Notice2024-26232
Movement of Organisms Modified or Produced Through Genetic Engineering; Notice of Additional Modifications Exempt Plants Can Contain
Primary source
Metadata and text below are from the Federal Register, a public-domain U.S. government work. Always verify the official published version before relying on it for any legal matter.
Published
November 13, 2024
Issuing agencies
Agriculture DepartmentAnimal and Plant Health Inspection Service
Abstract
We are adding modifications a plant may contain and qualify for exemption from regulations governing movement of organisms modified or produced using genetic engineering because the modifications are achievable through conventional breeding. An earlier notice proposed five types of modifications. Based on a review of public comments, we have been able to streamline and simplify our description of these modifications and are now finalizing two additional modifications a plant can contain and qualify for exemption. This action updates and clarifies the types of modifications that can be made to plants that qualify for exemption to reflect advances in science and technology, and what is achievable through conventional breeding methods to facilitate the application of biotechnology for the development of new crops.
Full Text
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[Federal Register Volume 89, Number 219 (Wednesday, November 13, 2024)]
[Notices]
[Pages 89569-89585]
From the Federal Register Online via the Government Publishing Office [<a href="http://www.gpo.gov">www.gpo.gov</a>]
[FR Doc No: 2024-26232]
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DEPARTMENT OF AGRICULTURE
Animal and Plant Health Inspection Service
[Docket No. APHIS-2023-0022]
Movement of Organisms Modified or Produced Through Genetic
Engineering; Notice of Additional Modifications Exempt Plants Can
Contain
AGENCY: Animal and Plant Health Inspection Service, USDA.
ACTION: Notice.
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SUMMARY: We are adding modifications a plant may contain and qualify
for exemption from regulations governing movement of organisms modified
or produced using genetic engineering because the modifications are
achievable through conventional breeding. An earlier notice proposed
five types of modifications. Based on a review of public comments, we
have been able to streamline and simplify our description of these
modifications and are now finalizing two additional modifications a
plant can contain and qualify for exemption. This action updates and
clarifies the types of modifications that can be made to plants that
qualify for exemption to reflect advances in science and technology,
and what is achievable through conventional breeding methods to
facilitate the application of biotechnology for the development of new
crops.
DATES: The APHIS website will be updated with these additional
modifications on November 13, 2024.
FOR FURTHER INFORMATION CONTACT: Dr. Neil Hoffman, Science Advisor,
Biotechnology Regulatory Services, APHIS, 4700 River Road, Unit 78,
Riverdale, MD 20737-1238; <a href="/cdn-cgi/l/email-protection#96d8f3fffab8d3b8def9f0f0fbf7f8d6e3e5f2f7b8f1f9e0"><span class="__cf_email__" data-cfemail="4a042f2326640f6402252c2c272b240a3f392e2b642d253c">[email protected]</span></a>; (301) 851-3877.
SUPPLEMENTARY INFORMATION: The regulations in 7 CFR part 340 govern the
movement (importation, interstate movement, or release into the
environment) of certain organisms modified or produced through genetic
engineering. The U.S. Department of
[[Page 89570]]
Agriculture's (USDA's) Animal and Plant Health Inspection Service
(APHIS) first issued these regulations in 1987 under the authority of
the Federal Plant Pest Act of 1957 and the Plant Quarantine Act of
1912, two acts that were subsumed into the Plant Protection Act (PPA, 7
U.S.C. 7701 et seq.) in 2000, along with other provisions. Since 1987,
APHIS has amended the regulations seven times, in 1988, 1990, 1993,
1994, 1997, 2005, and 2020.
On May 18, 2020, we published in the Federal Register (85 FR 29790-
29838, Docket No. APHIS-2018-0034) a final rule \1\ that marked the
first comprehensive revision of the regulations since they were
established in 1987. The final rule provided a clear, predictable, and
efficient regulatory pathway for innovators, facilitating the
development of organisms modified or produced using genetic engineering
(modified organisms) that are unlikely to pose plant pest risks.
