No description available.

Previous studies have shown that the mitochondrial orf224/ atp6 gene region is correlated with the Polima (pol) cytoplasmic male sterility (CMS) of Brassica napus, and that the effects of nuclear fertility restoration on orf224/ atp6 transcripts co-segregate with the pol restorer gene Rfp in genetic crosses. Results presented in this thesis indicate that the pol CMS restorer gene Rfp acts in an organ-specific manner to promote the processing of primary, dicistronic orf224/atp6 transcripts into new restorer-specific, monocistronic transcripts. The single 1.1 kb atp6 transcript of nap cytoplasm and some orfB transcripts of nap and pol cytoplasm B. napus mitochondria are shown to arise by removal of sequences from the 5' end of a longer precursor. Specific endonucleolytic cleavage of a precursor RNA, followed by non-specific 3' to 5' exonuclease action, may thus represent a common mechanism for tailoring transcripts in plant mitochondria. Northern analysis of a segregating F2 population indicates that the recessive rfp allele at the restorer gene locus, or a very tightly linked gene, acts as a dominant gene designated Mmt (modifier of mitochondrial transcripts). Mmt controls the presence of additional, smaller transcripts of the nad4 and ccl1-like (ccl1-l) genes. These results suggest that Rfp/Mmt is a novel nuclear genetic locus that affects the expression of multiple mitochondrial gene regions, with different alleles or haplotypes exerting specific effects on different mitochondrial genes. One of these genes, ccl1-l, is split in the B. napus mitochondrial genome into two widely separated and independently transcribed parts, contrasting with the situation in other plants where this gene is present as an uninterrupted ORF. Although transcripts of both parts are edited, no "trans-spliced" transcripts spanning both parts of the split ORF, that could be translated into a full sized protein product, were detected. Moreover, an antiserum directed against the product of t

Diacylglycerol acyltransferase (DGAT), an integral membrane protein of the endoplasmic reticulum, catalyses the final step in the sn-glycerol-3-phosphate pathway leading to triacylglycerol. Although DGAT has been cloned from a variety of species, including the major oilseed crop of Canada, canola (Brassica napus), little is known about the structure/function of the enzyme. BnDGAT1 is the major isoform of type I DGAT (DGAT-I) in microspore-derived cell suspension cultures of B. napus L. cv Jet Neuf, with the possible existence of a truncated form of BnDGAT1 known as BnDGAT2. In order to gain some insight into the topology of the enzyme, type I DGAT from B. napus was investigated using two approaches: (1) in vitro translation in the presence of microsomes and (2) immunochemical analyses of microsomes isolated from cell suspension cultures, both in combination with proteolytic mapping. Difficulties were encountered with the in vitro translation approach, possibly due to proper incorporation of the polypeptide into microsomal vesicles. Two cytocolic regions were identified in BnDGAT1, and one cytosolic region in putative BnDGAT2, using the immunochemical approach, thus providing some insight into the topology of B. napus DGAT-I. The results here support and nine and eight membrane-spanning topology for BnDGAT1 and BnGAT2, respectively. / xvii, 194 leaves ; 29 cm.

Cytoplasmic male sterility (CMS) is a widespread, maternally inherited trait that results in an inability of plants to produce functional pollen. The Ogura CMS system originated in radish, but has since been transferred to, and confers male sterility on, plants in the related genus Brassica . A gene which restores male fertility is needed for the Ogura CMS system to be exploited commercially for hybrid seed production in oilseed species such as Brassica napus. The restorer gene Rfo is a dominant radish nuclear gene that restores the male fertility to plants with Ogura cytoplasm. This gene has been transferred into Brassica napus through intergeneric crosses; however the introgressed segment of radish DNA contains an unknown number of genes, some of which confer undesirable traits, such as an elevated content of seed glucosinolates, antinutritive compounds that render the seed meal unusable as animal feed. A fine scale linkage map of the region in radish containing Rfo was constructed, and a map-based cloning approach relying on synteny between radish and Arabidopsis was used to clone Rfo. A radish gene encoding a 687 amino acid protein with a predicted mitochondrial targeting presequence was found to confer male fertility upon transformation into Ogura CMS B. napus . This gene, codes for a pentatricopeptide repeat (PPR)-containing protein with multiple, in this case 16, PPR domains. Two similar genes that do not appear to function as Rfo flank this gene. A transcript representing a non-functional allele (rfo) was detected in sterile radish plants. Comparison of the Rfo region with the syntenic Arabidopsis region indicates that a PPR gene is not present at the Rfo-equivalent site in Arabidopsis , although a smaller and related PPR gene is found about 40 kb from this site.

