Plant regeneration from isolated microspores of linseed (Linum usitatissimum L.)

Embryogenesis and plant regeneration were induced in isolated microspore culture of linseed (oilflax, Linum usitatissimum). Microspores underwent cell divison which led to either microcallus or embryoid formation, when they were cultured in a modified liquid Nitsch-Lichter-Nitsch medium (Lichter 1985) at two different incubation temperatures (30 and 35 °C); some embryoids and microcalli further developed to larger calli. After transfer of the microspore derived calli to a solid medium containing zeatin (Img 1^–1) shoot induction was achieved from 36 to 66% of the calli. The highest frequency of regenerated plants was obtained in microspore cultures of the hybrid Atalante x Szegedi 62 (F₁) at 30 °C, whereas for the second genotype Pedigree 2 x Kiszombori 41 (F₂) the higher incubation temperature seemed to be more efficient. Shoots could be successfully rooted on an indole acetic acid containing medium and then transplanted to vermiculite and finally to soil. Most of the plants survived the transfer into soil in the greenhouse, where they could be successfully grown to maturity.Abbreviations BAP 6-benzylaminopurine - 2,4D dichlorophenoxyacetic acid - IAA indole acetic acid - N6 Chu (1978) medium - NAA naphthaleneacetic acid - NLN Nitsch-Lichter-Nitsch (1985) - MS Murashige and Skoog (1962) medium - ZEA zeatin     

Serial analysis of combining ability in diallel and fractional diallel crosses in linseed

Summary An analysis of the nature of the gene action for seven characters influencing productivity and wide adaptation in Linum usitatissimum L. was undertaken in this investigation over three seasons based on diallel and fractional diallel crosses, among a set of ten genetically diverse parents. Estimation of combining ability and the nature of geno-type-environment interactions for the above characters was also examined.A comparison of the means of the parents and hybrids over three environments showed positive heterosis for height, number of tillers, fruit-bearing branches and capsules per plant while negative heterosis was observed for flowering time. The remaining two characters viz., height at branching and seeds per five capsules showed little differences due to mutual cancellation of the components of heterosis.A serial analysis of combining ability over three seasons using Griffing's model (1956) has revealed significant sea-sonal effects on the estimates of combining ability effects. Additive gene action was found to be predominant in most of the characters although non-additive variation due to s.c.a. was equally prominent for fruit-bearing branches and capsules per plant. On the basis of g.c.a. for all the characters, M. 10 and N. 55, both Peninsular types were the best followed by N.P. (R.R.) 45 and Mohaba Local.In the fractional diallel analysis the relative estimates g.c.a. and s.c.a. were similar for s=9 and 7. However, there was overestimation in favour of s.c.a. when s was reduced to 3. The serial analysis of fractional diallel had confirmed the results of full diallel. Significant genotype x environment interactions for flowering time, height, height at branching, fruit-bearing branches and capsules per plant were observed in this analysis also. The results have indicated that reduction of the diallel to a fraction with s=n/2 would vitiate the order and precision of estimates of combining ability.The analysis of genotype x environment indicated that the pattern of interactions among the parents of different geographical groups was not similar but varied with the character under study. Considerable interactions were observed for capsules per plant, fruit-bearing branches and flowering time in that order respectively. A comparison of the parental and hybrid populations for the degree of interaction for all the characters had revealed that heterozygosis might not at all be directly related to the degree of homeostasis. Actually, heterozygotes were found to show more interactions than homozygotes for flowering time, height at branching and tiller number and lower interactions for the other four characters. Significant seasonal effect on the degree of heterosis for vegetative as well as reproductive characters was observed in a majority of F₁'s with variable degree of dominance.

