This is a work product of the National Organic Standards Board (NOSB). It does not represent official National 

Organic Program (NOP) positions or policy.

National Organic Standards Board

GMO ad hoc Subcommittee

Discussion Document

Excluded Methods Terminology

February 6, 2013

Introduction

There is a fundamental contradiction at the heart of the regulatory approaches to organic 

agriculture and biotechnology. Organic certification is a process-based guarantee while 

biotechnology is subject to a product-based assessment (Caruso 2006). 

One of the strengths of the process-based regulation is that it takes into consideration the 

entire production system. If biotechnology were assessed on the basis of its impacts on the 

system rather than the narrow focus of whether or not a particular transgenic crop poses a 

“plant pest” risk, regulators would have to take into account the environmental impacts of 

increased herbicide applications, increased use of more volatile and toxic herbicides, 

impacts on pollinator populations, reduction of vegetative diversity, introduction of novel 

proteins into soil and water ecosystems, likelihood of selecting for resistance in weed and 

pest populations, negative socioeconomic impacts, and reduction of transgene-free 

germplasm availability.

There are contradictions concerning what are defined as “excluded methods,” the phrase 

used in the USDA organic regulations to describe certain products of biotechnology.

Furthermore, the concepts of "excluded methods" have widened to includequestions about 

what vaccines for livestock are being produced through use of “excluded methods”, issues 

around the use of micro-organisms in processed foods and the discussion leading up to a 

October 2012 NOSB recommendation to allow the use of biodegradable mulch film with a 

prohibition on the use of organisms or feedstock derived from “excluded methods”. Now 

that the USDA organic regulations have been fully implemented for 10 years, new issues 

and technologies in plant and animal breeding have been identified and it may be 

necessary to clarify the language in these regulations.

The purpose of this Discussion Document is to provide definitions of the terms included in 

the definition of “excluded methods” and to ask for stakeholder input on what changes may 

need to be made to the definition of “excluded methods” or what clarifications and 

interpretations may need to be made through guidance.

Background and Relevant areas in the Rule

Here is the definition of “excluded methods” in the National Organic Standards (7 CFR 

205.2; Terms Defined):

Excluded methods. A variety of methods used to genetically modify organisms or 

influence their growth and development by means that are not possible under natural 

conditions or processes and are not considered compatible with organic production. 

Such methods include cell fusion, microencapsulation and macroencapsulation, and 

recombinant DNA technology (including gene deletion, gene doubling, introducing a 
Page 2 of 20

foreign gene, and changing the positions of genes when achieved by recombinant 

DNA technology). Such methods do not include the use of traditional breeding, 

conjugation, fermentation, hybridization, in vitro fertilization, or tissue culture. (Federal 

Register / Vol. 65, No. 246 / Thursday, December 21, 2000 / Rules and Regulations p. 

80639)

Since the rule went into effect in 2002, no changes have been made to the definition of 

“excluded methods” in the regulations. Due to the lack of specificity in the definition, ACAs 

(Accredited Certifying Agents) have made their own determinations of how far back in the 

ingredient and input chain (or breeding line) to verify compliance with the prohibition on 

excluded methods in organic production and handling and interpret what these terms within 

the definition mean. This has caused confusion among stakeholders at times, where other 

techniques not mentioned in the definition are also considered by some as excluded, while 

some terms within the definition are open to multiple interpretations.

Three separate topic areas that came up in the last two years have caused this confusion 

to become more important to try to address. The NOSB Livestock Subcommittee has been 

grappling with the subject of excluded methods used in vaccines and, to guide their further 

deliberations, a Vaccines Working Group1 developed a parallel discussion of the terms 

relevant to vaccine production to solicit comment on what vaccine production methods may 

or may not be considered excluded. This discussion is posted as an Interim Report and the 

Working Group is seeking public input.2 In December 2011, the NOSB’s review of 

Docosahexaenoic acid (DHA) algal oil, an ingredient petitioned for use in organic handling,

prompted discussion over whether the form of mutagenesis that may be used to develop 

this ingredient should be considered an “excluded method”.

3 Lastly, the NOP recently 

issued Policy Memo 13-1 on February 1, 2013 concerning Cell Fusion Techniques used in 

Seed Production (discussed in the cell fusion section below) to clarify which cell fusion 

techniques should be considered use of an “excluded method”.

4

Some ACAs and groups such as Organic Materials Review Institute (OMRI) have 

developed their own decision making protocols for interpreting the definition of “excluded 

methods”. With advances in biotechnology since the time the “excluded methods” definition 

was first adopted by the NOSB in 1995 and innovations in product development from 

biotechnology, the decision making is getting more and more complicated.

Discussion

 1 In May 2012, the NOSB Livestock Subcommittee issued a resolution requesting that USDA provide 

information regarding which vaccine products were produced through the use of biotechnology. Available 

at: http://www.ams.usda.gov/AMSv1.0/getfile?dDocName=STELPRDC5098924. In response, the NOP 

initiated a Vaccines Working Group comprised of NOP staff, NOSB members, and staff from APHIS’ Center 

for Veterinary Biologics. The Working Group began meeting in July 2012 and provided an interim report to 

the Livestock Subcommittee in February 2012 on this issue. 2 Vaccines Working Group Interim Report: Identifying Vaccines Made with Excluded Methods. Available at: 

 http://www.ams.usda.gov/AMSv1.0/getfile?dDocName=STELPRDC5102576 3 December 2011 NOSB Recommendation on DHA algal oil. Available at: 

 http://www.ams.usda.gov/AMSv1.0/getfile?dDocName=STELPRDC5097102

4 National Organic Program, Policy Memo 13-1: Cell Fusion Techniques Used in Seed Production. Available 

 at: http://www.ams.usda.gov/AMSv1.0/getfile?dDocName=STELPRDC5102380
Page 3 of 20

The GMO ad hoc Subcommittee believes that strengthening and clarifying the definition of 

“excluded methods” in the USDA organic regulations will help all stakeholders with 

implementation of the regulations and strengthen processes behind keeping GMOs out of 

organic food. To do this, the GMO ad hoc subcommittee has prepared this discussion 

document to 1) examine the language currently in the “excluded methods” definition (Part 

A), and 2) outline other terms that could be added to the definition (Part B), providing 

context for regulatory definitions from organic standards of other countries (Appendix I). 

