Testing service For diagnosis of plant pathogens Edition 2011 CHF www.bioreba.com BIOREBA AG Your Partner in Agro-Diagnostics Christoph Merian-Ring 7 CH-4153 Reinach BL1 phone +41 61 712 11 25 admin@bioreba.ch Switzerland fax +41 61 712 11 17 www.bioreba.com
BIOREBA Contents Quality Management... 2 BIOREBA Testing Service... 3 Testing Methods... 3 Testing Procedure... 4 Sampling... 4 Service offer Potato Tests... 5 Grapevine Tests...6-7 Vegetable Tests...8-9 Field Crop Tests (cereals, beet, tobacco)... 9 Fruit Tree and Small Fruit Tests... 10 Ornamental Tests... 11 Prices... 12 Quality Management BIOREBA AG has been awarded the internationally recognized certificate of Quality Management System ISO 9001 and our testing service has been accredited according to ISO/IEC 17025 (Swiss Testing Service No. 541) for all ELISA tests and for most PCR tests offered in this brochure. STS 541 2
BIOREBA Testing Service BIOREBA Testing Service provides you its expertise in diagnostic assays for the detection of plant pathogens (viruses, bacteria, fungi and phytoplasma). More than 25 years of experience with analysis of samples of most agricultural sectors and employment of different state-of-the-art methods and technologies guarantee the highest possible accuracy of test results. Each case is treated in close communication with the customer to determine the optimal sampling and transport of the samples, as well as the arrangement of the most appropriate testing method and terms. This to make sure each customer gets the right service for its needs in the most cost effective and rapid way. Our testing service is in accordance to the strict rules and quality standards according to ISO/IEC 17025 and confidentiality is maintained throughout communications and data processing. Testing Service Testing Methods The serological method DAS-ELISA (double antibody sandwich enzyme-linked immunosorbant assay) is primarily used for the analysis of plant samples for the presence of specific pathogens. This method is economic and suitable in most cases and is based on the same BIOREBA reagents that are sold to our customers world-wide. We also offer the analysis of your samples with the molecular biological method polymerase chain reaction (classical or real-time PCR). DA S - ELISA. Antigen (pathogen) present in the sample is recognized by specific antibodies in microtiter plates. A color change indicates a positive result, whereas negative samples remain colorless. PCR. Pathogen-specific DNA or RNA is amplified and then detected as fluorescent band with specific size in a gel. Real-time PCR. Pathogen-specific DNA or RNA is amplified and recorded real-time as fluorescent increase. This method allows to quantify the pathogen concentration in the sample. 3
TestingProccedure Testing Procedure / Sampling 1) Please contact us by phone (+41 61 712 11 25) or email (admin@bioreba.ch) to specify kind of samples and test parameters, and to arrange conditions and timing. 2) We are providing special extraction bags for leaf samples. An import permission is required for customers abroad for certain type of samples. 3) Samples 1 : Leaf samples: Take approx. 0.25-0.50 g of plant material. A piece of 4 x 4 cm or several fragments are generally recommended. Place the sample in the rear compartment of the extraction bag at approx. one third of the heigth of the bag. Label the bag with the number corresponding to the sample. Tubers: Please ship tubers washed and dried (not wet) in a bag permeable to air. Wood samples: Cut canes or branches of 1-3 years old wood of 30-50 cm in length (or at least 2-3 internodes). Do not use dry (dead) wood samples. Label the samples with self-adhesive paper, or another appropriate permanent label according to your sample list. 4) Record your samples in a list consecutively numbered and send us this list electronically by email (e.g. as an Excel sheet). 5) Send the samples with priority (over night) shipping with the lowest delay as possible. Avoid shipping during week-ends or holidays. If required, attach a copy of the import permit issued by the Swiss Federal Office of Agriculture precedingly provided by BIOREBA by email or fax. 6) You will receive the written report of the test results within 5 working days after receipt of your samples by email, fax or letter mail. 1 Sampling: For the timing and selection of the appropriate test sample, it is important to bear in mind that the concentration of pathogens may vary according to season, plant tissue, meteorological conditions, variety of host plant and kind of pathogen. Furthermore, pathogen distribution can be systemic or locally restricted; hence, the appropriate and representative sampling is one of the most crucial factors to get reliable diagnostic results. We recommend to arrange the modality of sampling with us to ascertain that tests can be carried out under optimal conditions. 4
