Experiments to genetically engineer plants by rDNA/sNA methods may require registration with the APB (BL2-P or higher, see OBA – NIH Guidelines and Appendix C for additional information). To prevent release of transgenic plant materials to the environment, the guidelines provide specific plant biosafety containment recommendations for experiments involving the creation and/or use of genetically engineered plants. (Figure 3) Plant Biosafety levels are categorized into BL1-P to BL4-P (Table 4).

Table 3. Viral vectors and transgene containment.

Gene transfer vectora Host rangeb Insert or gene functionc Laboratory containment leveld
MMLV based –
gag, pol, and env deleted
Ecotropic
Amphotropic, VSV-G
pseudotyped
S, E, M, G, CC, T, MP, DR, R, TX,
Ov, Oc
S, E, M, MP, DR, T, G
Ov, Oc, R, CC
TX
BSL – 1*
BSL – 2
BSL2+
BSL – 3
Herpesvirus based –
nonlytic
Broad host range  S, E, M, MP, DR, T, G
Ov, Oc, R, CC
TX
BSL – 2
BSL2+
BSL – 3
Lentivirus based –
HIV, SIV, EIAV, FIV, etc.; gag, pol, env, nef, and vpr deleted
Ecotropic, amphotropic, VSV-G pseudotyped  S, E, M, MP, DR, T, G
Ov, Oc, R, CC
TX
BSL – 2
BSL2+
BSL – 3
Adenovirus based –
serotypes 2, 5 and 7; E1 and E3 or E4 deleted
Broad host range, infective for many cell types S, E, M, T, MP, DR, R, G, CC
Ov, Oc
TX
BSL – 2
BSL2+
BSL – 3
Alphavirus based –
SFV, SIN
Broad host range S, E, M, T, MP, DR, R, G, CC
Ov, Oc
TX
BSL – 2
BSL2+
BSL – 3
Baculovirus based Broad mammalian host cell range S, E, M, T, MP, DR, R, G, CC
Ov, Oc
TX
BSL – 1*
BSL2
BSL – 2+/BSL – 3
AAV based –
rep, cap defective
Broad host range; infective for many cell types, including neurons S, E, M, T, MP, DR, G
Ov, Oc, R, CC
TX
BSL – 1*
BSL2
BSL – 2+/BSL – 3
Poxvirus based –
canarypox, Vacciniae
Broad host range S, E, M, T, DR, MP, CC, R, G
Ov, Oc
TX
BSL – 2
BSL2+
BSL – 3
a Refers to the parental or wild-type virus and some of the common deletions used in viral vectors. MMLV, Moloney murine leukemia virus; SIV, simian immunodeficiency virus.
b Refers to ability of vector to infect cells from a range of species. Ecotropic generally means able to infect only cells of the species originally isolated from or identified in. Please note that the ecotropic host for HIV and HSV would be human cells, but the ecotropic host for MMLV would be murine cells. Amphotropic and VSV-G-pseudotyped virus host range includes human cells.
c Shown are general categories of cellular genes and functions. Please note that there are differences in the containment level for the same class depending on whether the viral vector integrates into the recipient genome at a high rate. The general categories are as follows: S, structural proteins (actin, myosin, etc.); E, enzymatic proteins (serum proteases, transferases, oxidases, phosphatases, etc.); M, metabolic enzymes (amino acid metabolism, nucleotide synthesis, etc.); G, cell growth, housekeeping; CC, cell cycle, cell division; DR, DNA replication, chromosome segregation, mitosis and meiosis; MP, membrane proteins, ion channels, G-coupled protein receptors, transporters, etc.; T, tracking genes such as those for green fluorescent proteins and luciferases and photoreactive genes; TX, active subunit genes for toxins such as ricin, botulinum toxin and Shiga and Shiga-like toxins; R, regulatory genes for transcription and cell activators such as cytokines, lymphokines and tumor suppressors; Ov and Oc, oncogenes identified via transforming potential of viral and cellular analogs, or mutations in tumor suppressor genes resulting in a protein that inhibits/moderates the normal cellular wildtype proteins. This does not include SV40 T antigen. SV40 T-antigen-containing cells should not be considered more hazardous than the intact virus. SV40 is considered a risk level 1 agent (the lowest level) according to the NIH Guidelines. The prevalence of SV40 infection in the U.S. population due to contaminated polio vaccine does not seem to have caused a statistically significant increase in the rate of cancers. However, the data from the various studies on SV40 association with cancer are equivocal (Strickler et al. 1998; Butel and Lednicky, 1999; Dang-Tan et al., 2004).
d This is a general assessment of containment levels for laboratory construction and use of these vectors for nonproduction quantities only based on the 4th edition of BMBL. This table cannot cover every potential use within a research or laboratory settings; as information is gained, risk assessments and containment levels may be changed. Local IBCs should use all available information and their best judgment to determine appropriate containment levels. BSL – 1* refers to the containment level based on parent virus risk group. However, most procedures involving the handling and manipulation of the viral vectors are done at BSL – 2 to protect cell cultures and viral stocks from contamination.
e Certain specific strains of poxviruses, such as MVA, NYVAC, ALVAC and TROVAC, are considered low-risk agents and can be handled at BSL – 1 in certain cases.

Table 4. Plant biosafety levels.

From Practical Guide to Containment: Plant Biosafety in Research Greenhouses, Revised Edition, page 13, D. Adair and R. Irwin.

Criteria Transgenic Plants Transgenic Microbes: Exotic Transgenic Microbes: Non-Exotic Transgenic Insects/Animals/Assoc. Microbes
Not a noxious weed or cannot outcross with one BL1-P
Not easily disseminated BL1-P
No detriment to environment BL2-P or BL1-P+ BL1-P BL2-P or BL1-P+
Noxious weed or can interbreed with weeds BL2-P or BL1-P+
Contains complete genome of non-EIA* BL2-P or BL1-P+
Contains genome of EIA BL3-P or BL2-P+
Treated with an EIA BL3-P or BL2-P+
Detriment to environment BL3-P-4** BL2-P or BL1-P+ BL3-P or BL2-P+
Involves EIA with detriment to environment BL3-P or BL2-P+
May reconstitute genome of infectious agent in plants BL3-P or BL2-P+
Contains Vertebrate Toxin BL3-P BL3-P BL3-P
*EIA—Exotic Infectious Agent
**BL4-P containment is recommended only for experiments with readily transmissible exotic infectious agents whether transgenic or not, such as air-borne fungi or viruses in the presence of their arthropod vectors that have the potential of being serious pathogens of major US crops.

 

Important Information

Plant Research Permits

Additional permits might be required from state and federal agencies before research with plants can be done. Contact Biosafety for information.