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  • 1
    ISSN: 1432-2145
    Keywords: Self-incompatibility ; S-locus genes ; Brassica napus ; Transgenic plants
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Self-compatible Brassica napus var ‘Westar’ was transformed with SLG, the S-locus-derived gene that encodes S-locus-specific glycoproteins (SLSG). Four allelic variants of SLG isolated from self-incompatible B. oleracea and B. campestris strains homozygous for different S alleles were used. We show that the transgenic plants synthesized SLSG with the same apparent charge, molecular weight, and antigenic properties as that produced by the corresponding self-incompatible strains from which the cloned SLG genes were isolated. In addition, transgene-encoded SLSG was detected specifically in the papillar cells of the stigma, and was correctly targeted to the papillar cell wall. However, SLSG was produced at reduced levels in transgenic plants relative to self-incompatible strains. The introduction of the SLG genes did not confer a self-incompatibility phenotype on the ‘Westar’ cultivar.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Journal of chemical ecology 18 (1992), S. 875-884 
    ISSN: 1573-1561
    Keywords: Aggregation pheromone ; olfactometer ; field trapping ; Coleoptera ; Chrysomelidae ; Phyllotreta cruciferae ; Brassica napus ; crucifer
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Laboratory olfactometer bioassays and field trapping experiments showed that the flea beetle,Phyllotreta cruciferae (Goeze), was highly attracted by oilseed rape(Brassica napus L.) when flea beetles were on the plant. This attraction was mediated by a flea beetle-produced aggregation pheromone based upon: (1) Oilseed rape damaged mechanically, or byP. cruciferae, or by diamondback moth,Plutella xylostella (L.), did not attractP. cruciferae. (2) Contact with the plants or feeding was required for the production of aggregation pheromone because oilseed rape alone was not attractive when separated from flea beetles by a screen. (3) Equal numbers of males and females were attracted.
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  • 3
    ISSN: 1573-5028
    Keywords: anther ; Arabidopsis thaliana ; Brassica napus ; tapetum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The Brassica napus cDNA clone A9 and the corresponding Arabidopsis thaliana gene have been sequenced. The B. napus cDNA and the A. thaliana gene encode proteins that are 73% identical and are predicted to be 10.3 kDa and 11.6 kDa in size respectively. Fusions of an RNase gene and the reporter gene β-glucuronidase to the A. thaliana A9 promoter demonstrated that in tobacco the A9 promoter is active solely in tapetal cells. Promoter activity is first detectable in anthers prior to sporogenous cell meiosis and ceases during microspore premitotic interphase. The deduced A9 protein sequence has a pattern of cysteine residues that is present in a superfamily of seed plant proteins which contains seed storage proteins and several protease and α-amylase inhibitors.
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    Plant molecular biology 19 (1992), S. 1049-1055 
    ISSN: 1573-5028
    Keywords: Brassica napus ; rapeseed ; gene expression ; nucleotide sequence ; storage proteins
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have isolated a five-member gene subfamily which encodes cruciferin, a legumin-like 12S storage protein of Brassica napus L., and have analyzed the structure and expression of the family members in developing embryos. Sequence analysis has shown that the coding regions of all five genes are highly similar, with the two most divergent members of the family retaining 89% sequence identity. The analysis of this cruciferin gene family's expression indicates that the developmental pattern of expression of each gene is similar, and the steady-state mRNA levels of each gene are approximately equivalent to each other at all developmental stages.
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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    Plant molecular biology 19 (1992), S. 1079-1083 
    ISSN: 1573-5028
    Keywords: oleosin ; embryogenesis ; cDNA ; Brassica napus ; oil-body protein
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Antibodies raised against purified rapeseed 19 kDa oleosin protein were used to screen an embryo-derived λgt11 expression library from Brassica napus. A near full-length cDNA clone, BnV, was isolated. The 781 bp cDNA contained an open reading frame of 549 bp followed by an untranslated region of 222 pb and a poly(A) region of 10 bp. Comparisons between this cDNA and a different oleosin cDNA previously isolated from the same library showed high degrees of sequence similarity in the central domain region and in the 3′ untranslated region. Sequence similarities between the derived protein sequence of this cDNA and all other known oleosin protein sequences are discussed.
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  • 6
    ISSN: 1573-5028
    Keywords: Brassica napus ; cruciferin ; seed globulin ; storage protein
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A gene encoding a subunit of the 12S storage globulin, cruciferin, in Brassica napus (oilseed rape) has been isolated and characterized. The gene consists of about 2200 bp including three short intervening sequences. Primer extension analysis showed that the major transcription start site is located 30 bp 5′ of the predicted ATG start codon. This gene belongs to one of three different major families encoding cruciferin subunits. By use of gene-family-specific probes and Southern blotting analysis the number of genes of the three different cruciferin subtypes in B. napus was estimated.
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  • 7
    ISSN: 1573-5028
    Keywords: acyl-(acyl carrier protein) thioesterase ; acyl carrier protein ; Brassica napus ; fatty acid synthesis ; rape ; seed development
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The level of two thioesterases, acyl-CoA thioesterase and acyl-ACP thioesterase was determined during seed maturation in oil seed rape. Both thioesterase activities rose markedly prior to the onset of lipid accumulation, but the induction kinetics suggest that the activities reside on distinct polypeptides. Acyl-ACP thioesterase (EC 3.1.2.14) was purified 2000-fold using a combination of ion exchange, ACP-affinity chromatogr aphy, chromatofocusing and gel filtration. Using native gel electrophoresis, and assays for enzymic activity, two polypeptides were identified on SDS-PAGE as associated with the activity. Cleveland mapping of these polypeptides, of 38 kDa component and 33 kDa respectively, demonstrated that they are related. An antibody was prepared against the 38 kDa component, and this also recognises the 33 kDa polypeptide in highly purified preparations. Western blotting of a crude extract identifies one band at 38 kDa consistent with the 33 kDa component being a degradation product generated during purification. The native molecule has a Mr of 70 kDa indicating a dimeric structure. The enzyme has a pH optimum of 9.5 and shows strong preference for oleoyl-ACP as substrate. The intact enzyme has an N-terminus blocked to protein sequencing. We also found that two other polypeptides co-purify with acyl-ACP thioesterase under native conditions. The N-terminal amino-acid sequence of these polypeptides is shown and their possible identity is discussed.