The Subcommittee is seeking public input on which of these terms belong in a revised 

definition for “excluded methods” under the USDA organic regulations, as well as the 

guiding principle(s) to use in crafting a new definition.

Part A

Examination of the exact language of the excluded methods definition at 7 CFR 205.2 will 

bring out the key issues.

1. A variety of methods used to genetically modify organisms or influence their growth and 

development by means that are not possible under natural conditions or processes and are 

not considered compatible with organic production.

The phrase “not possible under natural conditions or processes” has become problematic

in the context of “traditional” breeding methods that involve disruption of normal plant cell 

growth. For example, mutagenesis can be a process in which chemical or radiation stress 

is applied on a cell to force mutation to happen, but it also commonly occurs in nature and 

at least some of the mutagenesis chemicals are derived from nature. (More on 

mutagenesis under 5. traditional breeding). The concept of "natural" is not defined in any 

regulations and is very blurred after centuries of humans manipulating the environment and 

plants, animals and microbes.

This brings up the question, what exactly is it about a genetic modification process that is 

objectionable in the organic context? This larger question is what the GMO Subcommittee 

is re-visiting here in order for organic stakeholders to clarify the basis of objection to the 

technology because even acceptable breeding methods could very well not be 

possible under natural conditions. It may be that the species line is where people object 

to genetic exchange occurring. If this is the case, the terms interspecific (between species) 

and intraspecific (within species) or intergeneric (between genera) and intrageneric (within 

a genus) may come in handy. So many different techniques are used now that wording 

must be very carefully chosen or some crops already accepted in organic cultivation might 

be ruled out. Examples include triticale (created from breeding two different genera), 

bananas and seedless watermelon from somatic doubling, and more.

Dutch organic plant breeder Edith Lammerts van Bueren has proposed that organisms 

have “intrinsic value” and “integrity” represented by an intact genome (Lammerts van 

Bueren et al. 2003). While this argument may seem to have more of a philosophical than 

scientific basis to it, it may be a useful organizing statement in describing what the organic 

community finds acceptable means of plant and animal breeding. It also may be relevant to 

consider that while the prohibition on genetically modified organisms allows for certain 

traditional methods used to produce nonorganic seeds, (e.g. mutagenesis), the standards 

for organic plant breeding can be considered more restrictive, since the chemicals or 

irradiation techniques used are not permitted methods under organic standards. 
Page 4 of 20

2. Such methods include cell fusion

Cell fusion: The process in which two different cells fuse into one single cell. The resulting 

cell has all of the contents from the original cells and has one nucleus containing the 

genetic material from both of the original cells. This occurs naturally during sexual

reproduction, when gametes (eggs and sperm) fuse to produce the zygote, and can also 

occur under laboratory conditions between somatic cells (any cell other than a gamete). 

(Websters Online Dictionary5) 

Protoplast fusion

A technique in which protoplasts (plant cells from which the cell wall has been removed by 

mechanical or enzymatic means) are fused into a single cell. (National Academies 2004 

glossary)

or 

The fusing of two protoplasts (a bacterial or plant cell deprived of its cell wall but having an 

intact plasma membrane).- (CAN/CGSB-32.315-2004 Voluntary Labelling And Advertising 

of Foods That Are and Are Not Products of Genetic Engineering)

Also relevant here:

Somatic hybridization : “The technique of hybrid production of plants through the fusion of 

isolated somatic (body) protoplasts under in vitro conditions and subsequent development 

of their product (heterokaryon) to a hybrid plant is known as somatic hybridization.” 

(Chawla, H.S. 2001. Introduction to Plant Biotechnology, 2nd ed. Science Publishers, Inc.)

The NOP was asked to clarify its position on cell fusion because it has been used as an 

“acceptable” means of developing varieties of Brassica and citrus crops among others. Cell 

fusion has been used in traditional breeding and hybridization programs as well as in 

general propagation using tissue culture. It can be used within a genus or species or 

between very different species. 

So why might cell fusion be considered an excluded method? A form of cell fusion called 

somatic cell hybridization, somatic hybridization, or protoplast fusion involves destruction of 

cell walls using chemical or electrical stimuli, which then allows the genetic material to be 

fused. This approach has been used to develop both intraspecific and interspecific crosses 

(see selected citations A).

In 2012, the NOP received questions about whether seed varieties produced through cell 

fusion techniques are allowed in organic production. The issue for certifiers is complicated 

by the fact that it is not disclosed publically which varieties may have been bred using cell 

fusion and so enforcement of any prohibition is extremely problematic. Additionally the cell 

fusion event may have happened up to 30 years ago and the resulting trait simply passed 

to subsequent generations by traditional breeding methods.

