultivars (Willi, Duplo andTower) examined microspore survival at 8 d and embryo induction(42 d) on media containing either 8 % or 16 % sucrose and againrevealed much higher survival and induction at the higher concentration.The variation in response between the cultivars was also reducedby culture at the higher sucrose concentration. The beneficialeffect of the 16% level occurred regardless of the growth environmentof the donor plants and of the stage of pollen development atthe start of culture. However, macroscopic embryos emerged onlyfrom anthers on the 8 % sucrose concentration, suggesting thattransfer of anthers from a high to a normal sucrose concentrationduring culture would ensure that full advantage was taken ofthe much higher initial survival on the higher concentration Key words: Brassica napus, sucrose, microspore embryo production     

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.