Safety duplication of banana genetic resources

Contributors to this page: Bioversity International, France (Nicolas Roux), Bioversity International, Belgium (Ines Van den Houwe); IITA, Nigeria (Dominique Dumet, Badara Gueye).

When should it be used

• For all original germplasm collected by the genebank.
• For germplasm held only by the genebank.
• Safety duplicated material should be located in a different country.

See also the general page on safety duplication procedures.

Banana germplasm should be duplicated in a systematic way, consistently sharing information and germplasm to optimize the conservation system.

Material should be duplicated in more than one location and/or using more than one method (i.e. in the field, in vitro or tissue culture and cryopreservation).

Sample specifications

Minimum sample size

• Number of samples for testing
  Bioversity – three cryotubes per accession (experiment).
  IITA – 1-5 seedlings.
• Criterion for long-term storage
  At least 95% certainty that a minimum of one plant can be regenerated per experiment.

Container specifications

To ensure seed viability in the genebank and en route to the users.

Specification of packaging

• Glass test tubes covered and sealed.

Labeling

• Placement of label
  Bioversity – written directly on cryotubes using a pencil.
  IITA – placed on upper half.
• Labeling material
  Bioversity – not applicable.
  IITA – Barcode labels pasted on tubes or on polyethylene bags.
• Label information
  Bioversity – accession ID, freezing date, experiment number.
  IITA – accession number, line number and date of in vitro introduction and  of last introduction.

Storage specifications

• Type of storage:
  Bioversity – cryopreservation in liquid nitrogen tank.
  IITA – tissue culture slow growth.
• Minimal growth media: see composition here.
• Light: 12h light/24h and light intensity: 43 μmol/m²/s.
• Temperature:
  Bioversity –  -196°C
  IITA –  +18 ± 1 °C

Shipping method

• Prepare the documentation of outgoing plant material shipments with phytosanitary certificate, the import permit and other necessary documentation.
• Follow the same procedures for the plant material shipments from the genebank.
• Packages should be shipped using the safest, most reliable, most cost effective and fastest route and carrier possible, to avoid delays and deterioration of plant material quality during transfer.
• Avoid shipments during hot times of the year.

Transport

• Type of container
  Bioversity – dry shipper.
  IITA – plastic boxes.
• Method and duration
  Bioversity – Air courier or hand carried.
  IITA – car (4x4) 5 to 6 hours drive.
• Conditions
  Bioversity – frozen in liquid nitrogen.
  IITA – ambient temperature.
• Frequency of shipment
  Bioversity – initially 3-4 times a year; once a year thereafter.
  IITA – every 3-4 months.

Storage duration

• Life expectancy of clones
  Bioversity – indefinite.
  IITA – 4-6 months.

Legal arrangements

The genebank, host country and importing centre and country policies and practices for germplasm movement for black-box storage must be fulfilled. The documentation listed below should be included to facilitate customs clearance:

• MOU for safety back-up.
• Phytosanitary certificate.
• Certificate of origin.
• Certificate of no-commercial value.
• Electronic and hard copy of associated passport information.
• GMO-free certificate (if required).

IITA-Nigeria has the following procedure:

• An agreement with IITA-Cotonou to keep a black-box, including:
  • Special agreement with the Plant Quarantine Services of Benin to use only an import permit and no need for phytosanitary certificates.
  • Germplasm cannot be used in Benin or distributed elsewhere.
  • If material needs to be transferred to the field, should be first repatriated to Nigeria.

Bioversity follows slightly different procedures:

• An agreement with IRD, France & KULeuven, Belgium to keep banana samples maintained as safety duplicates in liquid nitrogen (cryopreservation).

Data arrangements

Bioversity – germplasm ID, inventory of black-box content sent with samples.
IITA – import permit stating list of accessions transferred, endorsed by PQS prior to departure. Report at boarder PQS office.
 

Provision for return of germplasm

Bioversity – loss of samples from LTS at Bioversity ITC; sample provided by the duplication site at the request of Bioversity, giving six-months' written notice (if less than 95% certainty that one minimum plant can be regenerated per experiment).
IITA – repatriation permit to request material when needed.

Recording information during safety duplication

The following steps should be taken regularly, due to the short-term duration of conserving clonal crops:

• Updated lists of existing germplasm should be regularly sent to the owner of the germplasm (every 3-4 months in the case of IITA).
• Lists should be checked and any new material (not duplicated in the recipient genebank) should be identified.
• Depending on seedling availability at the owner’s genebank, seedlings should start to be prepared and the recipient genebank informed to start preparing the relevant documentation.

The following information should be kept up-to-date:

• Number of seedlings per accession.
• Date of production of seedling.
• Date of safety duplication.
• Name of institute holding the safety duplicate.
• Box label where the sample is placed.

References and further reading

FAO/IPGRI. 1994. Genebank standards. Food and Agriculture Organization of the United Nations, Rome and International Plant Genetic Resources Institute, Rome. Available in English, Spanish, French and Arabic.