 5 http://www.websters-online-dictionary.org/definitions/Cell+Fusion
Page 5 of 20

In NOP Policy Memo 13-1, the type of cell fusion used to transfer traits such as male 

sterility within a plant family was determined to be possible with natural breeding 

techniques and compatible with organic production. 

"... the NOP further concludes that cell fusion (including protoplast fusion) is not 

considered an excluded method when the donor cells/protoplasts fall within the 

same taxonomic plant family, and when donor or recipient organisms are not derived 

using techniques of recombinant DNA technology."

This Policy Memo also explains that cell fusion techniques are considered an “excluded 

method” when the donor cells/protoplasts do not fall within the same taxonomic family. Cell 

fusion is also an “excluded method” when the donor or recipient organism is derived using 

techniques of recombinant DNA technology and techniques involving the direct introduction 

into the organism of hereditary materials prepared outside of the organism. 

Here again it seems as if the technology itself is not the issue so much as the crossing of 

the species or taxonomic family line.

3. microencapsulation and macroencapsulation

Micro-encapsulation: a process in which tiny particles or droplets are surrounded by a 

coating to give small capsules many useful properties. (Wikipedia)

Macroencapsulation: (cell and molecular biology) The envelopment of a large mass of 

xenotransplanted cells or tissue in planar membranes, hollow fibers, or diffusion chambers 

to isolate the cells from the body, thereby avoiding the immune responses that the foreign 

cells could initiate, and also to allow the desired metabolites (such as insulin and glucose 

for pancreatic islet cells) to diffuse in and out of the membrane.

(McGraw Hill Science and Reading Dictionary) 

http://www.answers.com/topic/macroencapsulation#ixzz2E2MQK67W

It may be time to remove these processes from the definition of “excluded methods” 

because they don’t involve recombining genes. They may be more appropriately classified 

as “nonagricultural” or “synthetic” materials or as a form of nanotechnology. Micro- and 

macroencapsulation appear to be cellular packaging mechanisms for engineered genes, 

food additives, or pesticides rather than a form of genetic engineering (see selected 

citations B). Specifying them as excluded methods is questionable and has been ridiculed 

by at least one academic commenter (Eisen 2012).

4. and recombinant DNA technology (including gene deletion, gene doubling, introducing a 

foreign gene, and changing the positions of genes when achieved by recombinant DNA 

technology).

DNA, Recombinant

Biologically active DNA which has been formed by the in vitro joining of segments of DNA 

from different sources. It includes the recombination joint or edge of a heteroduplex region 

where two recombining DNA molecules are connected.

Year introduced: 1977 (MeSH)

Gene Deletion
Page 6 of 20

A genetic rearrangement through loss of segments of DNA or RNA, bringing sequences 

which are normally separated into close proximity. This deletion may be detected using 

cytogenetic techniques and can also be inferred from the phenotype, indicating a deletion 

at one specific locus. Year introduced: 1993. (MeSH)

Genetic engineering

Changes in the genetic constitution of cells resulting from the introduction or elimination of 

specific genes via molecular biology (i.e., recombinant DNA) techniques. (National 

Academies 2004 glossary)

Recombinant DNA techniques

Procedures used to join together DNA segments. Under appropriate conditions, a 

recombinant DNA molecule can enter a cell and replicate there. (National Academies 2004 

glossary)

Recombination naturally occurs between chromosomes during the process of meiosis to 

form gametes for sexual propagation, in plants, animals and other organisms. 

Recombination naturally occurs during high frequency recombinant (Hfr) conjugation in 

which part of the chromosome from one bacterium is transferred to another bacterium, 

resulting in homologous recombination which genetically modifies the target bacteria. 

These are just two examples of genetic modifications through recombination events which 

may be allowed by the current definition of excluded methods.

This language seems valid and on point in addressing the main concerns most organic 

stakeholders have with transgenic technologies; however, the specifics could probably be 

updated to include other recombinant technologies and to remove phrasing that is not in 

common usage. For example, “gene doubling” is not often found in the literature.

Such methods do not include the use of

5. traditional breeding,

This term is assumed to include breeding methods that have been used prior to the 

emergence of transgenic technologies. It is not clear at which point traditional breeding 

techniques are divided from modern or non-traditional breeding techniques. Is there a time 

point at which all techniques before that time are considered traditional and all new 

techniques developed after that time are not considered traditional? The use of 

transposons (see below Part B) since the 1930's or chemical, physical, and biological 

mutagens since the 1940's are blurring the distinction between traditional breeding and 

biotechnology.

One form of traditional breeding that has not been formally defined and has been called 

into question is mutagenesis.

Mutagenesis (or mutation breeding)

A process whereby the genetic information of an organism is changed in a stable, heritable 

manner, either in nature or induced experimentally via the use of chemicals or radiation. In 
Page 7 of 20

agriculture, these genetic changes are used to improve agronomically useful traits. 

(National Academies 2004 glossary)

The “problem” with mutagenesis as an acceptable practice in organic breeding is that it 

sometimes relies on processes that would not occur naturally, but sometimes does involve 

naturally derived substances and processes. Additionally, some mutagenesis is now

accomplished by inserting DNA or other genetic material into a cell (insertional

mutagenesis). Because of its widespread usage in plant breeding since the 1940s, 

however, there may be a need to clarify which types of mutagenesis are acceptable under 

the organic standards. See selected citations C.

