Abundance, polymorphism and genetic mapping of microsatellites in oilseed rape (Brassica napus L.)

The objective of the present study was to estimate the abundance and degree of polymorphism of simple sequence repeat (SSR) markers in rapeseed. By screening about 45000 clones of a small inserts library of rapeseed total DNA the abundances of GA/TC and CA/TG simple sequence repeats in the rapeseed genome were estimated to be approximately one repeat every 100 kb and 400 kb, respectively. After sequencing 13 positive clones, primer pairs could be designed for 11 microsatellite loci. Seven of these primer pairs produced reproducible amplification products in a set of 31 rapeseed genotypes, with one pair amplifying two independent products, giving a total of eight amplified loci. The different microsatellite loci displayed between one and three visible alleles. At four loci, additional null alleles were observed. With up to four alleles, polymorphic microsatellite markers show significantly higher allele numbers in rapeseed than restriction fragment length polymorphism (RFLP) markers. Four of the eight microsatellite markers could be mapped on four different linkage groups of an RFLP map of the rapeseed genome.     

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.     

Impact of low linolenic acid content on seed yield of winter oilseed rape (Brassica napus L.)

An essential quality improvement of rapeseed oil can be obtained by reduction of its linolenic acid (C18:3) content from about 10% to less than 3% of the total fatty acids. Genotypes low in C18:3 have been developed by mutagenesis. The initial summer rapeseed mutant had been low yielding and highly susceptible to various diseases. It has been debated whether the low C18:3 character can be successfully combined with high seed yield for physiological reasons. Therefore, the low linolenic character of mutant M48 was transferred into high-yielding genotypes by repeated backcrossing to well-adapted low erucic acid, low glucosinolate (00-) winter rapeseed cultivars. Lines with low C18:3 content were selected from BC3 and BC4 generations and examined in 1990–95. Positive selection response for seed yield was shown to continue over the years. Presently, the best lines are yielding as well as the control cultivars being equivalent also in oil and glucosinolate contents. In order to test the effect of a low C18:3 content on seed yield, plants with low and with high C18:3 content, respectively, were selected from 16 segregating BC5-F₂ populations and bulked to form 32 F₃ populations. These ‘isogenic’ bulk populations were tested for field performance at four locations in 1995. The results show that C18:3 content of the seed oil is not associated with seed yield, oil content, beginning of flowering, plant height and disease resistance. Means of relative seed yield for the high and the low linolenic F₃ bulk populations were not significantly different with 88.0% and 86.9% of the control cultivars, respectively. There was a significant interaction between genotypes with high or low C18:3 content and location. This shows that under specific environmental conditions a low C18:3 content may be either favourable or unfavourable. The results indicate that the low C18:3 character of the original mutants per se does not cause a decrease in seed yield, oil content or general field performance.     

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.     

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.     

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.     

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.     

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.     

Marker-assisted increase of genetic diversity in a double-low seed quality winter oilseed rape genetic background

Generation of novel genetic diversity for maximization of heterosis in hybrid production is a significant goal in winter oilseed rape breeding. Here, we demonstrate that doubled haploid (DH) production using microspore cultivation can simultaneously introgress favourable alleles for double‐low seed quality (low erucic acid and low‐glucosinolate content) into a genetically diverse Brassica napus genetic background. The DH lines were derived from a cross between a double‐low quality winter rapeseed variety and a genetically diverse semisynthetic B. napus line with high erucic acid and high glucosinolates (++ quality). Twenty‐three low‐glucosinolate lines were identified with a genome component of 50–67% derived from the ++ parent. Four of these lines, with a genome component of 50–55% derived from the ++ parent, also contained low erucic acid. Heterosis for seed yield was confirmed in test‐crosses using these genetically diverse lines as pollinator. The results demonstrate the potential of marker‐assisted identification of novel genetic pools for breeding of double‐low quality winter oilseed rape hybrids.     

