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.     

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.     

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.     

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     

Parametric and nonparametric measures of phenotypic stability in linseed (Linum usitatissimum L.)

A combined analysis with three parametricand two nonparametric measures to assess G × E interactions and stability analyses toidentify stable genotypes of linseed across18 environments in Ethiopia wereundertaken. The combined analysis ofvariance for environments (E), genotypes(G) and G × E interaction was highlysignificant (p<0.01), suggestingdifferential responses of the genotypes andthe need for stability analysis. Theparametric stability measures ofcoefficient of variability and thestability variance showed that R12-N10D wasthe most stable genotype, whereascultivars' superiority measure indicatedChilalo to be the most stable cultivar.Like most of the parametric methods, thenon-parametric measures revealed thatR12-N10D had the smallest changes in ranksand thus was the most stable genotype incontrast to R12-D24C, which was unstableand the lowest yielder. A comparison of thefive stability measures showed that thecoefficient of variability, stabilityvariance and variance of ranks were similarin assessing the relative stability of thegenotypes, whereas cultivars' superioritymeasure deviated from the others. Thestability variance and variance of rankswere significantly rank correlated, andwere the best in determining thecomparative stability of linseed genotypes.The coefficient of variability was alsorelatively better than the cultivar'ssuperiority measure. Further studies ofrepeatability tests are, however, needed todetermine the best methods. The stabilitystatistics generally identified R12-N10D,followed by Chilalo, as the most stablevarieties, whereas R12-D24C and R11-M20Gwere the least stable varieties.     

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     

Isolation of induced mutants in linseed (Linum usitatissimum) having reduced linolenic acid content

Summary Seeds of Linum usitatissimum cv. Glenelg were treated with either gamma-rays or EMS in an attempt to induce mutations with a lower level of linolenic acid in linseed oil. Two mutant lines were identified in which linolenic acid constituted approximately 29% of the total fatty acid content compared with 43% in seed oil from untreated Glenelg plants. The reduced level of linolenic acid in the mutants is accompanied by an increase in the level of linoleic acid to 30% compared with 18% in Glenelg, but there was no change in the proportions of other fatty acids. These proportions of linolenic acid and linolenic acid are respectively the highest and lowest yet reported in stable genotypes of L. usitatissimum. The strong inverse relationship between these two fatty acids in these genotypes suggests that linolenic acid is synthesised by desaturation of linolenic acid and indicates that it may be possible to breed an edible linseed oil having both low levels of linolenic acid and high levels of linolenic acid.     

In vitro breeding of high-linolenic, doubled-haploid lines of linseed (Linum usitatissimum L.) via androgenesis

In the course of a breeding programme for high-linolenic-acid linseed (oilflax, Linum usitatissimum L.) doubled-haploid lines (DH-lines) of three F₁ hybrids were generated using an anther-culture technique. A total of 82 DH-lines were generated and multiplied in 1993, of which 39 were suitable for testing in a repeated field trial in 1994. Yield, fat content and fatty-acid composition were determined. The yield of the DH-lines was not significantly different from that of respective mid-parent values. Fatty-acid composition and fat content showed remarkable variation, and DH-lines exceeded cross parents in some cases for both fat content and linolenic-acid proportion. Superior DH-lines surpassed mid-parent linolenic-acid-content values by 4.5% and fat content by 1.9%. The results indicate that anther culture is a useful tool in linseed breeding.     

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.     

Genetic analysis of quantitative traits across environments in linseed (Linum usitatissimum L.)

Summary Fourteen lines of linseed (Linum usitatissimum L.) were evaluated for general and specific combining ability through line × tester cross analysis using five diverse testers in two different environments. Seventy F₁'s and nineteen parental genotypes were raised in completely randomized block design with two replications with respect to yield and yield related traits. Considerable genetic variation was observed for all the traits studied. The additive as well as non-additive gene effects played significant role in the inheritance of yield and related traits with preponderance of non-additive gene effects for all the traits studied. Higher proportion of general combining ability × environment interaction variance as compared to specific combining ability × environment estimates was recorded. Additive genetic variances were more sensitive than non-additive genetic variances to the changing environment. Low estimates of heritability (narrow sense) for primary branches per plant, capsules per plant, seeds per capsule, seed yield per plant, harvest index and oil content and medium heritability for days to 50% flowering, days to maturity, plant height and 1000-seed weight was observed. Among the female parents LCK-9816, Parvati, Himalini, KL-168 were good general combiners for yield and related traits along with oil content, whereas among male parents, Surbhi and KL-224 were good general combiners for yield related traits and oil content but KL-221 was good general combiner for seed yield and related traits.     