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\1\ To view the final rule and supporting documents, go to
<a href="https://www.regulations.gov/docket/APHIS-2018-0034">https://www.regulations.gov/docket/APHIS-2018-0034</a>.
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The May 2020 final rule described the scope or applicability of
regulations and stated that the regulations do not apply to plants with
modifications that are achievable through conventional breeding (85 FR
29790-29796). To ensure the regulations do not apply to plants that are
equivalent to those that could be developed through conventional
breeding, the May 2020 final rule established a regulatory exemption to
initially identify and continuously update modifications that are
achievable through conventional breeding and, thus, exempt from
regulation (85 FR 29791-29796; Sec. 340.1(b)).
Initially, APHIS identified three commonly known modifications
achievable through conventional breeding methods, including small
insertions/deletions at a single locus of a plant's genome (85 FR
29792; Sec. 340.1(b)(1) through (3)). Specifically, Sec. 340.1(b)
exempted plants that contain a single modification of one of the
following types, specified in Sec. 340.1(b)(1) through (3):
<bullet> The genetic modification is a change resulting from
cellular repair of a targeted DNA break in the absence of an externally
provided repair template; or
<bullet> The genetic modification is a targeted single base pair
substitution; or
<bullet> The genetic modification introduces a gene known to occur
in the plant's gene pool or makes changes in a targeted sequence to
correspond to a known allele of such a gene or to a known structural
variation present in the gene pool.
Knowing that it is impracticable to identify and list the universe
of modifications that are achievable through conventional breeding at
any given time because of advances in knowledge, technology and
conventional breeding methods, the May 2020 final rule also established
a process for listing additional modifications that plants can contain
while still being exempted from the regulations (85 FR 29793-29795;
Sec. 340.1(b)(4)). Thus, Sec. 340.1(b)(4) provides that the
Administrator may propose to exempt plants with additional
modifications, based on what could be achieved through conventional
breeding. Such proposals may either be APHIS-initiated or may be
initiated via a request that is accompanied by adequate supporting
information and submitted by another party. In either case, APHIS will
publish a notice in the Federal Register of the proposal, along with
the supporting documentation, and will request public comments. After
reviewing the comments, APHIS will publish a subsequent notice in the
Federal Register announcing its final determination. A list specifying
modifications a plant can contain and be exempt pursuant to Sec.
340.1(b)(4) is available on the APHIS website at <a href="https://www.aphis.usda/gov/biotech-exemptions">https://www.aphis.usda/gov/biotech-exemptions</a>.
On November 15, 2023, we published a notice in the Federal Register
(88 FR 78285-78291, Docket No. APHIS-2023-0022) proposing the five
modifications that plants could contain and be eligible for exemption:
First, we proposed that a diploid or autopolyploid plant with any
combination of complete loss of function modifications in one to all
alleles of a single genetic locus, or an allopolyploid plant with any
combination of complete loss of function modifications in one or both
alleles of a single genetic locus on up to four pairs of homoeologous
chromosomes, without the insertion of exogenous DNA, would qualify for
exemption (proposed 340.1(b)(4)(vi) (Additional Modification 1 (AM1)).
APHIS explained that this category was intended to apply to scenarios
involving targeted DNA breaks--through insertions, deletions, and other
types of modifications (such as a nick)--created using different
techniques that might not be expressly outlined in the initial
modifications APHIS described in the May 2020 final rule (namely,
paragraphs (b)(1) and (2) of Sec. 340.1), but functionally would
achieve the same end result--loss of function. In addition, it proposed
to extend loss of function mutations without the insertion of exogenous
DNA to polyploid plants.
Second, we proposed that any diploid or autopolyploid plant in
which the genetic modification is a single contiguous deletion of any
size, resulting from cellular repair of one or two targeted DNA breaks
on a single chromosome or at the same location(s) on two or more
homologous chromosomes, without insertion of DNA, or with insertion of
DNA in the absence of a repair template, would qualify for exemption
(proposed 340.1(b)(4)(vi)(AM2)). As proposed, allopolyploid plants with
additional modifications to homoeologous loci of homoeologous
chromosomes would not have qualified for exemption.
