Thesis (M.S.)--Virginia Polytechnic Institute, 1953. / Typewritten. Vita. References: leaves 88-89. Also available via the Internet.

Thesis (Ph. D.)--University of Michigan, 1922. / "Reprinted from American journal of botany, vol. XI, no. 3, March, 1924." Bibliography: p. 156-158.

Twenty-nine Rhizobium leguminosarum biovar trifolii strains were tested for acidity tolerance in acidified liquid medium. Only 41\ of the strains grew at pH 4.1. One acid-tolerant strain, USDA 2160, and one acid-sensitive strain, 162-X-103 from Nitragin Co., were inoculated on seeds of 'Nungarin', 'Seaton Park' and 'Clare' subclover cultivars. The inoculated and pelleted seeds were sown in potted Cluff soil with pH 5. 7. Three desiccation levels were imposed by delaying watering for 0, 15 or 30 days. Four gravimetric soil water contents (6.0, 6.6, 10.5 and 12.5\) were maintained under a greenhouse line-source sprinkler system for 7 weeks. The desiccation treatments were more detrimental to the survival of the acid-tolerant Rhizobium strain (USDA 2160) than they were for the acid sensitive strain (162-X-103). Symbiotic effectiveness, measured as shoot dry weight, was higher with strain 162-X- 103 than with strain USDA 2160 and was comparable to the Nfertilized control at the highest water level (12.5 %). At the lower water levels (6.0, 6.6 %) symbiotic N2-fixation was more affected than N-uptake. The Rhizobium strains were able to survive and grow even at the lowest soil water level. The number of rhizobia in the soil and nodulation of the subclover plants had a correlation of 0.56.

Perennial clovers are being evaluated for their potential to reduce groundwater levels in Australian cropping zones where many soils are considered too acidic for reliable lucerne nodulation. However, the release of effective inocula for perennial clovers into such areas where sub clover is the predominant legume, could potentially compromise nitrogen fixation from this valuable annual clover if the symbiosis between the new inoculants and sub clover is not optimal. Studies were therefore designed to increase our understanding of these symbiotic interactions to optimise the management of legume-rhizobia interactions to extend (rather than restrict) the use of legumes in new environments. To assist the understanding of interactions between clovers and their microsymbionts, a glasshouse-based study of the cross-inoculation characteristics of 38 strains of Rhizobium leguminosarum biovar trifolii (R. l. trifolii) associated with 38 genotypes of annual and perennial Trifolium spp. from world centres of diversity was undertaken. Rhizobial isolates and clovers were assembled from South and equatorial Africa, North and South America and the Euro-Mediterranean regions. There was substantial specificity amongst the African clovers for effective nodulation. No strain of rhizobia from the South American perennial T. polymorphum, or from the Ethiopian clovers, was able to nodulate sub clover effectively, whilst less than 33% of the 18 strains from these regions could form nodules with the less promiscuous Mediterranean annual T. glanduliferum. Seventy of 476 cross-inoculation treatments examined did not nodulate, whilst 81 treatments clearly demonstrated effective nodulation. The remainder of the crossinoculation pairings revealed only partially effective or ineffective nodulation. Two barriers to effective nodulation were identified from the cross- inoculation study: a geographic barrier representing the broad centres of clover diversity, across which few host- strain combinations were effective; and within each region, a significant barrier to effective nodulation between an isolate from an annual host on a perennial host, or vice versa. Clovers and their rhizobia from within the Euro-Mediterranean region of diversity were more able to overlap the annual/perennial barrier than genotypes from the other regions. The data indicate that it will be a substantial challenge to develop inocula for perennial clovers that do not adversely affect nitrogen fixation by sub clover and other annual clovers in commerce, especially if the perennial clovers originate from Africa or America. To investigate the management of legume-rhizobia interactions when introducing legumes into new environments, a study was initiated in Uruguay (Mediterranean annual clovers were introduced into a predominantly perennial clover setting) that could be considered opposite to the situation emerging within southern Australia (perennial clovers evaluated in a predominantly annual clover setting). The Uruguayan grasslands contain populations of indigenous R. l. trifolii that nodulate endemic T. polymorphum but form ineffective nodules on clovers originating from the Mediterranean region. Importantly in the Uruguayan setting, Government policy has facilitated the introduction of numerous varieties of annual Mediterranean clovers with the aim of improving overall winter production in their naturally managed grasslands. In an attempt to understand the rhizobial ecology of this scenario, a cross-row experiment was set-up in 1999 in a basaltic, acid soil in Glencoe, Uruguay, to follow the survival and symbiotic performance of nine exotic strains of R. l. trifolii. In this thesis I report on the ability of the introduced strains to compete for nodule occupancy of