Commission Implementing Regulation (EU) 2022/1195 of 11 July 2022 establishing measures to eradicate and prevent the spread of Synchytrium endobioticum (Schilbersky) Percival
(1) "specified pest" means Synchytrium endobioticum (Schilbersky) Percival;(2) "specified plants" means plants of Solanum tuberosum L. other than seeds.
1. The competent authorities shall designate a production site as infested by the specified pest where the presence of the specified pest in that site has been officially confirmed by the tests referred to in Article 3(2). 2. Specified plants grown in a production site designated as infested by the specified pest or which have been in contact with soil in which the specified pest has been found shall be officially designated as infected.
(a) an infested zone, including at least the production site designated as infested; and (b) a buffer zone, surrounding the infested zone.
(a) no specified plants shall be planted, grown or stored; (b) no other plants, intended for replanting outside the infested zone, shall be grown or stored, both in the ground or anywhere else; (c) soil shall be removed from plants other than those referred to in points (a) and (b), by appropriate methods ensuring that there is no identifiable risk of spreading the specified pest, before these plants are moved from the infested zone into the buffer zone, or out of the demarcated area, or immediately after; (d) machinery shall be cleaned from soil and plant debris, before or immediately after being moved out of the infested zone and before entering any production site located in the buffer zone or outside of the demarcated area; (e) any soil or debris originating from an infested zone may only be moved and used or deposited outside that zone under conditions ensuring that there is no identifiable risk of spreading the specified pest.
(a) they are transported for the purpose of removing soil from those plants by appropriate methods ensuring that there is no identifiable risk of spreading the specified pest; (b) the transport and the removal of soil take place under official supervision, and appropriate measures have been put in place to effectively prevent the spread of the specified pest.
(a) in the buffer zone, no plants intended for replanting outside the demarcated area are grown; (b) in the buffer zone, only specified plants are grown of a variety, which is resistant to the pathotypes of the specified pest found in the infested zone or to all pathotypes known to occur in their Member State, as provided for in Article 7, and other than for the production of specified plants for planting; and (c) any soil or debris originating from the buffer zone is moved and used or deposited outside the demarcated area under conditions, such that there is no identifiable risk of spreading the specified pest.
(a) soil sieving method, as described by Pratt (1976) Pratt MA. 1976. A wet-sieving and flotation technique for the detection of resting sporangia of Synchytrium endobioticum in soil.Annals of Applied Biology 82: 21 – 29.(b) soil sieving method, as described by van Leeuwen et al. (2005)van Leeuwen GCM, Wander JGN, Lamers J, Meffert JP, van den Boogert PHJF, Baayen RP. 2005. Direct examination of soil for sporangia of Synchytrium endobioticum using chloroform, calcium chloride and zinc sulphate as extraction reagents.EPPO Bulletin 35: 25 – 31.(c) zonal centrifuge technique for high throughput sample processing, as described by Wander et al. (2007)Wander JGN, van den Berg W, van den Boogert PHJF, Lamers JG, van Leeuwen GCM, Hendrickx G, Bonants P. 2007. A novel technique using the Hendrickx centrifuge for extracting winter sporangia of Synchytrium endobioticum from soil.European Journal of Plant Pathology 119: 165 – 174.
(a) morphological identification under a light microscope at 100x – 400x magnification; (b) conventional PCR using primers based on Lévesque et al. (2001) Lévesque CA, de Jong SN, Ward LJ & de Boer SH (2001) Molecular phylogeny and detection of Synchytrium endobioticum , the causal agent of potato wart.Canadian Journal of Plant Pathology 23: 200–201.et al. (2005)van den Boogert PHJF, van Gent-Pelzer MPE, Bonants PJM, de Boer SH, Wander JGN, Lévesque CA, van Leeuwen GCM, Baayen RP. 2005. Development of PCR-based detection methods for the quarantine phytopathogen Synchytrium endobioticum , causal agent of potato wart disease.European Journal of Plant Pathology 113: 47 – 57.(c) real-time PCR using primers and probes following van Gent-Pelzer et al. (2010)van Gent-Pelzer MPE, Krijger M, Bonants PJM. 2010. Improved real-time PCR assay for the detection of the quarantine potato pathogen, Synchytrium endobioticum , in zonal centrifuge extracts from soil and in plants.European Journal of Plant Pathology 126: 129 – 133.(d) real-time PCR using primers and probes following Smith et al. (2014) Smith DS, Rocheleau H, Chapados JT, Abbott C, Ribero S, Redhead SA, Lévesque CA, De Boer SH. 2014. Phylogeny of the genus Synchytrium and the development of TaqMan PCR assay for sensitive detection ofSynchytrium endobioticum in soil.Phytopathology 104: 422 – 432.
