Importance of sepals in fruit and seed development in linseed (Linum usitatissimum L.)

Summary The green sepals in linseed (Linum usitatissimum L.) persist till almost the time of fruit ripening. Since the plant retains very few leaves and the fruits are better exposed to receive sunlight, the green sepals may be important in fruit and seed development. Influence of the removal of 0, 1, 3 and 5 sepals on the day of anthesis, 7, 15 and 25 days after anthesis on fruit and seed development was examined. Excision of all the fine sepals on the day of anthesis stopped fruit and seed development in the varieties M 10 and NP (RR) 45 but in Mukta the reduction was about 55%. Removal of three sepals also decreased fruit growth in Mukta and NP (RR) 45. The effect of removal of sepals was more pronounced on the number of seeds per fruit than on seed weight. Sepals fixed ¹⁴CO₂ and translocated ¹⁴C to seeds. However, the ¹⁴CO₂ fixation was maximum at early stages of fruit development, whereas a higher percentage of the fixed ¹⁴C was translocated at later stages of growth. It is suggested that the sepal size in linseed could be a useful selection criterion.     

The application of chemical mutagenesis and biotechnology to the modification of linseed (Linum usitatissimum L.)

Summary In the early 1980s the phenomenon of somaclonal variation induced by cell culture was exploited to produce genetic variation in linseed. The linseed variety Andro, derived from the widely grown Canadian variety McGregor, was selected in saline culture and was released for production in Canada. Andro possesses traits very different from its parent, such as increased seedling vigour and tolerance to heat stress. Additional stable somaclonal variation in characters such as yield, days to maturity, seed weight and oil content were subsequently induced in McGregor. However, despite extensive screening of the somaclonal variants, no significant variation in the fatty acid profile was found.Chemical mutagenesis using ethyl methanesulphonate was, however, succesful in modifying the fatty acid profile of McGregor. Initial screening of M₂ seed by the thiobarbituric acid colourimetric procedure was followed by gas chromatography to select half-seeds with atypical fatty acid profiles. Two independent, partially dominant genes were identified that were responsible for reducing the linolenic acid (18 : 3) from 50% to 2% while increasing linoleic acid (18 : 2) to 70%. A single, partially dominant gene, inherited independently of the linolenic acid genes, increased palmitic acid (16 : 0) from 7% to 30% and palmitoleic acid (16 : 1) from trace amounts to 4%. Agrobacterium-mediated transformation of linseed has also been successful. Herbicide tolerance genes for glyphosate, sulfonylurea and phosphinothricin have been incorporated into Canadian varieties. Commercially useful levels of tolerance to sulfonylurea herbicides have been achieved with no adverse agronomic affect. It is expected that a transgenic variety containing this resistance will be registered for commercial production in Canada in 1994.Standard breeding techniques, the application of antisense technology and the overexpression of fatty acid synthesis genes are being used to further modify the fatty acid profile of linseed, as well as for the transfer of abiotic stress-related genes identified in bromegrass.     

Inheritance of resistance to Oidium liniSkoric in linseed (Linum usitatissimum L.)

Summary Resistance to powdery mildew (Oidium lini) in each of the four linseed cultivars, viz., LC 216, LC 255, LC 256 and LC 269 was found to be conditioned by one dominant gene. Crosses between the resistant parents indicated that the same gene was present in all the cultivars and is designated as Ol.     

Genetic analysis of the response of linseed (Linum usitatissimum L.) somatic tissue to in vitro cultivation

The response of linseed somatic tissue to in vitro cultivation can be improved if the gene action involved is known. Nine homozygous lines, seven F₁ and fifty F₂ plants were cultivated in MS basal medium with 0.1 mg/l BAP. Callogenesis and regeneration analyses were done using a random model in order to separate environmental and genotypic effects at 10, 20 and 30 days after culture initiation. Genetic variability for callogenesis and regeneration was observed, that could be used in a breeding program. The appropriate period for the evaluation of callogenesis was at 20 days after culture initiation. For regeneration, this moment would depend on the selection criterion adopted. To select genotypes on the basis of earliness in shooting at 10 days would be preferable, while for ability of producing shoots, 30days would be more suitable. For both characters the analyses of genetic action indicated that, non-additive effects were involved in the culture response, however the expression of callogenesis and regeneration ability had different genetic basis. This revised version was published online in August 2006 with corrections to the Cover Date.     

