The interrelationship between the accumulation of lipids, protein and the level of acyl carrier protein during the development of Brassica napus L. pollen

Lipid accumulation during pollen and tapetal development was studied using cryostat sections of unfixed anthers from Brassica napus (rapeseed). Diamidino-2-henylindole (DAPI), a DNA fluorochrome, was used to stain the pollen nuclei in order to identify ten stages of pollen development in Brassica. Storage lipids (i.e. triacylglycerides) were stained using the fluorochrome Nile red. Pollen coat lipids are formed in tapetal plastids between the mid-vacuolate and early maturation pollen stages. The pollen coat components, including lipids and a proportion of the proteins, are derived from the remnants of the tapetum, after its rupture, during the second pollen mitosis. Quantitative microfluorometric analyses demonstrated four phases of lipid body accumulation or depletion in the developing pollen cytoplasm. The majority of storage lipids found in the cytoplasm of the mature pollen grain accumulated during the late vacuolate and early maturation stages when the pollen is bicellular. The level of acyl carrier protein, a protein integrally involved in lipid synthesis, was also found to be maximal in the developing pollen during the bicellular pollen stages of development. This coincided with the most active period of lipid accumulation. These data could indicate that the lipids of the pollen are synthesized in situ, by metabolic processes regulated by expression of genes in the haploid genome.

Composição química e qualidade biológica da torta de colza (Brassica napus, L: )

A composição química e a qualidade biológica da torta de colza obtida da semente, variedade CTC-4, foram determinados. A farinha foi preparada me diante prensagem da semente e extração da gordura com solvente, lixiviada durante 2 horas, com água corrente na proporção de 1:5, e agitação constante, a fim de reduzir os níveis de glicosinolatos. O efeito tóxico foi observado através do peso dos órgãos e da histopatologia dos órgãos em experimento de 60 dias. A qualidade biológica da torta desintoxicada, ao nível de 5% e 10%, foi semelhante ao da caseína (PER controle 3,18 vs 3,08, 5% e 2,98, 10%). O PER da torta crua foi de 2,28. Os ratos que receberam a farinha crua tiveram órgãos maiores. As tireóides dos ratos submetidos as dietas 2 e 3, não demonstraram alterações que suspeitassem de um efeito bociogênico e as alterações ocorridas nas tireóides dos animais que receberam, D-4 foram semelhantes àquelas ocorridas nos grupos 2 e 3. Não houve danos célulares graves em relação ao fígado, os rins, o coração, baço e supra-renais para todos os grupos observados. O método de extração dos fatores tóxicos da colza foi eficiente de acordo com os resultados obtidos para o presente estudo.

Entomophilous pollination in rape (Brassica napus L. var. oleifera) in Brazil

The most frequent insect visitors to the flower were: Apis mellifera, 80.6%; Trigona spinipes, 12.8% and Dialictus sp, 6.6%. -from Authors

Allelopathic effects of aqueous extract of Brassica napus on germination of seeds of Phaseolus vulgaris

Secondary compounds produced by plants are considered an alternative method of weed suppression but can cause negative effects on crops in succession, especially in a no-tillage system, due to the degradation of crop residues with allelopathic potential. The objective of this work was to analyze the influence of foliar aqueous extracts of Brassica napus on the germination and initial development of seedlings of Phaseolus vulgaris. The extract was prepared as a stock 10 % weight/volume solution, and diluted into treatments of relative concentrations of 100 % (i.e. 10 % w/v stock), 75 %, 50 %, 25 % and 0 % (untreated control consisting of distilled water), in a completely randomized design. The seeds of Phaseolus vulgaris were moistened with the differing concentrated extracts and kept in a germination chamber at 25 °C, with a photoperiod of 12 h for nine days. The variables evaluated were: percentage germinating, first count of germination and germination velocity index, as well the root and hypocotyl length, and fresh and dry mass of the seedlings. The aqueous leaf extracts of Brassica napus did not influence the germination of Phaseolus vulgaris seeds, but did induce the growth of abnormal seedlings by inhibition of secondary roots and reduced prominence of the primary root.