(a) method of SASA (Science and Advice for Scottish Agriculture), consisting of the two following steps: (i) production of inoculum Old (brown) wart tissue shall be broken into smaller pieces and air dried at room temperature until it becomes hard. The hard tissue shall be ground, either manually or mechanically. The ground material shall be dry-sieved, collecting the fraction from 25 to 75 μm, and then extracted using the chloroform method of Pratt (1976) 1 ;(ii) production of fresh warts Approximately 10 mg of extracted resting spores shall be sprinkled onto the surface of 10 ml sterile distilled water in a small plastic Petri dish and incubated in the dark at 20 °C until germination. Potato tubers with small sprouts about 1 to 2 mm long shall be placed in transparent plastic boxes, lined with damp tissue paper with the marked sprouts facing up. The sprouts shall be ringed with melted Vaseline using a syringe. The ring shall be unbroken and high enough to hold the spore suspension without leaking. The 10 ml of germinating resting spores shall be diluted further to 20 ml with sterile water and placed within the rings using a pipette or a squeeze bottle until the sprout is completely submerged in spore suspension. The plastic boxes shall be covered with lids and incubated for 4 days at 10 °C, after which the boxes shall be opened, the inoculum and Vaseline rings shall be removed and the boxes shall be moved to a misted glasshouse at 15 to 18 °C (16 h light);
(b) method of Spiekermann & Kothoff (1924) Spieckermann A, Kothoff P. 1924. Testing potatoes for wart resistance. Deutsche Landwirtschaftliche Presse 51: 114 – 115.(c) method of Potoček et al. (1991)Potoček J, Krajíčková K, Klabzubová S, Krejcar Z, Hnízdil M, Novák F, Perlová V. 1991. Identification of new Synchytrium endobioticum (Schilb.) Perc. pathotypes in Czech Republic.Ochrana Rostlin 27: 191 – 205.(d) method of Glynne-Lemmerzahl (Glynne 1925 Glynne MD. 1925. Infection experiments with wart disease of potatoes. Synchytrium endobioticum .Annals of Applied Biology 12: 34 – 60.Lemmerzahl J. 1930. A new simplified method for inoculation of potato cultivars to test for wart resistance. Züchter 2: 288 – 297.Noble M, Glynne MD. 1970. Wart disease of potatoes. FAO Plant Protection Bulletin 18: 125 – 135.
| Cultivar | |||||
|---|---|---|---|---|---|
| 1(D1) | 2(G1) | 6(O1) | 18(T1) | 38(Nevşehir) | |
| Tomensa/Evora/Deodara | S | S | S | S | S |
| Irga/Producent | R | S | S | S | S |
| Talent | R | R* | R* | S | S |
| Saphir | R | S | R | R | S |
| Ikar/Gawin/Karolin/Belita | R | R | R | R | R |
| Category of potatoes | Total cropping area (ha) | Visual inspection of tubers | Laboratory testing | Other information | ||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Number of samples | Number of lots | Size of sample | Sampling period | No of suspicious | Number of samples | Size of samples | Kind of test | No of positive | ||||||
| Samples | Lots | Samples | Lots | |||||||||||
| Potatoes for the production of tubers for planting | ||||||||||||||
| Ware and processing | ||||||||||||||
| Other | ||||||||||||||
1. A minimum of 40 tubers or eye plugs per variety of the specified plant shall be tested. They shall be divided into two groups (replicates). 2. The test shall generally last for 2 years. Only in case that a variety shows to be extremely susceptible to a pathotype of the specified pest, the length of the test may be reduced to 1 year. 3. Before a testing season starts, the inoculum shall be tested for purity, using the methods described in Annex I. 4. A positive control, in the form of a variety of the specified plant, which is extremely susceptible to the pathotype of the specified pest to be tested, shall always be included in the test. 5. One of the following testing methods shall be used: (i) the Glynne-Lemmerzahl method (Glynne 1925, Lemmerzahl 1930, Noble & Glynne 1970); (ii) the Spieckermann method (Spieckermann & Kothoff 1924); or (iii) the SASA (Science and Advice for Scottish Agriculture) method, consisting of all of the following steps: tuber preparation: Tubers shall be removed from the cold store around 10 days before intended inoculation, washed gently, dried and stored in the dark at room temperature to induce sprouting. A highly susceptible variety ("Morene" or a variety with comparable susceptibility) shall be included in each inoculation to serve as positive control; germination of resting spores: Conditions to induce germination of resting spores shall be set up 21 days prior to inoculation. Approximately 10 mg of extracted spores shall be sprinkled onto the surface of 10 ml of sterile distilled water in small plastic Petri dishes and incubated in the dark at 20 °C until germination. The content of each Petri dish shall be diluted with another 10 ml of sterile distilled water for the inoculation; inoculation and incubation of sprouts: When the sprouts reach 1 mm in length, they shall be ringed with melted Vaseline. The Vaseline ring shall