Comparison of pedigree selection and single seed descent for oil yield in linseed (Linum usitatissimum L.)

Summary Four populations of linseed derived from five parents were advanced from the F₂ to the F₆ generation by SSD and to the F₅ by pedigree selection. These populations were used to compare the efficiency of the two methods in order to provide superior genotypes with respect to grain yield and oil content, i.e. oil yield. The results showed minimum differences between pedigree and SSD lines for grain yield, where in only one cross the SSD lines were significantly superior to the pedigree lines. Since pedigree selection was carried out for both, seed yield and oil content, a positive response to selection was expected. However, early selection for yield, a character with low heritability, was not successful. On the contrary, significant differences for oil content were detected between the two groups of lines in three of the four crosses studied. In these cases the pedigree lines were superior to the SSD lines. These results demonstrate that strict selection in early generations for oil content, a character with comparatively high heritability, is feasible and successful in linseed. However, selection for seed yield should be postponed to later inbred generations. Consequently, in breeding for maximum oil yield of linseed a two-step selection procedure is recommended.Abbreviations PS pedigree selection - SSD single seed descent - TGW thousand grain weight     

Genotypic and exogenous factors affecting shoot regeneration from anther callus of linseed (Linum usitatissimum L.)

Summary The objective of this study was to investigate factors affecting the regeneration capacity of linseed anther culture. Four different environmental conditions in a phytotron were tested with regard to their effects on anther donor plants of cv. Hella. Anther response and shoot regeneration from anther callus was maximal when donor plants were grown in a 16 hrs-day at 14°C day/8°C night temperature. Anthers of four linseed genotypes were cultured on different media. Maximum shoot regeneration was achieved when the induced calli were transferred onto a modified N6 medium containing zeatin (1 mg l^-1). Most of the calli regenerated shoots in the second subculture on regeneration media. Shoots were rooted on modified B5 or MS media containing NAA (0.1 mg l^-1). Cytological examinations of incubated anthers and root tips of regenerated plants indicated that the anther calli were derived from microspores.Abbreviations B5 Gamborg's (1975) medium - BAP 6-benzylaminopurine - 2,4D dichlorophenoxyacetic acid - N6 Chu's (1978) medium - NAA -naphthaleneacetic acid - MS Murashige & Skoog's (1962) medium - ZEA zeatin     

Breeding for Modified Fatty Acid Composition by Induced Mutations in Linseed (Linum usitatissimum L.)

Seeds of linseed (Linum usitatissimum L.) cultivars ‘Raulinus’ and ‘Bionda’ were treated mutagenically with ethyl methanesulfonate (EMS) in order to broaden the variation of fatty acid composition in the seed oil and to select mutants with reduced linolenic acid (C18: 3) content. The M₂ generation was evaluated for fatty acid composition of the oil. Genetic variation for unsaturated fatty acids was demonstrated to be extended in the M₂ generation as compared to the controls. Finally in the M₅ generation of cv. ‘Raulinus’, lines with reduced C18:3 (< 40% vs. 55%) and correspondingly increased linoleic or oleic acid contents, respectively, could be selected. These mutants proved to be genetically stable as demonstrated by parent-offspring (M₄/M₅) regression.     

Comparison of Mahalanobis's method and metroglyph technique in the study of genetic divergence in Linum usitatissimum L. germ plasm collection

Summary A group of fifty-seven varieties of Linum usitatissimum L. collected from different geographics sources was studied to understand the nature of genetic divergence using Mahalanobis's statistical distance (D²). Varieties having genetic similarity were grouped into fifteen clusters. Most of the clusters at individual level included varieties of different geographical regions. However, Argentine and Indian types formed separate and distinct constellations. The Indian group was situated at a maximum distance from Argentine and US groups, and was nearest to the Australian group. For improvement of Indian linseed, hybridization of indigenous types with Argentine and US types might result in superior recombinants.84.64% of the total divergence was attributable to plant height and seed size, which are based characteristics used in classifying the species into oil and flax types.Since it is difficult to compute D² values among large numbers of germplasm collections, another relatively simple and easy method, the metroglyph technique, was tried upon the same set of fifty-seven varieties. Comparisons between the results of the two analyses revealed the striking similarity as regards to group arrangement. It has been suggested that the metroglyph technique would be suitable for preliminary grouping prior to undertaking D² analysis.     

