ADP Ribosylation Factor 1 (ARF1) is a universal small GTP binding protein which has an important role in vesicular trafficking between endoplasmic reticulum and Golgi. ARF1 is a basic component of Coat Protein I (COPI) vesicles which have functions in both formation of coatomer complex and recruitment of cargo proteins. In this study, the expression ARF1 was analyzed in pea (P. sativum L. cv. Araka) grown at different developmental stages. Because of the differential hormonal levels at corresponding stages, the effects of hormones on ARF1 expression were also studied. The results of present research show that ARF1 expression in embryos and 2 days grown plants after germination is lower when compared to 6 days grown plants. In order to see the hormonal effect, 3 weeks old plants were supplied with 50&micro / M of each hormone for 3 times on alternate days. Protein extraction, cell fractionation,Western blot was carried out and immunoblot analysis was conducted with AtARF1 polyclonal antibodies. It was shown that, in pea shoots, abscisic acid and gibberellin increases the inactive GDP bound ARF1 by hydrolyzing ARF-GTP through activating ARFGTPase activating protein (ARF-GAP) or partially inhibiting ARF-Guanine Nucleotide Exchange Factor (ARF-GEF). In roots, ARF-GDP (cytosolic fraction), ARF-GTP (microsomal fraction) and total amount of ARF1 (13.000 x g supernatant fraction) were down regulated by ~11, ~19 and ~11 fold respectively with the application of gibberellin / and by ~11, ~7 and ~3 fold respectively with the application of abscisic acid / when compared to control plants. These results indicate the importance of plant hormones in the regulation of ARF1 in pea.

The arbuscular mycorrhizal (AM) association results from a successful interaction between the genomes of the two symbiotic partners. In this context, the aim of my research was to better characterize the role of the late stage symbiosis-related pea genes PsSym36, PsSym33 and PsSym40 in the functional AM (i) by investigating the effect of mutations in the three genes on fungal and plant gene responses and (ii) by creating conditions for the localization of two of the genes, PsSym36 and PsSym40, on the pea genetic map for future map-based cloning. The expression of a subset of ten fungal and eight plant genes,previously reported to be activated during mycorrhiza development, was compared in Glomus intraradices-inoculated roots of wild type and Pssym36, Pssym33 and Pssym40 mutant pea plants. Most of the fungal genes were down-regulated in roots of the Pssym36 mutant where arbuscule formation is defective, and several were upregulated with more rapid fungal development in roots of the Pssym40 mutant. Microdissection of mycorrhizal PsSym40 roots corroborated preferential expression of the three G. intraradices genes SOD, DESAT and PEPISOM in arbuscule-containing cells. Inactivation of PsSym36 also resulted in down regulation of plant genes whilst mutation of the PsSym33 and PsSym40 genes affected plant gene responses in a more time-dependent way. Results thus indicate an implication of the investigated pea SYM genes in the modulation of plant and fungal molecular interactions linked to signaling, nutrient exchange or stress response regulation during AM symbiosis formation and functioning. Conditions for localization of the PsSym36 and PsSym40 genes on the pea genetic map were developed for their future map-based cloning. Based on the molecular markers obtained, it was possible to conclude that localization of the PsSym40 gene most likely resides outside the linkage groups I, II, III or V of the genetic map of pea.

La majorité des plantes terrestres forment une symbiose racinaire avec des champignons mycorhiziens à arbuscules (MA). Les champignons fournissent à la plante de l'eau et des minéraux en particulier du phosphate (Pi), en échange les plantes leur apportent des éléments carbonés. La fertilisation phosphatée est connue pour inhiber l'interaction symbiotique, mais les mécanismes intervenant dans cette régulation sont inconnus. Nous avons montré que le Pi est capable d'inhiber presque totalement la mycorhization à un stade très précoce, avant même l'attachement du champignon à la surface de l'épiderme racinaire. Cette inhibition est liée aux teneurs en Pi dans les parties aériennes et fait intervenir une signalisation systémique. Par la suite, nous avons cherché à identifier les mécanismes impliqués dans la régulation de la mycorhization par le Pi. Les évènements précoces d'interaction ont été examinés avec un intérêt marqué pour les exsudats racinaires. Tout d'abord, nous avons analysé l'importance des strigolactones dans la régulation. Ces molécules sont sécrétées par les racines des plantes et stimulent le développement des champignons (MA). La production de strigolactones est elle aussi régulée de façon systémique par le Pi et les exsudats végétaux de plantes non carencés en Pi sont dépourvus de strigolactones. Cependant, un ajout exogène de ces molécules ne suffit pas pour lever l'inhibition associée à la présence de Pi. De plus, la part des exsudats végétaux en général dans cette régulation a été étudiée. Bien que les exsudats jouent un rôle pour favoriser (ou non) la mise en place de l'interaction, des mécanismes de contrôle supplémentaires existent au niveau de la racine elle-même. Parmi plusieurs mécanismes régulateurs hypothétiques nous avons testé si le Pi pouvait affecter la capacité des plantes à reconnaitre correctement leurs partenaires fongiques. Pour cela nous avons utilisé deux approches. (i) Il est connu que les racines répondent à la présence de champignon par des oscillations de concentrations en calcium dans les noyaux. Nous n'avons pas trouvé d'effet du Pi sur cette réponse. (ii) Nous avons aussi analysé si l'expression de gènes végétaux en réponse au champignon pouvait être régulée par le Pi. Quelle que soit la condition phosphatée testée, les plantes sont capables de répondre à la présence de champignon par l'induction de gènes de défense et de gènes " symbiotiques ". Cependant, le Pi affecte négativement l'expression de certains des gènes symbiotiques, laissant penser que les plantes seraient moins à même de répondre au champignon lorsqu'il y a du Pi. Pour finir, d'autres mécanismes de régulation possibles (tels que la composition des parois ou un effet hormonal des strigolactones) sont proposés et discutés.

