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The aim of the study was to determine the effect of genotype × environment interaction on the levels of α-, β-, γ- and δ-tocopherol (α-T, β-T, γ-T and δ-T, respectively) and plastochromanol-8 (PC-8) in seeds of 17 doubled haploids (DHs) obtained from the F1 hybrid derived from crossing black (DH H2 -26) × yellow (DH Z-114) seeds of winter oilseed rape.; Results: The content of tocopherols in the tested DH lines ranged from 415.6 to 540.1 mg kg-1 seeds, while PC-8 content ranged from 56.3 to 89.0 mg kg-1 seeds. The α-T/γ-T ratio reached a level of 0.78-1.29. Studies have shown that heritability for α-T, β-T, γ-T, total-T and PC-8 is mainly due to genotypic variation. For the δ-T homologue the level was dependent on environmental effect.; Conclusion: The obtained DH lines population of oilseed rape is characterized by high heritability coefficients for α-T, β-T, γ-T, total-T and PC-8 levels, which indicates a greater influence of genotype than the environment on the content of these compounds. Among all studied doubled haploids, seven DHs were selected which were characterized by stable contents of α-T, β-T, γ-T, δ-T and total-T with the simultaneous stable content of PC-8. © 2017 Society of Chemical Industry.; © 2017 Society of Chemical Industry.

Here we present a study on the genetic effects of agronomic and seed quality traits in double haploid (DH) populations, which were developed by microspore culture from reciprocal F1 hybrids produced between cultivar Californium and DH W-15. Each of the populations consisted of 25 DH lines. The field experiments were conducted in a randomized complete block design with three replications during three growing seasons. Field observations were recorded on seed yield and its structure: the number of branches and pods per plant, the number of seeds per pod, the 1000-seed weight, the content of fat in the seeds, and three unsaturated fatty acids (oleic, linoleic, linolenic) in the seed oil. In order to investigate the influence of cross direction on the studied traits, parental effects were evaluated on the basis of differences between reciprocal DH populations. The maternal effect was revealed on the number of seeds per pod and the effect of the paternal form on linolenic acid content. The occurrence of transgression effects also depended on the direction of crossing and this was particularly noticeable in terms of the number of seeds per pod. The use of multivariate statistical methods allowed for the simultaneous characterization and grouping of tested lines in terms of several traits. Graphic images of the distribution of DH lines in the space of the two first canonical variates showed a great variation between the two reciprocal populations, both in terms of yield and its components, as well as fat and those unsaturated fatty acids.

Resynthesized (RS) Brassica napus can be used to increase the genetic diversity of this important crop plant and to develop the heterotic gene pool required for successful hybrid breeding programmes. The level of heterosis in F1 hybrids depends on the individual performance of the parents and on the degree of genetic difference between them. However, RS forms obtained from crosses of B. rapa ssp. with B. oleracea ssp. possess many undesirable agronomic traits, such as low quality of seeds, low yield and seed oil content, high erucic acid level in the oil and high glucosinolate content in seed meal. Therefore, RS oilseed rape needs to be improved by crossing with natural double-low oilseed rape, leading to selected double-low quality semi-RS lines that can be used for breeding. In this study, we evaluated the seed yield potential of F1 hybrids derived from crosses between Ogura cytoplasmic male-sterility (CMS) lines and doubled haploid (DH) semi-RS restorer lines in three locations in Poland. The genotype by environment interaction (GE interaction) and general combining ability (GCA) of the restorer and CMS line effects, as well as the effects of heterosis, were also assessed. The results of the study provide the first insights into the use of semi-RS lines as components for the development of new hybrid cultivars. Even the introduction of 50% of the RS oilseed rape genotype to natural restorer lines resulted in a marked heterosis effect, with seed yield ranging from 4.56% to 90.17% more than that of the better parent. The yield of the best hybrid amounted to 108.6% of the seed yield of the open-pollinated cultivar Monolit and 94.4% of that of the hybrid cultivar Arsenal. The best DH semi-RS line, S1, which had a significantly positive GCA for seed yield, can be recommended as a possible parent for inclusion in breeding programmes aimed at developing new hybrid cultivars.

