Self-incompatibility as a pollination control mechanism for spring oilseed rape,Brassica napus L.

Summary Self-incompatibility was shown to be an effective method of pollination control in spring rapeseed (B. napus L. ssp. oleifera (Metzg.)) by comparing the yield of a Westar-Topas syn-1 produced by crossing two SI lines with the yield of the corresponding syn-1 produced by hand pollination. Although the trial showed high-parent heterosis in the syn-1s, there was insufficient replication to determine the level of heterosis.Abbreviations SI self-incompatible - SC self-compatible     

Superior performance of two homozygous diploid lines from naturally occurring polyhaploids in oilseed rape (Brassica napus)

Summary Homozygous diploids in oilseed rape were produced by pipetting drops of 0.1 per cent colchicine into the leaf axils of rooted cuttings from naturally occurring polyhaploids after removing the waxy covering with a scalpel. Trial results from spring and winter rape showed that occasional homozygous diploid lines yielded consistently more oil per ha than the parental varieties. Prospects for breeding oilseed rape as a self-fertilised crop are discussed.     

A genetic study of growth characters and yield characters of oilseed rape

Summary Analysis of single plants in parental, F₁ and F₂ populations of three crosses of oilseed rape indicated cytoplasmic effects in F₁ or F₂ generations for most of the observed characters. There was evidence of dominance for early flowering and early maturity. Heterosis was exhibited for yield and yield components. The heritabilities for the characters under study were generally very low.     

Relationships between seed yield and growth characters,yield components,seed quality of summer-type oilseed rape (Brassica napus L.)

Summary Five genotypes of rapeseed were observed in the field on a single plant and plot basis to evaluate growth characteristics, yield, yield components and seed quality. Observations were made in two years at two locations on three seeding date and three seeding rate treatments. Correlation of seed yield with growth characters demonstrated no consistent trend, indicating that an early maturity and high seed yield should be possible. Harvest index was strongly correlated with seed yield. Percent seed oil and protein were not directly related to seed yield.     

Hybridization of radish (Raphanus sativus L.) and oilseed rape (Brassica napus L.) through a flower-culture method

Summary Hybridization between radish and oilseed rape has been cumbersome, requiring elaborate embryo rescue techniques. With a modified flower culture method, we have achieved successful hybridization between radish and (transgenic) oilseed rape without the laborious and technically demandingin vitro ovule or embryo rescue techniques.The hybrid nature of the intergeneric hybrids was demonstrated using morphological traits, and DNA analyses. The described method will facilitate the generation ofRaphanobrassica hybrids useful for biosafety studies of the potential for transgenes to spread in weedyCruciferae as well as for breeding programs aimed at introducing useful radish genes, e.g. nematode resistance genes, into oilseed rape.     

Leaf glucosinolate profiles and their relationship to pest and disease resistance in oilseed rape

Summary Glucosinolates are sulphur-containing glycosides which occur within vegetative and reproductive tissue of oilseed rape. Following tissue damage, glucosinolates undergo hydrolysis catalysed by the enzyme myrosinase to produce a complex array of products which include volatile isothiocyanates and several compounds with goitrogenic activity. Many of these products have been implicated in the interaction betweenBrassica and their pests and pathogens and some may have a role in defence mechanisms. Low glucosinolate (00) oilseed rape cultivars have been shown to possess similar concentrations of leaf glucosinolates as high glucosinolate (0) oilseed rape cultivars. Likewise, despite considerable speculation to the contrary, 00 cultivars have been shown not to be more susceptible to pests and pathogens than 0 cultivars. The potential to enhance pest and disease resistance of oilseed rape by manipulating the leaf glucosinolate profile without reducing seed quality is discussed.     

Predictive study of the advantages of cleistogamy in oilseed rape in limiting unwanted gene flow

Summary The aim of this study was to evaluate the interest of beginning the selection process on a new genetic characteristic, cleistogamy, to manage gene flow in oilseed rape. The first step was to introduce this characteristic in an existing model of gene flow between oilseed rape populations in time and space, GeneSys-Rape. The second step was to evaluate the parameters of the model linked to this characteristic using field experimentations. Cleistogamous oilseed rape was shown to have an autogamy rate as high as 94% and to emit 10 times less pollen than an open-flowered oilseed rape in the same conditions. But the cleistogamous character was also shown to be unstable in the genotypes tested. The third step was to evaluate the interest of cleistogamy using simulations comparing several genotypes with or without cleistogamy in two different cropping systems. These simulations showed that an oilseed rape both dwarf and cleistogamous was interesting to limit gene escape and that a 99%-autogamous oilseed rape was interesting to limit both gene escape from and harvest contamination of the 99%-autogamous oilseed rape.     

