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Commission Directive 2008/61/EC of 17 June 2008 establishing the conditions under which certain harmful organisms, plants, plant products and other objects listed in Annexes I to V to Council Directive 2000/29/EC may be introduced into or moved within the Community or certain protected zones thereof, for trial or scientific purposes and for work on varietal selections (Codified version)
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The testing shall use appropriate laboratory methods and, where appropriate, indicator plants, including Citrus sinensis (L.) Osbeck, C. aurantiifolia Christm. Swing, C. medica L., C. reticulata Blanco and Sesamum L., in order to detect at least the following harmful organisms:
Citrus greening bacterium
Citrus variegated chlorosis
Citrus mosaic virus
Citrus tristeza virus (all isolates)
Citrus vein enation woody gall
Leprosis
Naturally spreading psorosis
Phoma tracheiphila (Petri) Kanchaveli & Gikashvili
Satsuma dwarf virus
Spiroplasma citri Saglio et al.
Tatter leaf virus
Witches' broom (MLO)
Xanthomonas campestris (all strains pathogenic to Citrus).
For diseases such as Blight and Blight-like for which there are no short-term indexing procedures the plant material must be subjected upon arrival to shoot-tip grafting onto seedling stock grown under sterile culture as set out in FAO/IPGRI Technical Guidelines, and the resulting plants subjected to therapy procedures according to point 1.
In the case of Fragaria L., irrespective of the country of origin of the plant material, the testing shall use appropriate laboratory methods and, where appropriate, indicator plants, including Fragaria vesca, F. virginiana and Chenopodium spp. for the detection of at least the following harmful organisms:
Arabis mosaic virus
Raspberry ringspot virus
Strawberry crinkle virus
Strawberry latent ‘C’ virus
Strawberry latent ringspot virus
Strawberry mild yellow edge virus
Strawberry vein banding virus
Strawberry witches' broom mycoplasm
Tomato black ring virus
Tomato ringspot virus
Colletotrichum acutatum Simmonds
Phytophthora fragariae Hickman var. fragariae Wilcox & Duncan
Xanthomonas fragariae Kennedy & King.
In the case of Malus Mill.:
where the plant material originates from a country which is not known to be free of any of the following harmful organisms:
Apple proliferation mycoplasm; or
Cherry rasp leaf virus (American),
the testing shall use appropriate laboratory methods and, where appropriate, indicator plants for the detection of the relevant harmful organisms, and
irrespective of the country of origin of the plant material, the testing shall use appropriate laboratory methods and, where appropriate, indicator plants for the detection of at least the following harmful organisms:
Tobacco ringspot virus
Tomato ringspot virus
Erwinia amylovora (Burr.) Winsl. et al.
In the case of Prunus L., as appropriate for each Prunus species:
where the plant material originates from a country which is not known to be free of any of the following harmful organisms:
Apricot chlorotic leafroll mycoplasm;
Cherry rasp leaf virus (American); or
Pseudomonas syringae pv. persicae (Prunier et al.) Young et al.,
the testing shall use appropriate laboratory methods and, where appropriate, indicator plants for the detection of the relevant harmful organisms; and
irrespective of the country of origin of the plant material, the testing shall use appropriate laboratory methods and, where appropriate, indicator plants for the detection of at least the following harmful organisms:
Little cherry pathogen (non-European isolates)
Peach mosaic virus (American)
Peach phony rickettsia
Peach rosette mosaic virus
Peach rosette mycoplasm
Peach X-disease mycoplasm
Peach yellows mycoplasm
Plum line pattern virus (American)
Plum pox virus
Tomato ringspot virus
Xanthomonas campestris pv. pruni (Smith) Dye.
In the case of Cydonia Mill. and Pyrus L. irrespective of the country of origin of the plant material, testing by appropriate laboratory methods, and, where appropriate, indicator plants, for detection of at least the following harmful organisms:
Erwinia amylovora (Burr.) Winsl. et al.
Pear decline mycoplasm.
Where the plant material originates in a country which is not known to be free of the following harmful organisms:
Ajinashika disease
The testing shall use an appropriate laboratory method. In the event of a negative result, the plant material shall be indexed on the vine variety Koshu and kept under observation during at least two cycles of vegetation.
Grapevine stunt virus
The testing shall use appropriate indicator plants, including the vine variety Campbell Early, and observation shall take place during one year.
Summer mottle
The testing shall use appropriate indicator plants, including the vine varieties Sideritis, Cabernet-Franc and Mission.