Diss. (sammanfattning) Umeå : Sveriges lantbruksuniv., 2007. / Härtill 3 uppsatser.

Diss. (sammanfattning) Uppsala : Sveriges lantbruksuniv., 2007. / Härtill 5 uppsatser.

The aim of the diploma thesis was to assess changes in pattern of legume storage proteins during germination. Four species of legumes were chosen for analyses ? Glycine max L., Lupinus angustifolius L., Pisum sativum L. and Vicia faba L. Seeds for analyses were sampled at the beginning, middle and end of germination. Proteins were extracted from lyophilised and homogenised material. These proteins were analysed by SDS-PAGE electrophoresis. The results proved that during seed germination the seed storage proteins cleave into smaller peptides, which forms new proteins. The intensity of protein bands in pea seeds was decreased in the area at around 48-45 kda and 40-36 kDa and the intensity of the proteins bands was increased at around the protein bands 25-23 kDa and 19-7 kDa. In lupine were not detected the protein bands over 39 kDa and during germination amount of protein bands in areas 15-7 kDa was increased

Made available in DSpace on 2014-06-11T19:29:43Z (GMT). No. of bitstreams: 0 Previous issue date: 2007-12-07Bitstream added on 2014-06-13T18:39:37Z : No. of bitstreams: 1 nogueira_dc_me_ilha.pdf: 439254 bytes, checksum: 81b60ae48567a86e242d01e1de2c9659 (MD5) / Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) / A região de cerrado tem como principais fatores limitantes da produção agrícola as condições climáticas e solos com baixa fertilidade natural. Neste sentido, são importantes estudos para entender os fatores que contribuem para o desempenho das culturas em condições de cerrado, como por exemplo a ervilha. O objetivo foi avaliar o efeito da aplicação foliar do boro, molibdênio e zinco na qualidade de grãos de ervilha e produtividade. A ervilha utilizada foi a variedade Utrillo, cujo cultivo foi realizado na área experimental da Fazenda de Ensino Pesquisa e Extensão-FE-UNESP-Campus de Ilha Solteira. Os tratamentos constaram das combinações entre aplicação de boro, molibdênio e zinco, (Testemunha, B, Mo, Zn, B+Mo, B+Zn, Mo+Zn, B+Mo+Zn). O Mo foi aplicado via foliar aos 20 dias após a emergência das plantas (DAE). O B e o Zn foram aplicados também via foliar aos 30 DAE. O delineamento experimental foi em blocos casualizados com quatro repetições, as médias comparadas pelo teste de Duncan a 5% de significância. Analisou-se produtividade e o conteúdo de nutrientes, de amido, de aminoácidos, de proteinas nos grãos e sementes e a qualidade fisiológica das sementes. Os resultados indicaram que os tratamentos alteram significativamente apenas o conteúdo de magnésio e de cobre nos grãos. Os demais fatores estudados não se alteraram devido ao tratamento imposto à cultura. / Some times the crops yield in the cerrado area is limited by weather conditions and the soil with a low fertility. The studies in this direction are important to understand the factors that contribute for the performance of the cultures in the cerrado conditions, for example pea’s crop. The objective was to evaluate the effect of the foliar application of boron, molybdenum and zinc on the quality of pea grains and grain yield. For experiment were used seeds of pea, cv. Utrillo, cultivated in the experimental area of the Farm of the Teach Research and Extension-FE-UNESP- Campus de Ilha Solteira”. The eight treatments consisted of the combinations among application of boron, molybdenum and zinc, (Control, B, Mo, Zn, B+Mo, B+Zn, Mo+Zn, B+Mo+Zn). 20 days after emergency (DAE) was applied Mo by foliar. B and Zn were applied by foliar at 30 DAE. The experimental delineation used was the randomized blocks with four repetitions; the averages had been compared by Duncan test at 5% of significance. It was analyzed the grain nutrient content, grain yield, starch, aminoacids and proteins content in the grains and seeds and physiological seed quality. The results had indicated that the treatments modify significantly only to the copper and magnesium content in the grains. The other studied factors they didn't alter due to the treatment imposed to the culture.

