USE OF PLANTS IN RESEARCH

Plant research is vital and can help to answer important questions related to agriculture, the environment, and basic life processes. In general, plant research does not generally pose a risk to human health. However, research involving plant diseases, genetically modified plants, or plant pests and organisms associated with plants can pose a hazard and be detrimental to agricultural crops and the environment. Therefore, specific levels of containment may be required to prevent any potential release of any modified plant or associated organisms. The movement, use, possession or release of exotic or potentially harmful plant-associated arthropods, biological control agents, plant pests, plant pathogens, noxious weeds and invasive plants are regulated by local, state, and federal agencies.

Regulation and Oversight

Transgenic plants and plant pests are subject to federal guidelines, regulations, and rules pertaining to their containment, movement, and release into the environment. In addition, States may have applicable regulations as well. 

NIH Guidelines

The NIH Guidelines describes and details risk assessment and containment measures for various biological experiments involving the use of recombinant or synthetic nucleic acid molecules (rsNAM).  These guidelines originally focused on experiments conducted with rsNAM microorganisms. However, the Guidelines have been expanded over the years to include a wide range of federally funded research projects, including plant research. Appendix L of the NIH guidelines provides information related to physical and biological containment (i.e. Plant Biosafety Levels BL1-P to BL4-P and growth chambers) for rsNAM research involving plants, plant-associated pests, animals, and plant pathogens in greenhouses. Any researcher conducting experiments with transgenic plants and/or plant associated organisms must receive IBC approval before beginning such work.

USDA-APHIS

The USDA Animal and Plant Health Inspection Service (APHIS) has authority under the Federal Plant Protection Act to protect U.S. agriculture from pests and disease.  This includes rsNAM-containing plants and potential plant pests. There are two operational programs within APHIS devoted to plants and their associated organisms. 

Plant Protection and Quarantine (PPQ) Focuses on risks associated with entry, establishment, or spread of animal and plant pests. Permits are required for the importation into the U.S. and transit through the U.S. of regulated plants and plant products for consumption or propagation.

Biotechnology Regulatory Services (BRS) Focuses on the risks associated with the introduction of organisms developed using genetic engineering that may pose a danger to U.S. agriculture and the environment. Permits are required for the importation, interstate movement, or environmental release of genetically engineered organisms that pose a risk to plant health.

More detailed infomation can be found at https://www.aphis.usda.gov/biotechnology-permits

HHS – USDA Select Agents and Toxins

Select Agents and Toxins (SAT) are biological agents that have been determined to have the potential to pose a significant threat to the health of humans, animals, plants, or animal and plant products.  The SAT program is jointly administered by the CDC (HHS) and APHIS (USDA). APHIS is the lead agency for regulating agricultural pests and products. Lists of select agents and regulations can be found at https://www.selectagents.gov/

Plant Biosafety Levels

Research involving plants or plant-associated pests, generally, does not pose a hazard to humans but that does not mean that there is no reason for concern. The main reason for biocontainment regarding plant research is to prevent the transfer of propagules and other organisms from a containment area to agricultural areas or the environment. Four levels of biocontainment are described in Appendix L of the NIH Guidelines for greenhouses and growth chambers. Biosafety Levels provide a description of a combination of administrative controls, work practices and procedures, equipment and facility features required to achieve a designed level of containment. Experiments may be assigned to one of four levels of biosafety containment (BL1-P, BL2-P, BL3-P, & BL4-P).

Biosafety Level 1 for Plants (BL1-P)

BL1-P is designed to provide a low level of containment for experiments involving transgenic plants where there is no evidence that a modified organism would be able to survive and spread in the environment and if accidentally released would not pose an environmental risk. 

Biosafety Level 2 for Plants (BL2-P)

BL2-P is assigned to experiments with transgenic plants and associated organisms, which if released could be viable in the surrounding environment and could have a negligible impact or could be readily managed.

Biosafety Level 3 & 4 for Plants (BL3-P & BL4-P)

BL3-P & BL4-P containment is designed to prevent accidental release of transgenic plants and plant associated organisms and non-GE plant research that involves exotic infectious agents that have the potential for significant detrimental impact on agriculture and the environment. Stony Brook University does not currently have BL3-P or BL4-P containment facilities.