Emergence and performance of volunteer oilseed rape (Brassica napus) in different crops

Volunteer oilseed rape (OSR, Brassica napus L.) causes various agronomic problems in crop rotations and can contribute to gene dispersal by pollen and by seed admixture. A 4-year field experiment (2008–2011) was set up in south-west Germany to investigate the performance of volunteers derived from two OSR cultivars with different levels of seed dormancy. Volunteers of a high-dormancy (HD) and a low-dormancy (LD) OSR cultivar were deliberately generated by spreading 10,000 seeds m⁻² on a field in August 2008 and 2009. Four different crops were grown on that area in the first year following the seed rain: winter wheat (Triticum aestivum L.), winter turnip rape (Brassica rapa L.), spring barley (Hordeum vulgare L.) and field pea (Pisum sativum L.). In the second year, maize (Zea mays L.) was sown uniformly across all plots. Numbers of OSR seedlings emerging in early autumn shortly after seed rain were not connected with the size of the soil seed bank in early spring of the following year. The seeds of the HD-cultivar formed a much greater soil seed bank (up to 14% of the initially spread seed number) compared with the LD-cultivar (up to 1.3%) in the soil layer of 0 to 30 cm in early spring 2009 and 2010). Across all crops, considerably more volunteers of the HD-cultivar than of the LD-cultivar were present at several survey dates in the first year following seed rain. The highest number of volunteers originated from the HD-cultivar with up to 11 volunteers m⁻² in winter turnip rape compared with a maximum of 0.48 plants m⁻² in the other crops. Cultivar-specific differences in volunteer density were observed as well in maize two years after OSR seed rain. Flowering and seed setting volunteers were only present in 2010 and the flowering time was crucially overlapping with that of sown winter OSR. The reproductive ability (seeds produced m⁻²) of the LD-volunteers was five times lower in winter turnip rape than of the HD-volunteer; a similar trend was observed for the OSR volunteers in the other host crops.Strategies to definitely reduce unwanted effects of OSR volunteers, such as gene flow, should include the use of LD-cultivars with a low potential to form a soil seed bank, particularly if selective herbicides are not available, for instance in broad-leaved crops, or if the volunteers are herbicide-tolerant.     

Genetic control of yield and yield components in winter oilseed rape (Brassica napus L.) grown under nitrogen limitation

Despite its high nitrogen absorption capacity, oilseed rape (OSR) has a low apparent nitrogen use efficiency (NUE), which makes its production highly dependent on nitrogen fertilization. Improving NUE in OSR is therefore a main target in breeding. The objectives of the present work were to determine the genomic regions (QTLs) associated with yield and to assess their stability under contrasted nitrogen nutrition regimes. One mapping population, AM, was tested in a French location for three growing seasons (2011, 2012 and 2013), under two nitrogen conditions (optimal and low). Eight yield-related traits were scored and nitrogen-responsive traits were calculated. A total of 104 QTLs were detected of which 28 controlled flowering time and 76 were related to yield and yield components. Very few genotype × nitrogen interactions were detected and the QTLs were highly stable between the nitrogen conditions. In contrast, only a few QTLs were stable across the years of the trial, suggesting a strong QTL × year interaction. Finally, eleven critical genomic regions that were stable across nitrogen conditions and/or trial years were identified. One particular region located on the A5 linkage group appears to be a promising candidate for marker assisted selection programs. The different strategies for OSR breeding using the QTLs found in the present study are discussed.     

Genetic variation for saturated fatty acid content in a collection of European winter oilseed rape material (Brassica napus)

Oil quality is an important issue in oilseed rape breeding programmes. Recently, the seed oil content of saturated fatty acids has received substantial interest, because in a number of industrialized countries dietary recommendations not only limit total fat intake but also intake of saturated fatty acids. The aim of the present study was to analyse the genetic variation for saturated fatty acid content in two sets of modern winter oilseed rape cultivars ( Brassica napus L.) tested in field experiments under typical German growing conditions. The results showed highly significant genetic differences among the cultivars for total saturated fatty acid content, which ranged from 6.8% to 8.1%, with a mean of 7.4%. For both sets medium to high genotypic heritabilities were found for most fatty acids and for oil and protein content. Close negative correlations between palmitic acid, the predominant saturated fatty acid, and oleic acid as well as oil content were found.     