Ileal amino acid digestibility in canola meals from yellow- and black-seeded Brassica napus and Brassica juncea fed to growing pigs

Twelve ileal cannulated pigs (30.9 ± 2.7 kg) were used to determine the apparent (AID) and standardized (SID) ileal digestibility of protein and AA in canola meals (CM) derived from black- (BNB) and yellow-seeded (BNY) Brassica napus canola and yellow-seeded Brassica juncea (BJY). The meals were produced using either the conventional pre-press solvent extraction process (regular meal) or a new, vacuum-assisted cold process of meal de-solventization (white flakes) to provide 6 different meals. Six cornstarch-based diets containing 35% canola meal as the sole source of protein in a 3 (variety) × 2 (processing) factorial arrangement were randomly allotted to pigs in a 6 × 7 incomplete Latin square design to have 6 replicates per diet. A 5% casein diet was fed to estimate endogenous AA losses. Canola variety and processing method interacted for the AID of DM (P = 0.048), N (P = 0.010), and all AA (P < 0.05), except for Arg, Lys, Phe, Asp, Glu, and Pro. Canola variety affected or tended to affect the AID of most AA but had no effect on the AID of Lys, Met, Val, Cys, and Pro, whereas processing method had an effect on only Lys and Asp and tended to affect the AID of Thr, Gly and Ser. The effects of canola variety, processing method, and their interaction on the SID values for N and AA followed a similar pattern as for AID values. For the white flakes, SID of N in BJY (74.2%) was lower than in BNY and BNB, whose values averaged 78.5%; however, among the regular meals, BJY had a greater SID value for N than BNY and BNB (variety × processing, P = 0.015). For the white flakes, the SID of Ile (86.4%), Leu (87.6%), Lys (88.9%), Thr (87.6%) and Val (84.2%) in BNB were greater than BNY and BJY. Opposite results were observed for the regular processing, with SID of Lys (84.1%), Met (89.5%), Thr (84.1%), and Val (83.6%) being greater in BJY, followed by BNB and BNY(variety × processing, P < 0.057). The SID of Met was greatest for the white flakes (90.2%) but least for the regular processing (83.0%) in BNY (variety × processing, P < 0.057). It was concluded that the AID and SID of N and AA of the CM tested varied according to canola variety and the processing method used. Overall, the SID values for Ile, Leu, Lys, Met, Thr, and Val averaged across CM types and processing methods were 81.8, 82.6, 83.4, 85.9, 80.8, and 78.4%, respectively.

Expression and distribution of a vacuolar aquaporin in young and mature leaf tissues of Brassica napus in relation to water fluxes

Recently, it has been shown that water fluxes across biological membranes occur not only through the lipid bilayer but also through specialized water-conducting proteins, the so called aquaporins. In the present study, we investigated in young and mature leaves of Brassica napus L. the expression and localization of a vacuolar aquaporin homologous to radish γ-tonoplast intrinsic protein/vacuolar-membrane integral protein of 23 kDa (TIP/VM 23). In-situ hybridization showed that these tonoplast aquaporins are highly expressed not only in developing but also in mature leaves, which export photosynthates. No substantial differences could be observed between different tissues of young and mature leaves. However, independent of the developmental stage, an immunohistochemical approach revealed that the vacuolar membrane of bundle-sheath cells contained more protein cross-reacting with antibodies raised against radish γ-TIP/VM 23 than the mesophyll cells. The lowest labeling was detected in phloem cells. We compared these results with the distribution of plasma-membrane aquaporins cross-reacting with antibodies detecting a domain conserved among members of the plasma-membrane intrinsic protein 1 (PIP1) subfamily. We observed the same picture as for the vacuolar aquaporins. Furthermore, a high density of gold particles labeling proteins of the PIP1 group could be observed in plasmalemmasomes of the vascular parenchyma. Our results indicate that γ-TIP/VM 23 and PIP1 homologous proteins show a similar expression pattern. Based on these results it is tempting to speculate that bundle-sheath cells play an important role in facilitating water fluxes between the apoplastic and symplastic compartments in close proximity to the vascular tissue.