Potato ELISA Tests Abbr. Pathogen Test samples PLRV Potato leafroll virus leaves, tubers, sprouts PVA Potato virus A leaves, tubers, sprouts PVM Potato virus M leaves, tubers, sprouts PVS Potato virus S leaves, tubers, sprouts PVX Potato virus X leaves, tubers, sprouts PVY Potato virus Y leaves, tubers, sprouts Service Offer AMV Alfalfa mosaic virus leaves, tubers, sprouts APLV Andean potato latent virus leaves, tubers, sprouts APMoV Andean potato mottle virus leaves, tubers, sprouts PMTV Potato mop-top virus leaves, tubers, sprouts PVV Potato virus V leaves, tubers, sprouts Rs Ralstonia solanacearum tubers, sprouts, stems Sss Spongospora subterranea f. sp. subterranea tubers, soil and peat substrates 1 TBRV Tomato black ring virus potato strain leaves, tubers, sprouts TNV Tobacco necrosis virus leaves, tubers, sprouts TSWV Tomato spotted wilt virus leaves, tubers, sprouts Verticillium Verticillium spp. stem base 1 Tests of soil and peat substrates is not included in the range of accredited services. PCR Tests Abbr. Pathogen Test samples Cms Clavibacter michiganensis subsp. tubers sepedonicus Rs Ralstonia solanacearum tubers, stem base Dsp 2 Dickeya spp. tubers, stem base Pa 2 Pectobacterium atrosepticum tubers, stem base Pcc 2 Pectobacterium carotovorum subsp. carotovorum tubers, stem base 2 PCR-Tests of Dsp, Pa und Pcc are not induded in the range of accredited services 5
Service Offer Grapevine ELISA Tests Group 1 Abbr. Pathogen Test samples ArMV Arabis mosaic virus leaves 1, canes 3 GFLV Grapevine fanleaf virus leaves 1, canes 3 ArMV+GFLV Arabis mosaic virus + Grapevine fanleaf virus leaves 1, canes 3 RpRSV-ch Raspberry ringspot virus-ch leaves 1, canes 3 RpRSV-g Raspberry ringspot virus-g leaves 1, canes 3 TBRV Tomato black ring virus leaves 1, canes 3 TRSV Tobacco ringspot virus leaves 1, canes 3 ToRSV Tomato ringspot virus leaves 1, canes 3 ToRSV-Ch Tomato ringspot virus-ch leaves 1, canes 3 SLRSV Strawberry latent ringspot virus leaves 1, canes 3 Group 2 Abbr. Pathogen Test samples GLRaV-1 Grapevine leafroll-assoc. virus 1 leaves 2, petioles 2, canes 3 GLRaV-1+3 Grapevine leafroll-assoc. virus 1+3 leaves 2, petioles 2, canes 3 GLRaV-2 Grapevine leafroll-assoc. virus 2 leaves 2, petioles 2, canes 3 GLRaV-3 Grapevine leafroll-assoc. virus 3 leaves 2, petioles 2, canes 3 GLRaV-4-9 Grapevine leafroll-assoc. virus leaves 2, petioles 2, canes 3 generic 4-9 GLRaV-6 Grapevine leafroll-assoc. virus 6 leaves 2, petioles 2, canes 3 GLRaV-7 Grapevine leafroll-assoc. virus 7 leaves 2, petioles 2, canes 3 GVA Grapevine virus A leaves 2, petioles 2, canes 3 GFkV Grapevine fleck virus leaves 2, petioles 2, canes 3 PCR Tests Abbr. Pathogen Test samples GVA 4 Grapevine virus A leaves 2, petioles 2, canes 3 GVB Grapevine virus B leaves 2, petioles 2, canes 3 RSPaV 4 Rupestris stem pitting assoc. virus leaves 2, petioles 2, canes 3 FD Flavescence dorée phytoplasma leaves 2, petioles 2 BN Bois noir phytoplasma leaves 2, petioles 2 1 Leaves from young shoots early in the growing season. 2 Well-developed mature leaves, especially petioles and veins, from the lower part of the plant late in the growing season. 3 Phloem tissue from mature canes during dormancy 4 PCR tests of GVA and RSPaV are not in the range of accredited service 6