6. conjugation, 

Conjugation, Genetic: A parasexual process in BACTERIA; ALGAE; FUNGI; and ciliate 

EUKARYOTA for achieving exchange of chromosome material during fusion of two cells. In 

bacteria, this is a uni-directional transfer of genetic material; in protozoa it is a bi-directional 

exchange. In algae and fungi, it is a form of sexual reproduction, with the union of male and 

female gametes. Year introduced: 1968 (MeSH) 

Conjugation can be used to transfer genetic information (via plasmids) between different 

genera of bacteria. Might this violate the guiding principle of not crossing taxonomic lines? 

(See Jones and Woods 1986 for background.)

7. fermentation, 

Fermentation: Anaerobic degradation of GLUCOSE or other organic nutrients to gain 

energy in the form of ATP. End products vary depending on organisms, substrates, and 

enzymatic pathways. Common fermentation products include ETHANOL and LACTIC 

ACID. (MeSH)

Should inclusion of fermentation on this list be reconsidered? While the process of 

fermentation can be used to multiply transgenic organisms and some fermentation 

processes are done with transgenic organisms, it is not a breeding technology. (See Jones 

and Woods 1986 for background on the use of microbes to manufacture solvents via 

fermentation.)

8. hybridization,

Hybridization, Genetic: The genetic process of crossbreeding between genetically 

dissimilar parents to produce a hybrid. (MeSH)

Hybrid

Progeny of genetically different parents, usually of the same species, that has enhanced 

productivity over either parent. Generally, the more genetically diverse the parent lines, the 

more hybrid vigor, or heterosis, is observed in the hybrid progeny. (National Academies 

2004 glossary)

Here are some other types of hybridization defined on the MeSH site for reference.
Page 8 of 20

Nucleic Acid Hybridization: Widely used technique which exploits the ability of 

complementary sequences in single-stranded DNAs or RNAs to pair with each other to 

form a double helix. Hybridization can take place between two complimentary DNA 

sequences, between a single-stranded DNA and a complementary RNA, or between two 

RNA sequences. The technique is used to detect and isolate specific sequences, measure 

homology, or define other characteristics of one or both strands. (Kendrew, Encyclopedia of 

Molecular Biology, 1994, p503). Year introduced: 1972(1971) (MeSH)

In Situ Hybridization

A technique that localizes specific nucleic acid sequences within intact chromosomes, 

eukaryotic cells, or bacterial cells through the use of specific nucleic acid-labeled probes.

Year introduced: 1993. (MeSH)

9. in vitro fertilization, 

Fertilization in Vitro

An assisted reproductive technique that includes the direct handling and manipulation of 

oocytes and sperm to achieve fertilization in vitro. Year introduced: 1979. (MeSH)

10. tissue culture

Tissue Culture Techniques

A technique for maintaining or growing TISSUE in vitro, usually by DIFFUSION, perifusion, 

or PERFUSION. The tissue is cultured directly after removal from the host without being 

dispersed for cell culture. Year introduced: 2005. (MeSH)

Another one that was defined long before the “year introduced”

Tissue culture does not “disperse the tissue for cell culture.” But cell culture is used in 

breeding and the process of culturing cells can stimulate genetic variability that can provide 

further breeding material. Cell culture can be a means of generating “natural” genetic

variability under “unnatural” conditions along the lines of mutagenesis. See selected 

citations D. Here are related definitions.

Cell Culture Techniques

Methods for maintaining or growing CELLS in vitro.Year introduced: 2005 (1996) (MeSH)

Primary Cell Culture

The initial culturing of cells derived directly from fresh TISSUES. Year introduced: 2012 

(MeSH)

Batch Cell Culture Techniques

Methods for cultivation of cells, usually on a large-scale, in a closed system for the purpose 

of producing cells or cellular products to harvest. Year introduced: 2012. (MeSH)

Part B

Contemporary breeding methods that may be candidates as “excluded methods”, but may 

not be.
Page 9 of 20

This section was compiled by looking through critiques of the current organic standards 

regarding transgenic technologies and from there following the discussions on the internet. 

A couple of other terms are relevant to the work of the Vaccines Working Group. One 

science blogger in particular (von Mogel 2010) dissects the rationale Jim Riddle presented 

in 2010 for why genetic engineering is incompatible with organic farming. Von Mogel 

includes a figure that purports to show relative risks of unintended consequences by 

breeding method (National Academies 2004). The figure is shown in Appendix II.

This is a list of other emerging breeding strategies that may need to be included in the 

recitation of “excluded methods.” These are:

• Gene silencing—occurs naturally and may also be engineered

Silencing

Shutdown of transcription of a gene, usually by methylation of C residues. (National 

Academies 2004 glossary)

• Embryo rescue

Embryo rescue

A sequence of tissue culture techniques used to enable a fertilized immature embryo 

resulting from an interspecific cross to continue growth and development, until it can be 

regenerated into an adult plant. (National Academies 2004 glossary)

This fairly common technique is used to clean plant tissue from viruses (such as potatoes) 

and is quite beneficial to organic agriculture.

• Microinjection—clearly an excluded method. Need to be specified?

Microinjection

Introduction of DNA into a cell by injection through a very fine needle. (National Academies 

2004 glossary)

• Biolistic transfer—already covered in the definition of excluded methods. Need to be 

specific? Also known as microprojectile bombardment.

Biolistic device

A device that bombards target cells with microscopic DNA-coated particles. Familiarly 

known as the Gene Gun, it was first developed in the early 1980s. (National Academies 

2004 glossary)

• Somaclonal variation (analogous to mutagenesis in that it is a form of natural 

genetic variation forced by unnatural conditions [cell culture]) 

Somaclonal variation

Epigenetic or genetic changes, sometimes expressed as a new trait, resulting from in vitro 

culture of higher plants. (National Academies 2004 glossary)

• Transposons — naturally occurring, double stranded DNA sequences with a defined 

structure. They are present in plant, animal and bacterial species.

