Variation and inheritance of erucic acid content in Brassica carinata germplasm collections from Ethiopia

Ethiopian mustard possesses a number of agronomic advantages over other oilseed crops with similar ecological adaptation in Ethiopia. However, its high erucic acid content is undesirable for a vegetable oil. Although efforts have been made to improve its quality, much has to be done to use natural variations that might exist within the species for fatty acid contents. This project was undertaken to study the variability of fatty acid contents, primarily erucic acid, in germplasm collections of Ethiopian origin, with an attempt to develop low (zero) erucic acid genotypes. The study used inbred lines as well as F₂ populations of 10 crosses between six parental lines. A wide variation in fatty acids was found. Oleic acid content varied from 5 to 34% and erucic acid content from 6 to 51%. Linoleic and linolenic acid contents were less variable. The high‐oleic genotypes exhibited not only low erucic but also higher linoleic (25%) and considerably lower linolenic acid (8%) contents. It was possible to classify the F₂ populations with the lowest erucic acid into three distinct classes. While the first class had an erucic acid content of 6–12%, the second and third classes had contents of 18–32% and 36–42%, respectively. The existence of a multiple allelic series of erucic acid in Ethiopian mustard would enable its fixation at zero levels without necessarily going into interspecific crossing.     

Genetics of the Erucic Acid Content in Interspecific Hybrids of Ethiopian Mustard (Brassies carinata Braun) and Rapeseed (B. napus L.)

Ethiopian mustard (Brassica carinata Braun) is a potential oil crop in which genes for low erucic acid content of the seed oil have not yet been found. In order to solve this problem the potential of rapeseed (B. napus L.) varieties as a source of these genes has been tested. Reciprocal F₁ hybrids between B. carinata and a low erucic acid variety of B. napus, F₂, and backcrosses with B. carinata were obtained. The fatty acid composition was determined in half seeds of F₁ and segregating generations from reciprocal interspecific crosses. The genetic analysis indicated that the erucic acid content of the seed oil of B. carinata is controlled by two genes with no dominance and additive in action.     

Increasing erucic acid content in Ethiopian mustard through mutation breeding

Ethylmethane sulphonate (EMS) was applied to seeds of the Ethiopian mustard (Brassica carinata A. Braun) line C-101. Bulk samples of M₃ seeds from 8331 M₂ plants were evaluated for the fatty acid composition of their oil by near-infrared reflectance spectroscopy (NIRS) and by further gas chromatography on selected samples. A putative mutant, N2-6230, showing very low oleic acid content (4.7% vs. average of 8.6% in C-101) and erucic acid content within the range of variation of the line C-101 (40-49.3%) was identified. The M₃ progeny of this mutant showed a wide segregation for erucic acid content (39.1-57.9% vs. 41.8-50.3% in C-101), and maintained levels of oleic acid lower than in line C-101. Selection for high erucic acid content in the M₃ and M₄ generations led to the fixation of this mutation in the M₅ generation (52.2-59.3% vs. 39.0-47.6% in C-101). This is the first high erucic acid line obtained in Brassica species through mutation breeding. Its utility in future programmes to develop very high erucic acid lines is discussed.     

Inheritance of reduced linolenic acid content in the Ethiopian mustard mutant N2-4961

The zero erucic acid Ethiopian mustard lines developed so far are characterized by an exceptionally high linolenic acid content in the seed oil. The mutant line N2‐4961, expressing low linolenic acid content in a high erucic acid background, was developed through chemical mutagenesis. The objective of this research was to study the inheritance of low linolenic acid content in this mutant. Line N2‐4961 was reciprocally crossed with its parent line C‐101 and the linolenic acid content of the reciprocal F₁, F₂ and BC₁ generations was studied. No maternal, cytoplasmic or dominance effects were detected in the analysis of F₁ seeds and F₁ plants from reciprocal crosses. Linolenic acid content segregated in 1: 2: 1 ratios in all the F₂ populations studied, suggesting monogenic inheritance. This was confirmed with the analysis of the reciprocal backcross generation. The simple inheritance of low linolenic acid content in N2‐4961 will facilitate the transference of this trait to zero erucic acid lines of Ethiopian mustard.     