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.     

Cytogenetic variation in somatic tissue cultures and regenerated plants of barley (Hordeum vulgare L.)

Summary Chromosome number of morphogenic and non-morphogenic calli and regenerated plants of barley were determined. Cultures were obtained from two kinds of explants, immature embryos and seedling leaves from three cultivars, Ingrid, Dissa and Golden Promise. Callus chromosome analyses were carried out during a 12 month period in a medium containing 2 mg/l of 2,4-D. Diploid cells were predominant in all cases; although in leaf-derived cultures, retraploid cells (2n=4x=28) showed a tendency to increase as time in culture increased and after more than six months in culture, diploid cells decreased to percentages of almost 70%. Aneuploid cells were generally infrequent in all cases. The source of explant has been more important than the genotype (cultivar) and the type of callus (morphogenic vs. non-morphogenic) in the chromosomal stability of cultures as time increases. From short term cultures, only 1.85% of the regenerated plants were tetraploid, the remaining were diploids. The ability of morphogenic calli to regenerate plants decreased before any significant reduction of diploid cells were observed.     

Plant regeneration from anther culture in Canadian cultivars of flax (Linum usitatissimum L.)

The effect of culture of anthers at 35 °C for one to four days prior to culture at 25 °C in darkness, genotype, anther orientation on callus induction and shoot regeneration in anther culture of flax was investigated. The influence of type and concentrations of cytokinins in the regeneration medium on shoot regeneration was also investigated. The results suggested that culture of anthers at 35 °C prior to continuous culture at 25 °C in darkness did not significantly improve the percentage of anthers producing calli. However, culture of anthers at 35 °C for one day significantly increased the overall efficiency of regeneration compared to no culture temperature treatment. Genotypic effects were significant for the percentage of anthers producing calli and the overall efficiency of regeneration. Anther orientation showed no significant differences. The regeneration medium containing 4.5 μM zeatin had significantly higher percentage of calli forming shoots than the same basal medium containing 0.01 μM TDZ. The importance of these findings for flax breeding was discussed. This revised version was published online in July 2006 with corrections to the Cover Date.     

Genetic analysis of in vitro wheat somatic embryogenesis

Summary A spring wheat genotype which produces somatic embryos in vitro, after short and long-term culture, was tested for its ability to sexually transmit this embryogenic trait. Reciprocal crosses were performed between a embryogenic line and a nonembryogenic variety.Immature embryos were cultured on Murashige and Skoog medium plus 2 mg/l 2,4-dichlorophenoxyacetic acid, gelled with 5.5 g/l agarose. Somatic embryogenesis was not expressed in the F₁'s. In contrast, from several hundred immature embryos of the F₂ generation of one cross, 10.7% and 1.6% expressed somatic embryogenesis in short and long-term cultures respectively. These percentages of embryogenic: non-embryogenic fits a model of a few complementary genes. The embryogenic capacity of the F₂ genotypes depends on the presence of recessive alleles at these gene loci. The long-term wheat somatic embryogenesis capacity requires a more complex mechanism than the short-term one.Abbreviations CS Chinese Spring - Aq Aquila - E Embryogenic - NE Nonembryogenic - SC Subculture     

Diallel analysis of callus formation ability in linseed anther culture

In linseed, the time needed to obtain pure lines can be reduced by using in vitro anther culture. The technique has been improved but there are no reports on the inheritance of the characters (callogenesis and regeneration) involved in the response. To shed some light on this aspect, anthers of four linseed varieties and their F₁ were cultivated in vitro. Callogenesis response was analysed according to a full diallel model. Among 7811 anthers, the percentage of callogenesis ranged from 27.40 to 82.04%, with the parental lines being around the average. The differences between specific hybrid combinations were significant and general and specific combining ability were highly significant. As well as dominant effects, additive and non‐additive effects are probable components of the heritability of this character, although additive effects would be more important than non‐additive. Differences between reciprocal crosses would indicate that cytoplasmic factors could also be involved in the determination of the character.     

