Sample details being recoreded on packets in the field Sampling of tree for fungal infection of the stem Leaves dried on absorbant paper ready for packaging Staining of vascular tissue caused by fungal infection


Sending material to the
Global Plant Clinic

If possible please contact the Global Plant Clinic before sending samples. Sample submission forms are available via the user login screen (see Forms for more details).

Sending plant material
 Packaging plant material
 Sending Culture specimens
 Packaging of Culture specimens
 Addressing specimens
 When to send specimens
 Dispatch checklist
 Use of Black Light to induce sporulation

 

SENDING PLANT MATERIAL

  • Specimens of fungi on host material should be thoroughly dried prior to dispatch to prevent the growth of unwanted saprophytes. It is recommended that leaves are pressed during drying to avoid curling. Fresh, damp material should not be sent as it rapidly deteriorates in transit
  • Never wrap herbaceous plant material in plastic. Exceptions include when nematodes are suspected, see nematode sending sheets
  • Roots: shake off excess soil and wrap each sample individually in newspaper
  • Stems and leaves: include healthy and diseased sections of stems. Wrap separately in paper. Avoid thin absorbent paper like tissues as these disintegrate and are difficult to remove
  • Soil: remember to double bag soil samples, with labels included with the soil
  • Delicate specimens are best stuck down onto card before being sent
  • Each sample should be placed inside a separate envelope and labelled with the sender's own reference number for the sample
  • Senders are advised to retain a representative collection of each sample, as material submitted for identification is not normally returned

PACKAGING PLANT MATERIAL

  • Pack carefully to avoid damage. Paper, cardboard or polystyrene granules cushion specimens
  • Package in a sturdy crush-proof container and pack with additional paper to prevent shifting
  • Samples should be insulated from wide temperature fluctuations
  • All samples sent abroad should be via airmail and include the necessary documentation to clear customs from your own country and into the UK
  • Before posting soil, please ensure you have included the import licence form
  • If sending killed and fixed nematodes in glass sample bottles, carefully wrap individual bottles and ensure they do not leak. Parafilm provides a good watertight seal
  • If sending mounted slides, a slide carrier, or slide box is best

SENDING CULTURE SPECIMENS

  • Send fresh isolates as soon as growth has been established in subcultures. Ideally they should not be more than 1-3 weeks old on arrival. Cultural mutants or non-sporulating strains cannot be identified. Non-sporulating cultures can sometimes be induced to sporulate by use of near-ultraviolet light (‘black light’)
  • Cultures should be grown on firm media. Suitable agars include Potato Carrot Agar, 2% Malt Agar or Oat Agar. Retain subcultures of the fungus sent for identification, as cultures are not generally kept at CABI Bioscience or returned after identification
  • Send cultures in: 30ml Universal containers; small McCartney bottles (either glass or plastic); small test tubes, (100 x 15 mm) or as freeze-dried ampoules. Petri dishes are not recommended for those sending cultures from overseas as these are easily damaged or contaminated in transit.
  • Check that bottle caps are not over-tightened. If the seal is too tight the fungi are likely to die and so cannot be subcultured on arrival
  • If using test tubes, they should be fitted with cotton-wool plugs which are best sterilized by flaming
  • Please avoid sending cultures in opaque plastic bottles and tubes as these do not permit clear observation of fungal colonies
  • Check for mite contamination and seal cultures. Mites introduce rapidly-growing contaminants that are easily spread. Assignments of cultures containing mites will be destroyed on arrival at CABI Bioscience. The cigarette paper sealing technique is a useful way of protecting cultures from mite invasion
  • Ensure all tubes are labelled with an adhesive label or with waterproof ink. Avoid grease pencil, as it easily rubs off
  • Culture tubes, ampoules and bottles should be wrapped separately and well packed in cotton wool or polystyrene granules. Do not use straw or material that may introduce contaminants into the cultures
  • We recommend that you keep a subculture of each isolate submitted in case there is a need to resubmit material, and so that you can examine your cultures after we have sent you our report

PACKAGING OF CULTURE SPECIMENS

  • When sending living cultures through the post it is strongly recommended that the international regulations be followed
  • Tubes, bottles or ampoules should be individually wrapped using strong absorbent material and placed in a primary leak-proof container. This primary container should not be metallic as the metal prevents the use of x-ray scanners at customs and have not been approved for use with certified postal packaging
  • The primary container should then be placed inside a secondary container which is approved for packing according to the United Nations Packing Instruction 602. Any further wrapping of the package should also meet these requirements. The primary container should have been passed and tested for use with the complete postal package and senders should check with their supplier if their primary container meets these specifications
  • Further information on the regulations covering postal packages can be obtained from IATA (International Air Transport Association) or can be found at http://www.dsmz.de/postregu.htm
  • These regulations do not apply when sending dead, dried material. However, sturdy packaging is necessary to avoid damage to the material in transit. Our experience has found that a padded envelope alone does not give sufficient protection. If using such an envelope the specimen(s) should be placed inside a folded sheet of stiff cardboard as well