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  • 8
    ISSN: 1573-5028
    Keywords: methionine enhancement ; seed proteins ; Brassica napus ; transgenic expression ; Brazil nut ; nutritional quality
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have increased the methionine content of the seed proteins of a commercial winter variety of canola by expressing a chimeric gene encoding a methionine-rich seed protein from Brazil nut in the seeds of transgenic plants. Transgenic canola seeds accumulate the heterologous methionine-rich protein at levels which range from 1.7% to 4.0% of the total seed protein and contain up to 33% more methionine. The precursor of the methionine-rich protein is processed correctly in the seeds, resulting in the appearance of the mature protein in the 2S protein fraction. The 2S methionine-rich protein accumulates in the transgenic seeds at the same time in development as the canola 11S seed proteins and disappears rapidly upon germination of the seed. The increase in methionine in the canola seed proteins should increase the value of canola meal which is used in animal feed formulations.
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  • 9
    ISSN: 1573-5028
    Keywords: acyl carrier protein ; Brassica napus ; lipid synthesis ; seed-specific expression ; transgenic tobacco ; 5′ flanking region
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Acyl carrier protein (ACP) is a key component of the fatty acid biosynthetic machinery in plants. A 1.4 kb 5′ flanking region of a Brassica napus ACP gene (ACP05) was transcriptionally fused to the reporter gene β-glucuronidase (GUS), and expression of the chimaeric gene monitored in transgenic tobacco. GUS activity was found to increase through seed development reaching a maximum value, coincident with the most active phase of storage lipid synthesis that was, on average, 100-fold higher than that observed in leaf. In control plants transformed with CaMV 35S-GUS constructs, GUS activity was similar in leaf and all stages of seed development. Based on average values, the level of GUS expression obtained via the ACP promoter was comparable to that obtained from the CaMV 35S promoter. We therefore conclude that the isolated 5′ ACP flanking sequence represents a strong promoter element involved in the developmental regulation of storage lipid synthesis in B. napus seed tissue. Putative regulatory elements in the 5′ upstream region of ACP05 were identified by dot matrix analysis and by sequence comparison with the upstream regions from a second seed-expressed rape ACP gene and from an Arabidopsis ACP gene.
    Type of Medium: Electronic Resource
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  • 10
    ISSN: 1573-5028
    Keywords: Brassica napus ; embryogenesis ; leucine-zipper motif ; oleosin ; oil-body protein ; seed development
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The sequence of an oleosin gene from Brassica napus has been determined. This gene contains a single intron of 437 bp and encodes a polypeptide of 195 amino acids. The oleosin gene product has an estimated molecular mass of 21.5 kDa and consists of a highly hydrophobic central domain flanked by relatively polar N- and C-terminal domains. The central domain is highly conserved between all oleosins sequenced to date and contains a run of periodically spaced leucine residues similar to that of a leucine-zipper motif. The gene has been shown to be expressed specifically in the embryo, maximally between 9 and 11 weeks after flowering, i.e. during the seed desiccation stage. Two transcriptional start sites have been mapped to -70 and -21 of the ATG and a putative ABA-responsive element and three repeated motifs have been identified in the promoter. These short promoter sequences could correspond to regulatory elements responsible for embryo-specific gene expression. Up to six genes exist in the oleosin gene family.
    Type of Medium: Electronic Resource
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  • 11
    Electronic Resource
    Electronic Resource
    Springer
    Plant molecular biology 18 (1992), S. 387-398 
    ISSN: 1573-5028
    Keywords: Arabidopsis thaliana ; Brassicaceae ; Brassica napus ; glucosinolate ; myrosinase ; Sinapis alba
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A full-length cDNA clone (MB3) and three partial clones (MA1, MB1 and MB2) which encode myrosinase (thioglucoside glucohydrolase, EC 3.2.3.1) were isolated from a Sinapis alba (white mustard) cDNA library. Nucleotide sequence analysis of these clones revealed that they are encoded by a gene family. Southern blot analysis with gene-specific probes showed that the gene family consists of a least two subfamilies (MA and MB) each with several members both in S. alba and in Brassica napus (oilseed rape). In Arabidopsis thaliana (wall cress) only three myrosinase genes seem to be present. Northern blot analysis indicated that all the myrosinase mRNA species have the same size, approximately 1.95 kb.
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  • 12
    ISSN: 1573-5028
    Keywords: Brassica napus ; chloroplast ; 3-isopropylmalate dehydrogenase ; molecular evolution ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Both insect and mammalian genes have previously been cloned by genetic complementation in yeast. In the present report, we show that the method can be applied also to plants. Thus, we have cloned a rape cDNA for 3-isopropylmalate dehydrogenase (IMDH) by complementation of a yeast leu2 mutation. The cDNA encodes a 52 kDA protein which has a putative chloroplast transit peptide. The in vitro made protein is imported into chloroplasts, concomitantly with a proteolytic cleavage. We conclude that the rape cDNA encodes a chloroplast IMDH. However, Southern analysis revealed that the corresponding gene is nuclear. In a comparison of IMDH sequences from various species, we found that the rape IMDH is more similar to bacterial than to eukaryotic proteins. This suggests that the rape gene could be of chloroplast origin, but has moved to the nucleus during evolution.
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  • 13
    Electronic Resource
    Electronic Resource
    Springer
    Theoretical and applied genetics 83 (1992), S. 476-479 
    ISSN: 1432-2242
    Keywords: Haploidy ; Brassica napus ; Yellow-seeded canola
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The use of haploidy to introgress recessive traits into Brassica napus canola is illustrated by describing the properties of doubled haploids obtained by microspore culture from crosses between a yellow-seeded rapeseed line (low erucic acid, high glucosinolate) and black-seeded canola. Of the 99 doubled haploid lines that were produced, 3 were yellow-seeded canola lines. This result was not significantly different than the predicted frequency of 1 in 64 for the homozygous recessive phenotype in a doubled haploid population segregating for six recessive genes. Thus, the study supports previous models of inheritance determined for yellow seededness and glucosinolate content in Brassica napus. Also, since the chances of obtaining a plant with the same characteristics in a F2 population are 1 in 4,096, the underscore results the advantages of using haploidy to introgress recessive traits into Brassica napus canola.
