No description available.

Submitted by Mario BC (mario@bc.ufrpe.br) on 2016-09-19T13:21:55Z No. of bitstreams: 1 Aline Medeiros de Arruda.pdf: 1077427 bytes, checksum: edf33aaa98a044c16135de656116b3d2 (MD5) / Made available in DSpace on 2016-09-19T13:21:55Z (GMT). No. of bitstreams: 1 Aline Medeiros de Arruda.pdf: 1077427 bytes, checksum: edf33aaa98a044c16135de656116b3d2 (MD5) Previous issue date: 2016-02-18 / Due to it adaptability and nutritive value, cowpea (Vigna unguiculata) is an important protein source in some regions of Brazil, with good perspectives in the Brazilian agribusiness. This pulse, in symbiosis with nitrogen fixing bacteria, has a high nitrogen fixation potential, and may dispense completely or partially nitrogen fertilizers, reducing production costs. The evaluation of native populations of these bacteria in cowpea cultivars allows the selection of efficient strains, adapted to local conditions, and leading to higher cowpea yields. This work aims to evaluate the symbiotic efficiency of rhizobial populations from Pernambuco State semiarid soils as a more efficient method for the selection of rhizobial strains for inoculant production, using three cultivars of cowpea (BRS Marataoã, BRS Guariba and IPA 206). Soil samples were collected from the 0-0.1 m depth layer in different Pernambuco municipalities from two groups of soil, one high and one low fertility. An experiment was done to collect nodules for isolation, and isolates were morphologically characterized in YMA media. Shannon-Weaver’s diversity, Simpson’s dominance, Margalef’s richness and Pielou’s uniformity indexes were calculated for each soil group. Rhizobial populations were evaluated in Leonard jars in a greenhouse. Shoot (MSPA), root (MSR) and nodules (MSN) dry masses, nodule number (NN), shoot nitrogen accumulation (ANPA) and relative efficiency (ER) were determined and submitted to analysis of variance and means comparison by the Scott-Knott test at 5% probability. Most of the more efficient isolates were from the low fertility soil, in special isolate 49. The lower diversity from the high fertility soils may be due to a reduced dependence of the legumes on biological nitrogen fixation, while the higher dependency of lower fertility soils may have applied a selective pressure towards more efficient strains. BRS Guariba had higher ER, showing good symbiotic specifity with the strains. / Devido às condições de adaptabilidade e valor nutritivo, o feijão-caupi (Vigna unguiculata) tornou-se importante fonte de proteína em algumas regiões do país, permitindo ampla perspectiva no agronegócio brasileiro. Esta leguminosa granífera possui alta capacidade de fixar N em simbiose com bactérias fixadoras de nitrogênio, da qual podem dispensar total ou parcialmente os fertilizantes nitrogenados, o que reduz os custos de produção, além de servir como alicerce ao programa de melhoramento vegetal. A avaliação de populações nativas destas bactérias em cultivares de feijão-caupi permite a seleção de estirpes eficientes, adaptados às condições locais, resultando no aumento de produtividade do feijão-caupi. Este trabalho tem como objetivo avaliar a eficiência simbiótica de populações rizobianas, provenientes de solos do semiárido pernambucano (Brazil), como método mais eficiente para seleção de estirpes rizobianas para recomendação para produção de inoculantes em três variedades de feijão-caupi: BRS Marataoã, BRS Guariba e IPA 206. Amostras de solos foram coletadas na camada de 0-0,1m de profundidade em diferentes municípios pernambucanos, provenientes de dois grupos de solos, um com alta fertilidade e outro de baixa fertilidade. Um experimento foi conduzido para a obtenção de nódulos para isolamento e os isolados foram caracterizados morfologicamente em meio YMA. Os índices de diversidade de Shannon-Weaver, dominância de Simpson, riqueza de Margalef e uniformidade de Pielou, foram calculados para cada grupo de solo. As populações de rizóbios foram avaliadas em vasos de Leonard em casa de vegetação utilizando o delineamento experimental de blocos casualizados com quatro repetições. As necessidades nutricionais do feijão-caupi foram supridas com solução nutritiva. Foram observadas e avaliadas as seguintes variáveis: Massa seca da parte aérea (MSPA) e raiz (MSR), número (NN) e massa de