In this study, the ability of various chemicals to be biotransformed into mutagens by plant cells was investigated. Two thiocarbamate herbicides, diallate and triallate, the sulfonylurea herbicide chlorsulfuron, and the aniline derivative m-phenylenediamine were tested for their ability to revert Salmonella typhimurium (strains TA100 and TA98) in the presence and absence of Nicotiana tabacum (TX1) cell cultures in liquid suspension. Chlorsulfuron and m-phenylenediamine were also tested in the presence and absence of Brassica napus cv. 'Topas' cells. Diallate was found to be activated by TX1 cells into a mutagen that induces base-pair substitution mutations. In the presence of the TX1 plant cell line, chlorsulfuron significantly increased the number of mutations on the strain TA98 of Salmonella. Tobacco TX1 cells did not activate triallate into a mutagen. m-Phenylenediamine was activated into a mutagen by TX1 and Brassica cells as detected by Salmonella TA98. This aniline derivative, in the absence of plant cells and at concentrations higher than 20 $ mu$ Moles/plate, was also able to significantly increase the number of TA98 revertants as compared to the control plants. Finally, Brassica napus cells activated chlorsulfuron into a mutagen that induces frameshift mutations.

Cytoplasrnic male sterility (CMS) is a maternally inherited defect in the production of pollen, the male gamete of the flower. This sterility can be suppressed by nuclear Restorer of Fertility (Rf) genes that normally downregulate the expression of the CMS-associated novel mitochondrial gene. In Brassica napus, nap CMS and pol CMS are associated with related chimeric mitochondrial genes orf222 and orf224, respectively. CMS in both nap and pol is associated with a polar loss of locule development, loss of synchronous locule development and clumping of sporogenous tissue away from the tapetal cell layer, as well as secondary effects on petal and bud formation. In nap CMS, early accumulation of orf222 transcripts in the locule regions of developing anthers is associated with sterility, while the absence of orf222 transcripts from the locules is associated with fertility restoration. Accumulation of novel antisense transcripts of atp6 in a cell specific manner which matches that of sense transcripts of orf222 and atp6 in nap CMS anthers may be indicative of a post-transcriptional regulatory mechanism associated with CMS in flower buds. / Restoration of fertility in Brassica napus nap and pol CMS is associated with nuclearly encoded genes Rfn and Rfp, respectively. These restorers are very closely linked to one another, and may be allelic. Further efforts to isolate Rfp have narrowed the genomic region to approximately 105 kb of a syntenic region in Arabidopsis thaliana. Cosmid clones isolated from a library of Brassica rapa genomic DNA introgressed with Rfp have been successfully sorted into contigs through the application of the amplified fragment length polymorphism technique. The region to which Rfp is mapped is syntenic to a region of Arabidopsis DNA that is a duplication of a second location at the 23 megabase region of chromosome 1 of that genome. This region contains pentatricopeptide (PPR) motif-encoding genes that are highly related to other restorers of fertility of other species. By inference, Rfp from Brassica napus may encode PPR motifs. The PPR genes related to these previously characterized restorers of fertility are often found alongside the restorer genes existing as mini-clusters of several PPR-encoding genes. This is likely caused by selective pressure acting on PPR-encoding genes that resulted in diversification and multiplication of these genes. In addition, the PPR genes of this duplicated region are not syntenically located, whereas the non-PPR-encoding genes maintain their syntenic locations. The same is true for orthologous comparisons between Arabidopsis and other plant species. PPR genes are therefore malleable and capable of alteration in response to changing environmental pressures, such as the evolution of sterility inducing genes.