The genetic diversity of two winter oilseed rape (Brassica napus L.) collections was investigated using amplified fragment length polymorphism marker technology. The first collection of 78 genotypes was analyzed using 11 primer-enzyme combinations and PCR products were resolved by 13.35% denaturing polyacrylamide gels. The second collection was investigated using seven fluorescent-labeled AFLP primer combinations and PCR products were separated using capillary electrophoresis. A total of 262 polymorphic AFLP markers were obtained for the first collection and 423 polymorphic markers for the second. On the basis of all markers, principal component analysis was performed for both collections separately. The unweighted pair group method with arithmetic method based on the coefficient of dissimilarity separated the genotypes in collection 1 into two clusters and those in the second collection into three distinct clusters. Resynthesized lines formed a cluster that was clearly distinct from 42 winter oilseed rape parental lines in the cytoplasmic male sterility ogura system. Analysis of molecular variance showed that 79% and 81% of the detected variation was found to be within the groups (in the two collections, respectively), while the variation between groups contributed, respectively, to only 21% and 19% of the variance. Our results indicate that AFLP technology can be useful for the creation of a gene pool of parental components of winter oilseed rape hybrid cultivars. Additionally, de novo resynthesized Brassica napus lines provide a significant opportunity for enrichment of the gene pool of winter rapeseed.

Background. In the study, an analysis of tocopherols, plastochomanol-8 and phytosterols was conducted using DH lines obtained from F1 hybrids of reciprocal crosses between yellow- and black-seeded lines. Material and methods. The biological material for the study consisted of two DH populations of winter oilseed rape obtained from F1 hybrids of reciprocal crosses between two DH lines: yellow- and black-seeded. Seed color was determined using a ColorFlex spectrophotometer. Fat content was determined via pulsed NMR. The levels of tocopherols, and plastochromanol-8 are analyzed using HPLC. Phytosterol contents and composition were determined by the GC method. Results. The fat content of the black-seeded parental line was 49% and this was higher than that of the yellow-seeded parental line (44%). The fat content of DH line populations ranged from 44 to 51%. Total tocopherol content ranged from 460 to 602 mg/kg and the α-T/γ-T ratio was from 0.66 to 1.09. In parental lines H 2 -26 and Z-114 the total tocopherol content was 534 and 525 mg/kg, but the α-T/γ-T ratios were 0.81 and 1.21, respectively. The yellow-seeded parental line (Z-114) was characterized by a higher PC-8 content (81 mg/kg) than the H 2 -26 black-seeded parental line (58 mg/kg). The largest part of the total phytosterol content in seeds of both populations was β-sitosterol from 976 to 2148 mg/kg, followed by campasterol, from 636 to 1364 mg/kg, and brassicasterol from 375 to 678 mg/kg. The total tocopherol content ranged from 462 to 595 mg/kg (population HxZ) and from 460 to 602 mg/kg (population ZxH). Signifi cantly positive correlations were observed between the seed color with α-T (r = 0.38, p < 0.01), γ-T (r = –0,34, p < 0.01) and PC-8 content (r = 0.29, p < 0.01). Correlations between the seed color with total tocopherol and total phytosterol content were not noted. Conclusion. Considering the range of genetic variation among doubled haploids of two populations, selected DH lines may be good parents for further breeding programs focused on increasing the amount and improving the quality of oilseed rapeseed oil. However, further studies will also be made to determine the infl uence of the environment on bioactive compounds in rapeseed oil. Cross direction of parental DH lines: yellow- and black-seeded has some infl uence, albeit not statistically signifi cant, on the diversity of doubled haploid in their populations with regard to average fat, tocochromanol and phytosterol content.