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  • 14
    Electronic Resource
    Electronic Resource
    Springer
    Theoretical and applied genetics 84 (1992), S. 403-410 
    ISSN: 1432-2242
    Keywords: Interspecific breeding ; Sinapis alba ; Brassica napus ; Isozymes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Isozyme analysis of Brassica napus cv ‘Topas’ and CGRC5006 as well as of Sinapis alba cv ‘Emergo’ revealed significant polymorphism between the two species for the isozymes, aconitate hydratase, glucose phosphate isomerase, and diaphorase. F1 hybrids between B. napus ‘5006’ and S. alba cv ‘Emergo’ were backcrossed to B. napus cv ‘Topas’, and the S1 progeny of the first two backcrosses were studied isozymically. At the backcross one level the frequency of S. alba or S. alba plus B. napus patterns observed ranged from 18% to 87% across the four lines studied. There were differences between lines for the frequency of S. alba patterns, which could have an impact on the efficiency of selection for subsequent backcrossing. By the backcross two generation in one of the two lines studied, GR86-24, the S. alba patterns for GPI and DIA had been lost, while in the other line, GR86-28, the S. alba pattern for ACO had been lost, resulting in lost opportunity for S. alba gene transfer. In a wide cross such as S. alba x B. napus, which requires an intensive effort to accomplish, the isozymes ACO, GPI, and DIA may serve as useful markers to ensure gene transfer between the two species has occurred. In addition, the identification of lines with divergent isozyme patterns from B. napus will provide the basis for establishing linkages between S. alba traits of interest and isozyme markers.
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  • 15
    Electronic Resource
    Electronic Resource
    Springer
    Theoretical and applied genetics 84 (1992), S. 303-306 
    ISSN: 1432-2242
    Keywords: Brassica napus ; Outcrossing rate ; Selfing rate ; Mixed mating
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Rapeseed (Brassica napus) is a predominantly selfpollinated crop with about one-third outcrossing. The outcrossing rate may be influenced by environmental factors, and hence changes in the heterozygosity level of a variety may occur during multiplication. In an investigation on environmental variation in outcrossing, we estimated the outcrossing rate in the Swedish spring rapeseed cv ‘Topas’ by isozyme analysis and found that outcrossing varied from 12% to 47% over five locations in Sweden, Denmark and Germany. Among flowers at different positions on the same plant, average outcrossing varied from 11% at the top to 39% at the bottom of the plant. In conclusion, environmental factors can greatly influence the outcrossing rate in rapeseed, and an investigation therefore merit further studies.
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  • 16
    Electronic Resource
    Electronic Resource
    Springer
    Theoretical and applied genetics 85 (1992), S. 222-228 
    ISSN: 1432-2242
    Keywords: Brassica napus ; Raphanus sativus ; Cytoplasmic male sterility ; Restorer ; Isozyme
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Co-segregation studies of isozyme markers and male fertility restoration showed that a restorer gene from radish was introduced into rapeseed along with an isozyme marker (Pgi-2). The radish chromosome segment carrying these genes was introgressed into rapeseed through homoeologous recombination, substituting for some of the rapeseed alleles. By crossing heterozygous restored plants to male-sterile lines and to maintainers, tight linkage was found between the restorer gene and the marker. The recombination fraction was estimated at 0.25 ± 0.02%. Although few restored plants lacked the radish isozyme marker, it was still possible to distinguish male-fertile from male-sterile plants by their PGI-2 patterns. Furthermore, homozygous and heterozygous restored plants could be separated by specific PGI-2 phenotypes. Thus, the Pgi-2 marker is now currently used in restorer breeding programs.
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  • 17
    Electronic Resource
    Electronic Resource
    Springer
    Molecular genetics and genomics 234 (1992), S. 379-389 
    ISSN: 1617-4623
    Keywords: Brassica napus ; Anther-specific ; Chalcone synthase ; RNA-PCR assay ; In situ hybridization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A cDNA library of Brassica napus (cv. Westar) was constructed using poly(A)+ RNA isolated from developing anthers of flower buds 2–3 mm in length. Differential hybridization, using cDNA probes complementary to poly(A)+ RNA from developing anthers or seedlings, was used for initial screening. In addition to Southern and Northern blot analyses of selected clones, RNA-PCR assays and in situ hybridization were used to study the temporal and spatial gene regulation in anthers at the transcriptional level. Five independent cDNA clones, showing no cross-hybridization to one another, were characterized, and their expression patterns could be grouped into three distinct categories. Two cDNA clones, BA112 and BA158, are tapetum-specific: the corresponding mRNAs accumulate in young anthers and decline as the tapetum cells degenerate later in anther development. The transcripts represented by BA54 and BA73 accumulate late in anther development and reach a maximum level in mature anthers prior to anthesis; BA54 has been confirmed to be pollen-specific. The third category, represented by BA42, is found to encode a protein sharing 64–67% amino acid similarity with chalcone synthase (CHS) from various plant species; the transcript is localized in the peripheral cells of the vascular bundle, tapetum, and developing microspores.
    Type of Medium: Electronic Resource
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  • 18
    Electronic Resource
    Electronic Resource
    Springer
    Plant and soil 142 (1992), S. 315-321 
    ISSN: 1573-5036
    Keywords: Canola/rapeseed ; Rhizoctonia solani AG2-1 ; cuticle ; epidermal cell wall ; Brassica napus ; Sinapis alba
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract A study was conducted to determine whether the cuticles in two genera of the family Cruciferae are effective barriers to infection by Rhizoctonia solani, and whether differences in cuticle and epidermal cell wall thickness and morphology of epicuticular wax exist between resistant and susceptible cultivars. As Canola/rapeseed (Brassica napus) and mustard (Sinapis alba) plants develop from 1 to 3 weeks of age, they become increasingly resistant to R. solani AG2-1 seedling root rot. Seven-day-old seedlings of S. alba cultivars are invariably more resistant than B. napus cultivars. Brassica napus cultivars do not show an obvious cuticle layer at 1 week but at 3 weeks the presence of a cuticle is seen through autofluorescence with a concomitant increase in resistance to R. solani. Removal of the cuticle from 3-week-old hypocotyls by chloroform treatment results in a decrease in cuticular autofluorescence and a significant increase in disease severity in both resistant and susceptible cultivars. Three-week-old plants of S. alba have a much lower percent disease rating and a significantly (p=0.05) thicker cuticle layer than similar-age plants of B. napus. The results suggest that the cuticle plays an important role in the resistance of S. alba and older plants of B. napus to infection by R. solani.