Canola, Brassics napus L. is a relatively new crop in South Africa. Insect pests have not yet been a major problem, but the notorious brassica specialist, diamondback moth (DBM), Plutella xylostella (L.) (Plutellidae) is establishing itself as a serious pest of this crop. DBM is the most important insect pest of plants from the family Brassicaceae throughout the world. It has developed resistance to all chemical pesticides used against it in the field and to toxins of the bacterium Bacillus thuringiensis. The pest status of DBM in South Africa is lower than in other countries with similar climates. However, due to indiscriminate use of pesticides, local populations of DBM are showing signs of resistance. An initial survey has indicated that in addition to DBM, canola is also attacked by aphids, Myzus persicae (Sulzer), Brevicoryne brassicae (L.), Lipaphis e1ysimi (Kaltenbach), (Aphiade), thrips, Thrips tabaci (Lindeman) and other pests mostly brassica specialists. The study was initiated to determine the composition of the community of insects found on canola, the seasonal phenology of DBM populations in canola, and the composition, relative abundance and seasonality of its parasitoids. Monitoring of the insects was carried out at weekly intervals for three years at Rietondale and Bapsfontein in Gauteng province of South Africa. Berlese funnels have been found to be useful in extracting insects from plants, and were used to indicate the presence ofDBM larvae and other insects found on canola. Adults of DBM were monitored with synthetic pheromone traps; larval and pupal populations were monitored by scouting canola plants. Samples of larvae, pupae and parasitoid cocoons were brought into the laboratory. Parasitoids that emerged were identified and their incidence recorded. Monolepta cf bifasciata (Chrysomelidae) and Listroderes costrirostris (Schoener) (Curculionidae) were the most abundant of the coleopteran pests. (DBM) and Heliothis armigera (H.) (Noctuidae) were most abundant lepidopteran pests of Canola. There was a high proportion of first and second instar larvae as indicated by the results of the Berlese funnels as compared to visual scouting in Bapsfontein. From May to August the infestation level of DBM was high, reaching the maximum of 0.25 larvae per plant in June 1996, then declined and remained low for the rest of the season in Rietondale. From September to December for all three years of the study, the population levels Of DBM were high, reaching a maximum of 9.6larvae per plant in September 1997, and remained low from January to August in Bapsfontein. The number of adult moths per trap per week ranged from 0 to 91 in Rietondale, peaking in January 1996 and September 1997. There was no correlation between infestation levels and the pheromone trap catches. In contrast to Rietondale, there was a high correlation between pheromone trap catches and subsequent larval infestations at Bapsfontein. Although DBM infestation levels were generally low, parasitism levels often reached 100% caused by a complex of parasitoids. During the period of study, the following hymenopteran parasitoids were recorded: Cotesia plutellae (Kurdjumov) and Apanteles eriophyes (Nixon), Braconidae), both larval parasitoids, Diadegma mollipla (Holmgren) (Ichneumonidae), and Oomyzus sokolowskii (Kurdjumov) (Eulophidae), larval-pupal parasitoids, Diadromus collaris (Gravenhorst) (Ichneumonidae) pupal parasitoid, and the hyperparasitoids Mesochorus sp. (Ichneumonidae) and Pteromalus sp. (Pteromalidae). Cotesia plutellae was the most abundant parasitoid occurring throughout the year..

No description available.