Resynthesized (RS) oilseed rape (Brassica napus L.) is potentially of great interest for hybrid breeding. However, a major problem with the direct use of RS B. napus is the quality of seed oil (high level of erucic acid) and seed meal (high glucosinolate content), which does not comply with double-low quality oilseed rape. Thus, additional developments are needed before RS B. napus can be introduced into breeding practice. In this study, RS oilseed rape was obtained through crosses between B. rapa ssp. chinensis var. chinensis and B. oleracea ssp. acephala var. sabellica. RS plant was then crossed with double-low (00) winter oilseed rape lines containing the Rfo gene for Ogura cytoplasmic male sterility (CMS ogu) system. Populations of doubled haploids (DH) were developed from these F1 hybrids using the microspore in vitro culture method. The seeds of semi-RS DH lines were analyzed for erucic acid and glucosinolate content. Among the populations of semi-RS DHs four 00-quality lines with the Rfo gene were selected. Using 344 AFLP markers to estimate genetic relatedness, we showed that the RS lines and semi-RS lines formed clusters that were clearly distinct from 96 winter oilseed rape parental lines of F1 hybrids.

Badano populację mapującą rzepaku ozimego złożoną z 78 linii podwojonych haploidów uzyskanych z mieszańców pochodzących ze skrzyżowania linii różniących się siłą kiełkowania. Siła kiełkowania nasion była automatycznie rejestrowana co 30 min przy pomocy aparatu fotograficznego umieszczonego w minikomputerze Raspberry Pi. Zdjęcia przeanalizowano przy pomocy oprogramowania stworzonego w John Innes Centre. W kolejnym etapie wykorzystując metodę sekwencjonowania całego genomu (Illumina® HiSeq) wszystkich linii populacji mapującej zidentyfikowano polimorfizmy pojedynczego nukleotydu (SNPs) w tych liniach. Dane te posłużą do mapowania genetycznego i identyfikacji genów regulujących proces kiełkowania.

Praca zawiera propozycję procedury obliczeniowej umożliwiającej ocenę hodowlaną mieszańców i ich form rodzicielskich (linii męskosterylnych (linii MS) i restorerów) na podstawie plonu mieszańców otrzymanych w wyniku niekompletnego schematu krzyżowania linia × tester w serii doświadczeń jednopowtórzeniowych z wzorcami. Proponowane podejście jest rozszerzeniem metody przedstawianej w pracy Wosia i in. (2010) dotyczącej oceny zdolności kombinacyjnej form rodzicielskich rzepaku ozimego na podstawie analizy mieszańców otrzymanych w wyniku kompletnego schematu krzyżowania linia × tester. Metodologia analizy prezentowana w wyżej cytowanej pracy jest oparta na ogólnej teorii analizowania wyników serii doświadczeń przeprowadzonych z tymi samymi genotypami w układzie bloków niekompletnych, podanej przez Kaczmarka (1986). Dla zilustrowania stosowanego w pracy podejścia została wykonana analiza statystyczna danych pochodzących z serii czterech doświadczeń jednopowtórzeniowych z 96 mieszańcami rzepaku ozimego replikowanymi jeden raz i dwoma wzorcami replikowanymi wielokrotnie. Dla przeprowadzenia analizy wariancji, wyznaczenia i testowania istotności ocen efektów głównych mieszańców, linii męskosterylnych i restorerów oraz ich interakcji ze środowiskiem wykorzystano programy komputerowe EKSPLAN (Krajewski i in., 2003) i SERGEN (Caliński i in., 1998).

In this report we characterized the ArabidopsisABI1 gene orthologue and Brassica napus gene paralogues encoding protein phosphatase 2C (PP2C, group A), which is known to be a negative regulator of the ABA signaling pathway. Six homologous B. napus sequences were identified and characterized as putative PP2C group A members. To gain insight into the conservation of ABI1 function in Brassicaceae, and understand better its regulatory effects in the drought stress response, we generated transgenic B. napus plants overexpressing A. thalianaABI1. Transgenic plants subjected to drought showed a decrease in relative water content, photosynthetic pigments content and expression level of RAB18- and RD19A-drought-responsive marker genes relative to WT plants. We present the characterization of the drought response of B. napus with the participation of ABI1-like paralogues. The expression pattern of two evolutionarily distant paralogues, BnaA01.ABI1.a and BnaC07.ABI1.b in B. napus and their promoter activity in A. thaliana showed differences in the induction of the paralogues under dehydration stress. Comparative sequence analysis of both BnaABI1 promoters showed variation in positions of cis-acting elements that are especially important for ABA- and stress-inducible expression. Together, these data reveal that subfunctionalization following gene duplication may be important in the maintenance and functional divergence of the BnaABI1 paralogues. Our results provide a framework for a better understanding of (1) the role of ABI1 as a hub protein regulator of the drought response, and (2) the differential involvement of the duplicated BnaABI1 genes in the response of B. napus to dehydration-related stresses. Electronic supplementary material The online version of this article (doi:10.1007/s11103-015-0334-x) contains supplementary material, which is available to authorized users.