Third, we proposed to allow the modifications described at Sec.
340.1(b)(2) and (3) to be made to all alleles of a genetic locus on the
homologous chromosomes of autopolyploids (proposed
340.1(b)(4)(vi)(AM3)). As proposed, allopolyploid plants with
additional modifications to homoeologous loci of homoeologous
chromosomes would not have qualified for exemption.
Fourth, we proposed that plants with up to four modifications, made
simultaneously or sequentially, of types that already qualify such
plants for exemption when made individually, and provided each
modification is at a different genetic locus, would be exempt from
regulation because such modifications are achievable through
conventional breeding methods (proposed 340.1(b)(4)(vi)(AM4)). It
proposed that allopolyploid plants could contain up to four of the
proposed complete loss of function modifications described or four
modifications described under Sec. 340.1(b)(2) and (3) or a
combination thereof, provided each modification is introduced into just
one allele; however, allopolyploid plants would not be exempt if they
contain a modification that is allowable only in diploid and
autopolyploid plants.
Fifth, we proposed that plants that have previously completed
voluntary reviews confirming the plants' exempt status as described in
Sec. 340.1(e), which provides the process by which developers can
request such a confirmation of exempt status, and that have been
produced, grown, and observed consistent with conventional breeding
methods appropriate for the plant species, could be successively
modified in accordance with any of the modifications listed under
paragraph 340.1(b) of the regulations (proposed 340.1(b)(4)(vi)(AM5)).
[[Page 89571]]
We initially took comments on the notice through December 15, 2023.
In a notice published in the Federal Register on December 27, 2023 (88
FR 89362, Docket No. APHIS-2023-0022), we reopened the comment period,
and extended it until January 19, 2024.
We received 6,477 comments by the end of the reopened comment
period. The comments were diverse and from interest groups, industry
representatives, industry trade organizations, private individuals,
scientists, plant breeders, and crop specialists.
Based on a review of public comments, we have made several
revisions to the five proposed modifications, simplifying and
consolidating them into two modification categories, AM1 and AM2. To
achieve this, APHIS consolidated the first and second proposed
modifications to create the AM1 described in this final notice. The
intent of the first and second proposed modifications was to provide
developers with greater flexibility in how they could generate targeted
breaks in a plant's DNA like those that occur through conventional
breeding methods. AM1, as finalized, carries through this intent by
building on the existing modification described at Sec. 340.1(b)(1),
which currently allows a single targeted break in DNA and self-repair
(i.e., a non-templated insertion, deletion, or a combination of
insertion and deletion (indel) to rejoin the DNA). AM1 now allows more
than one cut to make the targeted break and the use of external
templates in some circumstances. The finalized AM1 also carries through
the original intent of the proposal by allowing developers to use a
deletion of any size resulting from a targeted break, thereby
recovering the functionality APHIS originally included in the 2019
proposed rule (84 FR 26514-26541, Docket No. APHIS-2018-0034) but did
not expressly articulate in the May 2020 final rule, and which APHIS
proposed as additional modifications in the November 2023 notice (88 FR
78286, 88 FR 78288, Docket No. APHIS-2023-0022). Collectively, as
described in this final notice, AM1 allows plants with modifications
involving an insertion or deletion (indel), or contiguous deletion of
any size, made at a targeted location, with or without insertion of DNA
if generated without using a repair template, or without insertion of
DNA if generated using a repair template, to qualify for exemption.
Similarly, APHIS consolidated the third and fourth proposed
modifications to create the AM2 described in this final notice. The
intent of the third and fourth proposed modifications was to make
modifications that are already listed in the regulations (Sec.
340.1(b)(2) and (3)) available for use in polyploid plants and to
increase the number of modifications that can be made simultaneously or
sequentially to plants. AM2 carries through this intent by exempting
plants with up to 12 modifications, made simultaneously or
sequentially, if each modification occurs in a different gene and is of
a type listed under Sec. 340.1(b). By increasing the number of
modifications that can be made to a plant, AM2 also effectively allows
all modifications listed in Sec. 340.1(b) to be made in all
polyploids.
