UC Davis Policies and Practices for Work with Transgenic Plants and Plant Pathogens

Work with recombinant DNA organisms (transgenic plants, animal and microbes) and plant pathogens (local or exotic) conducted at UC Davis must be reviewed by the Institutional Biosafety Committee

Standard Microbiological Practices for Biosafety Level 1 (BSL1) Laboratories

1. Access to the laboratory is limited or restricted when experiments are in progress.

2. Persons wash their hands after they handle viable materials, after removing gloves, and before leaving the laboratory.

3. Eating, drinking, smoking, handling contact lenses, applying cosmetics, and storing food for human use are not permitted in the work areas.

4. Mouth pipetting is prohibited; mechanical pipetting devices are used.

5. Policies for the safe handling of sharps are instituted.

6. All procedures are performed carefully to minimize the creation of splashes or aerosols.

   (Biological safety cabinet is required at BSL2)

7. Work surfaces are decontaminated at least once a day and after any spill of viable material.

8. All cultures, stocks, and other regulated wastes are decontaminated before disposal by an approved decontamination method such as autoclaving. Materials to be decontaminated outside of the immediate laboratory are to be placed in a durable, leakproof container and closed for transport.

9. A biohazard sign can be posted at the entrance to the laboratory whenever human infectious agents are present. The sign may include the name of the agent(s) in use and the name and phone number of the investigator.

10. An insect and rodent control program is in effect.

11. It is recommended that laboratory coats, gowns, or uniforms be worn to prevent contamination or soiling of street clothes and dissemination of plant material or plant pathogens outside of the research area.

Tracking and identity of transgenic plants

All transgenic plants will be labeled as transgenic and be tracked by the researcher from creation to disposal. Tracking of plants should include complete and accurate records of where transgenic plants are being produced, maintained and disposed.

Principal investigators and researchers must inform greenhouse managers of the presence of transgenic plants and any special handling conditions or permits that apply to the crop or experiment. PI name and date should be on each plant or each flat of plants.

Transgenic plants will be adequately segregated from non‐transgenic plants unless experimental conditions or greenhouse space does not allow for such segregation. (Adequate segregation is separation that will not allow seeds or pollen for transgenic s to affect surrounding plants.) If non‐ transgenic plants are being kept closely located with transgenic plants, all plants must be treated as transgenic.

Maintenance of plants in laboratory

Plant will be maintained as described in the Biological Use Authorization including as applicable:

• Bagging of flowers
• Containment of fruit and seed
• Watering
• Staking and trimming
• Disposal
• Monitoring for pests
• Other conditions

Maintenance of plants in greenhouses

Transgenic plants maintained in greenhouses must have regular care and tending on a weekly or daily basis depending on the crop. If laboratory staff is not available for such work, it will be done by greenhouse staff and recharged to principal investigators or departments as necessary.

Plant growth should not extend beyond the borders of the assigned bench space unless specifically authorized by greenhouse staff. Plants should be trimmed and staked regularly as needed to accomplish containment. Fruit should be removed from the plant before senescence and must not be allowed to remain on benchs or floors, if dropped. Flowers should be removed or bagged to prevent pollen dispersal in open pollinated plants. Materials harvested from transgenic plants in the greenhouse must be contained in a sturdy leakproof container with a lid or other seal.

If significant plant growth is noted under benches assigned to transgenic plants, herbicide will be applied by greenhouse staff and charged to the owner of the plants. Abandoned or forgotten plants will tagged for disposal if they have gone without care or tending for one month or longer and may be destroyed at the discretion of the greenhouse manager if no further action is taken by the owner.

If an autoclave bag is being used for collection of plant material it must be removed from the greenhouse or contained within a sturdy container with a lid when work is not taking place. Autoclave bags full of transgenic plant waste are not to be left open on greenhouse floors or in common spaces when not in use.

Transport of plants outside of greenhouses, growth chambers or laboratory buildings

Plant material will be transported as described in the Biological Use Authorization including: Containment to prevent seed or pollen dispersal in transit (eg. container with lid or bagged plants in an open container)

Use of carts or vehicles for transport

Disposal of transgenic or infectious plant material

Laboratory solid waste (solid media, plates and plant material) is double‐bagged in clear autoclave bags with indicator tape attached and transported in a sturdy leakproof container to autoclave.

Tightly sealed bags will not allow steam to enter and will explode or break under autoclave pressure conditions. Bags should be sealed tightly enough to restrict plant material from escaping, but should also allow for steam to enter and exhaust without rupturing the outer bag. For some crops this may mean that the inner bag is sealed tightly with an outer bag with a looser seal.

Waste is autoclaved for a minimum of 30 minutes at 121C, 15 psi. It is recommended that autoclaves be tested and documented for effectiveness regularly by using spore indicator tests. Regulatory agencies (CDFA and APHIS) may also require such testing under certain agricultural quarantine conditions.

Liquid microbiological waste (bacterial cultures) should be brought up to a final volume of 10% bleach, allowed to sit for at least 30 minutes and then poured down the drain.

Soil from transgenic plants should be autoclaved after use as described for solid waste or treated (eg. sifting or fumigation) to eliminate any remaining pathogens, seeds or plant material from the soil.

Greenhouse plants are to be disposed of according to the specifications of the greenhouse site manager and the Biological Use Authorization (BUA). All transgenic materials are to be rendered biologically inactive before disposal to the landfill. This may be accomplished by depositing plants into the designated orange container at the greenhouse labeled “transgenic plants” for heat devitalization by greenhouse staff (8 hours steam heating in a sealed container). Alternately, plants may be bagged inside of the greenhouse and transported in a sturdy leak‐proof container to a lab building for autoclave treatment. If any other methods of biological inactivation are employed, these methods must be specified in the BUA.

Seeds, fruit, flowers, and vegetative propagules should be properly contained so as to prevent dispersal of plant material in transport to the orange transgenic plant bin. Small plants with great potential for seed dispersal should be bagged inside of the greenhouse before removal and transport to disposal bin. Loose plants and bagged material can be thrown into the orange bin.

Pots, trays and other hard plastic horticultural supplies in shared use should be thoroughly cleaned to eliminate and prevent survival of plant material or pathogens. This can be done by researchers or by greenhouse staff on a recharge basis. If it will be too labor intensive to eliminate seed or pathogens form material they can be treated as solid waste (bagged and autoclaved). Metal materials can be autoclaved.