Plant genotype affects the quality of oilseed rape (Brassica napus) for adults and larvae of the pollen beetle (Meligethes aeneus)

Plant quality is one of the main factors influencing the fitness of phytophagous insects. Plant quality can vary not only among genotypes of the same host plant species, but also relative to the insect sex or its life stage. In the present study, the performance of larvae and adults of the pollen beetle (Meligethes aeneus F., Coleoptera: Nitidulidae), a major insect pest of oilseed rape crops, is compared on six genotypes of oilseed rape (Brassica napus). All of the traits that are measured vary among genotypes, and comprise larval developmental duration, life span of unfed emerging adults and survival time of field-sampled adults fed with pollen from the different genotypes. No correlation is found between insect performance and quantity of food available, showing that the quality of the food (i.e. pollen) is the fitness determinant for this insect species. Additionally, the performance of larvae and adults is also not correlated despite use of the same plant genotypes, suggesting that the determinants of pollen quality differ at least partially between both life stages. It is hypothesized that this may be a result of extensive differences in diet breadth between the life stages: larvae are specialists of brassicaceous plants, whereas adults are generalists. Finally, it is suggested that the manipulation of plant quality to increase pollen beetle development time may comprise a valuable strategy for favouring biological control by natural enemies of this pest; for example, as a result of extending the vulnerability window of larvae to attack by parasitoids.

Uso de aguas residuales urbanas en el riego de colza (Brassica napus L.) con destino a biocombustible

El aceite de colza es comestible pero también puede utilizarse en la producción de biodiesel. Cuando el destino es el energético, el cultivo puede regarse con aguas residuales urbanas o cloacales. La mayor proporción del uso de éstas en el mundo ocurre en regiones áridas donde otras fuentes de agua son escasas, situación que se plantea en los oasis irrigados de Mendoza. En este trabajo se comparó el rendimiento de un cultivar invernal de colza regado con agua cloacal (AC) y agua subterránea (AS), y su potencial para producir biodiesel. La experiencia se llevó a cabo en una planta de tratamiento de agua cloacal de Obras Sanitarias en el departamento Tunuyán (33° 32’ 89’’ S; 69° 00’ 80’’ O; 859 m snm). El rendimiento de semilla de AC fue significativamente mayor que el de AS (7690 y 3886 kg/ha, respectivamente). La cantidad de biodiesel factible de producir por cada hectárea de cultivo asciende a 2800 kg en el tratamiento AC y a 1400 kg en AS. El uso de aguas residuales urbanas genera un nicho interesante para la producción de biocombustibles, utilizando un recurso hídrico con limitaciones para producir alimentos.

Parámetros biológicos y demograficos de áfidos (Hemiptera: aphididae) en variedades de colza canola (Brassica napus L.)

Myzus persicae (Sulz.), Brevicoryne brassicae L. y Lipaphis erysimi (Kalt.) constituyen plagas del cultivo de colza canola, Brassica napus L., y de otras crucíferas cultivadas. Los pulgones fueron colectados en un cultivo de canola y se multiplicaron en un insectario. Para el estudio se criaron 2 cohortes de 20 hembras neonatas, para cada especie de áfido y cultivar de colza, a 20 ± 1°C, 60-70% de humedad relativa y 14:10 horas de fotofase. Las principales diferencias entre especies de pulgones se observaron en la duración de los períodos reproductivo y post-reproductivo, la longevidad y la fecundidad. M. persicae fue el áfido más longevo y fecundo en Impact, mientras que en Teddy el más fecundo fue L. erysimi y el más longevo M. persicae. Con respecto a los parámetros poblacionales, la mayor tasa de incremento poblacional (R0) fue de 58,43 (♀/♀/generación) para M. persicae en Impact. En cambio en Teddy la mayor R0 (63,17) fue para L. erysimi. La tasa intrínseca de crecimiento natural (rm) más elevada fue para L. erysimi en Teddy (0,29 ♀/♀/día) y para M. persicae en Impact (0,24 ♀/♀/día). Estos estudios indican que, de acuerdo con el cultivar de colza empleado, teniendo en cuenta la tasa de crecimiento poblacional de las diferentes especies de pulgones, se podría inferir cuál será la especie dominante y el daño potencial que ocasionarían al cultivo.

