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Master of Science / Department of Entomology / C. Michael Smith / The Dectes stem borer, Dectes texanus LeConte, is a pest of soybean, Glycine max (L.) Merrill, in North America. Larval feeding weakens plant stems, triggering lodging of the infested plants and causing significant yield losses. D. texanus infestations in soybean fields are increasing across Kansas and other states, necessitating the development of effective tactics to control this pest. The use of D. texanus -resistant soybean cultivars is a desirable strategy to control this pest since cultural and chemical control options are lacking. In previous studies, the soybean plant introduction PI165673 was identified to be resistant to D. texanus. The objective of this research was to determine the inheritance of resistance to D. texanus in PI165673. F[subscript]2 progeny plants from crosses between the D. texanus susceptible genotypes KS5004N and K07-1544, and the resistant genotype PI165673 were tested in the field for resistance to D. texanus in 2011. Seeds from the cross K07-1544/PI165673 were advanced to the F[subscript]3 generation, and F[subscript]2:3 families were tested in the field for resistance to D. texanus in 2012. At 20 d after infestation with adults, the numbers of oviposition punctures and larvae on each plant were counted to estimate the oviposition puncture per larvae resistance ratio. Segregation for resistance to D. texanus and heritability estimates in the F[subscript]2 and F[subscript]2:3 populations indicated that resistance is controlled by more than one gene. Thirteen F[subscript]2:3 families had a higher (more resistant) resistance ratio than the susceptible parent K07-1544. Mean head capsule widths of larvae collected from K07-1544 and PI165673 plants in 2012 were similar, as was the percentage of larvae per larval instar. According to heritability estimates for each phenotypic trait, progress in breeding for D. texanus resistance using PI165673 will benefit from marker assisted selection. Identification of additional sources of D. texanus resistance besides that in PI165673 is needed to slow larval growth in the stem.

Plant diseases remain a major obstruction to meeting the world’s increased demand for soybean oil and protein. Reducing the losses caused by diseases in order to improve crop production is a high priority for agricultural research. The need for novel strategies for plant disease control cannot be overstated. In the present study, selected defense-related genes were silenced and/or overexpressed in soybean using a virus-based vector and the resultant plants were tested for their responses to pathogens. The first part of the study focused on Rps1k (Resistance to Phytophthora sojae) gene. The two conserved domains encoding ‘P-Loop NTPase’ and ‘PLN03210’ of Rps1k were independently overexpressed. Stem inoculation assays for the overexpressing plants showed significant resistance to virulent races; 90% standing plants compared to 10% in controls. Lesion length was greatly restricted only in case of plants overexpressing ‘PLN03210’. Simultaneous silencing of Rps1k-1 and Rps1k-2 resulted in remarkable susceptibility to avirulent races when tested by a detached-leaf assay. The second part of the study entailed silencing/overexpression of the chlorophyllase genes GmCLH1 and GmCLH2 and testing the responses of the silenced/overexpressing plants to the sudden death pathogen Fusarium virguliforme. Four weeks post root inoculation, GmCLH2-silenced plants showed enhanced resistance while the GmCLH2-overexpressing plants exhibited markedly increased susceptibility when compared to empty vector control. RT-PCR assay of PR genes revealed elevated expression of PR2 and PR4 in GmCLH2-silenced plants. In the third part of the study, soybean plants silenced for a leucine-rich repeat receptor-like kinase (GmRLK3) gene were examined for their responses to different pathogens. Silencing of GmRLK3 enhanced susceptibility to infection with Alternaria tenuissima or Sclerotinia sclerotiorum as revealed by rapid disease progress on treated leaves. Surprisingly, silencing of GmRLK3 in known susceptible soybean cultivars rendered the silenced plants resistant to P. sojae. The ensuing partial resistance to P. sojae was consistent with results of RT-PCR assays that showed a significant increase in the transcript level of the osmotin-encoding gene (PR5a) in the GmRLK3-silenced plants. PR5a is considered a marker for systemic acquired resistance.

Conselho Nacional de Desenvolvimento Científico e Tecnológico / Soybean stem white rot, caused by Sclerotinia sclerotiorum, occurs in more than one million hectares in Brazil. This disease should be managed by integrated control methods, including genetic resistance. This study evaluated and developed a reliable method for the selection of soybean genotypes resistant to stem white rot. The work was based on rating inoculations on the stem. The methods evaluated consisted of different inoculation sites in the plant, wounding or not or cutting the stem of the plants, and inoculating PDA disks containing fungal mycelium. The tests were conducted in two different environments, a controlled one, where plants were grown in a greenhouse and brought into the laboratory to be inoculated and incubated in a growth chamber where the temperature is favorable for fungal growth, and a field, where the pathogen is subjected to adverse environment conditions. Spearman´s coefficients of rank correlation was used to measure the correspondence between ranks of cultivar and ranks of the same cultivars in field experiments to establish the best inoculation method. It was observed that temperature growth conditions, and inoculation method may affect the selection of resistant soybean genotypes against stem rot. The best method to evaluate soybean resistance was by wound inoculating the middle third of the plant when the plants were at the end of flowering and early grain filling, with PDA disks and evaluating 14 days later. This method was rs = 0.86, and the variety that can be considered as resistance standard in other studies was Emgopa 316, while the susceptible standards were BRSMG Garantia and BRSMG68 Vencedora. / A podridão branca da haste da soja causada por Sclerotinia sclerotiorum ocorre mais de um milhão de hectares no Brasil. Esta doença deve ser manejada por métodos de controle integrado, entre eles a resistência genética que deve ser pesquisa. O objetivo do trabalho foi avaliar e definir um método confiável na seleção de genótipos de soja. Quanto ao método de inoculação, o trabalho se baseou em avaliar inoculações realizadas no caule. As metodologias diferiram quanto ao local de inoculação na planta, uso ou não de ferimentos, ou o uso do corte do ápice das plantas na inoculação, utilizando o disco de BDA contendo micélio do fungo. Os testes foram conduzidos em dois ambientes distintos, em ambiente controlado, onde plantas foram cultivadas em casa-de-vegetação e levadas para o laboratório para serem inoculadas e incubadas em câmara de crescimento onde a temperatura é extremamente favorável ao desenvolvimento do fungo e no campo onde a doença estará sujeito às condições ambientais adversas. Para definir qual o melhor método de inoculação, como também os genótipos que possuem maior resistência parcial, calculou correlações pelo coeficiente de Spearman entre os métodos avaliados e métodos com o ranking geral das cultivares. Observou-se que a temperatura, condições de cultivo, e o método de inoculação podem influenciar na seleção dos genótipos quanto à resistência à podridão da haste da soja. Definiu que o melhor método de avaliação à resistência de genótipos de soja foi a inoculação realizada com ferimento no terço médio da planta onde as plantas se encontravam no final do florescimento e início de enchimento dos grãos, com discos de BDA e avaliadas 14 dias após a inoculação, este método obteve rs=0,86, e a cultivar que pode ser considerado com padrão de resistência em outros estudos foi a Emgopa 316, e o padrão de suscetibilidade a BRSMG Garantia e BRSMG 68 Vencedora. / Mestre em Agronomia

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