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.     

Selection for fast germination in rapeseed (Brassica napus L. and B. Campestris L.)

Summary Eleven populations of Brassica napus L. and twelve populations of B. campestris L. were subjected to three cycles of selection for fast germination at 2°C and at 25°C. The seeds from the selected populations, and unselected control populations grown in the same environment as the selected populations, were examined for germination behaviour at 2°C and 25°C, and for growth behaviour at 10°C. The populations in both species responded differently in terms of germination behaviour to selection for fast germination. In most of the populations that did respond positively to selection, selection practised at 2°C was superior to selection at 25°C in improving percent germination at 2°C, and was as good as the selection at 25°C in improving germination rate at the higher temperature. Selection for fast germination had no effect on growth characteristics of B. napus and B. campestris populations grown at 10°C. Thus, selection for fast germination at one low temperature may lead to improvement in germination characteristics over a range of temperatures, but will not necessarily lead to improved growth performance of the selected populations.     

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.     

Inheritance of seed colour in Brassica campestris L. and breeding for yellow-seeded B. napus L.

Summary Seed colour inheritance was studied in five yellow-seeded and one black-seeded B. campestris accessions. Diallel crosses between the yellow-seeded types indicated that the four var. yellow sarson accessions of Indian origin had the same genotype for seed colour but were different from the Swedish yellow-seeded breeding line. Black seed colour was dominant over yellow. The segregation patterns for seed colour in F₂ (Including reciprocals) and BC₁ (backcross of F₁ to the yellow-seeded parent) indicated that the black seed colour was conditioned by a single dominant gene. Seed colour was mainly controlled by the maternal genotype but influenced by the interplay between the maternal and endosperm and/or embryonic genotypes. For developing yellow-seeded B. napus genotypes, resynthesized B. napus lines containing genes for yellow seed (Chen et al., 1988) were crossed with B. napus of yellow/brown seeds, or with yellow-seeded B. carinata. Yellow-seeded F₂ plants were found in the crosses that involved the B. napus breeding line. However, this yellow-seeded character did not breed true up to F₄. Crosses between a yellow-seeded F₃ plant and a monogenomically controlled black-seeded B. napus line of resynthesized origin revealed that the black-seeded trait in the B. alboglabra genome was possibly governed by two independently dominant genes with duplicated effect. Crossability between the resynthesized B. napus lines as female and B. carinata as male was fairly high. The sterility of the F₁ plants prevented further breeding progress for developing yellow-seeded B. napus by this strategy.     

Genetic and genotype × environment interaction effects analysis for erucic acid content in rapeseed (Brassica napus L.)

Genetic main effects and genotype × environment (GE) interaction effects for erucic acid content (EAC) in rapeseed(Brassica napus L.) were analyzed for two year data by using the genetic models and corresponding statistical approaches for quantitative traits of diploid plant seeds. Eight parents were included in adiallel mating design in two environments. It was found that the embryo, cytoplasmic and maternal main effects and their GE interaction effects could simultaneously affect the performance of EAC, especially for the cytoplasmic and maternal effects. Since the amount of genetic main effects from the expression of genes in different genetic systems accounted for about 64.1%of the total genetic effects, EAC of rapeseed was mainly controlled by genetic main effects. The total narrow-sense heritability for EAC was 83.6% with the general heritability being 51.9% and the interaction heritability being 31.7%. It was suggested by the predicted genetic effects, that Tower and Youcai 601 were better than other parents for reducing EAC in rapeseed quality breeding. This revised version was published online in July 2006 with corrections to the Cover Date.     

Nondestructive assessment of protein content in single seeds of rapeseed (Brassica napus L.) by near-infrared reflectance spectroscopy

The potential of near-infrared reflectance spectroscopy (NIRS) to detectwithin-plant differences for seed protein content was investigated. Fourhundred and fifty-one single seeds were scanned by NIRS using a specialadapter. After non-destructive NIRS scanning, the seeds were analysed forprotein content by the Dumas combustion method and a calibrationequation was developed. A validation set of 117 additional seeds fromthree individual plants from the cultivars Bristol, Lirajet and Maplus wasanalysed for protein content both by NIRS and combustion. The coefficientof determination between NIRS and combustion values in the validation setwas 0.94, with a standard error of performance (SEP) of 0.77% and aratio of the SEP to the standard deviation (SD) of the validation set of0.28. The coefficient of variation (CV) for seed protein content inindividual plants, as determined by the combustion method, was 11.7%for Bristol, 8.9% for Lirajet, and 9.5% for Maplus. The comparison ofsuch variation with the standard error (SE) of NIRS analysis, defined as thecombination of the SE of the combustion method and the SEP of NIRScalibration equation, revealed that the maximum explainable variance withinindividual plants that can be detected using NIRS analysis of proteincontent in single seeds was 0.86 for Bristol, 0.83 for Lirajet, and 0.85 forMaplus. These results demonstrated that NIRS is a powerful tool fornon-destructive assessment of within-plant variation for seed protein contentin rapeseed.     

