Stemphylium blight of lentil caused by <i>Stemphylium botryosum</i> Wallr., is a serious problem in Bangladesh, northeast India and Nepal causing more than 60 % yield losses under epidemic conditions. The pathogen started to appear on lentil in Saskatchewan in recent years and is widely distributed throughout western Canada but it is not well understood. An investigation of inheritance of resistance to stemphylium blight was done in the lentil cross Barimasur-4 × CDC Milestone. In order to develop a reliable indoor screening technique for this inheritance study, a suitable isolate of <i>Stemphylium botryosum</i>, a suitable culture medium for inoculum production and an appropriate plant age for indoor inoculation were identified. The maximum differential of disease severity was observed when lentil genotypes were inoculated at 14 days after planting (DAP). At 14 DAP, lentil plants rapidly defoliated but were capable of regrowth which caused variability in scoring for disease reaction. Inoculation at 42 DAP, close to the flowering stage, was found to be better for consistently scoring disease reaction. V8P medium was most suitable for inducing conidia production. Based on ability to sporulate, the isolate SB-19 from Saskatchewan was identified as suitable for conducting genetic studies of resistance to stemphylium blight. It was compared to isolate SB-BAN from Bangladesh for aggressiveness on two lentil cultivars. The SB-BAN isolate was found to be more aggressive. A preliminary screening of local and exotic germplasm done with the two isolates revealed considerable variability for disease resistance. Resistance to <i>S. botryosum</i> appeared to be quantitatively inherited in the cross Barimasur-4 × CDC Milestone according to both field and indoor screenings. The results of this study also confirmed that Precoz, one of the parents of Barimasur-4, was resistant to <i>S. botryosum</i>.

Pests and diseases have a significant economic impact on yield, quality and sale of processing carrots. In an effort to combat certain pests and diseases, the effects of photo, physical and mechanical (PPM) treatments on carrot (Daucus carota L. var. Sativus cv. Carson) were investigated. Generally, the incidence of certain pests and diseases was reduced through a variety of PPM treatments during two separate field experiments. However, not all of the reductions were statistically significant owing in large part to the low level of incidence of certain pests and diseases during the growth trials. To counteract this difficulty, experiments were conducted to determine the conditions necessary to induce blight in a controlled laboratory environment, with little success. In addition, HPLC-based detection of the defence hormone, salicylic acid, in carrot seedlings was established to help characterize the effects of PPM treatments from a biochemical perspective in future experiments.

Stemphylium blight is a defoliating fungal disease caused by <i>Stemphylium botryosum</i>. It has become more prevalent in Saskatchewan. Although not much is known about the biology of the fungus, increasing lentil (Lens culinaris) yield losses of up 62% have been reported in Bangladesh and India. The infection of lentil by <i>S. botryosum</i> was investigated under a range of temperatures (5 to 30°C), wetness periods (0 to 48 h) and wetness periods interrupted by dry periods of 6 to 24 h. The experiments involved testing the impact of environmental conditions on germination of conidia on glass slides and stemphylium blight infection on lentil (cv. CDC Milestone). Generalised linear models and non-parametric tests were used to determine the effects of these factors on conidial germination and disease development. Infection levels increased with increasing temperature and wetness duration. A latent period of 48 h was observed at 25°C and 30°C under continuous wetness. The duration of the latent period increased with decreasing temperatures and decreasing wetness duration. <i>S. botryosum</i> required warm temperatures (above 25°C) and a minimum wetness period of 8 h for optimal disease development. Low levels of infection were observed within the first 2 h of incubation at 10°C and increased with longer wetting periods up to 48 h and temperatures up to 30°C. The pathogen could maintain infectivity during interrupted wetness periods despite its requirement for prolonged wetness periods. Infection levels were not significantly affected by interrupting dry periods of 6 to 24 h although long dry periods (24 h) combined with higher temperatures (30°C) resulted in a decrease in stemphylium blight severity. Germination studies on glass slides supported these findings. Response surface models were developed that provided a good fit for the response of conidial germination to temperature and wetness duration. The coefficients of determination for the regression of observed against predicted effects ranged from 0.88 to 0.97. The general additive model could also be used to predict stemphylium blight severity responses to temperature and wetness duration (scaled deviance = 1.04). However, that model tended to overestimate infection levels especially at lower temperatures. The coefficients of determination for the observed against predicted effects at 5 to 30ºC ranged from 0.77 to 0.92 for the general additive model.

Stemphylium blight of lentil caused by <i>Stemphylium botryosum</i> Wallr., is a serious problem in Bangladesh, northeast India and Nepal causing more than 60 % yield losses under epidemic conditions. The pathogen started to appear on lentil in Saskatchewan in recent years and is widely distributed throughout western Canada but it is not well understood. An investigation of inheritance of resistance to stemphylium blight was done in the lentil cross Barimasur-4 × CDC Milestone. In order to develop a reliable indoor screening technique for this inheritance study, a suitable isolate of <i>Stemphylium botryosum</i>, a suitable culture medium for inoculum production and an appropriate plant age for indoor inoculation were identified. The maximum differential of disease severity was observed when lentil genotypes were inoculated at 14 days after planting (DAP). At 14 DAP, lentil plants rapidly defoliated but were capable of regrowth which caused variability in scoring for disease reaction. Inoculation at 42 DAP, close to the flowering stage, was found to be better for consistently scoring disease reaction. V8P medium was most suitable for inducing conidia production. Based on ability to sporulate, the isolate SB-19 from Saskatchewan was identified as suitable for conducting genetic studies of resistance to stemphylium blight. It was compared to isolate SB-BAN from Bangladesh for aggressiveness on two lentil cultivars. The SB-BAN isolate was found to be more aggressive. A preliminary screening of local and exotic germplasm done with the two isolates revealed considerable variability for disease resistance. Resistance to <i>S. botryosum</i> appeared to be quantitatively inherited in the cross Barimasur-4 × CDC Milestone according to both field and indoor screenings. The results of this study also confirmed that Precoz, one of the parents of Barimasur-4, was resistant to <i>S. botryosum</i>.