No description available.

La fixation symbiotique d’N par les légumineuses est très sensible aux ravageurs, provoquant des dommages sur les racines nodulées, avec un impact sur la fixation d’N et la croissance qui demeure mal connu. Nous avons alors analysé la réponse adaptative de la fixation symbiotique et de la croissance du pois Frisson sauvage et 3 de ses mutants hypernodulants P64, P118 et P121, respectivement mutés sur les gènes SYM28, SYM29 et NOD3, à une ablation de la moitié du système racinaire, en fin de phase végétative. La réponse adaptative a été mesurée 8 jours après ablation, dans des conditions d'alimentation en carbone par la photosynthèse variées. A 380 ppm, le mutant P118 a montré la plus faible diminution de l’activité spécifique de fixation (-17%) suite à l’ablation comparé au sauvage et aux 2 autres mutants (-36% à -62%) associé à une accélération chez les mutants P118 et P121 et un maintien (sauvage et P64) de la croissance des nodosités. A 150 ppm, suite à l’ablation, l’activité spécifique de fixation symbiotique par les nodosités a été diminuée (sauvage), maintenue (P64 et P118) ou augmentée (P121), associée à une accélération (sauvage et P121) ou un maintien (P64 et P118) de la croissance des nodosités. A 750 ppm, l’activité spécifique de fixation a diminué suite à l’ablation pour tous les génotypes, associée à un ralentissement (P64), un maintien (P118, sauvage) ou une faible accélération (P121) de la croissance des nodosités. Les résultats montrent une plus grande capacité de la fixation symbiotique des mutants hypernodulants (P118 et P121 essentiellement) à résister au stress provoqué par l’ablation. / Symbiotic N fixation of legumes is very sensitive to environmental stresses, like pea pests damaging nodulated roots. However, the impact on their N uptake capacity and plant growth has not been studied so far.We analyzed the adaptive response symbiotic N2 fixation and plant growth of pea wild type Frisson and hypernodulating mutants P64, P118 and P121 mutated respectively on genes SYM28, SYM29 and NOD3 to root pruning of half the root system at the end of the vegetative stage. The adaptive responses of pea: cv. Frisson and 3 of its hypernodulating mutants were compared under varying carbon supplies from photosynthesis.At 380 ppm, mutant P118 showed the lowest decrease of the specific activity of N fixation (-17%) following root pruning compared to the wild type and the 2 others mutants (-36% to -62%), associated to an acceleration (P118 and P121) and a maintained (wild type and P64) nodule growth. At 150 ppm, following root pruning, specific activity of N fixation of nodules decreased in wild type, was maintained in P64 and P118 and increased in P121. At 750 ppm, specific activity of N fixation of nodules decreased for all genotypes following root pruning, associated to a maintained nodule growth in wild type and P118, a slower growth in P64 and acceleration in P121.Our results showed a greater capacity of hypernodulating mutants P118 and P121 to withstand the stress induced by root pruning of half the root system.

Herbicides have become a key component in modern agricultural production. Meanwhile, there is a concern that some herbicides persist past the growing season of the treated crop, and negatively influence the production of the subsequently planted crops. Amongst various herbicides used in western Canada, acetohydroxyacid synthase (AHAS)-inhibiting herbicides warrant special attention given their residual properties and acute plant toxicity at low concentrations in soil. Soil residual AHAS inhibitors have the potential to influence both leguminous host plants and their bacterial symbiotic partners; consequently, the use of an AHAS inhibitor in a given year can negatively influence the inoculation success and grain yield of legumes cropped in the following year. <p>The present thesis project focused on one of the AHAS inhibiting herbicides (flucarbazone) and studied its potential for carryover injury and negative influence on the success of inoculation in field pea. A series of growth chamber and field experiments were conducted to test the following null hypothesis: the presence of residual flucarbazone in soil does not affect nodulation of field pea by inoculum rhizobia. <p>A growth chamber experiment clearly demonstrated the susceptibility of field pea to the presence of flucarbazone in soil where the lowest concentration of flucarbazone amendment (5 ìg kg1) significantly reduced the crop growth. In contrast, a field study failed to reveal any negative effects of flucarbazone use on crop growth and N2 fixation. <p>It was concluded that if the weather and soil conditions favour decomposition of flucarbazone as described in the present study, flucarbazone applied at the recommended field rate will not persist into the following season at high enough concentrations to negatively influence field pea growth, grain yields, and inoculation success. To ensure safety of rotational crops, it is important to strictly adhere to the herbicide application guidelines. Additionally, producers are cautioned to be particularly aware of the environmental and soil conditions that may reduce the rate of herbicide degradation.