CAPES / Os objetivos deste estudo foram avaliar o efeito dos extratos de canola (Brassica napus L.) e mostarda-da-índia (Brassica juncea), in vitro, sobre a severidade de Oídio (Oidium sp.) em discos cotiledonares de pepineiro e a germinação de conídios do fungo e comparar esses extratos com controle químico (enxofre inorgânico e tiofanato metílico + clorotalonil), leite cru de vaca a 10 % e óleo de neem, também in vitro, e no controle do Oídio em pepineiro, em casa de vegetação. Para isso, foram realizados três experimentos, in vitro. No primeiro, testaram-se três métodos de preparo (aquoso, macerado e infusão) dos extratos de canola e mostarda-da-índia na dose de 12% sobre a severidade do Oídio em discos cotiledonares de pepineiro e a germinação de conídios do fungo. Nesse experimento, todos os modos de preparo dos extratos de canola e mostarda-da-índia reduziram mais de 35% a severidade da doença nos discos cotiledonares e a germinação dos conídios do fungo. No segundo experimento, testaram-se as doses de 3 a 12% dos extratos das brássicas, preparados por maceração, sobre as mesmas variáveis do ensaio anterior. A dose de 12% dos extratos das brássicas reduziu a severidade da doença nos discos cotiledonares de pepineiro em mais de 59% e a germinação dos conídios do fungo em mais de 52%. No terceiro experimento, testaram-se os extratos macerados a 12% dessas brássicas, o leite cru de vaca a 10%, o óleo de neem e os fungicidas, também sobre as mesmas variáveis. Nesse ensaio, os tratamentos contento os extratos de brássicas reduziram mais de 80% a severidade do Oídio nos discos cotiledonares de pepineiro, em relação ao tratamento testemunha. Porém, no teste de germinação de conídios do fungo, apenas os tratamentos contendo os fungicidas foram eficientes, reduzindo em 65% a germinação dos conídios do fungo. Posteriormente foram realizados experimentos, em casa de vegetação, para comparar a eficiência desses tratamentos no controle do Oídio em pepineiro, em dois anos de cultivo. Os extratos das brássicas reduziram a severidade e a taxa de progresso da doença nos dois anos de cultivo. O extrato de mostarda-da-índia, em relação aos tratamentos testemunha seca e com água, reduziu a área abaixo da curva de progresso da doença (AACPD) da severidade de Oídio, em média, 68,7 e 57% e o extrato de canola reduziu, em média, 67,1 e 59,4%, nos anos 1 e 2, respectivamente. Quando os extratos de canola e mostarda-da-índia foram comparados aos tratamentos testemunha seca e com água, constatou-se que a redução da severidade final da doença foi superior a 70 e 65%, nos anos 1 e 2, respectivamente. Os extratos de canola e mostarda-da-índia apresentaram maior controle da doença, comparado ao obtido no tratamento contendo leite cru de vaca a 10%, e próximo ao controle do tratamento com óleo de neem. Os fungicidas apresentaram os melhores controles da severidade e da taxa de progresso da doença, com redução superior a 99% da severidade final da doença e da AACPD da severidade de Oídio. Conclui-se, portanto, que os extratos das canola e mostardada-índia têm potencial de controle de Oídio em pepineiro, em casa de vegetação. / The objectives of this study were to evaluate the effect of canola (Brassica napus L.) and mustard (Brassica juncea) extracts, in vitro, on the severity of powdery mildew (Oidium sp.) in cucumber cotyledon discs and on the germination of fungal conidia, and to compare these extracts with chemical controls (inorganic sulphur and methyl thiophanate + chlorothalonil), 10% cows milk and neem oil, in vitro and also in the control of powdery midew in greenhouse cucumbers. To achieve this, three experiments were carried out in vitro. In the first, three methods of preparation (aqueous, macerated and infusion) of the extracts of canola and mustard were evaluated for the dose of 12% on the severity of powdery mildew in cotyledon discs and the germination of fungal conidia. In this experiment, all modes of preparation of extracts of canola and mustard reduced the severity of the disease in the cotyledon discs and the germination of the fungal conidia by more than 35%. In the second experiment, doses from 3 to 12% of the brassica extracts, prepared by maceration, were tested on the same variables of the previous experiment. The 12% dose of the brassica extracts reduced the severity of the disease in cucumber cotyledon discs by more than 59% and the germination of fungal conidia by more than 52%. In the third experiment, the 12% dose of the macerated extracts of the two brassicas were tested, alongside 10% raw cow's milk, neem oil and fungicides, using the same variables as previously. In this trial, the treatments containing the brassica extracts reduced powdery mildew severity by more than 80% in the cucumber cotyledon discs, in relation to the control treatment. However, in the fungal conidia germination test, only the treatments containing the fungicides were efficient, reducing the germination of the fungal conidia by 65%. After this, experiments were carried out in a greenhouse to compare the efficiency of these treatments in the control of powdery mildew in cucumber, over a two year period of cultivation. The extracts of the brassicas reduced the severity and rate of disease progress over the two year period. The mustard extract, in relation to the dry and water control treatments, reduced the area under the disease progress curve (AUDPC) for powdery mildew severity, it by a mean of 68.7 and 57%, and the canola extract reduced it by a mean of 67.1 and 59.4%, in years 1 and 2, respectively. When the extracts of canola and mustard were compared to the dry and water control treatments, it was found that the reduction of the final severity of the disease was superior to 70 and 65%, in years 1 and 2, respectively. The extracts of canola and mustard showed greater control of the disease, compared with that obtained in the treatment containing raw cow's milk at 10%, and was close to the control of the treatment with neem oil. The fungicides presented the best control of disease severity and disease progression rate, with a reduction of final disease severity and in the AUDPC of the mildew severity by more than 99%. It is concluded, therefore, that the extracts of canola and mustard have the potential to control powdery mildew under greenhouse conditions.