Finally, the fifth proposed modification would have required
developers to complete a confirmation process to verify a plant's
exempt status before making sequential modifications and outlined
conditions to ensure that simultaneous or sequential modifications were
made in plants that had been produced, grown, and observed, consistent
with conventional breeding practices. APHIS has not finalized a
modification associated with this proposal. Instead, to stay true to
the voluntary nature of APHIS' confirmation request process and ensure
that plants are developed consistent with conventional breeding
practices, APHIS will only accept voluntary requests to confirm a
plant's exempt status for plants that have been produced. This means
APHIS will no longer accept confirmation requests involving plants with
hypothetical modifications because, if produced, the plants may not be
viable, may not have the intended phenotype, or have a different
genotype than originally requested.
We wish to highlight additional distinctions between AM1 and AM2
described in this final notice, and the modifications we initially
proposed. First, we are no longer restricting AM1 to loss of function
modifications if the gain of function (GOF) modification results from
natural DNA repair in the absence of a repair template. We received
comments and supporting literature during the comment period that such
GOF modifications can be accomplished through conventional breeding
techniques. Second, we are no longer making distinctions between
allopolyploids and autopolyploids when describing the modifications. We
received comments during the comment period indicating the distinction
between allopolyploids and autopolyploids was not necessary, with
documentation demonstrating that similar modifications can be made in
the two ploidy types by conventional breeding. Eliminating this
distinction was a key factor that enabled us to consolidate the
modifications from five to two and simplify our description of the
modifications overall. Third, we are increasing the number of
simultaneous or sequential modifications from 4 (as proposed) to 12 (as
described in this final notice). In the proposal we published in
November 2023, we noted that we welcomed comments from the public on
the number of individual modifications that are achievable
simultaneously or sequentially in plants based on conventional breeding
methods, and comments on the reasons for or against allowing for
simultaneous or sequential modifications in all plants. We received
comments during the comment period requesting an increase in the number
of simultaneous or sequential modifications covered by the exemption
and documentation that more than four modifications are possible by
conventional breeding. In our discussion below, we further describe
these comments and the literature references we received that show 12
simultaneous or sequential modifications are achievable through
conventional breeding. Fourth, we are no longer considering
hypothetical plants for confirmation requests based on comments we
received on AM5 suggesting the exclusion of hypothetical plants from
the scope of exemption would simplify the exemption. We are also
clarifying that any plant not subject to part 340 (because it is not
modified, meets the criteria for a regulatory exemption, or has
completed the regulatory status review process) may be modified in
accordance with the exemption.
Below, we first discuss the specific comments that resulted in the
changes to the modifications we proposed in the November 2023 notice.
We then discuss the other comments received on the notice.
Comment: Many commenters felt that we should not make a distinction
between Loss of Function (LOF) and GOF mutations in AM1. They noted
that the distinction greatly increases the complexity of the
modification descriptions.
Response: Proposed AM1 described LOF modifications in all alleles
of a single genetic locus in diploids and autopolyploids and on up to
four pairs of homoeologous chromosomes in allopolyploids. Our proposal
limited the modification to LOF mutations because GOF modifications are
statistically less common than LOF mutations, and we thought the same
GOF mutation would not be expected to occur across multiple alleles in
allopolyploids by conventional breeding. Based on
[[Page 89572]]
comments we received demonstrating proof of concept that GOF mutations
can occur across all subgenomes in allopolyploids (e.g., (Ostlie, et
al., 2015)), we are revising AM1 to allow GOF modifications that result
from the generation of insertions and deletions (indels) that occur
through DNA break and repair.
Because we are dispensing with distinctions between LOF and GOF and
allopolyploids and autopolyploids, we no longer consider it useful to
have a separate modification that allows for a deletion of any size
(proposed AM2). Instead, we have introduced this functionality into the
final AM1. Indels are typically modifications that are under 50 base
pairs (bp) whereas deletions of any size are a type of structural
variant (Mahmoud, et al., 2019).