Mediterranean clover hosts and show the impacts of the introduced strains on the productivity of the indigenous Uruguayan clover, T. polymorphum. Of the introduced strains, WSM1325 was a superior inoculant and remained highly persistent and competitive in forming effective symbioses with the Mediterranean hosts, T. purpureum and T. repens, in the Uruguayan environment over a 3 year period. T. purpureum and T. repens, when inoculated with the introduced strains, did not nodulate with any indigenous R. l. trifolii as typed from nodules of T. polymorphum. Conversely, there were no nodules on the Uruguayan host T. polymorphum that contained the introduced R. l. trifolii. These results revealed that there were effective symbioses between strains of R. l. trifolii and clovers, even though the soil contained ineffective R. l. trifolii for all hosts. This represents the first reported example of selective nodulation for an effective symbiosis in situ with annual and perennial clovers in acid soils. This phenomenon raised the question of whether this was restricted to the particular edaphic scenario in Glencoe, Uruguay. Glasshouse-based experiments in Australia were conducted to further understand the selection phenomenon. Two strains were selected for comparisons; strain WSM1325 isolated from an annual clover in the Mediterranean and WSM2304 isolated from the perennial clover T. polymorphum in Uruguay, South America. Variables that may have been specific to Glencoe were investigated. Thus, the effect of cell density and strain ratio at the time of inoculation, as well as soil pH, were examined on the two hosts (T. purpureum and T. polymorphum). Each was exposed to the same effective and ineffective micro-symbionts. In co-inoculation experiments at a cell density of 104 cells mL-1, each host nodulated solely with its effective strain, even when this strain was out-numbered 100:1 by the ineffective strain. However, the selection process ceased when the effective strain was out-numbered 1000:1. At higher basal cell concentrations of 105 - 108 cells mL-1, selection for WSM1325 to form effective nodules on T. purpureum was evident, but was significantly reduced as the ratio of ineffective cells in the inoculum increased above 4-fold. These results indicate that the selection mechanism is highly dependent upon the basal rhizobial cell density. Soil pH did not significantly alter the process, which could not be simply explained by the rate of strain growth, or extent of nodulation. Greater precision was sought in the terminology applied to nodulation outcomes where legumes have a choice of micro-symbiotic partners from within the same species of root-nodule bacteria. The nominated preferred terms are “nonselective”, “exclusive”, and “selective” nodulation. In view of the difference in host range between WSM1325 and WSM2304 and the selective nodulation process, a preliminary investigation into the genetic backgrounds of WSM1325 and WSM2304 was conducted. A selected range of gene regions were amplified by PCR from each strain and sequenced. Comparative analysis of the nucleotide sequences revealed that although the 16S rRNA sequences were identical, the atpD, GSII and nodD sequences contained distinct differences revealing disparity between the pSym replicons and between the chromosomal replicons of these strains. Of the genes sequenced, the highest degree of divergence was noted for the symbiotic NodD protein products, which are known to be critical determinants in the nodulation of specific hosts. An examination of the nodD gene region of WSM1325 and WSM2304 revealed a further contrasting feature; the regulatory gene nodR was present in the nodD gene region of WSM1325 but absent in WSM2304. Since NodR is known to be required for adding highly unsaturated fatty acyl groups onto the Nod-factor backbone, I could now hypothesise that the nodulation incompatibility observed between Trifolium hosts and micro-symbionts obtained from different geographical locations may result from differences in Nod-factor decoration. With the full genome sequence of the two strains WSM1325 and WSM2304 soon to be available, the role of nodR and any link to the selection phenomenon described in this thesis can be addressed.

The Rhizobia are a group of free-living soil bacteria known for their ability to symbiotically infect the roots of specific host plants as well as to produce siderophores in order to compete with other microorganisms for the limited availability of iron in the rhizosphere. In this study, Rhizobium leguminosarum ATCC 14479, which preferentially infects the red clover Trifolium pratense, was found to produce the trihydroxamate siderophore vicibactin (C 33H55N6O15) under iron restricted conditions. In addition, two other iron-binding, siderophore-like compounds: C20H36N4O10, C31H 55N6O15, were isolated and purified from the culture media. Due to the structural similarity of the latter compounds to vicibactin based on electrospray-mass spectrometry and nuclear magnetic resonance data, these heretofore unreported molecules are thought to be either modified or degraded products of vicibactin. Although vicibactin has previously been found to be commonly produced by other rhizobial strains, this is the first time it has been chemically characterized from a clover infecting strain of R. leguminosarum.