Allelic variation in linolenic acid content of high erucic acid Ethiopian mustard and incorporation of the low linolenic acid trait into zero erucic acid germplasm

Zero erucic acid germplasm of Ethiopian mustard is characterized by high levels of linolenic acid (about 21%). Two genetic sources of low linolenic acid (N2‐4961 and HF‐186, about 5%), have been developed in a high erucic acid background. The objectives of this research were to study the genetic relationship between the two low linolenic acid lines and to transfer the trait to zero erucic acid germplasm. F1 seed generations from crosses between both lines had higher average linolenic acid concentration than both parents. F2 seeds segregated for linolenic acid content following a continuous variation from 1.8 to 7.4%, exceeding the limits of the parental distribution ranges. Transgressive recombinants with very low linolenic acid concentration (0.7‐2.7%) were confirmed in the F3 seed generation. The results suggested that N2‐4961 and HF‐186 possess alleles for low linolenic acid at different loci. Transgressive low linolenic acid F3 plants were crossed with plants of a zero erucic acid line and a selection for zero erucic, low linolenic acid was conducted. As a result, a zero erucic acid F3:4 line containing 1.5 ± 0.7% linolenic acid was developed.     

Interspecific hybridization between Abelia × grandiflora ‘Francis Mason’ and A. schumannii via ovule culture

Ethiopian mustard (Brassica carinata Braun) is a potential oil crop for the Mediterranean area. The objective of this study was to develop an efficient system of mutagenesis using ultraviolet (UV) light irradiation of isolated microspores from Brassica carinata. From the survival curve based on embryo yield after irradiation of the microspores with UV light, the LD50 was estimated to be an exposure of 8 min. Total content of glucosinolates and fatty acid composition were analysed in the seeds of the doubled haploid homozygous plants with the purpose of selecting lines with modified glucosinolate and erucic acid contents. Three groups of doubled haploid lines exhibiting low and high glucosinolate contents, and high erucic acid content have been identified from a population of 270 doubled haploid lines. In eight lines, the content of glucosinolates was reduced from an average of 80.6 mol g^-1 seed to 37.5 mol g^-1 seed, whereas in four lines, the content of glucosinolates was increased up to 99.2 mol g^-1 seed. In six additional lines, the content of erucic acid was increased from 42.8% in the nontreated lines to 49.5% of the totalfatty acid composition in some of the mutant lines. All lines showed stablelevels of erucic acid in two generations, the M₂ and M₃.     

Selection for reduced linolenic acid content in Ethiopian mustard (Brassica carinata Braun)

Reducing linolenic acid content is one of the most important objectives for the development of Ethiopian mustard lines with high oil quality. This work was aimed at searching for variability of the fatty acid composition of oil within a germplasm collection of Ethiopian mustard. A total of 217 lines were analysed by gas-liquid chromatography (GLC) in 1991, and one was selected as having reduced values of both linolenic acid content (10.2% versus 14.0% of total fatty acids as the collection average) and linoleic acid desaturation ratio (LDR, 0.34 versus 0.45). After 3 years of pedigree selection for low linolenic acid content, this line showed, in 1995, average values of this fatty acid of 5.4% and 2.4% in two different environments, compared with 11.6% and 8.3%, respectively, in the control. The values of the LDR were 0.18 and 0.09, respectively, compared with 0.36 and 0.27 in the control line.     

Fatty acid composition of resynthesized Brassica napus and trigenomic Brassica void of genes for erucic acid in their A genomes

The fatty acid composition of seed oil of four interspecific hybrids, resulting from crosses between zero erucic acid Brassica rapa (AA), and high erucic acid Brassica alboglabra/Brassica oleracea (CC) and Brassica carinata (BBCC), void of erucic acid genes in their A‐genomes was examined. The erucic acid content in resynthesized Brassica napus (AACC) lines derived from these crosses was only about half that of the high erucic acid CC genome parents, indicating equal contributions of the two genomes to oil (fatty acid) synthesis and accumulation. The differences in C18 fatty acid synthesis between the parents were also evident in the resulting resynthesized B. napus plants. Hexaploid Brassica plants of the genomic constitution AABBCC, in which the AA genome was incapable of erucic acid synthesis, had lower erucic acid contents than the B. carinata (BBCC) parent. This is plausible considering the fact that the zero erucic acid AA genome contributes to oil synthesis in AABBCC plants, thus reducing erucic acid content.     