As noted previously, we are revising AM1 to: ``An indel or
contiguous deletion of any size, made at a targeted location, with or
without insertion of DNA if generated without using a repair template,
or without insertion of DNA if generated using a repair template.''
We wish to emphasize that AM1 is not prescriptive in how indel
modifications or contiguous deletions are made. It is based on the
outcome rather than any specific techniques used. We also wish to
resolve confusion around our use of the phrase ``without the insertion
of exogenous DNA.'' Our intent is to ensure exempt plants are free of
foreign DNA in the final product, but not to prohibit foreign DNA used
to make the final product. For example, CRISPR-Cas9, a foreign DNA,
could be used to make a modification and plants with the modification
and lacking CRISPR-Cas9 would still qualify for the exemption. To be
clear, to qualify for AM1, the final plant must not retain foreign DNA.
Lastly, although we initially defined GOF and LOF based on gene
activity, commenters noted they were confused, because LOF of a gene
can result in a GOF in phenotype and vice versa. Also, by our proposed
definition, promoter deletions that led to either increases or
decreases in the expression of a downstream gene could be GOF or LOF,
respectively. AM1, as described in this final notice, no longer makes a
distinction between LOF and GOF, thereby resolving this confusion and
incongruence and mooting these comments.
Comment: The language of the proposed modifications is complex and
can be simplified by not making a distinction between autopolyploids
and allopolyploids and loss of function and gain of function
modifications.
Response: After reading information provided in the comments
describing the types of modifications that can be made in
allopolyploids, APHIS agrees that our descriptions of modifications
that plants can contain and qualify for exemption can be simplified to
eliminate the distinction between autopolyploid and allopolyploids and
allow gain of function indels. More detail is provided in responses
below.
Comment: Many commenters felt the modifications should not make a
distinction between autopolyploids and allopolyploids and noted that
regulatory authorities in no other countries make this distinction.
Response: Although APHIS initially made a distinction between
allopolyploids (such as wheat) and autopolyploids (such as potato) in
the proposed modifications, based on our review of the comments and
cited literature, we agree that such distinction is not necessary.
For example, we originally proposed that AM4 would have allowed
multiple modifications involving single base pair substitutions and
insertions described in Sec. 340.1(b)(2) and (3), for autopolyploids
as homozygous modifications and for allopolyploids only as heterozygous
modifications. In the comments, we learned of two reasons to change our
view on this distinction. First, in some allopolyploids, such as wheat,
that are largely self-pollinating, homozygous modifications routinely
accumulate, and heterozygous alleles are less common (Rutkoski, et al.,
2022). Second, doubled haploids are commonly used in breeding to
generate homozygous alleles in a single generation in over 250 species
(Maluszynski, et al., 2003). Commenters provided 4 examples of 4-to-8
homozygous mutations pyramided in wheat and rapeseed (Tyagi, et al.,
2014; Zhang, et al., 2019; Zheng, et al., 2020; Luo, et al., 2021;
Wang, et al., 2023b). Given this new information, we have removed the
distinction between allopolyploids and autopolyploids in AM2 as
described in this final notice.
Similarly, as originally proposed, AM1, would have limited the
number of knockouts of a single genetic locus in allopolyploids to four
pairs of homoeologous chromosomes, consistent with the limit of four
modifications in proposed AM4, but counting modifications differently
in autopolyploids and allopolyploids. As described in more detail below
in our discussion of final AM2, which allows multiple modifications, we
will now count modifications in the same way in autopolyploids and
allopolyploids.