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  • 19
    Electronic Resource
    Electronic Resource
    Springer
    Euphytica 59 (1992), S. 221-229 
    ISSN: 1573-5060
    Keywords: Brassica napus ; simazine tolerance ; polygenic variation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Substantial variation in tolerance to the triazine herbicide simazine was observed among a wide range of Brassica napus cultivars grown in a hydroponic screening system. None of these cultivars was as tolerant of are simazine treatment as the Canadian cultivar Triton which carries cytoplasmic gene(s) which confer a high level of resistance through their modification of chloroplast membranes to which triazine herbicides normally bind. A field study showed that the most tolerant cultivar Haya had a significantly higher yield than Triton in both handweeded plots and plots treated with 1000 ml/ha simazine. This finding supported other observations that the yield potential of cultivars such as Triton is substantially reduced because of the impairment of photosynthesis by cytoplasmic gene(s) for triazine resistance. A slight reduction in the yield of Haya with an increase in simazine concentration from 500 to 1000 ml/ha suggested that further increases in simazine concentration required for optimal weed control are likely to have a detrimental effect on the yield of Haya and other tolerant cultivars. Genetic analyses of variation in simazine tolerance in populations derived from crosses among cultivars representative of the range in tolerance indicated that the narrow-sense heritability of tolerance was of sufficient magnitude to allow for isolation of genotypes tolerant of higher simazine concentrations. These could be utilized in developing high yielding cultivars in areas of southern Australia where simazine treatment is necessary for effective weed control in Canola crops.
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  • 20
    ISSN: 1573-5060
    Keywords: Brassica napus ; fertility ; interspecific hybridization ; self-incompatibility ; somatic hybridization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Brassica napus is a natural allotetraploid derived from the diploid species B. rapa L. (syn. campestris L.) and B. oleracea L. Somatic hybrids synthesized from highly heterozygous lines of these two diploid species were evaluated for fertility. The hybrids were obtained from two fusion experiments which differed in the B. rapa full-sibling parent used as the source of protoplasts. Both B. rapa siblings were lelf-incompatible (SI) yet contained different S-alleles; the B. oleracea species parent was self-compatible (SC). Eight tetraploid hybrids examined had very high female and male fertility; eight hybrids with higher ploidy had low fertility. Hybrids derived from one B. rapa sibling were self-incompatible, whereas those derived from the other B. rapa sibling were fully self-compatible. These data suggest that the different S-alleles of each B. rapa sibling displayed varying penetrance relative to the SC of the B. oleracea parent when combined in B. napus.
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  • 21
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    Plant cell, tissue and organ culture 31 (1992), S. 141-149 
    ISSN: 1573-5044
    Keywords: Brassica napus ; cryopreservation ; in vitro embryo ; microspore ; rapeseed
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Microspore cryopreservation is a potentially powerful method for long-term storage of germplasm for in vitro embryo production in plant species. In this study, several factors influencing embryo production following the ultra-low temperature (−196 °C in liquid nitrogen) storage of isolated microspores of rapeseed (Brassica napus L.) were investigated. Microspores were prepared in cryogenic vials and subjected to various cooling treatments before immersion in liquid nitrogen for varying periods. Efficiency of microspore cryopreservation was reflected by in vitro embryo production from frozen microspores. Of all the cooling treatments, microspores treated with a cooling rate of 0.25% °C/min and a cooling terminal temperature of −35 °C before immersion in liquid nitrogen produced the highest embryo yields (18% and 40% of unfrozen controls in two genotypes, respectively). Fast thawing in a 35 °C water bath was necessary to recover a high number of embryos from microspore samples being frozen at a higher cooling rate, while thawing speed did not affect samples after freezing at a slower cooling rate. The storage density of cryopreserved microspores affected embryo production. Storage at the normal culture density (8×104 microspores/ml) was less efficient for embryo production than at high densities (4×106 microspores/ml and 1.6×107 microspores/ml), although no significant difference was found between the high densities. Evaluation of plant lines derived from frozen microspores indicated no variation in isozyme pattern and no enhanced cold tolerance of these lines. Isolated microspores of B. napus could be stored for extended period for in vitro embryo production.
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  • 22
    ISSN: 1573-1561
    Keywords: Ceutorhynchus assimilis ; Coleoptera ; Curculionidae ; Brassica napus ; semiochemicals ; plant volatiles ; olfaction ; host plant ; attractant ; electroantennogram
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Electroantennograms (EAGs) were recorded from male and female cabbage seed weevils (Ceutorhynchus assimilis Payk.) in response to volatiles isolated and identified from the odor of oilseed rape (Brassica napus ssp.oleifera DC. cv. Ariana). Relatively large EAGs were obtained on stimulation with volatiles produced by the oilseed rape crop at the time when seed weevils were actively searching for host plants. Artificial rape odor without certain key volatile compounds was in most cases significantly less stimulatory than odor containing these volatiles. There were significant differences in the EAG response of the sexes ofC. assimilis to the green leaf volatiles of oilseed rape and several terpenes present in rape flower odor. The importance of the qualitative and quantitative composition of host-plant odor in host location byC. assimilis is discussed.
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  • 23
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    Springer
    Current genetics 19 (1991), S. 323-327 
    ISSN: 1432-0983
    Keywords: Brassica napus ; CMS ; ‘Polima’ cytoplasm ; Mitochondrial transcription
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We report here on RFLP and Northern blot analyses of the ‘Polima’ CMS-System in Brassica napus. Mitochondrial (mt)DNA and plastid (pt)DNA data indicate that the restorer, as well as the sterile and restored plants, possess a pol cytoplasm but that the restores has no Brassica mt-plasmid. An additional substoichiometric 3.3 kb EcoRI ptDNA fragment is present in the restorer. The transcription patterns of 12 mitochondrial-encoded genes of the pol cytoplasm have been determined for male-sterile and restored plants. Transcription patterns of the atp6 gene are correlated with the nuclear background. Restorer and restored plants exhibit two additional transcripts not detectable in sterile plants.