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Zusammenfassung Über einen Zeitraum von drei Vegetationsperioden wurde mit einem Material von 10 genetisch verschiedenen Eltern anhand von vollständigen und unvollständigen Diallelkreuzungen die Genwirkung bei 7 die Ertrags- und Anpassungsfähigkeit von Linum usitatissimum L. beeinflussenden Merkmalen geprüft. Außerdem wurde die Kombinationseignung dieser Merkmale und die Genotyp-Umwelt-Interaktion untersucht.Ein Vergleich der Durchschnittswerte der Eltern und Hybriden an drei verschiedenen Anbauorten ergab positive Heterosis bezüglich der Merkmale Höhe, Anzahl der Triebe, samentragende Zweige und Kapseln je Pflanze, negative Heterosis wurde dagegen bezüglich des Merkmals Blütezeit beobachtet. Die restlichen zwei Merkmale, Höhe der Verzweigung und Samenansatz von je 5 Kapseln, zeigten infolge gegenseitiger Aufhebung der Heterosiskomponenten geringe Unterschiede.Eine dreijährige Reihenanalyse hinsichtlich der Kombinationseignung nach dem Modell von Griffing (1956) ergab einen signifikanten Einfluß des Anbaujahres auf die Schätzung der Kombinationseignung. Bei den meisten Merkmalen konnte auf additive Wirkung der Gene geschlossen werden, obgleich nichtadditive Wirkung als Folge von spezieller Kombinationseignung bei den Merkmalen fruchtende Sprosse und Kapseln je Pflanze ebenso ausgeprägt war. Hinsichtlich der allgemeinen Kombinationseignung für alle untersuchten Merkmale erwiesen sich die beiden Halbinsel-Sippen, M 10 und N 55, als die besten, gefolgt von P.P. (R.R.) 45 und Mohaba Local.In den unvollständigen diallelen Analysen waren die relativen Schätzungen für allgemeine und spezielle Kombinationseignung für s=9 und 7 ähnlich. Wenn s jedoch auf 3 reduziert wurde, ergab sich eine Überschätzung der speziellen Kombinationseignung. Die Reihenanalysen des unvollständigen DiallelVersuchs bestätigten die Ergebnisse des vollständigen Diallel-Versuchs. Auch in diesem Fall wurden signifikante Genotyp-Umwelt-Interaktionen bezüglich Blütezeit, Höhe, Verzweigungshöhe, fruchtende Sprosse und Kapseln je Pflanze beobachtet. Die Ergebnisse deuteten an, daß eine Verringerung des Diallel-Versuchs auf einen Teil mit s=n/2 die Reihenfolge und Genauigkeit der Schätzung der Kombinationseignung beeinträchtigen würde.Die Analyse der Genotyp-Umwelt-Interaktion zeigte, daß die Art der Interaktion bei den Eltern verschiedener geographischer Gruppen nicht gleich war, sondern je nach dem untersuchten Merkmal variierte. Beträchtliche Interaktionen wurden für die Merkmale Kapseln je Pflanze, fruchtende Sprosse und Blütezeit — in dieser Reihenfolge — beobachtet. Ein Vergleich der Eltern- und Hybridenpopulationen bezüglich des Grades der Interaktion hat bei allen Merkmalen ergeben, daß Heterozygotie durchaus nicht unmittelbar mit dem Grad der Homeostasis in Beziehung zu stehen braucht. Tatsächlich wurde gefunden, daß Heterozygote bezüglich Blütezeit, Verzweigungshöhe und Zahl der Triebe größere Interaktionen und für die anderen 4 Merkmale geringere Interaktionen als Homozygote zeigen. Bei der Mehrzahl der F ₁-Nachkommen wurde sowohl für die vegetativen wie reproduktiven Eigenschaften ein signifikanter Einfluß des Anbaujahres mit verschiedenem Dominanzgrad auf den Grad der Heterosis beobachtet.

     

Combining ability analysis over F1-F5 generations in diallel crosses of bread wheat

Summary Combining ability studies for grain yield and its primary component traits in diallel crosses involving seven diverse wheat cultivars of bread wheat (Triticum aestivum L.) over generations F₁-F₅ are reported. The general and specific combining ability variances were significant in all generations for all the traits except specific combining ability variance for number of spikes per plant in the F₅. The ratio of general to specific combining ability variances was significant for all the traits except grain yield in all the generations. This indicated an equal role of additive and non-additive gene effects in the inheritance of grain yield, and the predominance of the former for its component traits. The presence of significant specific combining ability variances in even the advanced generations may be the result of an additive x additive type of epistasis or evolutionary divergence among progenies in the same parental array. The relative breeding values of the parental varieties, as indicated by their general combining ability effects, did not vary much over the generations. The cheap and reliable procedure observed for making the choice of parents, selecting hybrids and predicting advanced generation (F₅) bulk hybrid performance was the determination of breeding values of the parents on the relative performance of their F₂ progeny bulks.     