nódulos secos (MSN), acúmulo de nitrogênio na parte aérea (ANPA) e eficiência relativa (ER). Estas variáveis foram submetidas à análise de variância com as médias comparadas pelo teste de Skott Knott a 5% de probabilidade, utilizando-se o programa estatístico SISVAR 5.3. A maioria dos isolados mais eficientes foram oriundos do grupo de solo de baixa fertilidade, com destaque para o isolado 49. A menor diversidade rizobiana no grupo de solo de alta fertilidade pode ter sido devida a uma menor dependência destas plantas da fixação biológica de nitrogênio enquanto os solos menos férteis podem ter efetuado uma pressão seletiva, permanecendo os isolados mais eficientes. A cultivar BRS Guariba apresentou maior desempenho na eficiência relativa, demonstrando uma boa especificidade simbiótica entre as estirpes.

MSCAGR (Plant Production) / Department of Plant Production / Chickpea (Cicer arietinum L) is adapted to cool-seasons and its organs are of high nutritive value and serve as cheap sources of protein, especially in developing countries. Chickpea crop is mainly grown for human consumption, animal feed and for medicinal purposes. The introduction and promotion of chickpea to especially small-scale South African crop farmers has multiple objectives including the improvement of soil fertility. Small-scale farmer’s flounder to afford Nfertilizers, coupled with the challenges faced by programmes aimed at assisting them about soil fertility in their cropping fields that are still without enough N concentration to meet N demand. It is therefore important that other alternatives that can help improve the N status of soils be explored. The shoot δ13C is an indicator of WUE in C3 plants. However, shoot-WUE is affected by a variety of factors including genotypes, phosphorus fertilizer application and availability of native or introduced rhizobial bacteria. However, not much is known on whether application of phosphate fertilizer, seed inoculation with rhizobial strain affect the shoot C/N ratio of chickpea genotypes in South Africa. Therefore, field experiments were established at Thohoyandou and Syferkuil in Limpopo to assess the role of phosphorus fertilization and rhizobial inoculation on C assimilation, C/N ratio and shoot-WUE of chickpea genotypes. Field experiments were conducted during winter season in 2016 and 2017 (April to August). Treatments consisted of a factorial combination of two rates of phosphorus fertilizer (0 and 90 kg P ha-1 ), four desi chickpea genotypes (ACC#1, ACC#2, ACC#3 and ACC#5) and two rhizobial inoculation levels (bradyrhizobium strain and without rhizobial strain. In Thohoyandou, ACC#1 showed greater grain yield in 2016 and 2017. Which was associated with more branches and greater plant height. Furthermore, the interaction between genotypes, phosphorus fertilizer and rhizobial inoculation had significant effect on grain yield in 2016. ACC#1, 3 and 5 of chickpea genotypes fixed the most N compared to that of ACC#2. In addition, ACC#5 had the highest soil N-uptake in both seasons followed by ACC#3, while ACC#1 had the least value of soil N-uptake in both seasons. Phosphorus fertilizer application increased the fixation of N by 36.8% (P≤0.01), and similarly in soil N-uptake by difference of 59.9% compared to control in 2016. Furthermore, rhizobial inoculation increased N-fixed in 2016 and soil N-uptake in both seasons. ACC#5 had the highest N fixed at phosphorus-fertilized with bradyrhizobuim across two locations in both seasons. ACC#5 depended more on soil N-uptake than fixing its own N as compared to ACC#1. N fixation differed across seasons; however, ACC#3 had greater N-fixed in both locations. Moreover, chickpea genotype that fixed more N had least δ15N. This finding indicates that N fixation is exhibited by the genotypes that depend less on δ15N, because N2 fixation is inhibited by high soil N concentration or δ15N. Furthermore, ACC#2 and ACC#3 had greater δ13C at Thohoyandou in 2017; chickpea genotypes had significant effect on δ13C at P≤0.05 at Thohoyandou, 2016. The results showed that ACC#1 with phosphorus fertilizer application and no bradyrhizobium strain showed greater δ13C. Also, δ13C increased with a decrease in N-fixed (r=.1000), this indicates that there was a functional relationship between plant WUE and N fixation in chickpea, probably because improved water use in legumes supports N fixation. / NRF