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  • 24
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    Springer
    Sexual plant reproduction 4 (1991), S. 22-27 
    ISSN: 1432-2145
    Keywords: Brassica napus ; Cytoplasmic male sterility ; Microsporogenesis ; Temperature ; Transmission electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Under an intermediate temperature regime (23° C/18° C; day/night), microsporogenesis in stamens of the ogu cytoplasmic male-sterile (CMS) line of Brassica napus terminated by the tetrad stage, although in some cases degeneration of the sporogenous tissue occurred prior to meiosis. In most cases the tetrads were collapsed and bounded by a sparse exine, but contained many organelles. Also, the tapetum in CMS anthers was abnormal and often highly vacuolated by the tetrad stage. Under low temperature conditions (18° C/15° C; day/night), neither microsporogenous nor tapetal tissues were observed. In the normal stamens, no differences were observed under different temperature regimes. In conjunction with the adjoining paper, this study demonstrates that temperature conditions strongly affect the cytological processes associated with microsporogenesis in the CMS anthers.
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  • 25
    ISSN: 1432-2145
    Keywords: Brassica napus ; Colchicine ; Cytoskeleton ; Microspore-embryogenesis ; Pollen
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary An attempt has been made to manipulate the cytological processes regulating the switch from gametophytic to sporophytic development induced by culturing the microspores of higher plants. Previous studies have indicated that sporophytic development, which leads to the formation of haploid embryos, normally follows the symmetrical division of the microspore rather than the asymmetric mitosis characteristic of normal development. To determine whether symmetry of division is a key factor in the determination of subsequent development, cells were supplied with the antimicrotubule drug colchicine to disrupt elements of the microtubular cytoskeleton believed to be involved in nuclear positioning. The treatment resulted in a highly significant increase in the numbers of cells turning to sporophytic development; further, timed applications indicated that the cells were sensitive to the drug over a 12-h period immediately prior to pollen mitosis. The results suggest that alteration of division symmetry is sufficient to switch the developmental pathway from gametophytic to sporophytic. These findings are discussed in the perspective of current models proposed for the regulation of development in eukaryotic cells.
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  • 26
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    Sexual plant reproduction 4 (1991), S. 176-181 
    ISSN: 1432-2145
    Keywords: Pollen ; Brassica napus ; Mitoses ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Brassica napus pollen development during the formation of the generative cell and sperm cells is analysed with light and electron microscopy. The generative cell is formed as a small lenticular cell attached to the intine, as a result of the unequal first mitosis. After detaching itself from the intine, the generative cell becomes spherical, and its wall morphology changes. Simultaneously, the vegetative nucleus enlarges, becomes euchromatic and forms a large nucleolus. In addition, the cytoplasm of the vegetative cell develops a complex ultrastructure that is characterized by an extensive RER organized in stacks, numerous dictyosomes and Golgi vesicles and a large quantity of lipid bodies. Microbodies, which are present at the mature stage, are not yet formed. The generative cell undergoes an equal division which results in two spindle-shaped sperm cells. This cell division occurs through the concerted action of cell constriction and cell plate formation. The two sperm cells remain enveloped within one continuous vegetative plasma membrane. One sperm cell becomes anchored onto the vegetative nucleus by a long extension enclosed within a deep invagination of the vegetative nucleus. Plastid inheritance appears to be strictly maternal since the sperm cells do not contain plastids; plastids are excluded from the generative cell even in the first mitosis.
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  • 27
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    Sexual plant reproduction 4 (1991), S. 226-234 
    ISSN: 1432-2145
    Keywords: Male germ unit ; Sperm cells ; Isolation ; Pollen tubes ; Brassica napus ; Pollen-tube inner plasma membrane ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Sperm cells of pollen tubes grown both in vivo and in vitro form a male germ unit. Extensions from both sperm cells of each pollen tube are closely associated with the tube nucleus. A high yield (2.7 × 104. 20 mg−1 pollen grains germinated) of intact sperm cells was obtained following release by osmotic shock from pollen tubes grown in vitro. Structural integrity of isolated sperm was maintained by isolation at low temperature in an osmotically balanced medium. At 4° C many isolated sperm pairs were still enclosed within the pollentube inner plasma membrane. Sperm cells not enclosed within this membrane no longer remained connected as a pair. During isolation vesicles formed on the sperm cell surface from disruption of the fibrillar components bridging the periplasmic space. Both in the pollen tube and after isolation the sperm nucleus is in close association with at least one region of the sperm plasma membrane. Sperm isolated at room temperature showed the presence of nucleopores, and nuclei were euchromatic, instead of heterochromatic as in intact sperm in the pollen tube.
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  • 28
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    Biology and fertility of soils 12 (1991), S. 213-216 
    ISSN: 1432-0789
    Keywords: Soil management problems ; Multiply cropped paddy fields ; Mineral fertilizer ; Organic manure ; Sustainable agriculture ; Oryza sativa ; Trificum sativa ; Brassica napus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Multiple cropping of paddy fields has now been widely adopted in East China, in order to increase annual grain yields. The management practices recommended for these multiple cropped fields include the use of mineral fertilizers in combination with organic manure, to sustain a better soil nutrient balance, the adoption of rice-straw manuring, and use of zero or minimum tillage. This paper presents the results of a series of longterm experiments in multiple cropping and discusses the problems associated with these techniques.
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  • 29
    ISSN: 1573-5028
    Keywords: Gene transfer ; Arabidopsis thaliana ; Brassica napus ; transient expression ; pneumatic particle gun
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Successful transient expression of β-glucuronidase (GUS) in Arabidopsis thaliana leaves and roots and Brassica napus stems was obtained after gene delivery with a pneumatic particle gun driven by compressed air. Effects of the pneumatic pressure used to accelerate the particles (accelerating pressure; 85 to 200 kg/cm2) and of preculture periods of plant tissues (0 to 6 days) on the efficiency of gene delivery were studied. In A. thaliana leaves, best results were obtained at 115 kg/cm2 of accelerating pressure and 3 days of preculture. In A. thaliana roots, the optimum was at 200 kg/cm2 of accelerating pressure and 3 days of preculture. These results indicate that both preculture period and accelerating pressure are vital factors that determine the efficiency of gene delivery by particle gun.