Direct plant development from microspore-derived embryos of winter oilseed rape Brassica napus L. ssp. oleifera (DC.) Metzger

The influence of temperature/photoperiod treatment and gibberellic acid concentration (0, 0.1 or 1.0 mg/l) on direct conversion of microspore-derived embryos (MDEs) to plantlets of winter oilseed rape was investigated. Physiologically mature, 21-day-old MDEs were transferred to a solid B5 medium supplemented with gibberellic acid, and cultured at 24 ^°C, 4 ^°C or 1 ^°C for 14 days, and then at 24 ^°C for the next 21 days. Low temperature was linked with short photoperiod (8 h light/16 h dark), and high temperature was linked with long photoperiod (16 h light/8 h dark). The highest embryo conversion rate was at 1 ^°C with over 70%, compared to<20% at 4 ^°C. Two-way analysis of variance confirmed the significance of the effect of temperature/photoperiod treatment. By contrast, gibberellic acid concentration had no significant effect on stimulation of shoot development from apical meristems of MDEs. Roots developed from apical root meristems of MDEs very easily. The best obtained conversion rate of MDEs induced with cold treatment at1 ^°C for 14 days was 86.5%. Observations on morphological development of MDEs showed clear differences in reaction at various temperature/photoperiod treatments. This revised version was published online in August 2006 with corrections to the Cover Date.     

A comparison of traditional and haploid-derived breeding populations of oilseed rape (Brassica napus L.) for fatty acid composition of the seed oil

Summary Microspore embryogenesis technology allows plant breeders to efficiently generate homozygous micros-pore-derived breeding populations of oilseed rape (Brassica napus L.) without traditional generations of inbreeding. This study was conducted to compare the frequency distribution of microspore-derived population and single seed descent populations with respect to fatty acids of seed oil. Both microspore-derived populations and single seed descent populations were produced from each of three crosses made between selected parents containing contrasting amount of erucic, oleic, linoleic and linolenic acids. The fatty acid content of F₃ plants derived lines (F₅ seed) developed by single seed descent was compared to that of microspore-derived populations. The means, ranges and distribution pattern of seed fatty acid contents were similar in both populations for each fatty acid studied, although a few heterozygous lines were observed in the single seed descent populations. The results indicated that microspore-derived population form random, homozygous F₁ plant derived gametic arrays for all fatty acids evaluated. Selection for altered fatty acid composition in microspore-derived and single seed descent homozygous populations should be equally efficient, in the absence of linkage of traits investigated.     

Segregation and linkage analysis of DNA markers in microspore derived and F2 populations of oilseed rape (Brassica napus L.)

Summary The segregation of RFLP and RAPD markers was compared in two oilseed rape (Brassica napus L.) breeding populations from the cross Topas x R4, the latter being a low linolenic mutation line. A total progeny of 68 F₂ and 40 microspore derived plants were studied with 25 markers. The results indicated a significant excess of Topas alleles at five RAPD loci in the microspore derived population. This suggests that genomic regions which probably affect microspore culture ability do not have identical distribution in the two population types.     

Variation of prezygotic barriers in the interspecific hybridization between oilseed rape and wild radish

Escape of transgenes from genetically modified oilseed rape, Brassica napus, into wild radish, Raphanus raphanistrum, depends on sexual compatibility. The variation in prezygotic barriers of two different cultivars for interspecific hybridization with a population of wild radish was investigated by hand crossing and fluorescence microscopy of pistils. Significant differences were observed between oil seed rape cultivars in their ability to accept wild radish pollen germinating onto their stigma and the rate of fertilization of ovules. Some differences among the pollen donor plants were also detected. These results suggest that the rate of interspecific hybridization in the field would depend upon the oilseed rape cultivar and the genotype composition of the local wild populations. The implication of S-related genes, as revealed through identification by pistil tissue prints of class I and II S-types of SLG (S-Locus Glycoprotein) and SLR1 (S-Locus Related),and immuno-IEF, was not significant. This revised version was published online in August 2006 with corrections to the Cover Date.     

