Abstract
Developing hybrids, resistant to causal pathogen of Downy mildew (Plasmopara halstedii (Farl.) Berl. & de Toni), is one of the critical tasks in sunflower breeding. Molecular markers have advanced breeding practice in the past decades, however there are still unmet needs for reliable high-throughput (HT) selection of the pathogen resistant starting material and differentiation of the plants infected by different pathogens. In this study, we tested the known DNA marker (308 bp fragment from ribosomal DNA of P. halstedii) for detection of pathogen in different tissues of sunflower plant and at different stages of plant development. Specified DNA marker was re-validated in the total DNA, isolated from sporangium as well as from seedlings of infected pathogen resistant/susceptible inbred lines of Ukrainian breeding and 60 F2 crosses. An independent set of field grown plants with unknown resistance to P. halstedii, having symptoms of the bacterial/fungal/viral pathology were used for HT screening and genotypes, infected with downy mildew, were successfully identified. Pathogen appeared to be concentrated in the vessels of sunflower leaves, in contrast to parenchymal tissue. Our study demonstrates an addition to whole seedling inoculation technique of P. halstedii detection, which allows HT identification of the pathogen infected and non-infected sunflower plants.
References
Berg, T., Tesoriero, L., Hailstones, D.L., 2005. PCR-based detection of Xanthomonas campestris pathovars in Brassica seed. Plant Pathology 54: 416–427.10.1111/j.1365-3059.2005.01186.xSearch in Google Scholar
Bonants, P., Carroll, G., de Weerdt, M., van Browwershaven, I., Baayen, R., 2003. Development and validation of a fast PCR-based detection method for pathogenic isolates of the citrus ts P black spot fungus, Guignardia citricarpa. European Journal of Plant Pathology 109: 503–513.10.1023/A:1024219629669Search in Google Scholar
Dolgova, E.M., Аladina, Z.К., Мichailova, V.N., 1990. Express method for evaluation of sunflower resistance to Downy mildew. Breeding and Seed Production 68: 50–55 (In Russian).Search in Google Scholar
Doyle, J.J., Doyle, J.L., 1987. A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochemical Bulletin 19: 11–15.Search in Google Scholar
Ioos, R., Laugustin, L., Rose, S., van Browwershaven, I., Baayen, R., 2007. Development of a PCR test to detect the downy mildew causal agent Plasmopara halstedii in sunflower seeds. Plant Pathology 56: 209–218.10.1111/j.1365-3059.2006.01500.xSearch in Google Scholar
Jocic, S., Cvejic, S., Hladni, N., Miladinovic, D., Miklic, V., 2010. Development of sunflower genotypes resistant to downy mildew. In: Proceedings of International Symposium: Sunflower Breeding on Resistance to Diseases, Krasnodar, Russia, pp. 93–97.10.2298/HEL1053173JSearch in Google Scholar
Jocic, S., Miladinovic, D., Imerovski, I., Dimitrijevic, A., Cvejic, S., Nagl, N., Kondic-Spirka, A., 2012. Towards sustainable downy mildew resistance in sunflower. Helia 35: 61–72.10.2298/HEL1256061JSearch in Google Scholar
Тihonov, О.I., Zaichuk, V.P., Маslienko, А.V., 1981. Evaluation of sunflower resistance. Plant Protection 9: 47 (In Russian).Search in Google Scholar
© 2019 Walter de Gruyter GmbH, Berlin/Boston