Combining ability and heterosis in diallel crosses of maize

Summary The present investigation involved crosses among 20 elite yellow maize populations. These were evaluated in four environments in a randomized block design with four replications in each environment. Variety Cuba was observed to be the best general combiner for grain yield, followed by St Croix and Prolific. No clear association could be discerned between general combining ability (GCA) effects for grain yield and its components, and mean grain yield performance and GCA effects. Heterosis was observed in general and all the crosses involving Cuba yielded better than the midparent. The five hybrids, Kisan × Cuba, Antigua 3D × St Croix, Prolific × St Croix, Vijay × Antigua Gr. I and A 23 × Cuba, yielded more than the standard check. Significant yield superiority of these varietal hybrids over the best commercial composite (Jawahar) suggested the possibility of their commercial exploitation.Part of the thesis submitted by the senior author in the partial fulfilment of the requirements for the Ph.D. degree     

Growth of Glomus intraradices and its effect on linseed (Linum usitatissimum L.) in hydroponic culture

This paper presents a hydroponic system for culturing and maintaining the VAM fungus Glomus intraradices in symbiosis with linseed (Linum usitatissimum L.) under greenhouse conditions in pure nutrient solution. It was possible to obtain large quantities of mycorrhizal host plant roots as well as extramatrical mycelium and chlamydospores free of impeding residues of solid substrate components. Starting from linseed donor plants inoculated in sand and transferred to the nutrient solution, new infections arose within the fast growing root system, hyphae spread out into the liquid and infected mycorrhiza-free receptor plants. Data for infection rates and plant growth parameters are presented. In comparsion to other culture systems for VAM fungi, the advantages of this hydroponic system are discussed and potential uses suggested.     

Genomic regions underlying agronomic traits in linseed (Linum usitatissimum L.) as revealed by association mapping OA简

The extreme climate of the Canadian Prairies poses a major challenge to improve yield. Although it is possible to breed for yield per se, focusing on yield‐related traits could be advantageous because of their simpler genetic architecture. The Canadian flax core collection of 390 accessions was genotyped with 464 simple sequence repeat markers, and phenotypic data for nine agronomic traits including yield, bolls per area, 1,000 seed weight, seeds per boll, start of flowering, end of flowering, plant height, plant branching, and lodging collected from up to eight environments was used for association mapping. Based on a mixed model (principal component analysis (PCA) + kinship matrix (K)), 12 significant marker‐trait associations for six agronomic traits were identified. Most of the associations were stable across environments as revealed by multivariate analyses. Statistical simulation for five markers associated with 1000 seed weight indicated that the favorable alleles have additive effects. None of the modern cultivars carried the five favorable alleles and the maximum number of four observed in any accessions was mostly in breeding lines. Our results confirmed the complex genetic architecture of yield‐related traits and the inherent difficulties associated with their identification while illustrating the potential for improvement through marker‐assisted selection.     

Genomic regions underlying agronomic traits in linseed （Linum usitatissimum L.） as revealed by association mapping

The extreme climate of the Canadian Prairies poses a major chal enge to improve yield. Although it is possible to breed for yield per se, focusing on yield-related traits could be advantageous because of their simpler genetic architecture. The Canadian flax core col ection of 390 accessions was genotyped with 464 simple sequence repeat markers, and phenotypic data for nine agronomic traits including yield, bol s per area, 1,000 seed weight, seeds per bol , start of flowering, end of flowering, plant height, plant branching, and lodging col ected from up to eight environments was used for association mapping. Based on a mixed model （principal component analysis （PCA） t kinship matrix （K））, 12 significant marker-trait associations for six agronomic traits were identi-fied. Most of the associations were stable across environments as revealed by multivariate analyses. Statistical simulation for five markers associated with 1000 seed weight indicated that the favorable al eles have additive effects. None of the modern cultivars carried the five favorable al eles and the maximum number of four observed in any accessions was mostly in＆amp;nbsp;breeding lines. Our results confirmed the complex genetic architecture of yield-related traits and the inherent difficulties associated with their identification while il ustrating the potential for improvement through marker-assisted selection.     

Development and analysis of EST-SSRs for flax (Linum usitatissimum L.)