Isolation and characterization of neutral-lipid-containing organelles and globuli-filled plastids from Brassica napus tapetum

The monolayer tapetum cells of the maturing flowers of Brassica napus contain abundant subcellular globuli-filled plastids and special lipid particles, both enriched with lipids that are supposed to be discharged and deposited onto the surface of adjacent maturing pollen. We separated the two organelles by flotation density gradient centrifugation and identified them by electron microscopy. The globuli-filled plastids had a morphology similar to those described in other plant species and tissues. They had an equilibrium density of 1.02 g/cm3 and contained neutral esters and unique polypeptides. The lipid particles contained patches of osmiophilic materials situated among densely packed vesicles and did not have an enclosing membrane. They exhibited osmotic properties, presumably exerted by the individual vesicles. They had an equilibrium density of 1.05 g/cm3 and possessed triacylglycerols and unique polypeptides. Several of these polypeptides were identified, by their N-terminal sequences or antibody cross-reactivity, as oleosins, proteins known to be associated with seed storage oil bodies. The morphological and biochemical characteristics of the lipid particles indicate that they are novel organelles in eukaryotes that have not been previously isolated and studied. After lysis of the tapetum cells at a late stage of floral development, only the major plastid neutral ester was recovered, whereas the other abundant lipids and proteins of the two tapetum organelles were present in fragmented forms or absent on the pollen surface.

Embryo-Specific Gene Expression in Microspore-Derived Embryos of Brassica napus. An Interaction between Abscisic Acid and Jasmonic Acid1, 2

The induction of napin and oleosin gene expression in Brassica napus microspore-derived embryos (MDEs) was studied to assess the possible interaction between abscisic acid (ABA) and jasmonic acid (JA). Napin and oleosin transcripts were detected sooner following treatment with ABA than JA. Treatment of MDEs with ABA plus JA gave an additive accumulation of both napin and oleosin mRNA, the absolute amount being dependent on the concentration of each hormone. Endogenous ABA levels were reduced by 10-fold after treatment with JA, negating the possibility that the observed additive interaction was due to JA-induced ABA biosynthesis. Also, JA did not significantly increase the uptake of [3H-ABA] from the medium into MDEs. This suggests that the additive interaction was not due to an enhanced carrier-mediated ABA uptake by JA. Finally, when JA was added to MDEs that had been treated with the ABA biosynthesis inhibitor fluridone, napin mRNA did not increase. Based on these results with the MDE system, it is possible that embryos of B. napus use endogenous JA to modulate ABA effects on expression of both napin and oleosin. In addition, JA could play a causal role in the reduction of ABA that occurs during late stages of seed development.

(+)-Abscisic Acid Metabolism, 3-Ketoacyl-Coenzyme A Synthase Gene Expression, and Very-Long-Chain Monounsaturated Fatty Acid Biosynthesis in Brassica napus Embryos1

Microspore-derived embryos of Brassica napus cv Reston were used to examine the effects of exogenous (+)-abscisic acid (ABA) and related compounds on the accumulation of very-long-chain monounsaturated fatty acids (VLCMFAs), VLCMFA elongase complex activity, and induction of the 3-ketoacyl-coenzyme A synthase (KCS) gene encoding the condensing enzyme of the VLCMFA elongation system. Of the concentrations tested, (+)-ABA at 10 μm showed the strongest effect. Maximum activity of the elongase complex, observed 6 h after 10 μm (+)-ABA treatment, was 60% higher than that of the untreated embryos at 24 h. The transcript of the KCS gene was induced by 10 μm (+)-ABA within 1 h and further increased up to 6 h. The VLCMFAs eicosenoic acid (20:1) and erucoic acid (22:1) increased by 1.5- to 2-fold in embryos treated with (+)-ABA for 72 h. Also, (+)-8′-methylene ABA, which is metabolized more slowly than ABA, had a stronger ABA-like effect on the KCS gene transcription, elongase complex activity (28% higher), and level of VLCMFAs (25–30% higher) than ABA. After 24 h approximately 60% of the added (+)-[3H]ABA (10 μm) was metabolized, yielding labeled phaseic and dihydrophaseic acid. This study demonstrates that (+)-ABA promotes VLCMFA biosynthesis via increased expression of the KCS gene and that reducing ABA catabolism would increase VLCMFAs in microspore-derived embryos.