The Rhizobia are a group of free-living soil bacteria known for their ability to symbiotically infect the roots of specific host plants as well as to produce siderophores in order to compete with other microorganisms for the limited availability of iron in the rhizosphere. In this study, Rhizobium leguminosarum ATCC 14479, which preferentially infects the red clover Trifolium pratense, was found to produce the trihydroxamate siderophore vicibactin (C 33H55N6O15) under iron restricted conditions. In addition, two other iron-binding, siderophore-like compounds: C20H36N4O10, C31H 55N6O15, were isolated and purified from the culture media. Due to the structural similarity of the latter compounds to vicibactin based on electrospray-mass spectrometry and nuclear magnetic resonance data, these heretofore unreported molecules are thought to be either modified or degraded products of vicibactin. Although vicibactin has previously been found to be commonly produced by other rhizobial strains, this is the first time it has been chemically characterized from a clover infecting strain of R. leguminosarum.

Anthracnose, caused by the fungal pathogen Colletotrichum trifolii, is one of the most destructive diseases of alfalfa worldwide. Cloning and characterization of the host resistance (R) genes against the pathogen will improve our knowledge of molecular mechanisms underlying host resistance and facilitate the development of resistant alfalfa cultivars. However, the intractable genetic system of cultivated alfalfa, owing to its tetrasomic inheritance and outcrossing nature, limits the ability to carry out genetic analysis in alfalfa. Nonetheless, the model legume Medicago truncatula, a close relative of alfalfa, provides a surrogate for cloning the counterparts of many agronomically important genes in alfalfa. In this study, we used genetic map-based approach to clone RCT1, a host resistance gene against C. trifolii race 1, in M. truncatula. The RCT1 locus was delimited within a physical interval spanning ~200 kilo-bases located on the top of M. truncatula linkage group 4. Complementation tests of three candidate genes on the susceptible alfalfa clones revealed that RCT1 is a member of the Toll-interleukin-1 receptor/nucleotide-binding site/leucine-rich repeat (TIR-NBS-LRR) class of plant R genes and confers broad spectrum anthracnose resistance. Thus, RCT1 offers a novel resource to develop anthracnose-resistant alfalfa cultivars. Furthermore, the cloning of RCT1 also makes a significant contribution to our understanding of host resistance against the fungal genus Colletotrichum.

Submitted by Reginaldo Soares de Freitas (reginaldo.freitas@ufv.br) on 2017-04-03T12:25:22Z No. of bitstreams: 1 texto completo.pdf: 2716794 bytes, checksum: 6cbc55ddf42c47a8b94e0bca6edfe45c (MD5) / Made available in DSpace on 2017-04-03T12:25:22Z (GMT). No. of bitstreams: 1 texto completo.pdf: 2716794 bytes, checksum: 6cbc55ddf42c47a8b94e0bca6edfe45c (MD5) Previous issue date: 2004-01-29 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / O manejo de mosca minadora, Liriomyza trifolii (Burgess) (Diptera: Agromyzidae), é uma medida importante como forma de evitar a mudança de status desta praga na cultura do tomateiro. Este trabalho objetivou determinar a unidade amostral que melhor representa a variabilidade de minas com larvas na planta e propor planos de amostragem convencionais e seqüenciais de contagem e de presença-ausência para L. trifolii no tomateiro. A unidade amostral ideal foi determinada com o registro do tempo gasto para a contagem de minas com larvas nas unidades amostrais compostas por uma, duas e três folhas consecutivas ao longo do dossel da planta. Foram calculadas as variâncias relativas das unidades amostrais e realizadas análises de correlação e de regressão linear simples entre o as densidades absolutas (no de minas/terço do dossel e n de minas/planta) e densidades relativas (no de minas/unidade amostral e no de minas/terço do dossel). A unidade amostral com uma folha inserida entre os terços mediano e basal do dossel do tomateiro foi a que melhor representou a variabilidade e o número total de minas de L. trifolii na planta de tomate. Os números de minas de L. trifolii/folha e de folhas infestadas com densidades acima de zero, uma, duas e três minas/folha foram utilizados para se determinar os planos de amostragem. Os dados amostrais foram ajustados ao modelo de distribuição binomial negativo e obtido parâmetro de dispersão comum (Kcomum ). As proporções de unidades