Full Text
Table 1. Soil samples used for isolation of S. endobioticum spores
No. | Soil samples | Number of fungal spores per 100 g of soil |
1* | Artificial inoculation (loamy sand) | 5 |
2* | Artificial inoculation (loam) | 5 |
3* | Artificial inoculation (loamy sand) | 50 |
4* | Artificial inoculation (loam) | 50 |
5 | Artificial inoculation (loamy sand) | 500 |
6 | Artificial inoculation (loamy sand) | 5000 |
7 | Artificial inoculation (loamy sand) | 20000 |
8 | Moscow Region, old outbreak (peaty soil) | Unknown |
9 | Moscow Region, outbreak (loam) | Unknown |
10 | Voronezh Region, old outbreak (loamy sand) | Unknown |
11 | Negative control | 0 |
* The soil samples were used only for the direct DNA extraction method.
Table 2. Results of direct testing of soil samples for presence of S. endobioticum spores using carbon tetrachloride
No. | Number of fungal spores per 100 g of soil | Number of fungal spores isolated from 100 g of soil | Effectiveness, % |
5 | 500 | 180.87 ± 44.9 | 36.17 |
6 | 5000 | 492.64± 53.66 | 9.85 |
7 | 20000 | 849.45 ± 54.52 | 4.24 |
8 | Unknown | Not conducted | — |
9 | Unknown | 230.2 ± 44.7 | — |
10 | Unknown | 0 | — |
Negative control K | 0 | 0 | — |
Table 3. Analytical sensitivity of «Synchytrium endobioticum-RT»
DNA dilution | Concentration of fungal cells, cells/ml | Synchytrium endobioticumRT |
Cycle threshold Ct |
1 | 2 | 3 |
0 | 105 | 17.86 | 17.76 | 18.03 |
1 | 104 | 22.02 | 21.53 | 21.99 |
2 | 103 | 32.06 | 36.06 | 30.92 |
3 | 102 | 39.49 | — | — |
4 | 101 | — | — | — |
5 | 100 | — | — | — |
| K+ | 31.98 | 31.15 | 30.92 |
| Kv | — | — | — |
| Kch | — | — | — |
Table 4. DNA extraction from soil samples with Realtime PCR identification
Sample | Number of fungal spores per 100 g of soil | Cycle threshold Ct |
1 | 2 | 3 |
1 | 5 | — | — | — |
2 | 5 | — | — | — |
3 | 50 | 39.61 | — | — |
4 | 50 | 39.24 | — | — |
5 | 500 | 35.42 | 35.10 | 39.47 |
6 | 5000 | 34.16 | 32.21 | 33.49 |
7 | 20000 | 34.41 | 32.80 | 34.65 |
8 | Unknown | 38.15 | 28.74 | 33.88 |
9 | Unknown | 37.41 | 35.55 | 36.97 |
10 | Unknown | 41.61 | 33.20 | 33.56 |
11 | 0 | — | — | — |
K+* | — | 33.48 | 31.98 | 32.17 |
K+z** | — | 17.86 | 17.76 | 18.03 |
K-v | — | — | — | — |
K-ch | -— | — | — | — |
* Positive control of «Synchytrium endobioticum-RT» reagent kit.
** Positive control, DNA, extracted from a potato wart.
About the authors
All-Russian Plant Quarantine Center; Lomonosov Moscow State University
Author for correspondence.
Email: yutska@mail.ru
Junior Researcher, Mycology Laboratory, All-Russian Plant Quarantine Center; PhD student, Lomonosov Moscow State University, Faculty of Biology
32, Pogranichnaya st., Bykovo, Ramensky district, Moscow region, 140150, Russian Federation; 1/12 Leninskie gory, Moscow, 119991, Russian Federation
All-Russian Plant Quarantine Center
Email: yakovleva_va@mail.ru
PhD in Biological Sciences, Leading Researcher, Head of Phytosanitary Biology Department
32, Pogranichnaya st., Bykovo, Ramensky district, Moscow region, 140150, Russian Federation