Inheritance of long chain fatty acid content in rapeseed (Brassica napus L.)

A large industrial oil market has recently developed for high erucic acid (>500 g kg⁻¹) rape (Brassica napus L.) cultivars. This research was conducted to: (i) determine genetic effects for five fatty acids, (ii) determine if maternal effects influence fatty acid content of progeny, and (iii) estimate correlations among fatty acid contents in hybrid progeny. Lines with very high erucic acid content and very low erucic acid content were used to develop eight generations to estimate additive, dominance, and epistatic effects for fatty acid content using Generation Means Analyses. Mean oleic, linoleic, linolenic, eicosenoic and erucic acid content differed among generations and additive genetic effects were important for control of all five fatty acids, contributing from 84% to 97% of the total sums of squares for each fatty acid. Epistasis was observed in the inheritance of eicosenoic acid. Maternal effects were not detected.     

Sunflower mutant containing high levels of palmitic acid in high oleic background

A new sunflower mutant, CAS-12, was obtained, which has both high palmitic (≈30%) and high oleic acid contents, and also a substantial amount of palmitoleic acid (≈7%). The mutant was selected after X-ray irradiation of dry seeds of the inbred line BSD-2-423, which had normal palmitic (≈3%) and high oleic (≈88%) acid levels. The increase of palmitic and palmitoleic acids occurred at the expense of the oleic acid content, which decreased to around 55% in respect to the original line. Linoleic acid content is always under 5%. Palmitic and palmitoleic acid levels were similar to those of the high palmitic mutant CAS-5 obtained in a previous programme from a low oleic line isogenic to BSD-2-423 using a similar mutagenic treatment. In that previous programme we also selected three high stearic acid mutants using chemical mutagenic treatment on the same sunflower line (RDF-1-532). We attempted to obtain mutants in other lines but were unsuccessful. The isolation of similar mutants in isogenic parental lines illustrates the importance of the genetic background in the development of specific mutants with an altered seed oil fatty acid composition. The oil of this mutant will increase the range of potential uses of sunflower oil. This revised version was published online in July 2006 with corrections to the Cover Date.     

Erucic acid heredity in Brassica juncea - some additional information

Genetic studies were undertaken to reassess erucic acid heredity in Brassica juncea. Analysis of segregation in F₂ and BC₁ generations from two zero × high erucic acid crosses indicated that higher erucic acid in B. juncea was controlled by two dominant genes with additive effects, whereas segregation in a cross involving ‘CCWF 16′, a genotype having intermediate erucic acid (25.6%), and a zero erucic acid strain, indicated monogenic dominant control for intermediate erucic acid content. The B. juncea strain ‘CCWF 16’ was developed by hybridizing high‐erucic acid B. juncea cv.‘WF‐1’ with a ‘0’ erucic B. rapa cv.‘Candle’ followed by backcrossing with ‘WF‐1’ and half‐seed selection for low erucic acid in each backcross generation. This strategy resulted in substitution of the high erucic acid allele present in the A genome of B. juncea (AABB) by the zero erucic acid allele associated with ‘A’ genome of ‘Candle’. The intermediate erucic acid content in ‘CCWF 16’ was thus attributed to a gene present in the ‘BB’ genome. Experimental data clearly suggested that the gene (E₂) associated with the A genome had a greater contribution to the total erucic acid content in B. juncea than the gene (E₁) located on the B genome. This provided experimental evidence for a previous suggestion of unequal contributions of two dominant genes (E₁= 12%, E₂= 20%) to high erucic acid content in conventional digenomic Brassica species.     

Doubled haploid lines of Brassica carinata with modified erucic acid content through mutagenesis by EMS treatment of isolated microspores

Brassica carinata is a potential oilseed crop for the Mediterranean area. Chemical mutagenesis has been applied to microspores of B. carinata with the purpose of identifying lines with altered erucic acid content. From a population of nearly 400 doubled haploid plants recovered, nine lines have been identified that exhibit promising useful changes in erucic acid concentration in the seed oil. Three lines showed erucic acid contents below 25%, with a minimum of 17.1%, and in six lines the level of this fatty acid was greater than 52%. Changes in other fatty acids are also described and discussed.     