MSCAGR / Department of Plant Production / Chickpea (Cicer aritienum L.) is an ancient crop that originated in South-Eastern Turkey and belongs to the genus Cicer, tribe Cicereae, and family Fabaceae. Chickpea has the ability to fix atmospheric nitrogen (N) for its growth. However, chickpea productivity not only depends on N2 fixation or dry matter accumulation, but also the effectiveness of nutrient partitioning to seed, a key component to overall yield. There is a dearth of information on the effect of P with rhizobial inoculation in response of nutrients accumulation in the rhizosphere, shoots and grain of chickpea, especially when determined at different growth stages in the African continent. This study contributes knowledge on this crucial aspect which will likely lead to more other similar research reports in other settings. Therefore, the objectives of this study was to evaluate the effect of P fertilizer rates and rhizobial inoculation on yield and nutrients accumulation in two chickpea genotypes. Field experiments were conducted in winter 2017 and 2018 at University of Venda, Thohoyandou and University of Limpopo’s experiment farm, Syferkuil. Treatments consisted of a factorial combination of two rates of P fertilizer (0 and 90 kg P ha-1), two desi chickpea genotypes (ACC1 and ACC5) and two rhizobial inoculation levels (with and without rhizobiam strain). The treatments were laid out in a randomized complete block design (RCBD) and replicated three times on 22 April 2017 and 11 April 2018 (Syferkuil), 13 April 2017 and 29 April 2018 (Thohoyandou). Macronutrients including P, K, Ca, Mg were determined using the citric acid method. The total N concentration were determined by the micro-Kjeldahl method in both soil, shoots and grain. Zn was extracted using a di-ammonium ethylenediaminetetraacetic acid (EDTA) solution. The content of macronutrients (P, K, Ca, Mg, Ca, and Zn) in soil, shoots and grain was determined by first subjected to wet digestion (Mehlich, 1984). From the digest, various elements were read using relevant procedures. P contents was determined colorimetrically using a spectrophotometer. Yield and yield components were assessed at harvest maturity. Genotypes affected the accumulation of mineral elements in rhizosphere soil, shoots, grain and yield. Accession 5 performed better in most of nutrients elements compared to accession 1 in both seasons and sites. Application of phosphorus alone, and in combination with rhizobium inoculation increased the concentration of majority of nutrients in the rhizosphere. When the test accessions were grown at the Syferkuil and Thohoyandou study location in 2017, they showed significant differences in the concentration of N, P and K while Ca, Mg and Zn were similar in the rhizosphere. The concentrations of N, P and K were markedly higher in the rhizosphere of ACC5 compared to ACC1. In fact, the concentration of P was two-fold greater in the rhizosphere of ACC5 than ACC1. Accession 5 exhibited a markedly higher shoot dry weight, number and dry weight of pods, 100-seed weight, grain yield and harvest index compared to ACC1. P plus rhizobium inoculation, P, rhizobium inoculation affected grain yield and yield components of chickpea genotypes. This preliminary finding show that the combination of P and rhizobium inoculation affected the nutrients accumulation in the rhizosphere, shoots, grain, yield and yield components in both locations. Moreover, Thohoyandou had the highest nutrients accumulation on the rhizosphere, shoots, grain, yield and yield components compared to Syferkuil. / NRF

A greenhouse experiment was conducted to determine the interactive effects of water salinity, volume of water applied at each irrigation, sodium adsorption ratio and rhizobial strain upon 15 plant growth parameters of Phaseolus vulgaris L. Four levels of water salinity (ECW 0.4, 3.0, 6.0, or 9.0 mmho/cm), three levels of water volume (200, 400, or 800 ml per pot), three levels of SAR (4, 16, or 64) and three strains of Rhizobium phaseoli (K-17, K-47, or K-52) were used. The treatments were replicated with all possible combinations. Data were subjected to analysis of variance, multiple comparisons, and path coefficient analysis.