Recommendations for Grapevine Testing: Virus Tests We are employing efficient and reliable techniques for detection of viruses in grapevine. However, the virus concentration varies considerably according to tissue source, virus and physiological state of the host in function of time of the season. These facts have to be considered for obtaining reliable results. Service Offer Basically, all viruses can be detected in mature canes during dormancy. When testing leaves, however, viruses are separated into two distinct groups having each their specific requirements. Under climatic conditions of Switzerland and Middle Europe, following recommendations for tissue sampling can be given: For viruses of group 1 (see list on page 6) Preferable Leaves from young shoots and juicy bark early in the growing season as well as bark (phloem) scrapings from mature canes during dormancy. Not recommended Samples collected during the hot summer or old leaves in fall. For viruses of group 2 (see list on page 6) Preferable Well-developed mature leaves, especially petioles and veins, from the lower part of the plant late in the season as well as bark (phloem) scrapings from mature canes during dormancy. Not recommended Young tissue early in the growing season. For testing root stocks, wood of mature canes is the only reliable sample tissue. Phytoplasma Tests Phytoplasma (Flavescence dorée, Bois noir) are best detectable in veins and petioles of well-developed mature leaves late in the growing season, as for viruses of group 2. During the rest of the year, phytoplasma are preferably detected in root tissue. 7
Service Offer Vegetables ELISA Tests Abbr. Pathogen Test plants (examples) Aac Acidovorax avenae subsp. citrulli watermelon, cucurbits BCMNV Bean common mosaic necrosis virus bean BCMV Bean common mosaic virus bean BGMV* Bean golden moasic virus bean BNYVV Beet necrotic yellow vein virus beet, spinach CaMV Cauliflower mosaic virus cabbage, cauliflower, broccoli, turnip, radish etc. CMV Cucumber mosaic virus tomato, bean, celery, cucurbits, pepper, spinach, etc. CYSDV Cucumber yellow stunting cucurbits, etc. disorder virus GCLV Garlic common latent virus garlic, leek, and other Allium spp. ErLV Erysimum latent virus various LMV Lettuce mosaic virus lettuce, pea, spinach, etc. LYSV Leek yellow stripe virus leek, rarely in other Allium spp. MiLBVV Mirafiori lettuce big-vein virus lettuce OYDV Onion yellow dwarf virus garlic, leek, onion, shallot, and other Allium spp. PepMV Pepino mosaic virus tomato PMMoV Pepper mild mottle virus pepper Poty Potyvirus various group PRSV Papaya ringspot virus (=watermelon mosaic virus 1) cucumber, squash, watermelon, other cucurbits, papaya, etc. PVX Potato virus X potato, tomato PVY Potato virus Y pepper, potato, tomato RaMV Radish mosaic virus cabbage, arugula SLCV* Squash leaf curl virus bean, cucurbits SLV Shallot latent virus garlic, leek, onion, shallot, and other Allium spp. SqMV Squash mosaic virus cucumber, squash, other cucurbits TBSV Tomato bushy stunt virus tomato, pepper, artichokes TCSV** Tomato chlorotic spot virus tomato TNV Tobacco necrosis virus bean, cucurbits, tomato ToMoV* Tomato mottle virus Tomate, Bohne ToMV Tomato mosaic virus tobacco, tomato, etc. ToRSV Tomato ringspot virus cucumber, rhubarb * detected wiht the TYLCV reagent ** detected with the TSWV reagent 8
TSV Tobacco streak virus bean, pea, pepper, tomato, etc. TSWV Tomato spotted wilt virus bean, cucurbits, lettuce, spinach, tomato, etc. TuMV Turnip mosaic virus Brussels sprout, cabbage, cauliflower, turnip, etc. TYLCV Tomato yellow leaf curl virus tomato, bean, squash TYMV Turnip yellow mosaic virus turnip, cabbage Verticillium Verticillium spp. tomato, celery WMV 1 Watermelon mosaic virus 1 see PRSV Service Offer WMV 2 Watermelon mosaic virus 2 cucumber, pumpkin, squash, watermelon, etc. ZYMV Zucchini yellow mosaic virus cucumber, watermelon, zucchini, squash, etc. Field crops (cereals, beet, tobacco) ELISA Tests Abbr. Pathogen Test plants (examples) AMV alfalfa mosaic virus alfalfa, bean, pea, potato, tomato, etc. BNYVV beet necrotic yellow vein virus beet, spinach, etc. BYDV-B barley yellow dwarf virus-b barley, oat, wheat, triticale, maize BYDV-F barley yellow dwarf virus-f barley, oat, wheat, triticale, maize BYDV-RPV barley yellow dwarf virus-rpv barley, oat, wheat, triticale, maize CaMV cauliflower mosaic virus cabbage, cauliflower, broccoli, turnip, radish etc. CMV cucumber mosaic virus bean, clover, beet, cucurbits, potato, tobacco, etc. CYDV-RPV cereal yellow dwarf virus-rpv synonym: BYDV-RPV MDMV maize dwarf mosaic virus maize, sorghum (foodder, grain and weed) PeAMV petunia asteroid mosaic virus hop, spinach Poty group poty virus various PVY potato virus Y tobacco RaMV radish mosaic virus rape, mustard, cabbage, arugula etc. SCMV sugarcane mosaic virus sugarcane, occasionally sorghum, maize and other Gramineae TMV tobacco mosaic virus bean, beet, tomato, tobacco, etc. TNV tobacco necrosis virus bean, beet, cucurbits, potato, soybean, tomato, tobacco, etc. TSV tobacco streak virus alfalfa, bean, cotton, soybean, tomato, tobacco, etc. TSWV tomato spotted wilt virus cucurbits, peanut, potato, soybean, tobacco, etc. TYMV turnip yellow mosaic virus turnip, cabbage, mustard, rape 9
