Methods for detection of Phytophthora fragariae var. rubi on raspberry
2009
Koprivica, M., Ministry of Agriculture, Forestry and Water Management of the Republic of Serbia, Belgrade (Serbia). Plant Protection Directorate | Dulic-Markovic, I., FAO SEEDEV | Jevtic, R., Institute of Field and Vegetable Crops, Novi Sad (Serbia) | Cooke, D.E.L., Scottish Crop Research Institute, Dundee (United Kingdom)
Phytophthora fragariae var. rubi, a causal agent of raspberry root rot, is a serious soil-borne pathogen listed by EPPO as an A2 quarantine pest. Root samples were collected from badly diseased raspberry plants showing a variety of characteristics and often dramatic symptoms during surveys carried out in western Serbia during 2002. Identification of the causal agent was performed in collaboration work with the Scottish Crop research Institute, Dundee (UK). Nectrotic roots were planted on selective French bean agar. Detection of isolates was based on cultural and morphological features compared with referent cultures. DNA was extracted directly from the sampled roots using extraction buffer, purified by multi spin separation columns or in 24:1 mixture of chlorophorm- iso-amyl alcohol and amplified by nested PCR. Diluted DNA extracts were also amplified by conventional PCR with modified universal Phytophthora primers and digested with Msp1. Digestion patterns of the universal primers PCR product from infected roots matched those of Scottish strains. Phytophthora fragariae var. rubi occurred on 8 out 14 sites. Our results indicate that nested PCR or digestion of the universal Phytophthora primers PCR product for detection of P. fragaria var. rubi are more sensitive and less time-consuming and therefore recommended for use.
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