s To determine the effect of genotype and the environment on tocopherol (T) and plastochromanol-8 (PC-8) contents in winter oilseed rape (Brassica napus L.), a three-year study was performed. The plant material consisted of 25 doubled haploid (DH) lines derived from the F1 hybrid obtained from the cross between two DH lines (Z × H), (Z-114- yellow seeds and H2-26- black seeds). Quantitative contents of α-T, β-T, γ-T, δ-T, and PC-8 were determined by HPLC. Total tocopherol content ranged between 40.62 and 51.20 mg/100 g and PC-8 5.43 – 8.92 mg/100 g. The α-T/γ-T ratio ranged from 0.77 to 1.23. The heritability in the broad sense was examined for the following components: PC-8, α-T, β-T, γ-T, δ-T, α/γ-T, and γ-T contents in oil. Heritability levels were 0.60, 0.58, 0.53, 0.50, and 0.43, respectively. This suggest the superiority of the effect of genotype on tocopherol contents. The h2 values for δ-T and β-T were 0.36 and 0.32, which demonstrates a great influence of environmental conditions. Among all analyzed genotypes, eleven were selected as satisfactory and stable regarding tocopherol or PC-8 contents.

The aim of the study was to determine the effect of genotype × environment interaction on the levels of α-, β-, γ- and δ-tocopherol (α-T, β-T, γ-T and δ-T, respectively) and plastochromanol-8 (PC-8) in seeds of 17 doubled haploids (DHs) obtained from the F1 hybrid derived from crossing black (DH HThe content of tocopherols in the tested DH lines ranged from 415.6 to 540.1 mg kgThe obtained DH lines population of oilseed rape is characterized by high heritability coefficients for α-T, β-T, γ-T, total-T and PC-8 levels, which indicates a greater influence of genotype than the environment on the content of these compounds. Among all studied doubled haploids, seven DHs were selected which were characterized by stable contents of α-T, β-T, γ-T, δ-T and total-T with the simultaneous stable content of PC-8. © 2017 Society of Chemical Industry.

Resynthesized (RS) Brassica napus can be used to increase the genetic diversity of this important crop plant and to develop the heterotic gene pool required for successful hybrid breeding programmes. The level of heterosis in F1 hybrids depends on the individual performance of the parents and on the degree of genetic difference between them. However, RS forms obtained from crosses of B. rapa ssp. with B. oleracea ssp. possess many undesirable agronomic traits, such as low quality of seeds, low yield and seed oil content, high erucic acid level in the oil and high glucosinolate content in seed meal. Therefore, RS oilseed rape needs to be improved by crossing with natural double-low oilseed rape, leading to selected double-low quality semi-RS lines that can be used for breeding. In this study, we evaluated the seed yield potential of F1 hybrids derived from crosses between Ogura cytoplasmic male-sterility (CMS) lines and doubled haploid (DH) semi-RS lines with restorer gene in three locations in Poland. The genotype by environment interaction (GE interaction) and general combining ability (GCA) of the restorer and CMS line effects, as well as the effects of heterosis, were also assessed. The results of the study provide the first insights into the use of semi-RS lines as components for the development of new hybrid cultivars. Even the introduction of 50% of the RS oilseed rape genotype to natural restorer lines resulted in a marked heterosis effect, with seed yield ranging from 4.56% to 90.17% more than that of the better parent. The yield of the best hybrid amounted to 108.6% of the seed yield of the open-pollinated cultivar Monolit and 94.4% of that of the hybrid cultivar Arsenal. The best DH semi-RS line S1, which had a significantly positive GCA for seed yield, can be recommended as a possible parent for inclusion in breeding programmes aimed at developing new hybrid cultivars.

