Detection of Clavibacter michiganensis subsp. sepedonicus the causal agent of potato ring rot, in the breeding and propagation materials of the three-stage control process

To maintain Clavibacter michiganensis subsp. sepedonicus-free (Cms-free) potato genetic resources, a three-stage process of checking limited lots of initial and new breeding potato materials was designed for small breeders in terms of costs, personnel and facilities. For an intensified evaluation of the presence of Cms in different vegetative stages within one growing season a mixture of 10 randomly selected artificially infected and healthy tubers of individual cultivars and breeding materials were prepared. In the first stage, a visual assessment of the vascular vessels of mother tubers was performed and at least 20% of each material was deemed positive, indicating that it should be discarded. In the second stage, one half of mother tubers were grown in the greenhouse and the other in a netting house. Plants in bloom and daughter tubers were evaluated. Within the second stage 1.4%–28% of each individual potato material could be excluded based on double-antibody sandwich enzyme-linked immunosorbent assays and SYBR Green real-time PCR assays, including a melting point analysis using PSA 1/R and CelA F/R  primer sets and biological tests on aubergines. In the third stage, in vitro plants transferred from potato resources that tested negative in the previous stages were evaluated using the methods used in the second stage. The specificity of the antibodies and the two primer sets, PSA1/R and CelA F/R, were evaluated with 100% success

Methodology for assessing potato varieties for resistence to soft rot bacteria using slice test

The methodology is to assessment the level of resistance of potato genotypes to soft rot bacteria of Dickeya, Pectobacterium and Pseudomonas genera using slice test

Determination of causal agent of ring rot of potato, Clavibacter michiganensis subsp. sepedonicus bacteria in breeding and multiplicated materials

The methodology includes a method of artificial infection of plants and potato tubers as a prerequisite for the study of migration and localization of the pathogen in tubers and within the plant vascular tissues, time schedule and recommended size of samples of individual parts of plants and potato tubers during the growing season, depending on external conditions and potato genotype, how to deal with samples of the plants and potato tubers prior to treatment, method of processing samples of individual plant parts and tuber depending on the detection method and the optimization of methods for detection of the pathogen Clavibacter michiganensis subsp. sepedonicus. In compliance with the proposed methodological procedure should be interrupted vertical transmission of the pathogen in the process of breeding, the possibility of detection of the pathogen in breeding and propagation material and reduction of suspicious samples should be increased and the late positive determination of ring rot in the official phytosanitary and company checks abroad should be reduced