A set of 146,611 expressed sequence tags (ESTs) were generated from 10 flax cDNA libraries. After assembly, a total of 11,166 contigs and 11,896 singletons were mined for the presence of putative simple sequence repeats (SSRs) and yielded 806 (3.5%) non-redundant sequences which contained 851 putative SSRs. This is equivalent to one EST-SSR per 16.5 kb of sequence. Trinucleotide motifs were the most abundant (76.9%), followed by dinucleotides (13.9%). Tetra-, penta- and hexanucleotide motifs represented <10% of the SSRs identified. A total of 83 SSR motifs were identified. Motif (TTC/GAA)n was the most abundant (10.2%) followed by (CTT/AAG)n (8.7%), (TCT/AGA)n (8.6%), (CT/AG)n (6.7%) and (TC/GA)n (5.3%). A total of 662 primer pairs were designed, of which 610 primer pairs yielded amplicons in a set of 23 flax accessions. Polymorphism between the accessions was found for 248 primer pairs which detected a total of 275 EST-SSR loci. Two to seven alleles were detected per marker. The polymorphism information content value for these markers ranged from 0.08 to 0.82 and averaged 0.35. The 635 alleles detected by the 275 polymorphic EST-SSRs were used to study the genetic relationship of 23 flax accessions. Four major clusters and two singletons were observed. Sub-clusters within the main clusters correlated with the pedigree relationships amongst accessions. The EST-SSRs developed herein represent the first large-scale development of SSR markers in flax. They have potential to be used for the development of genetic and physical maps, quantitative trait loci mapping, genetic diversity studies, association mapping and fingerprinting cultivars for example. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Location of a gene conditioning seed coat colour in linseed (Linum usitatissimum L.) using chromosomal interchanges

The geneG, conditioning seed coat colour was found to be located at 15.37±3.05 map units from the breakage point of one of six newly induced interchanges in linseed. Since no information about the karyotype in linseed is available, the chromosome on whichG is located, cannot be specified.Part of a thesis submitted by the second author at the Indian Agricultural Research Institute, New Delhi, for a Ph.D. degree.     

Technologically indicative properties of straw fractions of flax, linseed (Linum usitatissimum L.) and fibre hemp (Cannabis sativa L.)

In this study of the behaviour of the fractions of unretted and frost-retted fibre straws in damp air, a production scale method to separate fibre and shive from fibre plants was introduced and tested on bast fibre plants (Linum usitatissimum L. and Cannabis sativa L.). The method consists of optional drying of stalks, unloading bales, milling the straws with a hammer mill, separating the fractions from air stream with a cyclone and finally separating fibres from shives with a screening drum. Fractions were characterized focusing on technologically indicative properties such as equilibrium moisture content, ash and microbiological quality. Unretted fractions of the bast fibre plant stem reached higher equilibrium moisture contents than the retted fractions, and hemp fibres absorbed more moisture from air than did the Linum fibres. In very humid air, all fractions began to lose weight due to moulding. The weight decrease during the first week was lower in frost-retted than in unretted fractions. The frost-retted fractions appeared to be more resistant to humidity in the short term. The total number of microbes and especially the numbers of yeasts and moulds can be used as a criterion of hygienic level. For green fractions, the mould level was similar in fibres and in shives, but frost-retted shives contained more moulds than the unretted shives. The mould content of a fraction had no direct correlation with the moulding tendency of the fraction. The ash contents of fibres were somewhat higher than those of shives, due to a probable soil contamination. Ash content did not have significant correlation with microbiological quality, although ash is a possible risk factor for hygienic quality. According to the results of this study it is highly important to study the quality of the production chain of bast fibre plants to ensure the quality of industrial products. From the producer's point of view, raw material with defined quality can be directed to the most suitable application. The behaviour of fractions in various ambient atmospheres, and other quality aspects such as hygienic level can be used as criteria for defining the most appropriate product applications.     