The introgression of S alleles into forage rape,Brassica napus L. from turnip. Brassica campestris L. ssp. Rapifera

Summary Forage rape. B. napus, is self-compatible, the work described illustrates the introgression of functional incompatibility alleles into B. napus from turnips, B. campestris, in two generations. By grading seed, produced by backcrossing turnip/rape hybrids to rape, the frequency of 2n=38 semi articial B. napus recovered, amply justifies the cytological screening involved in such breeding programmes.     

Gene dispersal from genetically modified oil seed rape in the field

Summary A field trial was carried out with oil seed rape plants that had been genetically modified to contain genes coding for marker characters (-glucuronidase, kanamycin resistance and asulam resistance). The aim of the experiment was to examine a method of studying gene dispersal in an agricultural environment. The central area of the field plot comprised 150 genetically modified plants and 450 plants not expressing the marker genes. These were surrounded by additional non-expressing plants. The plants were allowed to freely cross pollinate and a sample of the resultant progeny from non-expressing plants were screened for expression of the marker characters and for the presence of the introduced genes (by PCR). Limited gene dispersal was detected and the frequency of modified seedlings amongst the progeny of a plant appeared to have been strongly influenced by the genotype of the immediately adjacent plants.     

Heterosis and epistasis in rapeseed estimated from generation means

Summary Heterosis and epistasis in spring oilseed rape (Brassica napus L.) was analysed by comparing generation means for ten agronomic traits. Parents, F₂, F₃ and F₆ generations of four crosses with Swedish and French material were investigated. The F₂ was 11% higher in yield, earlier in flowering time, and slightly later in maturation when compared with the parents. Randomly derived single seed descent lines had an 8% lower yield, were later flowering and maturing than the parents in F₆. This poorer average performance of recombinant lines is explained by the loss of favourable epistatic interactions present in the parents.     

The transfer of triazine resistance from Brassica napus L. to B. oleracea L. I. Production of F1 hybrids through embryo rescue

Summary Triazine resistant Brassica napus ssp. oleifera and ssp. rapifera were hybridized to cultivars of B. oleracea ssp. italica, ssp. botrytis, ssp. capitata and ssp. fimbriata. The interspecific embryos did not survive in vivo but could be rescued in vitro using a culture medium developed by Monnier (1973). The embryos did not grow directly into normal plants but were successfully regenerated using the protocol developed by Keller (1984). Hybridization efficiency ranged from 0 to 2.64 hybrids per pollination. Interspecific embryo abortion may be related to abnormal endosperm development.     

The transfer of characters from Brassica campestris L. to Brassica napus L.: Production of clubroot-resistant oil-seed rape (B. napus ssp oleifera)

Summary Four oil-seed rape lines were crossed with a clubroot resistant Brassica campestris line from the European Clubroot Differential sct. The allotriploid hybrids were backcrossed to the rape lines to introgress clubroot resistance into oil-seed rape. Using a combination of screening for disease resistance and chromosome number, a high proportion of 38-chromosome, clubroot resistant selections were obtained.     

Genetic analyses of agronomic and seed quality traits of doubled haploid population in Brassica napus through microspore culture

Summary The results showed that the F1 genotype from the cross (Brassica napus cv. Zheshuang 758 × cv. Z-4115) had good response to embryogenesis, and their embryo yield and rate of plant regeneration reached 69.8 embryo/bud and 46.9%, respectively. Characters from the doubled haploid (DH) populations in B. napus were analyzed and it was showed that the means of each agronomic trait were between their parents, but they were nearer to the paternal in 6 agronomic traits (plant height, branch position, number of pods in the main raceme, length of pod, number of pods/plant and number of seeds/pod). The number of genes controlling each agronomic trait was analyzed based on the DH populations. The results showed that the number of genes controlling number of pods in the main raceme was the highest (15.6), and the least number of genes was involved for stem width (only 7.9). According to estimated coefficients of skewness and kurtosis of the traits tested, gene interaction was found to be absent for stem width, plant height, length of main raceme, number of primary and secondary branches, pod density in the main raceme and seed weight/plant. Complementary interaction was also observed in five agronomic traits (number of pods in the main raceme, length of pod, number of pods/plant, number of seeds/pod and 1000-seed weight). A significantly positive correlation was observed between seed yield/plant and four agronomic traits (length of main raceme, length of pod, number of pods/plant and 1000-seed weight). The experiment also showed that the erucic acid, glucosinolate, oil and protein contents of DH populations were 34.23%, 87.09 μmol/g, 44.09% and 42.67%, respectively. The numbers of genes controlling each quality trait were 7.8, 9.7, 9.4 and 8.7, respectively. Partial correlations between the seed quality traits and the agronomic characters of DH populations were analyzed. In this experiment, the partial correlations among seed quality traits were also analyzed and it was found that the oil content had a negative correlation with the other three seed quality traits.     