This study presents the results of experiments concerning: (1) interspecific hybridization of Brassica oleracea × Brassica rapa via application of in vitro placental pollination and (2) embryological analysis of the process of resynthesis of Brassica napus. In order to overcome certain stigma/style barriers, B. rapa pollen was placed in vitro on an opened B. oleracea ovary (with style removed). Pollinated ovaries were cultured on Murashige and Skoog (MS) medium. After 24-d culture, the developing embryos were isolated from immature seeds and transferred onto MS medium supplemented with 0.47 μM kinetin, 0.49 μM 1-naphthaleneacetic acid, and 10% (v/v) coconut water. When the embryos had turned green, they were immediately placed onto MS medium with 100 μM kinetin. After development of the seedling, plantlets were transferred to soil. Chromosome doubling was achieved after another week. Cytometric analysis of nuclear DNA confirmed the hybrid nature of the plants. Resynthesis of B. napus can be performed through interspecific hybridization of B. oleracea × B. rapa followed by embryo rescue and genome doubling.

Celem prezentowanych badań było określenie wpływu kierunku krzyżowania na ekspresję koloru nasion i cechy struktury plonu oraz wyselekcjonowanie jasnonasiennych linii DH o wyższych parametrach cech struktury plonu i zawartości tłuszczu w porównaniu do żółtonasiennej linii rodzicielskiej. Materiał do badań stanowiły dwie populacje linii DH otrzymane z mieszańców F1 krzyżowania odwrotnego pomiędzy czarnonasienną linią DH H2-26 a żółtonasienną linią DH Z-114. Podwojone haploidy wysiano w doświadczeniu polowym w jednym powtórzeniu z systematycznie rozmieszczonymi wzorcami (obie linie rodzicielskie). Wykonano pomiary biometryczne dla cech struktury plonu, zmierzono zawartość tłuszczu i oceniono barwę nasion. Analiza wariancji przeprowadzona dla 6 cech, umożliwiła wyznaczenie charakterystyk statystycznych dla każdej z obu populacji linii DH i populacji rodzicielskich oraz zbadanie istotności różnic między nimi. Podział 4 badanych populacji na grupy jednorodne, istotnie różniące się między sobą, dla każdej omawianej cechy, pozwolił na dokonanie oceny wpływu kierunku krzyżowania. Dodatkowo przeprowadzona ocena kontrastu pomiędzy daną linią DH a lepszym z wzorców umożliwiła wyodrębnienie linii transgresyjnych. Na podstawie testowania porównań między jasnonasiennymi liniami DH a linią rodzicielską Z-114 wyróżniono dwie linie: DH HZ-6 i DH ZH-54 o istotnie wyższej liczbie łuszczyn na roślinie.

Traditional breeding methods based on inbreeding are difficult to implement in the case of Sinapis alba (white mustard) because this plant displays high levels of self-incompatibility. More rapid progress in breeding could be possible if biotechnological methods and in vitro cultures were used. However, white mustard is not readily amenable to biotechnological treatment. Seeds of traditional S. alba cultivars (e.g., Nakielska) are characterized by high levels of glucosinolates and erucic acid. However, a new Polish variety of white mustard (Bamberka) possesses low erucic acid content in the oil. The main goal of the study was elaboration of a plant regeneration system via in vitro culture of hypocotyl and cotyledon explants from low and high erucic acid-containing white mustard cultivars. In these experiments, a simple system for in vitro regeneration of white mustard was developed, with the aim to promote maximum formation of shoots within a short period of time. Traditional and improved cultivars of S. alba showed comparable capacity for shoot development from hypocotyl-derived and cotyledon-derived explants. The two types of cultivars were characterized by essentially equivalent shoot regeneration responses, being slightly higher in hypocotyl than the cotyledonary explants. A greater influence on shoot regeneration from hypocotyl explants was observed on medium supplemented with 4.4 μmol 6-benzylaminopurine, 0.57 μmol indole-3-acetic acid, and a low concentration of kinetin (4.6 μmol). This technique will allow for rapid generation of sufficient plant material for further use in a variety of white mustard breeding projects.