Genetic variability and association of characters in linseed (Linum usitatissimum L.) plant grown in central Ethiopia region

Linseed is one of the most important oil seed crop in the central highlands of Ethiopia for which yield enhancement is the major breeding purposes and genotypic variability is important for selection in any breeding programs. However, shortage of improved varieties’ that provides optimum seed yield is one of the major constraints of the crop. Therefore, this study was carried out to assess the genetic variability and association among quantitative traits of 36 linseed genotypes. The experiment was conducted in 2018 main cropping season by using simple lattice design. The analysis of variances reveled highly significant difference among the genotype for most of traits considered in present study. High phenotypic and genotypic coefficient of variation was recorded for tiller per plant, harvest index, oil yield (kg ha⁻¹), and seed yield (ton ha-1) number of capsules per plant. High heritability along with genetic advance was observed for seed yield (tones ha-¹), oil yield (kg ha-¹) harvest index which indicates selection of these traits at early generation would be effective. Oil yield (kg ha⁻¹) harvest index and number of capsules plant ⁻¹ showed highly significant positive with seed yield (ton ha⁻¹). Cluster analysis revealed that 36 linseed genotypes were grouped into two clusters and four genotypes remain ungrouped. The maximum inter clusters distance was observed between clusters II and the local check. The data set was reduced into four significant principal components (PCs) that comprise (80%) of the variance. The first PC accounted for 34% of the variances that implies greater proportion of variable information explained by PC1. The traits, which contributed more to PC1, were seed yield per plant, primary branches per plant, secondary branches per plant and plant height showed positive association and had positive direct effect on seed yield. This indicates that any improvement of oil yield and harvest index would result in substantial increase on seed.     

An assessment of somaclonal variation in linseed (Linum usitatissimum)

Somaclonal lines of linseed from the parent cultivar Norlin were produced from a callus-based in oitro regeneration system (the R₀ generation). In field trials conducted over two seasons, 47 R₁ (plants produced from the R₀ generation) and 20 R₂ somaclonal lines (plants produced from the R₁ generation) were compared to the parent cultivar Norlin for quantitative characters. Irrespective of the genotype, traits in R₁'s and R₂'s were assessed on the basis of regression analysis as showing heritabilities of between 28% and 64%. Generally, the somaclonal variation assessed during these early generations revealed some detrimental traits, e.g. lower seed yield than the parent (control) cultivar and reduced 1000 seed weights, but a few lines were identified which had early or late flowering dates, improved seed yield and increased 1000 seed weights. It is concluded that somaclonal variation could be of value as an adjunct to classical breeding.     

Isolation and characterisation of RNA from flax (Linum usitatissimum L.)

A method for isolation of flax RNA is described; properties of the isolated RNA are given. RNA degradation was held to a minimum through a high pH (9.5) extraction buffer, diethylpyrocarbonate (4%) as a nuclease inhibitor, a high concentration (1.5%) of sodium dodecylsulphate, 2 mm Mg²⁺, and separation of the RNA from contaminating materials on Sephadex G-50.     

A note on the application of diallel crosses for the analysis of genetic variation in natural populations

Summary The complete diallel cross among homozygous lines can be a useful tool to analyze the genetic architecture of natural populations. However, it represents the natural population only approximately, in particular if the number of lines is small and the analyzed traits exhibit inbreeding depression or other forms of directional dominance. Some incorrect expected mean squares that can be found in the literature suggest tests for genetic variance components that can be misleading under such circumstances. Expected mean squares for a factorial analysis and for a modified Hayman analysis are presented and the effect of the number of lines and directional dominance is discussed.     

A lipoxygenase-divinyl ether synthase pathway in flax (Linum usitatissimum L.) leaves

Incubation of linoleic acid with an enzyme preparation from leaves of flax (Linum usitatissimum L.) led to the formation of a divinyl ether fatty acid, i.e. (9Z,11E,1'Z)-12-(1'-hexenyloxy)-9,11-dodecadienoic [(omega5Z)-etheroleic] acid, as well as smaller amounts of 13-hydroxy-9(Z),11(E)-octadecadienoic acid. The 13-hydroperoxide of linoleic acid afforded the same set of products, whereas incubations of alpha-linolenic acid and its 13-hydroperoxide afforded the divinyl ether (9Z,11E,1'Z,3'Z)-12-(1',3'-hexadienyloxy)-9,11-dodecadienoic [(omega5Z)-etherolenic] as the main product. Identification of both divinyl ethers was substantiated by their UV, mass-, (1)H NMR and COSY spectral data. In addition to the 13-lipoxygenase and divinyl ether synthase activities demonstrated by these results, flax leaves also contained allene oxide synthase activity as judged by the presence of endogenously formed (15Z)-cis-12-oxo-10,15-phytodienoic acid in all incubations.