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.     

Isolation of induced mutants in Ethiopian mustard (Brassica carinata Braun) with low levels of erucic acid

Ethiopian mustard (Brassica carinata Braun) is a potential oil crop for the rain-fed Mediterranean area. However, its usage is limited by the high erucic and high glucosinolate content of the oil and meal, respectively. In the course of a mutagenesis programme, an agronomically good line of Ethiopian mustard was treated with EMS in order to widen the natural variability of nutritional traits in this species. As a result of this programme several low erucic mutants were isolated; two of these mutants showed erucic acid values in the M4 generation in the range 5–10% of total fatty acids. Near-infrared reflectance spectroscopy (N1RS) was successfully applied as a rapid screening method for erucic acid in this breeding programme.     

Embryonic heterosis in the linolenic acid content of Matthiola incana seed oil

Summary The fatty acid composition of seed-oil of breeding lines and F₁ hybrids of Matthiola incana was analyzed, using direct esterification and gas chromatography. The breeding lines tested differed significantly with respect to the levels of palmitic, oleic, linoleic and linolenic acids. Embryonic-stage heterosis in linolenic acid concentration was demonstrated by F₁ hybrid seeds, derived from mating horticulturally different lines of M. incana. Linolenic acid content was negatively correlated with both oleic acid content (r=–0.85) and linoleic acid content (r=–0.66). None of the breeding lines or the F₁ hybrids significantly passed the limit of 67% linolenic acid. Possible genetic and biochemical explanations for the above phenotypic data are discussed.     

The effect of T-DNA on the agronomic performance of transgenic flax plants

Summary Registration and commercial release of crop cultivars improved using recombinant DNA technologies will require extensive field testing of the transgenic lines. In addition to testing the efficacy of the transferred gene(s), regulatory bodies and farmers need data from large scale agronomic studies to provide assurance that the foreign DNA does not adversely affect normal agronomic productivity or quality of the crop. Here, five transgenic sulfonylurea resistant flax (Linum usitatissimum) lines were included in a multiple location cultivar registration trial, evaluated and compared with four standard commercial cultivars and 24 other (conventionally produced) breeding lines. The transgenic lines did not appear to be affected by T-DNA for any quality or agronomic parameter tested, including yield.     

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.     

Identification of RAPD markers linked to linolenic acid genes in rapessed

Summary The inheritance of the low linolenic acid content (derivated from mutant lines) in rapeseed was investigated. Molecular techniques of gene mapping through RAPD markers were applied on a microspore-derived progeny from a high × low linolenic acid F₁ hybrid. Bulked segregant analysis made it possible to test rapidly number of RAPD primers. Two linkage groups of 6 markers (72.7 cM and 75.6 cM) were determined. Each corresponded to a major QTL which explained 24% and 30.7% of the total phenotypic variation of the linolenic acid content. It was confirmed that two independant mutations were implied in the low linonenic acid content.     

Genetic analysis of the high oleic acid content in cultivated sunflower (Helianthus annuus L.)

Summary Sunflower lines breeding true for very high oleic acid content in their oil (average levels higher than 85%) were crossed with standard sunflower lines with mean oleic acid levels of 30%. Analysis of the oil of F₁ seeds indicated dominance for high oleic levels and control of the genotype of the embryo. Segregating generations were obtained selfing heterozygous high oleic BC_nF₁ plants from several generations of a backcrossing program to incorporate the high oleic character to standard inbred lines and testcrossing these plants to low oleic material. Analysis of F₂ and testcrossed seeds showed three kind of segregations, in both F₂ and testcrossed populations, with different proportions of low, intermediate and high oleic types. Genetic analysis of these data supported the hypothesis, that the high oleic character is controlled by three dominant complementary genes OL₁, OL₂ and OL₃. Additional data showing F₁ seeds with intermediate oleic content and segregations for high oleic in progenies of intermediate types, suggest the presence of major factors modifying high oleic acid content.     