Praca zawiera wyniki badań zdolności kombinacyjnych linii DH przeprowadzonych na podstawie analizy statystycznej trzyletniej serii doświadczeń linia × tester z rzepakiem ozimym. Podstawą badań serii była analiza wariancji oparta na modelu mieszanym obserwacji. Została ona przeprowadzona dla serii doświadczeń, których obiektami były mieszańce uzyskane ze skrzyżowania 7 linii z 4 testerami. Każde z doświadczeń założone było w układzie losowanych bloków w trzech replikacjach. W pracy przedstawione są wyniki analizy efektów ogólnej (GCA) i specyficznej (SCA) zdolności kombinacyjnej linii rodzicielskich ze względu na zawartość kwasów tłuszczowych (oleinowego, linolowego i linolenowego) u mieszańców F2 linia × tester. Dla każdego z badanych kwasów wyznaczono oceny średnich efektów GCA i SCA linii DH i testerów oraz dokonano testowania ich istotności. Podstawowym celem pracy był wybór tych linii DH rzepaku ozimego, które spełniałyby wymagania dotyczące odpowiedniej zawartości kwasów tłuszczowych w oleju nasion przydatnych w produkcji biopaliw oraz farb i lakierów.

Oilseed rape (rapeseed, Brassica napus L.) is one of the most important crop plants, taking the third place after palm oil and soybean among oil plants and the fifth among economically important crops, following rice, wheat, maize and cotton (Snowdon et al., 2007). Oilseed rape is cultivated primarily in temperate regions, where climatic conditions prevent growth of most other oilseed crops. Spring cultivars of oilseed rape dominate in the USA, Canada, Australia and China, while winter varieties are cultivated mainly in northern European countries (Great Britain, France, Germany, Poland, Scandinavia, etc.).

With 5 figures and 1 table Abstract One of the goals in oilseed rape breeding is the identification of genotypes with low linolenic acid content in seed oil. Here, we present new genetic markers for mutant alleles of a chemically induced low linolenic rapeseed line DH219/05. Genomic clones comprising fatty acids desaturase 3 (FAD3) genes from the mutant and wild-type rapeseed lines were sequenced. Two statistically important single nucleotide polymorphisms were detected: (1) a C to T substitution in the third position of the sixth codon of the seventh exon in the BnaA.FAD3 gene and (2) a G to A transition in the 5′ splice donor site of the sixth intron in the BnaC.FAD3 gene. Allele-specific SNP markers were designed involving detection of the wild-type and mutant alleles by SNaPshot analysis and locus-specific PCR primers. Strong negative correlation between the presence of mutant alleles in the A and C genomes and linolenic acid content was revealed by analysis of variance. Sequence analysis of transcript variants confirmed predictions on possible negative effects of mutations on FAD3 gene expression.

Haploid microspore-derived embryos (MDEs) constitute a unique material for the introduction of new traits into winter oilseed rape (Brassica napus). MDEs have been transformed by using Agrobacterium tumefaciens strains EHA105 and LBA4404, both carrying the binary vector pKGIB containing the uidA gene encoding beta-glucuronidase (GUS) and the bar gene as a marker of resistance to phosphinotricin. Transformed embryos expressed GUS and regenerated plants that were resistant to herbicide Basta, as confirmed by a leaf-painting test. Progeny plants of the transformant T-39 were all transgenic, as they inherited T-DNA from their doubled haploid parental plant. Southern-blot analysis confirmed the integration and transmission of T-DNA into T1 plants. Transformation of MDEs facilitates the obtaining of winter oilseed rape homozygous for the introduced genes.