ADDRESSING THE MATERIAL

  • Material ideally should be collected and dispatched on the same day
  • Packages should be clearly marked with the following text: The Global Plant Clinic Diagnostic & Advisory Service, CABI Bioscience, Bakeham Lane, Egham, Surrey TW20 9TY, United Kingdom. Perishable Biological Material. Keep material cool but do not refrigerate (see address labels).
  • Complete section 1 and if necessary section 6 of the MAFF plant health licences and include it with your samples:
    MAFF plant health licence no. PHL 199B/5745(11/2007) fungi and bacteria
    MAFF plant health licence no. PHL 251D/5785(03/2008) viruses
    MAFF plant health licence no    . PHL 199B/5596(03/2007) soil
    MAFF plant health licence no. PHL 199B/5746(11/2007) nematodes
    (MAFF licence numbers subject to change    )
  • Those sending material from other European Union countries must complete boxes 1 and 13; those from non-EU countries need only complete box 1
  • The enquiry number if known
  • Viral material can be sent to the Global Plant Clinic or directly to:
    Dr. Phil Jones, Tropical Virus Unit, Plant Pathogen Interactions DIV, IACR - Rothamsted, Harpenden, Herts, AL5 2JQ (see address labels).
  • The material should be marked: Perishable Biological Material. Keep material cool but do not refrigerate
  • Include the MAFF plant health licence no. (Found on the import licence section 11) and the enquiry number if known

WHEN TO SEND SPECIMENS

  • Try to avoid sending samples over the weekend or before a public holiday.
2007 UK public holidays
April 6th
April 9th
May 7th
May 28th
August 27th
Christmas/New Year Holiday December 24th - January 3rd 2005 (latest date for receiving plant material is 13th December)

DISPATCH CHECK LIST

  • Have you completed the sample submission form (user login) including all relevant information and included a print out with your samples?
  • Have you printed out, completed and included the correct Import licence?
  • Have you completed and included all the necessary forms for dispatching the samples from your country including local permits?
  • Have you contacted the Global Plant Clinic to let them know when the samples are due to arrive?
  • Have you packaged the samples in appropriate materials that will protect the samples during postage and conform to the regulations covering postal packages?
  • Have you checked the dates when the clinic will be closed?
  • If in any doubt about the above instructions and before sending samples please contact the Global Plant Clinic

OUR OBLIGATION

  • To the Convention on Biological Diversity (the Rio Convention) means that CABI Bioscience must ask you if any of the material you send for identification was collected after December 1993. If that is the case then you need to inform us if prior consent was received to collect it. This should have been obtained from the land owner and/or the relevant national authority.

USE OF BLACK LIGHT TO INDUCE SPORULATION

  • Some fungi require irradiation by near ultraviolet light to induce sporulation in culture. Near ultraviolet light is often referred to as ‘black light’ (wavelength 300-380nm). Although black light may affect such factors as pigmentation, spore morphology and the gross morphology of the colony, these effects are not sufficient to interfere with identification
  • In our laboratories the black light unit consists of three 1.22m fluorescent tubes fixed beneath a benchtop and connected to a timing mechanism. A near ultraviolet light tube (Phillips TL 40 W/08) is placed between two cool white tubes (Phillips MCFE 40 W/33) and the cultures are supported on a shelf 320mm below the light source. A wooden shield extending 15cm below the benchtop, is fitted around the unit to prevent light rays from directly striking a person. An alternating cycle of 12 h UV and 12 h darkness should be established as some fungi require a dark period in order to sporulate
  • The fungi are grown in plastic Petri dishes or plastic universal bottles. American 'Pyrex' Petri dishes can also be used. Glass however, is not suitable as it does not allow adequate transmission of near ultraviolet light
  • Many fungi sporulate most successfully on nutritionally weak media such as PCA (Potato Carrot Agar), OA (Oat Agar) or TWA (Tap Water Agar) Unless recommended for particular genera, sugar rich media should be avoided as this promotes excessive amounts of mycelium. The dishes are sealed with electrical tape or sticky tape to prevent rapid drying of the culture and infestation by mites. Cultures should be checked regularly for signs of sporulation
  • Irradiation should start 3-4 days after inoculation. Except in the case of thermophilic or psychrophilic fungi, the optimum temperature range for growth under black light is usually between 21-28°C. Temperatures in excess of 30°C should be avoided as the effects of black light can be lost at high temperatures

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