Studies on the amylose content and gelatinisation temperature in certain local cultivars and induced grain shape mutants in rice

Summary Ninety local cultivars and 124 induced grain shape mutants were screened for their amylose content and gelatinisation temperature as indexed by the alkali disintegration patterns. The amylose content was high in most of the local cultivars and ranged from 19.2 to 32.0% and from 12.3 to 33.2% in the mutants. Wide variation was also observed for the alkali disintegration values and most of the varieties had low alkali values. A positive but low correlation between alkali values and amylose content and a negative correlation between alkali values and protein content was observed. The preliminary studies suggest that the amylose content at different milling levels increased with an increase in the level of milling. Some of the local cultivars and induced mutants were found to be superior with high amylose and high gelatinisation temperature and could be utilized in breeding programmes directed towards improving the cooking quality of high yielding varieties.     

Genetics of induced dwarf mutants in pearl millet,Pennisetum americanum (L.)Leeke

Summary Inheritance pattern of seven dwarf mutants revealed each of them to be controlled by a single recessive gene. Tests for allelism indicated existence of two groups of dwarfs, group 1 comprising six mutants and group 2 a single dwarf. The dwarfs of group 1 were found to be allelic to D₂ of the dwarf types already known in this crop and the dwarf of group 2 was allelic to D₁ of these types.Botany Department, R.V.V.N. College, Dharanikota, India.     

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.     

Effects of seed irradiation on yield and quality of essential oil in palmarosa (Cymbopogon martinii Stapf.)

Summary Dry seeds of palmarosa (Cymbopogon martinii Stapf) var. motia were subjected to 12 doses of gamma rays ranging from 5 kr to 100 kr to find out the effects of gamma irradiation on growth parameters, herbage yield and yield and quality of essential oil. Several biological parameters such as seedling growth, pollen fertility, mitotic chromosome behaviour, productive tillers/plant, leaf area index, leaf/stem ratio (by fresh weight) and total herbage yield/ /plant together with yield and quality of oil have been studied in the first (M₁) generation. Marked stimulatory effects of gamma radiation in 10 kr and 15 kr doses have been compared with the control for herbage yield, oil percentage and total alcohols (free geranyl and geranyl acetate).The two yield contributing traits, herbage yield/plant and leaf area index were remarkably increased in 15 kr, which indicates that in palmarosa the 15 kr dose is more effective than the higher doses in producing microlevel mutations. The yield and quality of oil at 10 kr and 15 kr doses were enhanced significantly. The results on high yield and quality of oil as induced by gamma-irradiation were discussed from the point of view that a gene may have been altered by mutagen treatment to produce a metabolic block between geraniol and geranyl acetate on the biosynthetic pathway or that modifier genes controlling the expression of a key precursor molecule may have been further modified to give rise to a geraniol rich chemotype in palmarosa.     

Inheritance of high oleic acid content in safflower

Safflower (Carthamus tinctorius L.) oil with high oleic acid content (>75%) has a great value for both food and non-food uses. The trait has been reported to be environmentally stable and controlled by recessive alleles at one single gene Ol, even though the influence of modifying genes has been suggested. Additionally, germplasm with higher oleic acid content (>85%) has been reported. The objective of the present research was to study the inheritance of high oleic acid content in genetic sources with both levels of high oleic acid content (>75 and >85%, respectively). A genetic study was conducted by crossing the nuclear male-sterile line CL1 (18% oleic acid) and the high oleic acid lines CR-6 (80%) and CR-9 (87%). The evaluation of the F₁ and F₂ seed generations of the crosses CL1 × CR-6 and CL1 × CR-9 indicated that in both cases the high oleic acid trait was controlled by partially recessive alleles at a single locus. The observation of F₂, F₃, and F₄ segregants with high oleic acid phenotype but lower oleic acid levels than the parents revealed the presence of modifying genes affecting the trait. Crosses between the two high oleic acid lines produced no transgressive segregation other than that caused by the mentioned modifying genes, suggesting that the high oleic acid lines CR-6 and CR-9 share the same alleles at the Ol locus. Differences for oleic acid content between both lines were hypothesized to be produced by the accumulation of genes with a minor effect on the trait.