Transposons
Page 10 of 20

Each end of the transposon includes inverted repeats. In prokaryotes, the internal structure 

includes at least one gene for transposase and may contain many more depending upon 

the type of transposon. When the transposase gene is expressed, the protein binds to the 

inverted repeats of the transposon, cleaves the genomic DNA and excises the transposon. 

Transposase can then cleave the genomic DNA at another spot and recombine the 

transposon into a new position in the genome. By moving from one location to another in 

the genome, transposons can cause gene deletions or change expression patterns through 

gene deletion, resulting in changed phenotypes. (Ivics, Z. and Z. Ivsvak. 2010; MeSH)

Transposons were initially identified as jumping genes by Barbara McClintock in research 

on variegation of corn kernels in the 1930's (Pray & Zhaurova, 2008). More recently 

researchers have used transposons as a vector for inserting specific foreign genes into the 

genomes of various species. The transposon system called "Sleeping Beauty" was used to 

genetically modify swine cells with genes from rice (Carlson et. al., 2011). The Vaccine 

Working Group has more detail on transposons as used to produce vaccines.

• Transduction— while this theoretically could occur in nature, the specific purpose of 

its intentional use is in biotechnology applications.

Transduction

The process through which the genomes of bacteria can be modified with the use of 

bacterial virus, called a phage. Some types of phage attach to the bacterial cell wall and 

insert the viral genome into the cell. The viral genome may then be inserted into the 

bacterial genome through a recombination event which is part of the lysogenic cycle. After 

receiving a trigger, the viral genome will be excised and the lytic cycle will be triggered

(MeSH).

This method can stably introduce genetic mutations into the new bacteria. This technique 

is widely used to create gene deleted vaccine products.

Discussion Questions

The GMO ad hoc Subcommittee is seeking response from the organic community on the

issues presented in this discussion. A few of the questions to be addressed are:

1. Does the definition of "excluded methods" in the Organic Rule need to be revised? 

Please provide reasoning for either a "yes" or a "no" answer.

2. On what general principle(s) should practical and consistent distinctions be made 

between “excluded” and permitted methods of breeding that could apply to plants, animals 

and micro-organisms? Under such general principles should we further define or replace 

terms such as "natural conditions" and "traditional breeding"?

3. Are there other terms beyond those discussed here that should be addressed in the 

context of excluded methods?

4. Of the terms and practices discussed here, which ones should be in the definition of 

excluded methods and which not excluded? Why?
Page 11 of 20

5. How far back into the development or manufacture of a substance, or in the development 

of vaccines, or in the lineage of a breeding line, should the excluded methods prohibition 

apply? How far back is practical and verifiable?

Subcommittee Vote

Motion to adopt the proposed Discussion Document on Excluded Methods

Motion by: Jean Richardson Second: Colehour Bondera

Yes:7 No:0 Absent:0 Abstain:0 Recuse:0
Page 12 of 20

Appendix I. Other GE and GMO definitions 

USDA Advisory Committee on Biotechnology and 21st Century Agriculture (AC21)

http://www.usda.gov/documents/ac21_report-enhancing-coexistence.pdf

―Genetically Engineered is meant to include biotechnology-derived organisms produced 

through the application of 1) in vitro nucleic acid techniques, including recombinant 

deoxyribonucleic acid (DNA) and direct injection of nucleic acid into cells or organelles or 2) 

fusion cells beyond the taxonomic family, that overcome natural physiological reproductive 

or recombinant barriers and that are not techniques used in traditional breeding and 

selection.

Codex Alimentarius Commission, International Food Standards

http://www.codexalimentarius.org/standards/list-of-standards/en/

CAC/GL 44-2003

PRINCIPLES FOR THE RISK ANALYSIS OF FOODS DERIVED FROM MODERN 

BIOTECHNOLOGY

Adopted in 2003. Amendments 2008, 2011

SECTION 2 – SCOPE AND DEFINITIONS 

7. The purpose of these Principles is to provide a framework for undertaking risk analysis 

on the safety and nutritional aspects of foods derived from modern biotechnology. This 

document does not address environmental, ethical, moral and socio-economic aspects of 

the research, development, production and marketing of these foods3. 

8. The definitions below apply to these Principles: 

“Modern Biotechnology” means the application of: 

i) In vitro nucleic acid techniques, including recombinant deoxyribonucleic acid 

(DNA) and direct injection of nucleic acid into cells or organelles, or 

ii) Fusion of cells beyond the taxonomic family, that overcome natural physiological 

reproductive or recombinant barriers and that are not techniques used in traditional 

breeding and selection4

.

3 

This document does not address animal feed and animals fed such feed except insofar 

as these animals have been developed by using modern biotechnology. 

4 This definition is taken from the Cartagena Biosafety Protocol under the Convention on 

Biological Diversity. 