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.     

Heterosis and inbreeding depression in forage rape (Brassica napus L.)

Summary Heterosis and inbreeding depression for total fresh and dry weights were studied over two seasons in 15 crosses of six cultivars of forage rape (Brassica napus L.). Some of the hybrids exhibited considerable heterosis over both mid-parent and better parent; followed by depression in the F₂. The cross Windal × Canard had 62% and 25% greater dry matter yield over the best cultivar used in this experiment in 1978 and 1979, respectively. Possibilities of the utilization of this hybrid superiority are discussed.     

Inter-plot competition in yield trials of swedes (Brassica napus ssp. rapifera L.)

Summary Two experiments were done to evaluate the use of single-row plots for assessing dry-matter yield of swedes. In the first experiment the correlation between the dry-matter yields of 20 cultivars in unguarded single-row plots and those in guarded double-row plots was poor (r=0.39). It was improved (r=0.58) by adjusting the single-row plot fresh-weight yields for inter-plot competition, through an analysis of covariance in which the covariate was the mean of the adjacent plot yields, and then estimating the pure stand dry-matter yields. However, the correlation was still lower than that between unguarded double-row plots and the guarded double-row plots (r=0.73), and this latter correlation was also improved (r=0.82) by adjusting the fresh-weight yields of the unguarded plots for inter-plot competition.The second experiment provided an explanation for the partial success of the covariance adjustment. The individual neighbour effects of six cultivars were determined. Doon Major and Ruta Øtofte decreased the yields of adjacent cultivars whereas Dryden and Melfort increased them. Angela and Magres had little effect. The correlation between the dry-matter yield neighbour effects of cultivars and their fresh-weight yields was high (r=–0.81), but not complete. So although competitive ability was associated with root fresh-weight yield, other factors must also be involved.It was concluded that a two-replicate trial with unguarded double-row plots is preferable to a four-replicate trial with unguarded single-row plots for dry-matter yield assessment of early generation families in a pedigree swede breeding programme.     

A comparison of methods for hybrid seed production using self-incompatibility in swedes (Brassica napus ssp. napobrassica)

Procedures for producing seed of hybrid swedes using self-incompatibility were examined. Single-cross, double-cross and modified double-cross hybrids were compared in isolation plots using natural pollinators and in polythene tunnels using blow-flies. With good coincidence of flowering and the same flower colour, nearly 100% hybrid seed was produced by natural pollinators with the single-crosses, the double-cross and one of the two modified double-cross hybrids; the other modified double-cross hybrid produced 87%hybrid seed. With poor coincidence of flowering and different flower colours the proportion of hybrids dropped to 61%. Using different flower colours and blow-flies as pollinators in polythene tunnels, higher levels of outcrossing were produced than in isolation plots with natural pollinators; the opposite result was obtained when the same flower colour was used. This revised version was published online in July 2006 with corrections to the Cover Date.     

Prediction of heterosis using QTLs for yield traits in rapeseed (Brassica napus L.)

Three double low (erucic acid and glucosinolates) self-incompatible lines and 22 varieties from different origins were selected to produce 66 hybrids according to a NC II mating design. Field experiments for identification of hybrid performance and heterosis were conducted in two successive rapeseed growing seasons in Wuhan, China. After heterosis identifications, SI-1300 and Eagle were chosen to construct an F₂ segregating population. One hundred and eighty four F_2:3 lines were planted at Wuhan and Jingmen to test yield traits. F₂ plants and the 25 parents were analyzed using simultaneously AFLP (amplified fragment length polymorphism) and SSR (simple sequence repeat) markers. A total of 270 and 718 polymorphic loci were detected in the F₂ population and among the 25 parental lines, respectively. Of the 718 polymorphic loci, 178 were significantly correlated to yield traits. With the use of one-way ANOVA, 84 common QTLs were detected for 12 traits at two trial locations. Although the genetic distances based on general/specific heterozygosities and single-locus QTLs showed significant correlations with hybrid performance and heterosis for some yield traits, the determination coefficients were low. The results suggested that neither heterozygosities nor QTLs for yield traits were suitable to predict hybrid performance and heterosis in Brassica napus.     

Self-incompatibility interactions in Brassica napus

Summary The interactions of nine S-gene lines of Brassica napus were examined. Seven different S-genes were involved in the crosses, with three lines of different origin having the same S-gene. In general, the reactions on selfing the heterozygotes were as expected from the results of test crosses with the homozygous parent lines. All types of dominance relationships were found, including reversal of dominance in pollen and pistil. Weakening of activity of one or both genes was found in several cases. Background genotype was found have to some effect and reciprocal differences in reactions were also observed.     