Les plantes possèdent de multiples systèmes de défense contre les insectes phytophages. Dans un contexte agronomique, manipuler ces défenses au moyen de la sélection pourrait contribuer à réduire les dommages causés par les insectes ravageurs en augmentant la résistance naturelle des plantes. Cette stratégie se heurte cependant à des contraintes très fortes lorsqu'il s'agit de l'appliquer aux insectes. Après avoir détaillé ces contraintes, nous proposons une démarche alternative aux méthodes classiques. Celle-ci vise à identifier des traits-clés de la plante qui modulent son interaction avec le ravageur. Si de tels traits sont identifiés et validés expérimentalement, ils permettront ensuite de conduire la sélection sans nécessiter d'insecte. Nous avons testé cette démarche dans un système composé du colza (Brassica napus) et du méligèthe Meligethes aeneus, un ravageur majeur de cette culture. Le méligèthe est un coléoptère pollinivore dont les adultes sont généralistes mais ne pondent que sur certaines brassicacées. Les dégâts agronomiques sont causés par ces adultes qui, avant que la floraison ne démarre, détruisent les boutons floraux pour atteindre le pollen qu'ils contiennent. Quatre étapes cruciales de l'interaction ont été étudiées : l'attraction à distance, l'alimentation des adultes, la production et la ponte des œufs, et le développement larvaire. Six génotypes de colza ont été comparés dans une série d'expérimentations au laboratoire. La mise en relation des résultats de préférence/performance de l'insecte avec des profilages métaboliques larges de tissus floraux a permis d'identifier des traits-clés candidats. Les conclusions principales de ce travail sont (i) que la composition biochimique du périanthe est déterminante dans la stimulation de l'alimentation des adultes, et que cette stimulation pourrait être largement sous l'influence d'un petit nombre de composés dont le saccharose ; (ii) que cette stimulation détermine de façon majeure, par un effet domino, la production d'œufs en contraignant l'ovogenèse ; (iii) que la qualité nutritionnelle du pollen impacte à la fois les larves et les adultes, et que cette qualité pourrait être déterminée en bonne partie par la concentration en amidon et en certains glucosinolates (des métabolites secondaires typiques de quelques familles végétales dont les brassicacées). La combinaison des différents résultats obtenus permet également de proposer des hypothèses plus générales, parmi lesquelles le fait que le contexte agronomique dans lequel l'interaction a lieu ait largement influencé, voire perturbé, l'interaction qui liait le méligèthe et les brassicacées sauvages avant que les cultures de colza ne se généralisent. Cette thèse a montré qu'une nouvelle voie était peut-être envisageable pour contribuer à protéger les cultures de façon durable contre les insectes ravageurs, en particulier pour les systèmes agronomiques où les dégâts sont causés à un stade temporairement sensible de la culture. / Plants display multiple defense systems against phytophagous insects. Manipulating these defenses by means of selection could contribute to decrease damages caused by insect pests, by increasing natural resistance of crops. This strategy faces great constraints when applied to insects. We first detail these constraints and then propose an alternative approach to classical methods. It consists in identifying key plant traits that determine the intensity of the interaction between the plant and the pest. If such traits are identified, selection could be conducted on the basis if these sole traits, without needing any insect. We tested this approach in a system composed of oilseed rape (Brassica napus) and the pollen beetle (Meligethes aeneus), a major pest of this culture. This coleopteran generalist pollen feeder lays eggs only on certain brassicaceous plants. Agronomical damage arise before plant flowering and are caused by adults, which destroy flower buds to get the pollen they contain. Four crucial steps of the interaction were studied: attraction, adult feeding, egg production and oviposition, and larval development. Six oilseed rape genotypes were compared in a series of experiments conducted in the laboratory. By linking insect preference/performance to large metabolic profiling of bud tissues, we identified candidate key traits. Main conclusions of this work are (i) that biochemical composition of the perianth, especially a few compounds among which sucrose, is determinant for feeding intensity; (ii) that feeding stimulation has an important impact on egg production by constraining oogenesis; (iii) that pollen nutritional quality, probably mostly determined by starch and some glucosinolates, interacts with both pollen beetle larvae and adults. Combination of several results also allows drawing more general hypotheses about the oilseed rape - pollen beetle interaction. One of these is that the agronomical context in which the interaction takes place may have largely influenced, or even disturbed, the interaction that linked this insect and wild brassicaceous plants before oilseed rape cultivation. This thesis showed that a new way might be possible to protect cultures against insect pests. It could be both efficient and sustainable, especially in systems where agronomical damage is caused at a temporary vulnerable plant stage.