Energy balance of winter oilseed rape (Brassica napus L.) cropping as related to nitrogen supply and preceding crop

Data from a field experiment (1995–2000) conducted on a fertile sandy loess in the Hercynian dry region of central Germany were used to determine the energy efficiency of winter oilseed rape (Brassica napus L.) as affected by previous crop and nitrogen (N) fertilization. Depending on the previous crop, winter oilseed rate was cultivated in two different crop rotations: (1) winter barley (Hordeum vulgare L.)–winter oilseed rape–winter wheat (Triticum aestivum L.), and (2) pea (Pisum sativum L.)–winter oilseed rape–winter wheat. Fertilizer was applied to winter oilseed rape as either calcium ammonium nitrate (CAN) or cattle manure slurry. The N rates applied to winter oilseed rape corresponded to 0, 80, 160 and 240 kg N ha⁻¹ a⁻¹.Results revealed that different N management strategies influenced the energy balance of winter oilseed rape. Averaged across years, the input of energy to winter oilseed rape was highly variable ranging from 7.42 to 16.1 GJ ha⁻¹. Lowest energy input occurred when unfertilized winter oilseed rape followed winter barley, while the highest value was obtained when winter oilseed rape received 240 kg N ha⁻¹ organic fertilization and followed winter barley. The lowest energy output (174 GJ ha⁻¹), energy from seed and straw of winter oilseed rape, was observed when winter oilseed rape receiving 80 kg N ha⁻¹ as organic fertilizer followed winter barley. The energy output increased to 262 GJ ha⁻¹ for winter oilseed rape receiving 240 kg N ha⁻¹ as mineral fertilizer followed pea. The energy efficiency was determined using the parameters energy gain (net energy output), energy intensity (energy input per unit grain equivalent GE; term GE is used to express the contribution that crops make to the nutrition of monogastric beings), and output/input ratio. The most favourable N rate for maximizing energy gain (250 GJ ha⁻¹) was 240 kg N ha⁻¹, while that needed for minimum energy intensity (91.3 MJ GE⁻¹) was 80 kg N ha⁻¹ and for maximum output/input ratio (29.8) was 0 kg N ha⁻¹.     

Inheritance of progoitrin and total aliphatic glucosinolates in oilseed rape (Brassica napus L)

Inheritance of progoitrin and total aliphatic glucosinolate concentrations were investigated in oilseed rape, using parental, F₁, F₂ and first backcross generations, derived from a cross between resynthesized spring rape and a double-low spring rape cultivar. Progoitrin and total aliphatic glucosinolate concentrations were measured in mature seeds of single plants from these generations, using micellar electrokinetic capillary chromatography. For progoitrin, an additive/dominance model of gene action adequately explained the variation among the generation means, but for total aliphatic glucosinolate concentration, non-allelic interactions were also detected. Predictions based on estimates of the genetic parameters indicated that recombinant inbred lines, rather than second cycle hybrids, appeared to offer a better prospect of reducing glucosinolate concentrations in this material. Estimates of the minimum number of genes controlling these two characters were broadly in line with the number required for the known stages of their biosynthesis. This revised version was published online in July 2006 with corrections to the Cover Date.     

Molecular characterization of novel resynthesized rapeseed (Brassica napus) lines and analysis of their genetic diversity in comparison with spring rapeseed cultivars*

Resynthesized (RS) rapeseed generated from interspecific hybridization between suitable forms of Brassica rapa L. (syn. campestris; genome AA, 2n = 20) and B. oleracea L. (CC, 2n = 18) represents a potentially important resource to expand genetic diversity in the narrow gene pool of oilseed rape (B. napus L., AACC, 2n = 38). In this study 165 RS rapeseed lines originating from crosses between an Indian Yellow Sarson (B. rapa ssp. trilocularis) and five different cauliflower (B. oleracea convar. botrytis) cultivars were studied using amplified fragment length polymorphism (AFLP) markers and their genetic diversity was compared in relationship to an assortment of 40 diverse spring oilseed and fodder rape varieties. Using three AFLP primer combinations, a total of 467 polymorphic bands were scored. Cluster analysis allowed differentiation among the different RS lines, which, as expected, were genetically highly divergent from the cultivars. The genetic diversity of the material is discussed in relation to its morphological variability with a view to the implementation of RS lines in oilseed rape breeding.     