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.     

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.     

甘蓝型油菜DH群体苗期抗旱性的评价

中国油菜主产区常常受到秋冬旱和春旱危害而影响产量及品质。本文通过盆栽试验,以株高(PH)、根长(RL)、地上部干重(SDW)、根干重(RDW)、根冠比(R/S)、总干重(TDW) 6个性状的抗旱系数作为抗旱性评价指标,对甘蓝型油菜DH(doubled haploid)群体的118个株系及其亲本进行苗期抗旱性评价,筛选极端抗旱DH系。结果表明,与对照相比,干旱严重抑制了甘蓝型油菜苗期的生长,6个评价指标均表现出显著差异,其中根干重的变异系数最大;在对照和干旱条件下,群体各株系各性状均表现出超亲连续分离,大部分呈正态分布,抗旱条件下的分离更为明显;相关性分析表明,地上部干重、根干重和总干重抗旱系数可作为甘蓝型油菜苗期抗旱性的主要评价指标;     

In vitro selection for oleic and linoleic acid content in segregating populations of microspore derived embryos of Brassica napus

Microspore derived embryos (MDEs) in Brassica napuscontain large amounts of storage lipids which show a genotype specific fatty acid composition (FAC). One cotyledon of regenerating emblyos can be dissected at an early stage during the in vitro culture and used for fatty acid analysis. Thus, in breeding programmes to modify oil quality, only MDEs having the desired FAC need to be regenerated to plantlets and transferred to the greenhouse. In the present study the applicability of this method for the selection of a high oleic acid content and a low linoleic acid content in the seed oil has been tested by crossing a Brassica napus mutant line having a high oleic acid (C18:1) content in the seed oil (75%) with a wild type doubled haploid line with 62% C18:1 in the seed oil. Microspore culture was applied to the F1 plants. In total 59 MDEs were obtained, from which 31 were cultured with and 28 without 15μM abscisic acid for 3 weeksin vitro. One cotyledon was dissected under aspetic conditions and used for fatty acid analysis. The remaining part of the embryos were further regenerated to plantlets and transferred to the greenhouse to obtain seeds after self pollination. Seeds harvested from the doubled haploid lines in the greenhouse were used for fatty acid analysis and also for growing in the field. The abscisic acid treatment of the MDEs generally improved the correlations for linoleic and oleic acid between the MDEs and the seeds harvested in the greenhouse and the field. The correlations ranged from 0.68** to 0.81**.This indicates that selection for high oleic acid can be started already during an early stage of the in vitro culture. This revised version was published online in July 2006 with corrections to the Cover Date.     

油菜不同产量类型品种氮素吸收与利用特性研究

摘要: 【研究目的】探讨甘蓝型油菜不同产量类型品种氮素吸收与利用特性。【方法】在不同土壤肥力条件下以甘蓝型油菜品种（2006-2007年度73个、2007-2008年度98个）为材料,成熟期测定各器官干物重、氮素含量,采用组内最小平方和动态聚类方法对供试品种产量进行聚类。研究不同产量类型品种氮素积累与分配差异。【结果】结果表明：供试品种间产量差异很大,类型间差异显著。随着产量增加,氮素吸收总量、氮素籽粒生产效率增加,籽粒氮素积累量增加,茎枝、果壳氮素分配比例下降,籽粒氮素比例增加。土壤肥力高,植株吸氮总量增加,氮素籽粒生产效率降低。【结论】增加氮素吸收总量,促进氮素向籽粒中输送,使得高产和高氮素利用效率统一。     

Identification of rape seed cultivars (Brassica napus) by starch gel electrophoresis of enzymes

Summary Isozyme patterns were used to characterize ten commercial rape seed (Brassica napus) cultivars. Extracts of cotyledons were made 4–7 days after germination and separated by electrophoresis on starch gels. A sample of more than 100 plants of each variety was analysed and stained for 4 polymorphic enzyme loci (Lap, Gpi, Acon, and Sdh). Therewith, it was possible to distinguish all ten rape varieties qualitatively by at least one typical enzyme pattern. Further, it could be demonstrated that all pairs of varieties showed clear qualitative differences in isozyme patterns, when only tow loci were screened (Acon, Sdh). Using ² test of homogeneity all pairs of varieties differ significantly in their frequencies of isozyme patterns for Acon and in all but one for Sdh.     

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.     

The effects of insect pollination on canola (Brassica napus L. cv. O.A.C. Triton) seed germination

Summary The presence of pollinators, e.g. honey bees, on canola (cv. O.A.C Triton) increases the germinability of resulting seeds from 83% to 96%.