European Union

Directive 2001/18/EC of the European Parliament and of the Council of 12 March 2001

Deliberate Releases into the Environment of Genetically Modified Organisms 

http://eurlex.europa.eu/LexUriServ/LexUriServ.do?uri=CONSLEG:2001L0018:20080321:EN:PDF

Article 2 
Page 13 of 20

Definitions - (2) ‘genetically modified organism (GMO)’ means an organism, with the 

exception of human beings, in which the genetic material has been altered in a way that 

does not occur naturally by mating and/or natural recombination;

Within the terms of this definition:

(a) genetic modification occurs at least through the use of the techniques listed in Annex I 

A, part 1;

(b) the techniques listed in Annex I A, part 2, are not considered to result in genetic 

modification;

ANNEX I A

TECHNIQUES REFERRED TO IN ARTICLE 2(2)

Techniques of genetic modification referred to in Article 2(2)(a) are inter alia:

(1) recombinant nucleic acid techniques involving the formation of new combi- nations 

of genetic material by the insertion of nucleic acid molecules produced by whatever 

means outside an organism, into any virus, bacterial plasmid or other vector system 

and their incorporation into a host organism in which they do not naturally occur but in 

which they are capable of continued propagation;

(2) techniques involving the direct introduction into an organism of heritable material 

prepared outside the organism including micro-injection, macro- injection and microencapsulation;

(3) cell fusion (including protoplast fusion) or hybridisation techniques where live cells 

with new combinations of heritable genetic material are formed through the fusion of 

two or more cells by means of methods that do not occur naturally.

PART 2

Techniques referred to in Article 2(2)(b) which are not considered to result in genetic 

modification, on condition that they do not involve the use of recombinant nucleic acid 

molecules or genetically modified organisms made by techniques/methods other than those 

excluded by Annex I B:

(1) in vitro fertilization,

(2) natural processes such as: conjugation, transduction, transformation,

(3) polyploidy induction.

ANNEX I B

TECHNIQUES REFERRED TO IN ARTICLE 3

Techniques/methods of genetic modification yielding organisms to be excluded from the 

Directive, on the condition that they do not involve the use of recombinant nucleic acid 

molecules or genetically modified organisms other than those produced by one or more of 

the techniques/methods listed below are:

(1) mutagenesis,

(2) cell fusion (including protoplast fusion) of plant cells of organisms which can exchange 

genetic material through traditional breeding methods.

Canadian Standards:

CAN/CGSB-32.315-2004, Voluntary Labelling And Advertising of Foods That Are and Are 

Not Products of Genetic Engineering

http://www.tpsgc-pwgsc.gc.ca/ongc-cgsb/programme-program/normesstandards/internet/032-0315/index-eng.html
Page 14 of 20

Genetic engineering (Génie génétique): Refers to techniques by which the genetic 

material of an organism is changed in a way that does not occur naturally by multiplication 

and/or natural recombination. Examples of the techniques used in genetic engineering 

include but are not limited to the following:

1. recombinant DNA (rDNA) techniques that use vector systems 

2. techniques involving the direct introduction into the organism of hereditary materials 

prepared outside the organism 

3. cell fusion (including protoplast fusion) or hybridization techniques that overcome 

natural physiological, reproductive, or recombination barriers, where the donor 

cells/protoplasts do not fall within the same taxonomic family 

Unless the donor/recipient organism is derived from any of the above techniques, examples 

of excluded techniques include but are not limited to the following:

1. in vitro fertilization 

2. conjugation, transduction, transformation, or any other natural process 

3. polyploidy induction 

4. mutagenesis 

5. cell fusion (including protoplast fusion) or hybridization techniques where the donor 

cells/protoplasts fall within the same taxonomic family. 

Note: (Descriptions of most of these techniques are found in Appendix A.)

Appendix A

Cell fusion (Fusion cellulaire): The fusing of two cells to form a single cell.

Macroinjection (Macro-injection): The introduction of larger molecules into single cells.

Microencapsulation (Micro-encapsulation): The enclosure of small DNA molecules into a 

capsule, which could be any fatty, fibrous, or membranous structure.

Microinjection (Micro-injection): The introduction of DNA or other compounds into single 

cells with a microscopic needle.

Mutagenesis (Mutagenése): The induction of genetic mutation through chemical, physical, 

or radiation treatment, causing nucleotide(s) of the exposed organism's DNA to be altered. 

This occurs naturally at a very low rate of occurrence, or can be accelerated with in vitro 

methods.

Plasmid (Plasmide): A circular DNA molecule found in bacteria. Plasmids can transfer 

genes between bacteria and are important transformation tools.

Polyploidy (PolyploÏdie): The condition where more than two copies of chromosomes are 

present within a cell - this is caused either by the prevention of cell division or by 

reproduction of extra copies of chromosomes.

Protoplast fusion (Fusion de protoplastes): The fusing of two protoplasts (a bacterial or 

plant cell deprived of its cell wall but having an intact plasma membrane).

Recombinant DNA (rDNA) techniques (Techniques de l'ADN recombinant): The transfer, 

in vitro, of spliced genes between different organisms of the same or different species, or 

the transfer of synthetic genes, which in turn changes the heritable traits of the organism. 

Such transfer of genes can be accomplished using vector systems or by direct introduction 

using a number of techniques including but not limited to chemoporation, electroporation, 

liposome fusion, macroinjection, microencapsulation, microinjection, and transduction.

Taxonomic family (Famille taxonomique): An orderly classification of living organisms 

according to their presumed natural relationships, in which a group of related living 

organisms form a category ranking above a genus and below an order, and usually 

comprising several to many genera.
Page 15 of 20

Transduction (Transduction): The transfer of DNA from one micro-organism to another via 

a virus that infects bacteria.

Transformation (Transformation): A process whereby a cell incorporates foreign DNA into 

its genome.

Vector (Vecteur): An organism, plasmid, or virus that is used to deliver selected foreign 

DNA into a host cell.
Page 16 of 20

Appendix II. Graph from National Academies study assessing the safety of genetically 

engineered foods, p. 64. (National Academies 2004). 