Service Offer Fruit trees & small fruits ELISA tests Abbr. Pathogen Test plants (examples) ACLSV apple chlorotic leaf spot virus apple, pear, plum/prune, cherry, peach, quince, apricot ApMV apple mosaic virus apple, hop, plum/prune (very rare), ApP apple proliferation phytoplasma apple ArMV arabis mosaic virus cherry, peach, elderberry, hop, raspberry, strawberry ASGV apple stem grooving virus apple ASPV apple stem pitting virus apple, pear CLRV cherry leaf roll virus cherry, peach, walnut, elderberry CTV citrus tristeza virus citrus PDV prune dwarf virus cherry, peach, plum/prune PeAMV petunia asteroid mosaic virus cherry, plum PNRSV prunus necrotic ringspot virus cherry, plum/prune, apricot, peach, hop Poty poty virus various group PPV plum pox virus (Sharka) plum, apricot, peach, P. mahaleb and other Prunus spp., sour cherry PRSV papaya ringspot virus papaya RBDV raspberry bushy dwarf virus raspberry, black raspberry RpRSV raspberry ringspot virus cherry, plum/prune, raspberry, strawberry, redcurrant SLRSV strawberry latent ringspot virus plum/prune, peach, cherry, elderberry, raspberry, strawberry, blackberry, blackcurrant, redcurrant SMYEPV strawberry mild yellow edge strawberry potexvirus ToRSV tomato ringspot virus apple, peach, cherry, plum/prune, raspberry TRSV tobacco ringspot virus cherry, blueberry, papaya TSV tobacco streak virus strawberry, raspberry (red and black) Verticillium Verticillium spp. apple, rose, maple Xfr Xanthomonas fragariae strawberry PCR tests Abbr. Pathogen Samples Phytoplasma Phytoplasma general in leaf veins, petioles, or roots of pomaceous, stone and small fruits PPV plum pox virus (Sharka) in leaves of stone fruits (Prunus spp.) 10
Ornamentals ELISA Tests Abbr. Pathogen Test plants (examples) Aacat Acidovorax avenae subsp. orchids cattleyae AMV alfalfa mosaic virus lupine, Petunia ApMV apple mosaic virus periwinkle, Petunia, rose, other Rosaceae spp. ArMV arabis mosaic virus forsythia, narcissus, Petunia, Prunus spp., rose, etc. CbMV calibrachoa mottle virus Calibrachoa, Petunia CMV cucumber mosaic virus begonia, Chrysanthemum, geranium, gladiolus, Impatiens, lily, orchids, Petunia, etc. ErLV erysimum latent virus various INSV impatiens necrotic spot virus begonia, carnation, Chrysanthemum, dahlia, geranium, gladiolus, Impatiens, narcissus, orchids, Petunia, etc. PeAMV petunia asteroid mosaic virus Petunia PFBV pelargonium flower break virus geranium PLCV pelargonium leaf curl virus geranium PLPV pelargonium line pattern virus geranium PNRSV prunus necrotic ringspot virus Petunia, rose Poty group poty virus various PVA potato virus A Petunia PVX potato virus X Petunia PVY potato virus Y dahlia, Petunia Rs Ralstonia solanacearum geranium SLRSV strawberry latent ringspot virus gladiolus, narcissus, rose TMV tobacco mosaic virus geranium, Petunia TNV tobacco necrosis virus tulip ToMV tomato mosaic virus Petunia ToRSV tomato ringspot virus Chrysanthemum, carnation, geranium, gladiolus, Impatiens, orchids, Petunia, rose, etc. TRSV tobacco ringspot virus begonia, Chrysanthemum, geranium, gladiolus, Impatiens, lily, narcissus, Petunia, rose, etc. TSV tobacco streak virus dahlia, Impatiens, Petunia TSWV tomato spotted wilt virus begonia, carnation, Chrysanthemum, dahlia, geranium, gladiolus, Impatiens, orchids, Petunia, etc. Verticillium Verticillium spp. Chrysanthemum, dahlia, rose Xcp Xanthomonas campestris pv. pelargonii geranium Service Offer 11
Prices of ELISA tests Price per test (CHF) Prices No. samples for 1 pathogen per sample for each additional pathogen per sample 1-5 25. 12. 6-20 19. 10. 21-60 16. 8. 61-139 14. 6.50 ab 140 12. 5. Additional costs: A basic fee of CHF 150. is added to each order. For extraction of wood, bulbs, or onion, an additional fee of CHF 3. /sample will be charged. Additional costs for testing tubers on request. Prices are in CHF, VAT not included (but not charged for customers outside Switzerland). For large or standing orders ask for a special quote. Prices of PCR tests On request. In function of host, pathogen(s), and number of test samples. 12 Version: 5-15.08.2011