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  • 30
    ISSN: 1573-5028
    Keywords: Brassica napus ; microscope development ; pectin esterase ; pollen-specific gene ; promoter sequences
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Differential screening of a Brassica napus genomic library led to the isolation of the clone named Bp 19 containing a gene which is highly expressed during microspore development. The accumulation of Bp 19 mRNA starts in uninucleate microspores, increases during development reaching a peak in the late stages but declines considerably in mature pollen. The nucleotide sequence of the entire coding region and of extended portions of the 5′ and 3′ flanking regions was determined. Several homologous cDNA clones were also isolated and sequenced. The Bp 19 gene contains a single intron of 137 bp and gives origin to a mRNA of ca. 1.9 kb which codes for a polypeptide of 584 amino acids. Bp 19 protein has an estimated molecular weight of 63 kilodaltons and has a highly hydrophobic amino terminal region which shows features of a signal peptide. The carboxy half of the Bp 19 protein, starting at amino acid 269, has striking sequence similarity to the pectin esterases of tomato and of the plant pathogen Erwinia chrysanthemi. Four short domains are extremely well conserved in all the three proteins and therefore could represent catalytic sites responsible for enzyme activity. Comparison of the 5′ flanking region of the Bp 19 gene with the sequence of other pollen-specific promoters revealed the presence of several conserved regions. These short promoter sequences could correspond to regulatory elements responsible for pollen-specific gene expression.
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  • 31
    ISSN: 1573-5028
    Keywords: acetolactate synthase ; Brassica napus ; cDNA ; expression ; nucleotide sequence
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
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  • 32
    ISSN: 1573-5028
    Keywords: Brassica napus ; embryogenesis ; napin gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract S1 nuclease analysis and sub-family-specific oligonucleotide probes were used to characterize the expression during embryogenesis of the napin storage protein gene family ofBrassica napus (oilseed rape). The expression of one sub-class represented by the napin gene gNa peaks and declines earlier than the other members of the family. This sub-class was highly expressed representing ca. 20% of napin mRNA at 26 days after anthesis.
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  • 33
    ISSN: 1573-5028
    Keywords: acyl carrier protein ; Brassica napus ; enoyl-ACP reductase ; fatty acid synthesis ; seed development ; nuclear-encoded chloroplast proteins
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The onset of storage lipid biosynthesis during seed development in the oilseed crop Brassica napus (rape seed) coincides with a drastic qualitative and quantitative change in fatty acid composition. During this phase of storage lipid biosynthesis, the enzyme activities of the individual components of the fatty acid synthase system increase rapidly. We describe a rapid and simple purification procedure for the plastidlocalized NADH-dependent enoyl-acyl carrier protein reductase from developing B. napus seed, based on its affinity towards the acyl carrier protein (ACP). The purified protein was N-terminally sequenced and used to raise a potent antibody preparation. Immuno-screening of a seed-specific λgt11 cDNA expression library resulted in the isolation of enoyl-ACP reductase cDNA clones. DNA sequence analysis of an apparently full-length cDNA clone revealed that the enoyl-ACP reductase mRNA is translated into a precursor protein with a putative 73 amino acid leader sequence which is removed during the translocation of the protein through the plastid membrane. Expression studies in Escherichia coli demonstrated that the full-length cDNA clone encodes the authentic B. napus NADH-dependent enoyl-ACP reductase. Characterization of the enoyl-ACP reductase genes by Southern blotting shows that the allo-tetraploid B. napus contains two pairs of related enoyl-ACP reductase genes derived from the two distinct genes found in both its ancestors, Brassica oleracea and B. campestris. Northern blot analysis of enoyl-ACP reductase mRNA steady-state levels during seed development suggests that the increase in enzyme activity during the phase of storage lipid accumulation is regulated at the level of gene expression.
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  • 34
    ISSN: 1573-5028
    Keywords: Brassica napus ; extensin genes ; GUS gene fusions ; phloem-specific expression ; transgenic plants
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The organs and tissues where the Brassica napus extA extensin gene is expressed have been identified. The extA gene with 3.75 kb of 5′ flanking sequence was transferred to tobacco via disarmed Agrobacterium tumefaciens vectors and transgenic plants regenerated. The gene was found to be inactive in transgenic tobacco leaf, but was active as measured by RNA transcript assays in both stem and root tissues. To determine the cell-specific expression pattern of the extA gene, a promoter-reporter gene fusion construct was made consisting of 1.0 kb of 5′ extA sequence fused to the coding region of the glucuronidase (GUS) gene. This fusion construct was introduced into B. napus via Agrobacterium rhizogenes, and expression of GUS in transgenic rape hairy roots was examined. GUS activity was only seen in the vascular tissues of the rape root, and was found to be specifically localised in the phloem.
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  • 35
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    Plant molecular biology 17 (1991), S. 295-299 
    ISSN: 1573-5028
    Keywords: Brassica napus ; microspore-specific ; microsporogenesis ; DNA sequence ; peptide motif
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The I3 cDNA isolated from a library made from 2–4 mm (immature) anthers of Brassica napus shows microspore-specific expression. Homologous transcripts are detected in buds and anthers of male-fertile plants, but not in green tissues, roots, or in cytoplasmic male-sterile buds. High expression of the transcript is limited to microspores entering and undergoing mitosis. The predicted peptide sequence of the cDNA shows an unusual repeated alanine/proline motif at the C-terminus, which may be of importance in the native protein structure.
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  • 36
    ISSN: 1573-5028
    Keywords: Brassica napus ; DNA-binding proteins ; light-inducible gene ; tissue specificity ; transmembrane protein
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Using a fractionated genomic bank, we have cloned and characterized a Brassica napus gene (rbcSF1) encoding the small sub-unit of ribulose 1,5-bisphosphate carboxylase. The promoter of this gene contains a 29 bp direct repeat capable of forming a single or a double hairpin loop, and three elements that are recognized by leaf nuclear proteins in vitro. The most upstream are the S-box, a small A/T-rich sequence between −516 and −512, and the F-box between −492 and −475. Finally, we have also observed binding to the G-box, a regulatory element common to numerous plant promoters. The promoter of rbcSF1 also has a 113 amino acids open reading frame (ORF113) in the non-coding strand. When used to probe a northern blot of leaf RNA, this ORF hybridizes to a 1.5 kb transcript. The protein encoded by ORF113 contains a transmembrane domain.