Inheritance and agronomic performance of an apetalous flower mutant in Brassica napus L

A spontaneous mutant with apetalous flowers was obtained from a hybrid progeny in Brassica napus. The result of genetic analysis showed that the apetalous character was controlled by only one gene locus, petalous flower exhibited incomplete dominance over apetalous flower and that its expression was not affected by cytoplasmic factors. Sixteen agronomically important characteristics of the apetalous line Apet33-10 were compared with those of its petalled near-isogenic line Pet33-10. Results from 4 years of tests indicated that there was no difference between Apet33-10 and Pet33-10 in all tested agronomic characteristics, except for the pod number of main inflorescence and second-order branches at low sclerotinia disease incidence. The Sclerotinia sclerotiorum disease severity index of Apet33-10 was significantly lower than that of Pet33-10, and correspondingly, the plot yield of Apet33-10 was increased obviously in comparison to that of Pet33-10 if sclerotinia disease was serious. The pod number of main inflorescence of Apet33-10 was significantly lower than that of Pet33-10. On the other hand, the pod number of second-order branches of Apet33-10 was significantly higher than that of Pet33-10, hence no overage difference of the pod number per plant was observed between the two lines.     

Rapid and efficient screening of phosphinothricin tolerant oilseed rape (Brassica napus) with a novel germination test

A novel screening test is described for the discrimination of transgenic phosphinothricin tolerant oilseed rape from non transgenic rape seedlings. The method is based on the germination of rape seeds on filter paper soaked with a 0.005% phosphinothricin solution. Under these conditions inhibition of seedling development by the herbicide can be observed after 10 days. The germination test gains an advantage over the routinely used herbicide spraying, because it is rapid, needs little space and allows efficient screening of huge numbers of seeds. The assay has been successfully applied to the screening of different transgenic and non transgenic rapeseed varieties/lines and has been compared to other methods such asBasta® spray test, drop test, ELISA-technique and PCR-amplification of the pat gene. This test allows on one hand large screening programmes to monitor the foreign gene in the environment and on the other quality control of seedlots before market introduction of herbicide tolerant oilseed rape.     

Evaluation of cytoplasmic effects on agronomic and seed quality traits in two doubled haploid populations of Brassica napus L.

Cytoplasmic effects have been occasionally implicated in the inheritance of several traits in oilseed rape (Brassica napus L.), including linolenic acid concentration (18:3) in the oil. It is important that these be considered when choosing the direction of cross for producing new breeding populations. To study this phenomenon, a reciprocal cross was made between two genotypes of oilseed rape, Reston and LL09, which differed for their erucic and linolenic acid concentrations in the seed oil. Two DH populations, which were produced by microspore culture from reciprocal F₁ plants, were evaluated in the growth room for one generation and in the field at two locations in Southern Ontario in 1993and 1994. Field notes were taken on days to flower, days to maturity,plant lodging, plant height and, seed quality traits. In the growth room study, the phenotypic distribution of 18:3 differed significantly between the two reciprocal DH populations. In the field, significant reciprocal differences between the population means were detected for 18:3,flowering date and protein content in both years and for days to maturity and oil content in 1993 only. To further study the parental lines,chloroplast (cp) and mitochondrial (mt) DNA from parental lines were isolated and subjected to RFLP and RAPD analysis. Several random primers revealed reproducible DNA polymorphism (RAPD) between the parental mt DNA. It is concluded that the direction of cross should be taken into consideration by oilseed rape breeders relying solely on doubled haploids for developing genotypes with modified seed quality traits in Brassica napus L. This revised version was published online in July 2006 with corrections to the Cover Date.