Dry Matter Partitioning in Linseed (Linum usitatissimum L.)

A pot experiment was conducted at University of Wales Aberystwyth, UK, Frongach field station to evaluate the effects of sowing dates on the dry matter partitioning of linseed. Root, stem and capsule (boll) weight increased progressively as sowing was delayed up to third sowing (19 April) but showed a gradual decrease when sowing was further delayed. The contribution of each of the three parts to total weight did not respond to sowing date in any consistent manner. The total dry matter per plant followed a similar pattern to the dry matter of plant parts, which increased with sowing up to third sowing (19 April) and then decreased when sowing was further delayed. Dry matter accumulation per day degree > 4 °C also increased as sowing was delayed with a peak at third sowing (19 April). Dry matter accumulation per day degree is concluded to be directly related to the total day degrees received during the growth period.     

Comparison of genetic diversity of flax (Linum usitatissimum L.) between Canadian cultivars and a world collection

This study reports results of phenotypic measurements of genetic diversity in the world collection of flax maintained by Plant Gene Resources of Canada (PGRC) and compares the range of diversity in the world collection with the diversity observed in 19 Canadian registered flax cultivars. Morphological and seed‐oil characters were used to describe the phenotypic diversity in 2331 flax accessions. The plants were grown by PGRC at Saskatoon, Saskatchewan, Canada, in 1998 and 1999. The comparison between the Canadian cultivars and the world collection was based on single characters, as well as on character complexes by application of an existing intraspecific classification for the species. Considering single quantitative or qualitative character expressions, the Canadian cultivars represented a wide range in diversity for the species. The variation of characters Canadian plant breeders have selected for (e.g. plant height, seed weight, seed colour, petal colour, oil content) was reduced further than those characters not focused on by plant breeders (e.g. dotting of the sepals, style colour, ciliation of capsule septa, oil quality characters). A comparison of diversity based on the intraspecific classification proposed for flax by Kulpa and Danert, who described 28 botanical varieties, showed that all Canadian cultivars belong to two botanical varieties. This study demonstrates the usefulness of agrobotanical characterization of genebank collections for plant breeding and illustrates the application of the traditional method of intraspecific classification for comparison of gene pools.     

Evaluation of Fibre Flax (Linum usitatissimum L.) Performance under Minimal and Zero Tillage in Eastern Canada

The feasibility of producing fibre flax in minimum tillage or zero tillage (ZT) systems was investigated. The results were variable between the sites, which differed in soil type and previous cropping history. Tillage regime had no impact on fibre flax phenological development including number of days to emergence, days to flowering and days to harvest at either site. However, tillage regime had a significant effect on mean stem diameter, dry matter content and plant height at all three sampling dates at the sandy loam site, and a significant effect on mid‐season plant height at the clay site. At the sandy loam site, ZT plots had the highest populations of plants with the finest stem diameters, the lowest branching ratios, but the shortest plants by the end of the season. Overall, plant densities were greater, stem diameters thinner, and biomass production higher in the heavier soil of the clay site. These results indicate that under the growing conditions present in 1998, fibre flax can successfully be grown in minimum or ZT systems on different soil types in Eastern Canada. Moreover, the growth of fibre flax in these systems does not compromise the proportion of tall plants having thin stems and minimal branching, a critical quality parameter for fibre flax production.     

亚麻遗传连锁图谱的构建

利用DIANE (纤用亚麻栽培种)和宁亚17 (油用亚麻栽培种)为杂交亲本,构建30个F₂单株作为作图群体,选用71对SRAP和24对SSR共显性标记构建了全长为546.5 cM,含12个连锁群(LGs)的亚麻遗传连锁图谱,标记均匀分布于12个连锁群,每个连锁群有4~15个标记,标记间平均距离为5.75 cM。结果表明,SRAP标记和SSR标记中共显性标记适合于亚麻遗传连锁图谱的构建,但该图谱覆盖的基因组范围较小,需继续图谱的完整性工作。本研究为今后的亚麻在分子生物学方面的研究提供了基础信息。