Along these lines, as originally proposed, AM3 would have allowed
single nucleotide substitutions (also known as base pair substitutions)
to all alleles of a single genetic locus in autopolyploids, but not
allopolyploids. In response to this proposal, commenters provided
references to published scientific data to demonstrate the use of
conventional breeding to produce an identical homozygous single
nucleotide substitution across all three subgenomes of wheat (Ostlie,
et al., 2015). This modification, a cytosine to thymine (C/T)
transition that converted valine at amino acid 2004 to an alanine,
created resistance to ACCase type inhibitors (Ostlie, et al., 2015) and
the researchers enhanced their chances of finding the desired
modification by using selection with ACCase inhibitors. To evaluate
whether the single nucleotide substitution across all three subgenomes
could be found without selection, we examined the EMS generated mutant
collection (Krasileva, et al., 2017) that is publicly available through
the EnsemblPlants database (<a href="https://plants.ensembl.org/index.html">https://plants.ensembl.org/index.html</a>). The
technology created by (Krasileva, et al., 2017) makes it possible to
identify mutations across multiple genomes. Plants with the desired
mutations can then be crossed to generate plants with the identified
mutations across three genomes. Using this source, we identified 11
cases where wheat lines had C/T mutations that resulted in identical
mutations in ACCase in all 3 subgenomes (D53N; G55D; V212M; A321T;
G543D; G655E; S708N; G1377D; A1848T; G1984E; E2203K) and 2 cases where
wheat lines had G/A mutations that resulted in the identical ACCase
mutation in all three subgenomes (P647S and L1003F). This finding
demonstrated to us that the Krasileva mutagenesis library could be used
to identify plants with the identical single nucleotide substitution
across all three subgenomes even in the absence of selection. This is a
proof of concept that single nucleotide substitutions across subgenomes
can be isolated using ordered mutant libraries prepared from
allopolyploids.
Mutagenized lines tend to create specific types of DNA
modifications. For example, ethyl methanesulfonate (EMS) mutagenesis
preferentially converts the base guanine (G) to adenine (A) and the
base cytosine (C) to thymine (T) (Leitao, 2012). A similar mutagen,
methyl methanesulfonate (MMS) preferentially converts A to T, T to A, A
to G, and T to C (Leitao, 2012). Radiation mutagenesis by gamma
radiation or fast neutron bombardment
[[Page 89573]]
preferentially results in deletions (Wyant, et al., 2022).
Historically, breeders have created collections of lines based on
naturally occurring variation to be used for their breeding pool.
Naturally occurring mutations have been shown to occur at comparable
frequencies for all 12 combinations of nucleotide substitutions (Weng,
et al., 2018). A recent trend is to characterize the collection by
whole genome sequencing (genotyping by sequencing) to facilitate
identification of specific mutations. Sequenced collections of
diversity panels are available in Arabidopsis (The 1001 Genomes
Consortium, 2016), maize (Bukowski, et al., 2018), rice (Zhao, et al.,
2021), soybean (Torkamaneh, et al., 2021), cotton (He, et al., 2021),
canola (Hurgobin, et al., 2018), tobacco (Thimmegowda, et al., 2018),
strawberry (Qiao, et al., 2021), alfalfa (Shen, et al., 2020), sorghum
(Jensen, et al., 2020), and wheat (Brinton, et al., 2020), to name a
few. In some cases, second releases are available with more sequenced
lines covering greater variation than the original. We can expect these
community resources to include more species and details over time.
Genotyping by sequencing is generally applicable to any species.
Given the new information about the availability, for breeding
purposes, of naturally occurring and mutagenized collections genotyped
through sequencing, APHIS concludes that it is possible to identify and
introduce single nucleotide substitutions and deletions across the
subgenomes of allopolyploids by conventional breeding.
As originally proposed, AM3 would have also allowed a modification
that introduces a gene known to occur in the plant's gene pool or makes
changes in a targeted sequence to correspond to a known allele of such
a gene or to a known structural variation present in the gene pool for
autopolyploids, but not for allopolyploids. In the comments, we were
made aware of an example where homozygous copies of a cellulose
synthase-like F6 gene were introduced into all three subgenomes of
wheat (Danilova, et al., 2019). This new information demonstrates that
sequences from the gene pool can be introduced into all subgenomes of
allopolyploids by conventional breeding.