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  • 37
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    Plant molecular biology 17 (1991), S. 1-8 
    ISSN: 1573-5028
    Keywords: Agrobacterium rhizogenes ; Brassica napus ; genetic transformation ; hairy roots ; regenerated shoots
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Genetically transformed root clones of rapeseed (Brassica napus) were obtained afterin vitro infection of excised hypocotyl segments with a wild type strain ofAgrobacterium rhizogenes and two strains ofA. rhizogenes harbouring kanamycin resistance. The ability of hairy root formation was affected by light and was highly dependent on the location of the infection site at the hypocotyl. Inoculation of decapitated hypocotyls with an intact root system gave rise to direct shoot formation from the site of inoculation. Histological sections showed that several meristems were initiated at the inoculation site. Root and shoot clones were isolated and subcultured axenically in hormone-free liquid MS medium. Identification of transformed root and shoot clones was based on opine assays. Further selection was carried out in kanamycin-enriched medium. All opine-positive root clones showed NPT II (neomycin phosphotransferase) activity. Nearly half of the shoot clones expressed a strong NPT II activity while the rest gave a weak or no NPT II response.
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  • 38
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    Plant foods for human nutrition 41 (1991), S. 69-88 
    ISSN: 1573-9104
    Keywords: selenium ; nutrition ; plant spraying ; multiple use ; Helianthus tuberosus ; Medicago sativa ; Brassica napus ; polarography
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The technique of spraying plants with inorganic forms of selenium can be employed for phytochemical production of organic selenium compounds. Fractionation of the plant material makes it possible to produce a highly concentrated and well defined selenium supplement with potential use in animal and human nutrition. The fractionation also gives opportunities to combine production of organic selenium compounds with other products, for example plant fibres. Multiple use of plants can contribute to a more efficient utilization of land area (in comparison to monocultures solely adapted to food production). It also gives the opportunity to develop systems suitable for long term fixation of carbon, as long as the plant material is not reoxidised to carbon dioxide. Plant fibres could provide raw material for the production of paper or building materials in combination with the production of organic selenium compounds preferentially accumulated in another fraction of the processed plant.
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  • 39
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    Euphytica 58 (1991), S. 31-35 
    ISSN: 1573-5060
    Keywords: Brassica napus ; SSD lines ; heterosis ; epistasis ; yield ; quality traits
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Heterosis and epistasis in spring oilseed rape (Brassica napus L.) was analysed by comparing generation means for ten agronomic traits. Parents, F2, F3 and F6 generations of four crosses with Swedish and French material were investigated. The F2 was 11% higher in yield, earlier in flowering time, and slightly later in maturation when compared with the parents. Randomly derived single seed descent lines had an 8% lower yield, were later flowering and maturing than the parents in F6. This poorer average performance of recombinant lines is explained by the loss of favourable epistatic interactions present in the parents.
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  • 40
    ISSN: 1573-5060
    Keywords: Brassica napus ; rapeseed fatty acid accumulation ; microspore-derived embryo ; zygotic seed ; triacylglyceride
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Microspore culture of rapeseed (Brassica napus L.) has provided a powerful tool not only for breeding but also in developmental studies. In this study, microspore-derived embryos (MDE) of B. napus were evaluated as a model in seed for studying accumulations of triacylglyceride (TAG) fatty acids in both a low and high erucic acid rapeseed line; and accumulations of TAG and free fatty acids (FFA) in a high erucic acid rapessed line. The accumulation patterns confirmed that MDE had a similar TAG fatty acid profile to seed during the embryo development within each genotype. The oil accumulation in MDE after 36 days in culture (DIC) approached levels similar to those in zygotic seed 25 days after flowering (DAF). Significant differences were detected in contents of both total free fatty acids and specific free fatty acids between MDE and seed. During the developmental period, total free fatty acids changed from 16% to 2.1% in MDE, but from 10.5% to 0.1% in seed. MDE had much higher percentage of free linolenic and erucic acids than seed, particularly during the late developmental stages. The current study indicated that MDE can be used as a model to study TAG and TAG fatty acids in seed but caution must be taken to study free fatty acid metabolism.
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  • 41
    ISSN: 1573-5060
    Keywords: Brassica napus ; donor plant age ; embryogenesis ; haploid ; microspore culture ; microspore stage
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Effect of age of donor plants and age of inflorescence on embryogenesis in microspore culture of B. napus was examined. Microspores isolated from buds of older plants had a higher embryo yield than those of younger ones. The effect of the age of inflorescence showed a different pattern. In older plants, a higher embryogenesis response was observed in microspores isolated from buds of new inflorescences, while in young plants, microspores isolated from buds of old inflorescences showed high embryo yield. These different responses were considered to be attributable to a difference in the developmental stage of pollen at the time of microspore isolation. Our results indicated that microspores collected from older inflorescences and older plants have sufficient embryogenic potential when the optimum developmental stage of pollen was used. Frequency of embryo to plant conversion was influenced by the size of embryos subcultured, but not by donor plant age or the age of the inflorescence.
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  • 42
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    Plant cell, tissue and organ culture 24 (1991), S. 43-47 
    ISSN: 1573-5044
    Keywords: Brassica napus ; protoplasts ; purification
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The effects of three different general purification protocols have been assessed quantitatively using mesophyll protoplasts of Brassica napus. Within the initial sample two distinct sub-populations were determined. The methods used influenced the ratio of the vacuolated to chloroplastic type protoplast sub-populations. Overall recovery rates of the initial sample varied according to the method used from 38% to 27%, but the relative recovery of the sub-populations varied considerably with a purified ratio of between 1.0:0.78 to 1.0:7.0. Size distribution profiles of the initial and purified populations are also presented.
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  • 43
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    Plant cell, tissue and organ culture 26 (1991), S. 195-201 
    ISSN: 1573-5044
    Keywords: Brassica napus ; cell culture ; diffusion ; liquid medium ; oxygen availability ; protoplast culture
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A method for estimating the oxygen availability in plant cell cultures grown in stationary liquid media (e.g. many protoplast cultures) was developed. The method is based on short-term measurements of respiration rate versus oxygen concentration on a sample of cells, suspended in liquid media. From such data it is possible to estimate the oxygen concentration at the bottom of a stagnant liquid culture, by calculating the amount of oxygen reaching the cells by diffusion. As an example, rape (Brassica napus L. cv. Omega) hypocotyl protoplasts were grown with different oxygen concentrations at the site of the cells, obtained by varying the cell density, the height of the liquid layer and the oxygen content of the gas phase. The number of surviving calli was positively correlated with the estimated oxygen availability in the range between 60 and 350 μM O2, below 60 μM all cells died. This indicates that oxygen availability can be a limiting factor in the range usually encountered in protoplast cultures, and that the method can be useful when designing optimal growth conditions for stationary cultures of plant cells.