Les légumineuses sont capables de s’associer en symbiose avec des bactéries du sol Rhizobium. Cette interaction culmine par la formation d’un nouvel organe racinaire appelé nodule, à l’intérieur duquel les bactéries différentiées fixent l’azote atmosphérique sous une forme assimilable par la plante hôte. La mise en place de cette association repose sur un dialogue moléculaire entre les deux partenaires, faisant intervenir des signaux bactériens lipo-chitooligosaccharidiques appelés Facteurs Nod (FNods). Chez la légumineuse modèle Medicago truncatula, la perception de ces signaux symbiotiques au niveau de l’épiderme racinaire, initie une voie de signalisation qui conduit à des oscillations calciques nécessaires pour l`activation de gènes de la plante hôte, tel le gène marqueur ENOD11. Il a été montré que les facteurs de transcription ERF ERN1/ERN2, étroitement apparentés, agissent comme des activateurs directs de la transcription d’ENOD11, via leur liaison à la séquence cis régulatrice NFbox. Le mutant ern1 est de manière cohérente requis pour l’activation d’ENOD11 en réponse aux FNods mais également au cours des étapes suivantes d’infection et de développement nodulaire. Cependant, ce mutant présente un phénotype symbiotique partiel soulevant la question d’une redondance fonctionnelle, qui pourrait être attribuée à la présence du facteur ERN2, étroitement apparenté. Ainsi, au cours de ma thèse, j’ai étudié la relation fonctionnelle entre les facteurs ERN1/ERN2 par de approches diverses visant à déterminer leur expression et fonctions relatives au cours de la symbiose rhizobienne. Mon travail de thèse a dans tout d’abord porté sur l’étude des profils d’expression spatio-temporels de gènes ERN au cours de la symbiose rhizobienne, corroborée par la dynamique de localisation cellulaire des protéines de fusions ERN. Ces facteurs sont exprimés de manière séquentielle mais aussi conjointe aux cours de la signalisation FNods et l’infection rhizobienne. Par la suite, des expériences de complémentation croisée, dans le fond mutant ern1, ont montré qu’ERN2 peut remplacer ERN1 pour l’induction d’ENOD11 en réponse aux FNods et pour la formation de nodules, dès lors qu’il est exprimé sous le contrôle du promoteur d’ERN1. Ceci indique que ces facteurs ont des activités biologiques similaires et suggère que l’absence de complémentation d’ern1 par le facteur endogène ERN2 est probablement liée à une régulation transcriptionnelle différentielle de la part de leurs promoteurs. Enfin, nous avons initié la caractérisation phénotypique de nouvelles lignées mutées au niveau du gène ERN2, dans le but d’identifier les fonctions spécifiques de ce facteur au cours de la nodulation. A travers l’analyse d’une lignée Tilling (ern2.1) présentant une mutation ponctuelle dans le domaine de liaison à l’ADN, nous avons mis en évidence un rôle d’ERN2 dans la progression des cordons d’infection au niveau du cortex racinaire. Des études moléculaires ont permis de montrer que l’acide aminé muté est un résidu conservé et important pour la topologie du domaine de liaison à l’ADN, mais également pour l’activité transcriptionnelle d’ERN2 sur ENOD11. Contrairement à ern1, le mutants ern2.1 et ern2.2 (mutant d’insertion) sont capables de former des nodules. Néanmoins, l’infection nodulaire apparait dans les deux cas parfois défectueuse, conduisant à une sénescence précoce. Ces résultats démontrent qu’ERN2 remplit aussi des rôles spécifiques au cours de la nodulation, qui ne sont pas entièrement complémentés par ERN1. Il semblerait donc que les facteurs ERN contrôlent des étapes communes et divergentes de l‘infection rhizobienne, ERN1 ayant un rôle prépondérant dans l`initiation et progression de l’infection tandis qu’ERN2 aurait un rôle secondaire, plus centré dans la progression des cordons. La lignée double mutant ern1ern2.1, ouvre de nouvelles perspectives pour l’étude de la redondance fonctionnelle entre ces deux facteurs