Inoculation Techniques, Development of Brassica Napus Breeding Lines and Identification of Markers Associated with Resistance to Sclerotinia Sclerotiorum (Lib.) De Bary

Sclerotinia stem rot caused by Sclerotinia sclerotiorum (Lib.) de Bary is an economic disease affecting canola (Brassica napus L). Since expression of sclerotinia stem rot symptoms shows much variability and the trait is quantitative in nature, reliable phenotypic evaluation methods for characterization are needed. The three major objectives of this dissertation were to: i) evaluate eight different inoculation methods to discriminate between S. sclerotiorum-resistant and susceptible B. napus germplasm; ii) develop breeding lines with resistance to multiple diseases, and; iii) to identify QTL associated with resistance to sclerotinia stem rot using association mapping (AM). The eight methods evaluated were the detached leaves, detached stems, petiole inoculation (PIT), straw-inoculation, stem-piercing with toothpick, mycelial spray (MSI), petal inoculation and oxalic acid assay. MSI and PIT can better discriminate between the isolates and germplasm. Breeding lines resistance to S. sclerotiorum, Leptosphaeria maculans, and Rhizoctonia solani were developed from a cross between two moderately sclerotinia stem rot resistant plant introductions (PI). F2 seedlings were screened for sclerotinia stem rot using PIT. Surviving plants were self pollinated and their progeny screened again. This process was repeated until the F6 generation. In addition, F5 seedlings were evaluated for their reaction to R. solani and F5 and F6 seedlings for their reaction to L. maculans. Eight lines were identified as moderately resistance to these three pathogens. The genomes of a group of 278 B. napus plant introductions were screened using Diversity Array Technology to detect QTL associated with resistance to sclerotinia stem rot. The population was classified into nine sub-populations and 32 significant markers each explaining between 1.5 and 4.6% of the variation were identified. Blastn search indicates that similar nucleotide sequences are distributed throughout the genomes of B. oleracea, B. rapa, and A. thaliana. Results of these studies suggest the PIT and MSI are reliable screening tools to evaluate materials for resistance to sclerotinia stem rot; materials identified as resistant to S. sclerotiorum were also moderately resistant against R. solani and L. maculans and could be valuable sources for canola improvement programs; and AM allowed us to identify QTL associated with resistance to sclerotinia stem rot

Evaluating Responses of Sugar Beet Cultivars to Fusarium Species in Greenhouse and Field Conditions

Fusarium yellows of sugar beet (Beta vulgaris L.) is becoming an important disease in Minnesota and North Dakota in the United States. One of the best ways to manage Fusarium yellows is to develop and use resistant cultivars. Responses of eight sugar beet cultivars to three Fusarium species were determined in the greenhouse and compared to the responses of the same eight cultivars grown in a field already infected by Fusarium yellows. There were significant and appreciable relationships between greenhouse and field studies for responses of eight sugar beet cultivars on Fusarium yellows. The estimated correlation coefficient for area under disease progress curve (AUDPC) between observations in fields and those in greenhouses was 0.987 (p<0.01). The mean AUDPC were significantly different among cultivars (p<0.01) in the greenhouse and in the field studies. Of the cultivars, Van der Have 46177 was the most susceptible, but Crystal R434 the most resistant. Crystal 820, Van der Have 66561, and Beta 4797R were resistant, and Beta 4818R, Seedex Magnum, and Hilleshog 2463Rz were moderately resistant. There was a strong negative relationship between the AUDPC and recoverable sucrose yield in the field experiments and the estimated coefficient of determination was 0.939 (p<0.01). It was concluded that greenhouse screening can act as a useful and reliable means to evaluate and select beet germplasms and/or accessions for resistance to Fusarium yellows.Peer reviewe

Comparative Pathogenicity and Virulence of Fusarium Species on Sugar Beet

In all, 98 isolates of three Fusarium spp. (18 Fusarium oxysporum, 30 F. graminearum, and 50 Fusarium sp. nov.) obtained from sugar beet in Minnesota were characterized for pathogenicity and virulence on sugar beet in the greenhouse by a bare-root inoculation method. Among the 98 isolates tested, 80% of isolates were pathogenic: 83% of the F. oxysporum isolates, 57% of the E graminearum isolates, and 92% of the Fusarium sp. nov. isolates. Symptoms varied from slight to moderate wilting of the foliage, interveinal chlorosis and necrosis, and vascular discoloration of the taproot without any external root symptoms. Among the pathogenic isolates, 14% were highly virulent and 12% were moderately virulent. Most of the highly virulent isolates (91%) and moderately virulent isolates (89%) were Fusarium sp. nov. All pathogenic isolates of F. graminearum and most pathogenic isolates (87%) of F. oxysporum were less virulent. In general, more-virulent isolates induced first foliar symptoms earlier compared with less-virulent isolates. This study indicates that both F. oxysporum and Fusarium sp. nov. should be used in greenhouse and be present in field studies used for screening and developing sugar beet cultivars resistant to Fusarium yellows complex for Minnesota and North Dakota.Peer reviewe