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.     

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.     

Overwintering of winter cereals in Hungary in the case of global warming

Summary Under phytotronic conditions investigations were made on the effect of important environmental factors, such as temperature, water and an increasing concentration of atmospheric CO₂, on the hardening of young cereal plants.In all the varieties derived from the major wheat growing regions of the world the hardening process was favourably influenced by a doubling of atmospheric CO₂ content, so that a significantly larger number of plants survived the frost test than for plants of the same variety raised under normal conditions.A reduction in freezing temperature and an increase in soil moisture content caused a slight reduction in survival % for varieties with excellent frost resistance and a great reduction for those with medium or poor frost resistance.Predictions suggest that in Central Europe, as the result of global climatic changes, there will be a reduction in the quantity of winter precipitation, a considerable rise in winter temperatures and an increase in atmospheric CO₂ concentration. Judging by the experimental results, these changes could improve the overwintering of winter cereals; at the same time, however, a number of factors (mainly the reduction of precipitation) leading to yield losses must be expected during the vegetation period.     

氮钾配施对油菜产量及氮素利用的影响

实际生产中氮钾肥投入不平衡严重限制了氮肥肥效及作物的产量潜力。为了探明不同施氮量下钾肥施用对油菜产量及氮素利用的影响,于2016—2017年及2017—2018年在湖北省武穴市开展连续2年的田间试验,采用氮钾两因素完全试验设计,设氮0、90、180、270 kg N hm-2和钾0、60、120、180 kg K2O hm-2各4个水平。在油菜成熟期取样测定产量、地上部氮钾积累量以及氮肥利用率。结果表明,在钾供应不足时(K0和K60),冬油菜施用氮肥的平均增产率为113.7%,而在钾供应充足的条件下(K120和K180),施用氮肥的平均增产率高达172.9%;与K0处理相比, K120处理冬油菜氮肥回收利用率平均提高了16.6%,继续增施钾肥对不同施氮量下冬油菜氮肥回收利用率的进一步提高无显著影响;达到区域平均产量时,钾供应充足较低钾(K60)投入平均降低33.9%的氮肥用量。综上所述,氮钾配施显著提高了冬油菜产量和氮肥利用率,在冬油菜实际生产中除了重视氮肥施用外,应增加钾肥投入,通过优化氮钾肥配施比例可进一步提高油菜产量,实现冬油菜高产和养分高效。     

Genetic diversity of oilseed Brassica napus inbred lines based on sequence-related amplified polymorphism and its relation to hybrid performance

Significant heterosis for seed yield in oilseed rape has created interest in the development of hybrid cultivars. The DNA‐based marker protocol, sequence‐related amplified polymorphism (SRAP) was used to determine genetic diversity among oilseed rape maintainer and restorer lines. This measure was used in an attempt to establish an association between genetic distance and heterosis in hybrids for various agronomic traits. A total of 118 polymorphic loci were generated by 18 SRAP primer combinations. Based on the polymorphism generated by the markers, calculated similarity index values ranged from 0.46 to 0.97. Cluster analysis grouped 10 maintainer and 12 restorer lines into three groups, with the exception of two maintainer lines, PM5 and PM9, which fell outside these groups. The grouping of the lines was largely in agreement with the available pedigree data on their origin and agronomic performance. Analysis of variance among inbred lines and their resulting F₁ hybrids over two locations revealed significant differences for plant height, days to maturity and seed yield, but not for oil content. Substantial mid‐parent heterosis was observed only for seed yield, and ranged from 26% to 169%. All hybrids surpassed their respective inbred lines for this trait, except for a single cross combination of related lines. In general, crosses of lines located in different clusters yielded more than those from the same clusters. Regression analysis revealed a statistically significant relationship between the genetic distance of the parents and seed yield in their hybrid, and their derived mid‐parent and high‐parent heterosis. The correlation coefficient between genetic distance and yield (0.64) indicated a moderately strong relationship, so it is possible that some of the SRAP markers might be linked to quantitative trait loci for seed yield.