Irrespective of the country of origin of the plant material, the testing shall use appropriate laboratory methods and, where appropriate, indicator plants for the detection of at least the following harmful organisms:
Blueberry leaf mottle virus
Grapevine Flavescence dorée MLO and other grapevine yellows
Peach rosette mosaic virus
Tobacco ringspot virus
Tomato ringspot virus (strain ‘yellow vein’ and other strains)
Xylella fastidiosa (Well & Raju)
Xylophilus ampelinus (Panagopoulos) Willems et al.
Bacteria
Clavibacter michiganensis (Smith) Davis et al. ssp. sepedonicus (Spieckermann et Kotthoff) Davis et al.;
Ralstonia solanacearum (Smith) Yabuuchi et al.
Viruses and virus-like organisms
Andean potato latent virus,
Potato black ringspot virus,
Potato spindle tuber viroid,
Potato yellowing alfamovirus,
Potato virus T,
Andean potato mottle virus,
Common potato viruses A, M, S, V, X and Y (including Yo, Yn and Yc) and potato leaf roll virus.
However, in the case of true seed of potato, the indexing procedures shall be carried out in order to detect at least the viruses and virus-like organisms listed above at (a) to (e).
Bacteria
For tubers, test the heel end of each tuber. The standard sample size shall be 200 tubers. However, the procedure can be applied for samples with less than 200 tubers.
For young plants and cuttings, including micro-plants, test the lower sections of the stem and, where appropriate, the roots, for each unit of the plant material.
For testing of progeny tubers, or of stem bases for non-tuber forming species, one normal cycle of vegetative growth after the testing referred to in points 1 and 2, is recommended.
For the material referred to in point 1, the testing method for Clavibacter michiganensis (Smith) Davis et al. ssp. sepedonicus (Spieckermann et Kotthoff) Davis et al. shall be the Community method set out in Annex I to Council Directive 93/85/EEC(1). For the material referred to in point 2, this testing method could be applied.
For the material referred to in point 1, the testing method for Ralstonia solanacearum (Smith) Yabuuchi et al. shall be the test scheme set out in Annex II to Council Directive 98/57/EC(2). For the material referred to in point 2, this testing method could be applied.
Viruses and virus-like organisms, other than potato spindle tuber viroid
The minimum testing for vegetative material (tubers, young plants and cuttings, including micro-plants) shall include a serological test done at or near flowering for each of the specified list of harmful organisms other than potato spindle tuber viroid, and followed by a biological test of material testing negative in the serological test. For Potato leaf roll virus, two serological tests shall be done.
The minimum testing for true seed shall be a serological test or a biological test if no serological test is available. Retesting of a proportion of negative samples and testing of borderline results by another method is highly recommended.
The serological and biological testing referred to in points 1 and 2 shall be done on glasshouse grown plants, sampled from at least two positions on every stem, including a young fully expanded leaflet at the top of each stem and an older leaflet from a midway position; each stem shall be sampled because of possible non-systemic infection. In the case of the serological testing, no bulking of leaflets from separate plants shall be done, unless the bulking rate has been validated for the method of use; leaflets from each stem may however be bulked to make up the sample from each plant. In the case of the biological testing, the maximum bulking is up to five plants with inoculation of a minimum of duplicate indicator plants.
The appropriate indicator plants to be used for the biological testing referred to in points 1 and 2 shall be those listed by the European and Mediterranean Plant Protection Organization (EPPO), or other officially approved indicator plants, which have been shown to detect the viruses.
Only material which has been directly tested shall be released from quarantine. Where eye indexing has been done, only the progeny of the tested eye may be released. The tuber should not be released because of possible problems with non-systemic infection.
Potato spindle tuber viroid
For all material, glasshouse grown plants shall be tested, as soon as they are well established but prior to flowering and pollen production. Testing on tuber sprouts/in vitro plants/small seedlings shall only be regarded as a preliminary test.
Samples shall be taken from a fully expanded leaflet at the top of each stem of the plant.
All material for testing shall be grown at temperatures not less than 18 °C (preferably at temperatures higher than 20 °C) and with at least a 16-hour photo-period.
Testing shall be by radioactive or non-radioactive labelled cDNA or RNA-probes, return-PAGE (with silver staining) or RT-PCR.
The maximum bulking rate for probes and return-PAGE is 5. Use of this or higher bulking rates must be validated.
OJ L 259, 18.10.1993, p. 1. Directive as amended by Commission Directive 2006/56/EC (OJ L 182, 4.7.2006, p. 1).
OJ L 235, 21.8.1998, p. 1. Directive as amended by Commission Directive 2006/63/EC (OJ L 206, 27.7.2006, p. 36).
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