au cours des symbioses racinaires / Legumes are able to associate in symbiosis with Rhizobia bacteria in the soil, which culminates in the formation of a new organ referred to as the root nodule, within which differentiated bacteria fix nitrogen to the benefit of the host plant. The establishment of this association relies on a molecular dialogue between the two partners, involving bacterial lipo-chitooligosaccharide signals called Nod factors (NF). In the model legume Medicago truncatula, the perception of these symbiotic signals in the root epidermis, initiates a signaling pathway that leads to calcium oscillation responses required for the activation of downstream genes such as the well-characterized ENOD11. Previously, ERN1 and the closely-related ERN2 transcription factors (TFs) were reported as direct activators of ENOD11 via binding to the NFbox regulatory unit. In addition, phenotypic analysis of the ern1 knockout mutant has confirmed the importance of ERN1 not only during NF signaling but also throughout subsequent infection and nodule development stages. Nevertheless, the ern1 mutant displays a less severe phenotype compared to plants mutated in other NF signaling genes, raising the question of a possible functional redundancy with the endogenous closely-related ERN2 factor. My PhD project was focused on the study of the functional relationship between ERN1 and ERN2 TFs. By using a variety of strategies we aimed at determining both ERN expression profiles and relative functions during nodulation. We first examined the spatio-temporal expression profiles of these genes during rhizobial symbiosis and correlated this with the dynamics of cellular localization of ERN fusion proteins. These analyses revealed that these factors possess both common and distinct expression profiles, correlated with cell-type specific and dynamic in vivo protein accumulation, tightly associated with rhizobial pre-infection and subsequent infection stages in M. truncatula. Further cross-complementation studies in the ern1 mutant background showed that, when ERN2 is expressed under the control of the ERN1 promoter, it can fully restore the ern1 phenotype regarding NF-elicited ENOD11 activation and nodule formation. This indicates that these factors have similar biological activities and suggests that the incapacity of endogenous ERN2 to complement the ern1 mutant is mainly due to differences in their promoter activities. Finally, we also initiated a phenotypic characterization of M. truncatula ern2 mutant lines, in order to get a better insight into ERN2 specific functions during nodulation. The phenotypic analysis of a Tilling line (ern2.1) carrying a point mutation in a conserved amino acid in the ERN2 DNA binding domain, revealed a role for ERN2 during infection thread progression in the root cortex. Further molecular studies demonstrated that this mutated amino acid in the Tilling line is conserved and required for optimal DNA binding domain topology and transcriptional activity of ERN2 on its target ENOD11 gene. In addition, the ern2.1 line and a second ern2.2 insertional mutant line are both capable of forming nodules, in contrast to the ern1 mutant. Nevertheless, these nodules are partly infection defective leading to premature senescence. These findings provide evidence that ERN2 possesses specialized functions during nodulation that cannot be fully complemented by ERN1. This suggests that ERN possess common and divergent functions, ERN1 having a predominant role in rhizobial infection initiation and progression while ERN2 having a secondary and more centered role during infection thread progression. The ern1ern2.1 double mutant line, recently generated during my PhD, opens new perspectives to further study the functional relationship between ERN TFs during root endosymbioses.

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.

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.