References

Carlson, DF, JR Garbe, W Tan, MJ Martin, JR Dobrinsky, PB Hackett, KJ Clark and SC 

Fahrnekrug. 2011. Strategies for selection marker-free swine transgenesis using the 

Sleeping Beauty transposon system. Transgenic Res 20(5): 1125

Caruso, D. 2006. Intervention: confronting the real risks of genetic engineering and life on a 

biotech planet. San Francisco: The Hybrid Vigor Institute.

Eisen, M. 2012. Senators Boxer and Sanders fill Agriculture Bill with anti-GMO nonsense. 

http://www.michaeleisen.org/blog/?p=1153

Ivics, Z. and Z. Ivsvak. 2010. The expanding universe of transposon technologies for gene 

and cell engineering. Mobile DNA. 1:25

Jones, D.T., and D.R. Woods. 1986. Acetone-Butanol Fermentation Revisited. 

MICROBIOLOGICAL REVIEWS, Dec. 1986, p. 484-524. 

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC373084/pdf/microrev00055-0136.pdf

Lammerts van Bueren, E.T., P.C. Struik, M. Tiemens-Hulscher, and E. Jacobsen. 2003. 

The concepts of intrinsic value and integrity of plants in organic plant breeding and 

propagation. Crop Sci. 43:1922-1929. 

MeSH (Medical Subject Headings), the NLM [National Library of Medicine] controlled 

vocabulary thesaurus used for indexing articles for PubMed. 
Page 17 of 20

http://www.ncbi.nlm.nih.gov/mesh

National Academies Committee on Identifying and Assessing Unintended Effects of 

Genetically Engineered Foods on Human Health, National Research Council. "3. 

Unintended Effects from Breeding." Safety of Genetically Engineered Foods: Approaches 

to Assessing Unintended Health Effects. Washington, DC: The National Academies Press, 

2004. http://www.nap.edu/openbook.php?record_id=10977&page=1

Pray, L. & Zhaurova, K. 2008 Barbara McClintock and the discovery of jumping genes 

(transposons). Nature Education 1(1)

Sooby, Jane 2012. personal communication and assistance preparing the first draft of this 

document.

von Mogel, K.H. 2010. Ten bad reasons why GE is incompatible with organic. 

http://www.biofortified.org/2010/05/ten-bad-reasons/

Selected citations and abstracts

A. cell fusion, somatic cell hybridization, somatic hybridization, protoplast fusion

“Arabidobrassica”: A novel plant obtained by protoplast fusion. Yury Yu. Gleba, Franz 

Hoffmann. Planta July 1980, Volume 149, Issue 2, pp 112-117.

http://link.springer.com/article/10.1007%2FBF00380870?LI=true#page-1

“The results represent the first case of intergeneric-intertribal hybridization of flowering 

plants.”

Protoplast Fusion Technology and Its Biotechnological Applications. (India) 

http://www.aidic.it/IBIC2008/webpapers/96Verma.pdf

Somatic hybrids produced by protoplast fusion between S. tuberosum and S. brevidens: 

phenotypic variation under field conditions. S. Austin, M. K. Ehlenfeldt, M. A. Baer and J. P. 

Heigeson. Theor Appl Genet (1986) 71:682-690.

http://download.springer.com/static/pdf/805/art%253A10.1007%252FBF00263264.pdf?auth

66=1354679330_b79ef433d91a5620b0caed8b8612b6fd&ext=.pdf

Somatic hybrids between Solarium brevidens and Solarium tuberosum: Expression of a 

late blight resistance gene and potato leaf roll resistance. (USA 1986)

http://download.springer.com/static/pdf/146/art%253A10.1007%252FBF00269122.pdf?auth

66=1354678213_aa1aef7574f60c3e2fecc6febb8df18b&ext=.pdf

Somatic hybridization in citrus: An effective tool to facilitate variety improvement.

J. W. Grosser, P. Ollitrault, O. Olivares-Fuster. In Vitro Cellular & Developmental Biology -

Plant November–December 2000, Volume 36, Issue 6, pp 434-449

Download PDF (354 KB)

Summary: Citrus somatic hybridization and cybridization via protoplast fusion has 

become an integral part of citrus variety improvement programs worldwide. Citrus 

somatic hybrid plants have been regenerated from more than 200 parental combinations, 

and several cybrid combinations have also been produced. Applications of somatic 

hybridization to citrus scion improvement include the production of quality tetraploid 
Page 18 of 20

breeding parents that can be used in interploid crosses to generate seedless triploids, and 

the direct production of triploids by haploid + diploid fusion. ....Several allotetraploid somatic 

hybrid rootstocks are performing well in commercial field trials, and show great promise for 

tree size control. Seed trees of most of these somatic hybrid rootstocks are producing 

adequate nucellar seed for standard propagation. Somatic hybridization is expected to 

have a positive impact on citrus cultivar improvement efforts.

B. microencapsulation, macroencapsulation

Microencapsulation: Methods and Industrial Applications. By Simon Benita. 

http://books.google.com/books?hl=en&lr=&id=sz669oFo6AC&oi=fnd&pg=PP1&dq=pesticide+microencapsulation&ots=_x0RTSoIAI&sig=mJ

QZ6Ox-CfpRkBpASH8YzS6CJ90#v=onepage&q=pesticide%20microencapsulation&f=false

Chapter 2. Advances in the Technology for Controlled-Release Pesticide Formulations

Microencapsulation: Is listed under Drug Compounding (MeSH)

The preparation, mixing, and assembling of a drug. (From Remington, The Science and 

Practice of Pharmacy, 19th ed, p1814) 

Sher et al. 1999. Microencapsulation of pesticides by interfacial polymerization utilizing 

isocyanate or aminoplast 

chemistry.http://onlinelibrary.wiley.com/doi/10.1002/%28SICI%291096-

9063%28199812%2954:4%3C394::AID-PS829%3E3.0.CO;2-S/abstract - fn1

Summary: Interfacial polymerization microcapsulation processes based on isocyanate or 

aminoplast chemistry, where all wall-forming reactants are placed in the dispersed oil 

phase are described. 