Orychophragmus violaceus, a Potential Edible-oil Crop

Orychophragmus violaceus, a member of the Cruciferae family, has been found to have a high oil quality with high contents of palmitic (14.3 %) and oleic (20.3 %) acids, and lower contents of linolenic (4.8 %) and erucic (0.9 %) acids. Plants of O. violaceus exhibit a high number of branches, pods per plant, and seeds per pod, which contributes to the high yield potential of this plant. Individual selection was made in the original population of O. violaceus, and a few early, disease-tolerant and high-yielding lines were obtained. Intergeneric hybridization was performed between B. napus and O. violaceus and several hybrid plants (F₁) were obtained. After treatment with colchicine, amphidiploid plants developed. O. violaceus shows great potential for becoming an edible oil crop or being used as genetic material in a rapeseed breeding programme.     

Evaluation of microspore-derived embryos as models for studying lipid biosynthesis in seed of rapessed (Brassica napus L.)

Summary Microspore culture of rapeseed (Brassica napus L.) has provided a powerful tool not only for breeding but also in developmental studies. In this study, microspore-derived embryos (MDE) of B. napus were evaluated as a model in seed for studying accumulations of triacylglyceride (TAG) fatty acids in both a low and high erucic acid rapeseed line; and accumulations of TAG and free fatty acids (FFA) in a high erucic acid rapessed line. The accumulation patterns confirmed that MDE had a similar TAG fatty acid profile to seed during the embryo development within each genotype. The oil accumulation in MDE after 36 days in culture (DIC) approached levels similar to those in zygotic seed 25 days after flowering (DAF). Significant differences were detected in contents of both total free fatty acids and specific free fatty acids between MDE and seed. During the developmental period, total free fatty acids changed from 16% to 2.1% in MDE, but from 10.5% to 0.1% in seed. MDE had much higher percentage of free linolenic and erucic acids than seed, particularly during the late developmental stages. The current study indicated that MDE can be used as a model to study TAG and TAG fatty acids in seed but caution must be taken to study free fatty acid metabolism.     

Effect of Nitrogen and Sulphur Application on Yield and Fatty acid Composition of Mustard (Brassica juncea L.) Oil

The effects of nitrogen and sulphur on the yield and fatty acid composition of mustard ( Brassica juncea L.) oil were studied in a field experiment. Significantly higher grain and oil yields were obtained with N and S application. Applications of nitrogen up to 60 kg ha⁻¹ and sulphur up to 40 kg ha⁻¹ favourably influenced the grain yield. Increasing levels of N decreased the oil content while application of sulphur improved the oil content. The contents of linoleic and linolenic acid were maximum (16.82 and 8.73%, respectively) with 60 kg N along with 40 kg S ha⁻¹. No use of fertilizers led to higher contents of undesirable fatty acids such as palmitic (hypercholesterimic) and erucic (do not have food value) acids.     

Morphological evaluation of olive germplasm present in Tuscany region

Undesirable characteristic of rapeseed oil is a relatively high level of linolenic acid (18:3), which is easily oxidized leading to rancidity and a shortened shelf life of the oil. Previous attempts to reduce linolenic acid levels in rapeseed oil through breeding have been impaired by complex genetics and strong environmental sensitivity of this trait. Therefore, our objective was to develop molecular markers for low linolenic acid that could facilitate the breeding of low linolenic rapeseed. Bulked segregant analysis was employed to identify two RAPD markers associated with 18:3 in a doubled haploid population segregating for linolenic and erucic acid levels. Based on analysis of individual DH lines, the markers RM350 and RM574, representing two independent loci, accounted for a total of 39% of the genetic variability in this population. This marker RM350 alone accounted for 25% genetic variation for this trait with no evidence of recombination. Significant interlocus interaction found between the markers RM350 and RM574 suggested that epistasis was involved in the genetic control of 18:3 level in this population. Another marker designated as RM322, which was independent of the other two, was found significantly associated with the erucic acid level and oil content. RAPD markers identified in this study should be a useful tool for the early detection of low linolenic, or low or high erucic acid genotypes in rapeseed breeding programs based on doubled haploids. This revised version was published online in July 2006 with corrections to the Cover Date.     