Plant Containment Practices and Handling

Appropriate selection of the following biological containment practices may be used to meet containment requirements for a given organism. The present list is not exhaustive; there may be other ways of preventing effective dissemination that could possibly lead to establishment of the organism or it’s genetic material in the environment resulting in deleterious consequences to manage natural ecosystems. 

Biological Containment Practices (Plants)

Effective dissemination of plants by pollen or seed can be prevented by one or more of the following procedures:

• Cover the reproductive structures to prevent pollen dissemination at flowering and seed dissemination at maturity.
• Remove reproductive structures by employing male sterile strains or harvest the plant material prior to the reproductive stage.
• Ensure that experimental plants flower at a time of year when cross-fertile plants are not flowering within the normal pollen dispersal range of the experimental plant.
• Ensure that cross-fertile plants are not growing within the known pollen dispersal range of the experimental plant.

Biological Containment Practices (Microorganisms)

Effective dissemination of microorganisms beyond the confines of the greenhouse can be prevented by one or more of the following procedures:

• Confine all operations to injections of microorganisms or other biological procedures (including genetic manipulation) that limit replication or reproduction of viruses and microorganisms or sequences derived from microorganisms and confine these injections to internal plant parts or adherent plant surfaces.
• Ensure that organisms which can serve as hosts or promote the transmission of the virus or microorganism, are not present within the farthest distance that the airborne virus or microorganism may be expected to be effectively disseminated.
• Conduct experiments at a time of year when plants that can serve as hosts are either not growing or are not susceptible to productive infection.
• Use viruses and other microorganisms or their genomes that have known arthropod or animal vectors, in the absence of such vectors.
• Use microorganisms that have an obligatory association with the plant.
• Use microorganisms that are genetically disabled to minimize survival outside of the research facility and whose natural mode of transmission requires injury of the target organism or assures that inadvertent release is unlikely to initiate productive infection of organisms outside of the experimental facility.

Biological Containment Practices (Macroorganisms)

Effective dissemination of arthropods and other small animals can be prevented by using one or more of the following procedures:

• Use non-flying, flight-impaired, or sterile arthropods.
• Use non-motile or sterile strains of small animals.
• Conduct experiments at a time of year that precludes the survival of escaping organisms.
• Use animals that have an obligatory association with a plant that is not present within the dispersal range of the organism.
• Prevent the escape of organisms present in run-off water by chemical treatment or evaporation of run-off water. Containment for arthropods is described in the Arthropod Containment Guidelines and Addendum 1 Containment Practices for Arthropods Modified with Engineered Transgenes Capable of Gene Drive

Green House Growth Chambers

Growth chambers are specially designed rooms, chambers or equipment that allow researchers to control the environmental conditions (i.e. humidity, light, temperature, etc.) when studying plants. Growth chambers can offer many advantages over the use of field plots and greenhouses and answer important questions about plant development and growth related to climate change and plant pests and pathogens. According to the NIH Guidelines growth chambers may be used for containment at BL1-P and BL2-P levels under certain conditions.

Field Work

Field work with transgenic plants requires permits from APHIS prior to the start of work. The permit process allows for the introduction (importation, interstate movement, or environmental release) of certain genetically engineered organisms under conditions determined and described in 7 CFR part 340. If you have any plans to do field work with genetically engineered plants or associated organisms, inquire about permits first.

Biohazardous Plant or Plant Associated Organism Waste

Experimental organisms and materials (i.e. plants, seeds, soil, fruit, etc) utilizing BL1-P or BL2-P containment must be rendered biologically inactive by appropriate methods (i.e. autoclave or chemical inactivation, etc.) before disposal outside of the greenhouse facility. In addition, pots, trays, and other hard plastic horticultural supplies in shared use should be thoroughly cleaned and decontaminated to  eliminate and prevent survival of plant material or pathogens.

Should you need further assitance or have questions or concerns regarding plant research conducted at Stony Brook University please contact the Biological Safety Officer, Christopher.kuhlow@stonybrook.edu