Pests and diseases have a significant economic impact on yield, quality and sale of processing carrots. In an effort to combat certain pests and diseases, the effects of photo, physical and mechanical (PPM) treatments on carrot (Daucus carota L. var. Sativus cv. Carson) were investigated. Generally, the incidence of certain pests and diseases was reduced through a variety of PPM treatments during two separate field experiments. However, not all of the reductions were statistically significant owing in large part to the low level of incidence of certain pests and diseases during the growth trials. To counteract this difficulty, experiments were conducted to determine the conditions necessary to induce blight in a controlled laboratory environment, with little success. In addition, HPLC-based detection of the defence hormone, salicylic acid, in carrot seedlings was established to help characterize the effects of PPM treatments from a biochemical perspective in future experiments.

Disease problems in crop plants are still a major threat to the agro-industry worldwide. Various strategies have been developed and evaluated in recent years. One strategy is to use naturally-occurring antipathogen factors such as lysozymes and chitinases in transgenic plants. In the present study, transgenic plants containing chick-egg white lysozyme (Lys 75) have been evaluated for lysozyme production in planta, subcellular localisation, and resistance to some potato pathogens, including Phytophthora infestans and Erwinia carotovora subsp. atroseptica, the two major potato pathogens worldwide. In addition, the evaluation of resistance was also undertaken for transgenic plants carrying other naturally-occurring antipathogen factors including a bean chitinase gene (BCH 35) and a snowdrop lectin gene (GNA 74). In order to accurately quantify the lysozyme production in Lys 75 plants, the turbidimetric lysozyme enzyme assay was optimised. Also, a modified substrate for the enzyme has been developed by covalently linked the Micrococcus lysodeikticus cell wall with a dye, remazol brilliant violet 5R to enable a colorimetric assay of the enzyme. In order to quantitatively assess resistance levels of the transgenic plant, a new method (leaf-bridge bioassay) for conducting and evaluating resistance in planta has been developed. All transgenic plants in tissue culture were tested for resistance using this technique. Evaluation of the progress of infection in detached leaves of Lys 75 showed that lysozyme gave some degree of protection against the bacterial pathogen, Erwinia carotovora subsp. atroseptica and the fungal pathogen, Fusarium sulphureum. Analysis of intercellular fluid from the Lys 75 leaves showed that more than 80% of the total lysozyme expressed in the leaf was located in the intercellular space which is a strategic place to combat pathogen attack. In contrast, the levels of protection in BCH 35 plants were relatively low compared with Lys 75. The progress of infection was delayed in BCH 35 leaves challenged with F. sulphureum only. No resistance at all was observed in GNA 74 to all the pathogens used. All the transgenic potato lines were susceptible to P. infestans. Recently, a new strategy to combat disease problems has been suggested based on a 'durable resistance'. Potato variety Stirling' which shows durable resistance in the field has been used to study the early biochemical and molecular events during elicitation of 'Stirling' cell suspension cultures with an elicitor mix derived from infective units of a compatible strain of P. infestans. For comparison, an elicitor mix from an incompatible strain of P. infestans was also prepared and used. The mixed elicitor comprising zoospore extract, culture filtrate and mycelium homogenate induced defence responses in 'Stirling' cell suspension cultures as judged by the increase in PAL enzyme activity. PAL activity in 'Stirling' ceUs elicited with an elicitor mix derived from an incompatible strain of P. infestans was twice the activity in the compatible interaction. The peak levels in both types of interaction were at 6 h post-elicitation. An oxidative burst was demonstrated also in both types of interactions indicated by rapid release of H(_2)O(_2) into the culture medium. The H(_2)O(_2) level peaked at 2 h post-elicitation in both interactions before being reduced to its normal level at 4 h. The H(_2)O(_2) released during incompatible interaction was twice the levels monitored in the compatible. A subtracted cDNA library of differentially expressed mRNAs during elicitation of 'Stirling' cell suspension cultures with the elicitor mix from a compatible strain of P. infestans was constructed using suppression subtractive hybridisation. Two cDNA clones, STS 42 and STS 52, relevant to the present study were identified and characterised. STS 42 showed high degree of similarity to potato leucine aminopeptidase gene which is induced in response to wounding. Gene expression studies using RT-PCR showed that the mRNA levels of STS 42 increased gradually throughout the 18 h elicitation. STS 51 was identified as a member of the ribonuclease T2 histidine proteins. It showed some degree of similarity to plant ribonucleases involved in self-incompatibility reactions during pollination. It has a site for tyrosine kinase phosphorylation at the hydrophilic region of the sequence and could possibly be involved in phosphorylation during signal transduction. mRNA levels of STS 51 were increased during the first 12 h of elicitation.

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Thesis (Ph. D.)--University of Adelaide, Dept. of Applied and Molecular Ecology, 1999. / Includes bibliographical references (leaves 182-217).

Thesis (M.S.)--West Virginia University, 2008. / Title from document title page. Document formatted into pages; containsvii, 73 p. : ill. (some col.). Vita. Includes abstract. Includes bibliographical references (p. 66-68).