Combination of resistance to Verticillium longisporum from zero erucic acid Brassica oleracea and oilseed Brassica rapa genotypes in resynthesized rapeseed (Brassica napus) lines

Resynthesized (RS) forms of rapeseed (Brassica napus L.; genome AACC, 2n = 38) generated from interspecific hybridization between suitable genotypes of its diploid progenitors Brassica rapa L. (syn. campestris; genome AA, 2n = 20) and Brassica oleracea L. (CC, 2n = 18) represent a potentially useful resource to introduce resistance against the fungal pathogen Verticillium longisporum into the gene pool of oilseed rape. Numerous cabbage (B. oleracea) accessions are known with resistance to V. longisporum; however, B. oleracea generally has high levels of erucic acid and glucosinolates in the seed, which reduces the suitability of resulting RS rapeseed lines for oilseed rape breeding. In this study resistance against V. longisporum was identified in the cabbage accession Kashirka 202 (B. oleracea convar. capitata), a zero erucic acid mutant, and RS rapeseed lines were generated by crossing the resistant genotype with two spring turnip rape accessions (B. rapa ssp. olerifera) with zero erucic acid. One of the resulting zero erucic acid RS rapeseed lines was found to have a high level of resistance to V. longisporum compared with both parental accessions and with B. napus controls. A number of other zero erucic acid RS lines showed resistance levels comparable to the parental accessions. In the most resistant RS lines the resistance and zero erucic acid traits were combined with variable seed glucosinolate contents. Erucic acid‐free RS rapeseed with moderate seed glucosinolate content represents an ideal basic material for introgression of quantitative V. longisporum resistance derived from B. oleracea and B. rapa into elite oilseed rape breeding lines.     

Genetic control of fatty acid composition in seed oil of Sinapis alba L.

Summary Inheritance of fatty acid composition was studied in an F1 diallel cross in Sinapis alba. Crosses were made among accessions having contrasting amounts of oleic (C18:1) and erucic (C22:1) acid. Concentrations of oleic, linoleic (C18:2), eicosenoic (C20:1) and erucic (C22:1) acids were determined by gas-chromatography for each mating combination. Genetic analysis confirmed that the composition of the fatty acids was controlled mainly by the nuclear genes of the embryo. Additive gene action with partial dominance for the reducing alleles was noted for oleic and linoleic acids, while erucic acid showed an additive mode of inheritance with partial dominance for the enhancing alleles. Positive heterosis was demonstrated for eicosenoic acid content. Erucic acid content was strongly negatively correlated with oleic acid, suggesting a genetic interdependence between the two fatty acids. Broad-sense and narrow-sense heritability estimates for each of oleic, linoleic and erucic acids were very high, due to low between-plants non-genetic component of variance.Contribution No. 3662-E, 1992 series.     

工业专用型高芥酸油菜新品种选育

高芥酸油菜品种是在工业上具有广泛用途的专用型品种。本研究通过两个常规芥酸品种杂交,采用以单株和单粒筛选相结合对芥酸含量正向选择为核心的技术,育成了芥酸含量达60%的甘蓝型高芥酸油菜新品种高芥1号。同时阐明了在高芥酸含量背景下各种脂肪酸间的相关性,为相关育种提供了—些理论依据。     

Independent Inheritance of Erucic Acid Content and Flower Colour in the C-Genome of Brassica napus L.

The synthetic Brassica napus L. line No7076 was obtained from a cross between yellow-flowered and zero-erucic turnip rape (B. campestris) Sv85-38301 and white-flowered and high-erucic (41.4%) B. oleracea ssp. alboglabra No6510. This synthetic B. napus is pale-flowered and has an average erucic acid content of 25.8 %. It was crossed with the yellow-flowered and zero-erucic B. napus line SvS4-2S053 and segregation of the erucic acid content and flower colour was studied in F₁ and F₂ generations. The high erucic acid content was controlled by a single gene in the C-genome and was additively inherited. Strong evidence was obtained in support of independent segregation of the erucic-arid content and the flower colour characters controlled by the C-genome of B. napus.