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  • 44
    ISSN: 1573-1561
    Keywords: Sinaibin ; glucosinolate ; resistance ; Sinapis alba ; Brassica napus ; Phyllotreta cruciferae ; Coleoptera ; Chrysomelidae ; Mamestra configurata ; Lepidoptera ; Noctuidae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Sinalbin was identified as a chemical component of insect anti-xenosis and antibiosis resistance mechanisms in seedlings ofSinapis alba by DEAE-Sephadex chromatography, HPLC, treatment with sulfatase and myrosinase, various feeding tests using artificial and natural substrates, and by measuring sinalbin concentrations in cotyledons and leaves during seedling development. The effects of sinaibin on feeding were dependent upon the insect species and upon the rapidly changing profile of sinaibin concentrations in the developing seedling. The high concentrations of sinalbin found in young cotyledons (up to 20 mM) and leaves (up to 10 mM) deterred the feeding of the flea beetle,Phyllotreta cruciferae Goeze and larvae of the bertha armyworm,Mamestra configurata Walker. The protection that sinalbin confers upon the vulnerable, newly emerged seedling (and upon tiny, young leaves) appears critical for the first few days of survival ofS. alba under feeding pressure from flea beetles in the field. The lower concentrations of sinaibin found in older cotyledons and leaves (2–3 mM) offer little or no protection againstP. cruciferae and may actually stimulate the feeding of this crucifer specialist. These concentrations of sinaibin, however, are still effective in reducing the level of feeding by larvae of the more generalist feederM. configurata.
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  • 45
    ISSN: 1573-5060
    Keywords: Agrobacterium ; Brassica napus ; CaMV 35S promoter ; mas promoter ; gene expression ; risk assessment
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Gene fusions between the β-glucuronidase (GUS) reporter gene and the promoters of the cauliflower mosaic virus 35S RNA transcript (CaMV 35S) and the mannopine synthase (mas) genes were introduced into rapeseed varieties via Agrobacterium-mediated transformation. Fluorometric assay of β-glucuronidase activity indicated different expression patterns for the two promoters. In seedlings, the CaMV 35S promoter had maximum activity in the primary roots, while the mas promoter was most active in the cotyledons. Etiolated seedlings cultured in the dark showed reduced activity of the mas promoter. Before vernalization at the rosette stage, both promoters were more active in older plant parts than in younger ones. At this stage the highest activity was recorded in cotyledons. After the plants had bolted reduced promoter function was detected in the upper parts of the transformed plants. Both promoters were found to be functional in the majority of the studied organs of transgenic rapeseed plants, but the promoter activity varied considerably between the organs at different developmental stages. The ability of pollen to transfer the introduced genes to other varieties and related species (e.g. Brassica napus and Diplotaxus muralis) by cross-pollination was studied in greenhouse experiments, and field trials were carried out to estimate the distance for biologically-relevant gene dispersal. In artificial crossing, the introduced marker gene was transferable into other varieties of Brassica napus. In field trials, at a distance of 1 metre from the source of transgenic plants, the frequency of an outcrossing event was relatively high (10-3). Resistant individuals were found at 16 and 32 metres from the transgenic pollen donors, but the frequency of an outcrossing event dropped to 10-5.
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  • 46
    ISSN: 1573-5060
    Keywords: asymmetric somatic hybridization ; Brassica napus ; Brassica nigra ; disease resistance transfer ; dot blot analysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Asymmetric somatic hybrid plants between Brassica napus L. (oilseed rape genome AACC) and a transgenic line of Brassica nigra L. Koch (black mustard genome BB) were tested for their resistance against rapeseed pathogens Phoma lingam (black leg disease) and Plasmodiophora brassicae (club root disease). The transgenic B. nigra line used (hygromycin-resistant, donor) is highly resistant to both fungi, whereas B. napus (recipient) is highly susceptible. The asymmetric somatic hybrids were produced using the donor-recipient fusion method (with X-irradiation of donor protoplasts) reported by Zelcer et al. (1978) for the production of cybrids. Using hygromycin-B for selection, a total of 332 hybrid calli were obtained. Regenerants, resistant or susceptible to both diseases, were selected. Many hybrids expressed resistance to only one pathogen. Dot blot experiments showed that the asymmetric hybrid plants contained varying amounts of the donor genomic DNA. Furthermore, a correlation was detected between the radiation dose and the degree of donor DNA elimination.
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  • 47
    ISSN: 1573-5060
    Keywords: Brassica napus ; disease tolerance ; oxalic acid ; oxalate oxidase ; Sclerotinia sclerotiorum ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Oxalic acid is thought to have a primary role in the pathogenicity of several plant pathogens, notably Sclerotinia selerotiorum. A gene coding for the enzyme oxalate oxidase was isolated from barley roots and introduced into oilseed rape as a means of degrading oxalic acid in vivo. This report describes the production of several transgenic plants of oilseed rape and the characterisation of these plants by Southern, Western and enzyme activity assays. Plants were shown to contain an active oxalate oxidase enzyme and were tolerant of exogenously supplied oxalic acid.
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  • 48
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    Euphytica 85 (1955), S. 323-327 
    ISSN: 1573-5060
    Keywords: Brassica napus ; fatty acids ; gas chromatography ; Lunaria annua ; protoplast regeneration ; somaclonal variation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary A programme of research was designed to investigate methods for the modification of the fatty acid profiles of high performance lines of oilseed rape (Brassica napus L.) in an attempt to produce lines with enhanced levels of industrially useful fatty acids. The methodology employed to achieve these objectives was based on the exploitation of somaclonal or protoclonal variation, and targeted somatic hybridization using wild cruciferous germplasm as fusion partners. A range of somaclonal lines was produced from shoot regeneration protocols. These lines underwent replicated, randomised glasshouse trials for morphological assessment followed by gas chromatographic analysis to monitor any changes in fatty acid profile. It was found that a small number of lines exhibited potentially useful changes in oleic acid and polyunsaturated fatty acid content. Protoplast regeneration and electrofusion protocols for a range of winter oilseed rape lines were developed, and methods for the isolation and fusion of protoplasts of the wild crucifer Lunaria annua (chosen for its high nervonic acid content) established.
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