amostrais infestadas com densidades acima de zero, uma, duas e três minas/folha foram determinadas com o modelo de probabilidade de distribuição binomial negativa. Os planos de amostragem foram obtidos com o valor de Kcomum igual a 0,835, nível de controle igual a 3,0 minas/folha e os valores das proporções de unidades amostrais infestadas. Os planos convencionais de contagem e de presença-ausência foram determinados aos níveis de erro de 5, 10, 15, 20 e 25%. Os planos de amostragem seqüencial de contagem e de presença-ausência foram determinados através do teste da razão de probabilidade seqüencial de Wald. Foram requeridas 24 e 96 amostras/talhão para a amostragem convencional ao nível de erro de 25% pela contagem do número de minas e avaliação da presença- ausência, respectivamente. Os planos de amostragem seqüencial, através da contagem de minas e da avaliação de presença-ausência foram consistentes, precisos e requereram o máximo de 15 e 27 amostras para tomada de decisão correta, respectivamente. O plano seqüencial de contagem proporcionou uma economia de 37,5% no tempo de amostragem em relação ao plano convencional. Verificaram- se economias de 93, 84, 75 e 66% no tempo de amostragem com os planos seqüenciais de presença-ausência em relação aos planos convencionais com densidades de infestação acima de zero, uma, duas e três minas/folha, respectivamente. Em lavouras com baixas infestações, recomenda-se os planos seqüenciais de presença-ausência com densidades de infestação da amostra acima de zero mina/folha. / The management of leaf miner, Liriomyza trifolii (Burgess) (Diptera: Agromyzidae), is an important tool to avoid the change of status of this insect in the tomato crop. This work aimed to determine the sample unit that best represents the variability of mines with larvae in the plant and to propose conventional and sequential sampling plans with counting and binomial evaluation for L. trifolii in tomatoes. The ideal sample unit was determined considering the time spent for counting mines with larvae in the sample units composed by one, two and three consecutive leaves inserted in the canopy. The relative variance of the sample units were calculated and subjected to correlation and linear regression analyses among the absolute densities (number of mines/ canopy section and number of mines/plant) and relative densities (number of mines/ sample unit and number of mines/ canopy section). The leaf inserted between the medium and basal section of the canopy was the sample unit that best represented the variability and the total number of mines of L. trifolii in the tomato plant. The number of mines of L. trifolii/leaf and the number of leaves infested with densities above zero, one, two and three mines/leaf were used to determine the sampling plans. The sample data were adjusted to binomial negative frequency model and the parameter of common dispersion (K common) was obtained. The proportion of sample units infested with densities above zero, one, two and three mines/leaf were determined using the binomial negative distribution model. The sampling plans were obtained with the Kcommon value of 0,835, economic injury level equal to 3.0 mines/leaf and the proportion values of the sample units infested. The conventional plans of counting and of presence- absence were developed at the error levels of 5, 10, 15, 20 and 25%. The sequential sampling plans of counting and of presence-absence were developed through the Wald Sequential Probability Ratio Test. Twenty four and 96 samples/crop were required for the conventional sampling at the level of error of 25% through the counting of the number of mines and through the evaluation of the presence-absence, respectively. The sequential sampling plans based in the counting of mines and the presence-absence evaluation were consistent, precise and required the maximum of 15 and 27 samples for making correct decisions, respectively. The sequential sample plan by counting provided an economy of 37.5% in the sampling time in relation to the conventional sampling plan. Economies in the sampling time of the 93, 84.75 and 66% were verified with the sequential sampling plan for presence-absence compared with the conventional sampling plans with densities infestations above zero, one, two and three mines/leaf, respectively. In crops with low infestations, the sequential sampling plan based in the presence-absence with densities of sample infestation above zero mines/leaf should be preferred. / Não foi localizado o cpf do autor.