be unbroken to hold the spore suspension without leaking and high enough for the suspension to cover the sprout. A single sprout or a single cluster of sprouts shall be ringed on each tuber. The tubers shall be placed in plastic boxes, lined with damp tissue paper with the ringed sprouts facing upwards. The Vaseline rings shall be filled with spore suspension, using a pipette or a squeeze bottle until the sprout is completely submerged. The plastic boxes shall be covered with lids and incubated for 4 days at 10 °C in the dark, after which the Vaseline rings shall be removed and the boxes shall be placed open in a glasshouse at 15–18 °C under periodic misting (3 times per day for 30 min). In cases where the infection failed, for example because the sprout rotted or failed to develop, the tuber may be retested using another sprout; assessment: Sprouts shall be examined for infection 28 days after the inoculation, using a stereo microscope with 10–15x magnification and a light microscope. Reactions of score 4 or 5, as set out in the table, shall be observed on the positive control on at least 80 % of tubers. At least one tuber shall show a score of 5.
6. All tubers shall be assessed and given a resistance ranking score from 1 to 5, as set out in the table. 7. Each tested variety shall be placed in a resistance group ("highly resistant", "resistant", "slightly susceptible", or "extremely susceptible"), according to the range of scores observed within the respective population of individual tubers or eye plugs tested: (i) a variety shall be considered "highly resistant", if all tubers in all replicates have a score of 1; (ii) a variety shall be considered "resistant", if all tubers in all replicates have a score between 1 and 3; (iii) a variety shall be considered "slightly susceptible", if one or more tubers score 4 (if only one tuber scores 4, the test may be repeated, in order to exclude impurity in the variety lot); (iv) a variety shall be considered "extremely susceptible", if at least one tuber in one replicate scores 5. Standard scoring notations for potato testing populations Standard score Group of resistance Resistance description Description 1 R1 Extremely resistant Early defence necrosis; no visible sorus formation. 2 R1 Resistant Late defence necrosis; sorus formation partially visible, sori immature or necrotic before maturity. 3 R2 Weakly resistant Very late defence necrosis; single ripe sori or sorus fields developed, but completely surrounded by necrosis; up to five non-necrotic summer sori permitted, clear necrosis in other zones of the same tuber piece. No formation of warts or resting spores. To decide between groups 3 and 4, it may be necessary to prepare thin slides of infected tissue: if there are no resting spores, the score shall be 3. 4 S1 Slightly susceptible Scattered infections; sori or sorus fields non-necrotic, few in number; late necrosis can be present on other infection sites on the sprout; the sprout can be slightly malformed (thickened). Resting (winter) sporangia are present. To decide between group 3 and 4, it may be necessary to prepare thin slides of infected tissue: if resting spores are present, the score shall be 4. 5 S2 Extremely susceptible Dense infection fields, numerous ripe non-necrotic sori and sorus fields, fields with dense non-necrotic infection sites, predominant wart formation.
no signs of infection with the specified pest have been discovered in two bioassays (as described in point (3) with susceptible potato cultivars; or no signs of infection with the specified pest have been discovered in 1 bioassay (as described in point (3) with susceptible potato cultivars and no viable resting spores have been found during a direct examination of the soil from the infested zone by microscope following an extraction of spores with one of the methods provided for in point 2 of Annex I.
the infested zone shall be divided into units of 0,33 ha each; 60 subsamples shall be taken from each unit to a depth of 20 cm and evenly distributed throughout the area or pooled according to known infested foci; the subsamples shall be thoroughly mixed, so as to obtain 3 samples per ha.
(a) no signs of infection with the specified pest shall be discovered in two bioassays, as described in point 3, with susceptible potato cultivars; or (b) no signs of infection with the specified pest have been discovered in one bioassay, as described in point 3, with susceptible potato cultivars and less than 5 viable resting spores per gram of soil have been found during a direct examination of the soil from the infested zone by microscope following an extraction of spores with one of the methods provided for in point 2 of Annex I.
the infested zone shall be divided into units of 0,33 ha each; 60 subsamples shall be taken from each unit to a depth of 20 cm and evenly distributed throughout the area or pooled according to known infested foci; the subsamples shall be thoroughly mixed, so as to obtain 3 samples per ha.