Pesticide microcapsule formulations can be used to reduce mammalian toxicity and extend 

activity, to control evaporation, to reduce phytotoxicity, to protect pesticide from rapid 

environmental degradation, to reduce leaching and to reduce pesticide levels in the 

environment.

http://onlinelibrary.wiley.com/doi/10.1002/(SICI)1096-9063(199812)54:4%3C394::AIDPS829%3E3.0.CO;2-S/abstract

Microencapsulation is a cell-based method of gene therapy, using genetically-modified cells 

to provide a novel protein for the treatment of various inherited or somatic diseases. In 

contrast to conventional gene therapy using viruses to transfer therapeutic genes into the 

patients’ own cells, this method uses universal cell lines genetically engineered to secrete 

high levels of the therapeutic gene product. These cells are implanted within immunoprotective microcapsules into patients to act as a continuous source of the desired gene 

product, thus providing a potentially safer, reversible and more economic treatment than 

viral gene therapy. http://fhs.mcmaster.ca/gene/

The in vivo delivery of heterologous proteins by microencapsulated recombinant cells.

Trends Biotechnol. 1999 Feb;17(2):78-83.

http://www.ncbi.nlm.nih.gov/pubmed/10087608?ordinalpos=2&itool=EntrezSystem2.PEntre

z.Pubmed.Pubmed_ResultsPanel.Pubmed_RVDocSum

Macroencapsulation—MeSH search gives no results
Page 19 of 20

Patrick Aebischer (http://len.epfl.ch/) is cited as the developer of macroencapsulation, 

which appears to be a packaging technology for genetic therapies. From Neuronal 

Degeneration and Regeneration: From Basic Mechanisms to Prospects. By F W Van 

Leeuwen. 

http://books.google.com/books?id=516KNxhxd8UC&pg=PA518&lpg=PA518&dq=Macroenc

apsulation+genetic&source=bl&ots=eoaX_4ZZv7&sig=nqahF3GMkDNcpOXNk6tedviHcXE

&hl=en&sa=X&ei=qki8UKPCJ6mu0AGgrYDwDg&ved=0CE4Q6AEwBDgU#v=onepage&q=

Macroencapsulation%20genetic&f=false

C. Mutagenesis

Wieczorek, A. M. & Wright, M. G. (2012) History of Agricultural Biotechnology: How Crop 

Development has Evolved. Nature Education Knowledge 3(10):9. Cites FAO as stating that 

“More than 2,500 plant varieties (including rice, wheat, grapefruit, lettuce and many fruits) 

have been developed using radiation mutagenesis.” 

http://www.nature.com/scitable/knowledge/library/history-of-agricultural-biotechnology-howcrop-development-25885295

Falk, R. 2010. Mutagenesis as a Genetic Research Strategy. Genetics. August; 185(4): 

1135–1139. http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2927745/

Alonso et al. 2003. Genome-Wide Insertional Mutagenesis of Arabidopsis thaliana. 

Science 301 (5633): 653-657. Abstract online at 

http://stke.sciencemag.org/cgi/content/abstract/sci;301/5633/653

D. cell culture, somaclonal variation

Somaclonal variation — a novel source of variability from cell cultures for plant 

improvement. P. J. Larkin, W. R. Scowcroft. Theoretical and Applied Genetics

16. X. 1981, Volume 60, Issue 4, pp 197-214. 

Abstract: It is concluded from a review of the literature that plant cell culture itself 

generates genetic variability (somaclonal variation). Extensive examples are discussed 

of such variation in culture subclones and in regenerated plants (somaclones). A number of 

possible mechanisms for the origin of this phenomenon are considered. The phenomenon 

may be employed to enhance the exchange required in sexual hybrids for the introgression 

of desirable alien genes into a crop species. It may also be used to generate variants of a 

commercial cultivar in high frequency without hybridizing to other genotypes. 

http://link.springer.com/article/10.1007%2FBF02342540?LI=true

Somaclonal variation - Genetic basis and breeding applications. David A. Evans. 

Somaclonal variation, the recovery of genetic changes in plants regenerated from tissue 

culture, offers an opportunity to uncover natural variability and to use this variability for the 

development of new varieties. This review focuses on the unique variation generated by 

this technique and the current use of somaclonal variation to develop new plant varieties. 

Abstract—pay for full text. 

http://www.sciencedirect.com/science/article/pii/0168952589900218

Somaclonal variation as a tool for crop improvement. Angela Karp. Euphytica
Page 20 of 20

February 1995, Volume 85, Issue 1-3, pp 295-302. Abstract: Somaclonal variation is a tool 

that can be used by plant breeders. The review examines where this tool can be applied 

most effectively and the factors that limit or improve its chances of success.... Somaclonal 

variation is cheaper than other methods of genetic manipulation. At the present time, it is 

also more universally applicable and does not require ‘containment’ procedures. It has 

been most successful in crops with limited genetic systems and/or narrow genetic bases, 

where it can provide a rapid source of variability for crop improvement.

Full article online at http://link.springer.com/article/10.1007%2FBF00023959?LI=true