Submitted by (edna.saturno@ufrpe.br) on 2016-11-28T13:24:21Z No. of bitstreams: 1 Marcos Aurelio Araujo Lima.pdf: 265280 bytes, checksum: ad282a74478ab0633f0cbad41f436c79 (MD5) / Made available in DSpace on 2016-11-28T13:24:21Z (GMT). No. of bitstreams: 1 Marcos Aurelio Araujo Lima.pdf: 265280 bytes, checksum: ad282a74478ab0633f0cbad41f436c79 (MD5) Previous issue date: 2008-11-03 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / In the last years the leafminer Liriomyza trifolii (Burgess) has become a serious pest on melon crops. To develop a pest management program it is important to know some biological aspects of this insect species. Thus, the objective of this work was to study the biology of L. trifolii, to determine its thermal requirements and its fertility life table reared in melon under different temperatures. The temperatures studied were 20, 22, 25, 28, 30 e 33ºC. Egg hatching occurred within 2.8 (30ºC) to 6.6 (20ºC) days. Larval period ranged from 2.4 to 5.9 days, at 33 and 20ºC, respectively. A variation of 7.8 (33ºC) to 19.7 (20ºC) and 13.1 (33ºC) to 32.2 (20ºC),respectively, was observed for pupal stage and from egg to adult emergence. Larvae viability was always higher than 79%, while pupae viability was always lower than 83%, reaching 35% at 20ºC. The lower threshold temperature for eggs, larvae, pupae and egg-adult periods was 12.46,13.04, 13.20 and 12.95ºC, respectively. Thermal requirement (K) for egg, larva, pupa and from egg laying to adult emergence was 47.98, 41.54, 120.19 and 211.86 degree-days. Based on thermal requirements determined for L. trifolii and average temperatures for Mossoró, Apodi and Jaguaruana were estimated that it’s able to complete up to 23 generations per year in these areas. The mean duration of one generation (T) was 23.68 and 18.15 days, respectively, at thetemperatures of 25 and 28ºC. The net reproductive rate (Ro) was higher at 25ºC, corresponding to an increase of 86.20 times at each generation. The intrinsic rate of population increase (rm) was 0.18 (25ºC) and 0.15 (28ºC) and the finite ratio of population increase ( ) was 1.21(25ºC) and 1.17 (28ºC). The best performance of L. trifolii was at 25ºC. / Nos últimos anos as áreas cultivadas com meloeiro têm apresentado diminuição em sua produtividade devido à ocorrência da mosca-minadora, Liriomyza trifolii (Burgess). Para que se possa realizar um manejo adequado é necessário que se conheçam alguns aspectos biológicos desse inseto. Esta pesquisa teve como objetivos estudar a biologia, determinar as exigências térmicas e tabela de vida de fertilidade de L. trifolii com base nos dados obtidos do desenvolvimento deste inseto nas temperaturas de 20, 22, 25, 28, 30 e 33ºC. A duração do período embrionário variou de 2,8 (30ºC) a 6,6 (20ºC) dias. A duração da fase de larva foi de 2,4 a 5,9 dias, nas temperaturas de 33 e 20ºC, respectivamente. Ocorreu uma variação de 7,8 (33ºC) a 19,7 (20ºC) e 13,1 (33ºC) a 32,2 (20ºC), respectivamente, para a fase de pupa e tempo total de desenvolvimento. A viabilidade da fase de larva foi sempre superior a 79%. A viabilidade pupal foi sempre inferior a 83% e chegando a 38,1% na temperatura de 20ºC. Para as fases de ovo, larva, pupa e ovo-adulto os valores das temperaturas base foram de 12,46, 13,04, 13,20 e 12,95ºC. A constante térmica (K) para as fases de ovo, larva, pupa e tempo total de desenvolvimento foram de 47,98; 41,54; 120,19 e 211,86 graus-dia. Baseados nas exigências térmicas determinadas para L. trifolii e nas normais térmicas para Mossoró, Apodi e Jaguaruana estimou-se que a mesma pode completar mais 23 gerações/ano em ambas as localidades. A duração média da geração (T) foi igual a 23,68 e 18,15 dias, respectivamente, nas temperaturas de 25 e 28ºC. A taxa líquida de reprodução (Ro) foi maior na temperatura de 25ºC, correspondendo a um aumento de 86,20 vezes a cada geração. A taxa líquida de crescimento populacional (rm) foi de 0,18 (25ºC) e 0,15 (28ºC) e a razão finita de aumento ( ) 1,21 (25ºC) e 1,17 (28ºC). Diante de todos os parâmetros biológicos estudados, a melhor temperatura para o desenvolvimento de L. trifolii foi 25ºC.