Outlook We are constantly working to optimize the real-time PCR method. This involves continuously verifiying that the primers used in the PCR reaction for the detection of Verticillium nonalfalfae can still detect all mild and aggressive versions that occur in the Hallertau.
References Borza, T., Beaton, B., Govindarajan, A., Gau, X., Liu, Y., Ganga, Z., Wang-Pruski, G. (2018): Incidence and abundance of Verticillium dahliae in soil from various agricultural fields in Prince Edward Island, Canada. Eur J Plant Pathol 151, 2825—2830. https://doi.orgll 0.1007/310658—017—1408-1 EPPO Bulletin (2020) PM 7/78 (2) Verticillium nonalfalfae and V. dahliae: 50 (3): 462-476. Guček, T., Stajner, N., Radišek, S. (2015): Quantification and detection of Verticillium albo-atrum in hop (Humulus lupulus) with real-time PCR. Hop Bulletin 22, 26-39. Maurer, K.A., Radišek, S., Berg, G., Seefelder, S. (2013): Real-time PCR assay to detect Verticillium albo-atrum and V. dahliae in hops: development and comparison with a standard PCR method. Journal of Plant Diseases and Protection, 120 (3), 105–114. Seigner, E, Haugg, B, Hager, P., Enders, R., Kneidl, J. & Lutz, A. (2017): Verticillium wilt on hops: Real-time PCR and meristem culture – essential tools to produce healthy planting material. Proceeding of the ScientificTechnical Commission of the International Hop Growers´ Convention, Austria, 20-23. Wei, F., Fan, R., Dong, H.-T., Shang, W.-J., Xu, X.-M., Zhu, H.-Q., Yang, J.-R., and Hu, X.-P. (2015): Threshold microsclerotial inoculum for cotton Verticillium wilt determined through wet-sieving and real-time quantitative PCR. Phytopathology 105:220-229. Weller, S.A., Elphinstone, J.G., Smith, N.C., Boonham, N., and Stead, D.E. (2000): Detection of Ralstonia solanacearum strains with a quantitative, multiplex, real-time, fluorogenic PCR (TaqMan) assay. Appl Environ Microbiol. 66(7), 2853-8.
6.8
Meristem culture for the production of healthy seedlings
Project Management: Team: Collaboration:
Dr. E. Seigner, A. Lutz B. Haugg P. Hager, R. Enders, IPZ 5c Dr. L. Seigner, and the Virus Diagnostics Team, IPS 2c
Objectives Infestations of Verticillium, viruses, and viroids can lead to dramatic losses in yield and quality. Since these diseases cannot be combated with pesticides, a biotechnological method, such as meristem culture, is used to produce Verticillium- and virus-free material. In 2020, the main focus was on improving the method for eliminating apple mosaic virus (ApMV), which is often more stubborn than the hop mosaic virus and can still be detected in regenerated plants, even after a meristem culture intervention.
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