Based on the comments and information we collectively received
related to the proposed modification described as AM3, and as discussed
in the above paragraphs, we are removing the proposed limitation to
autopolyploids. The modifications described in Sec. 340.1(b)(2) and
(3) apply to a single modification. As a result, they were effectively
limited to a single pair of homologous chromosomes in polyploids
species. As discussed more fully below, based on the comments and
literature in this final notice, we will allow up to 12 such
modifications in plants (now AM2). This means modifications can now be
made across subgenomes of polyploids and the plants can qualify for
exemption from regulation, further removing distinctions involving
ploidy plants.
As originally proposed, AM2 would have allowed a modification
consisting of a single contiguous deletion of any size in diploids and
autopolyploids. Given the proof of concept for using an ordered mutant
collection to identify single nucleotide substitutions across
subgenomes of allopolyploids, we considered whether a similar approach
could be used to identify similar deletions across subgenomes such that
allopolyploids would also qualify for proposed AM2. (Krasileva, et al.,
2017) identified just 1268 deletions in their mutant collection, which
is not surprising based on observations that EMS primarily creates
point mutations (Gilchrist and Haughn, 2010). Fast neutron or gamma
radiation mutagenesis, however, predominantly creates deletions
(Gilchrist and Haughn, 2010; Kumawat, et al., 2019) and mutant
population resources using these techniques have been reported (Anai,
2012; Du, et al., 2021). It is likely that ordered mutant collections
prepared by fast neutron bombardment or gamma radiation mutagenesis can
be used to isolate similar, but not identical, deletions across
subgenomes. Given this, and for simplicity, the functionality described
in the modification proposed AM2, is now included in the modification
described as AM1 in this final notice.
Comment: Many commenters felt that proposed AM4 was overly limiting
because breeders routinely combine many more favorable genes, alleles,
or quantitative trait loci (QTL) than four during a breeding project.
One commenter suggested there should be no upper limit following the
lead of other countries such as Canada. Another noted that a complex
trait such as flowering time may require the combination of 50 to 100
QTLs.
Response: In the May 2020 final rule, when USDA first adopted the
exemption for plants with modifications achievable through conventional
breeding, APHIS explained:
``There are many biological and practical factors that affect a
plant breeder's ability to develop a new crop variety by introducing
genetic variation and intentionally selecting for desired traits. These
include the number of targeted loci and type of desired genetic
changes, the genetic distance between the desired changes, generation
time, breeding system (sexual or asexual), ploidy type and level and
genomic complexity, resource availability (time, money, labor, and
genomic resources), extent of domestication, and other factors. These
factors, and thus the extent of intentionally selected genetic
variation that can be introduced, vary widely among plant species.
Moreover, new plant breeding techniques can make possible more complex
combinations of genetic modifications than can practically be achieved
through conventional breeding methods.
Initially, the exemptions will apply only to plants containing a
single targeted modification in one of the categories listed. APHIS
anticipates scientific information and/or experience may, over time,
allow APHIS to list additional modifications that plants can contain
and still be exempted from the regulations so that the regulatory
system stays up to date and keeps pace with advances in scientific
knowledge, evidence, and experience. This may include multiple
simultaneous genomic changes.'' (U.S. Department of Agriculture Animal
and Plant Health Inspection Service, 2020c).
Since APHIS initially adopted its exemption 4 years ago, there has
been steady introgression of desired genes, alleles, and QTLs in
several crops through modern conventional breeding methods. Genomic
assisted breeding, genetic mapping and studies, high through-put
genotyping, speed breeding, multi-parent advance generation inter-
crosses, and pyramid breeding strategies, to name a few, have advanced
quickly and are now affordable for many crop types. New methods, like
OutcrossSeq (Chen, et al., 2021), are consistently emerging to improve
and accelerate breeding methods for difficult to breed crops, like
those for which no inbred lines are available for genetic study and
breeding because they are self-incompatible, clonally propagated, or
have a long generation time, making the identification or integration
of agronomically important genes difficult, particularly in crops with
a complex autopolyploid genome or with predominant asexual
reproduction.
We also considered the progress made in breeding potato, a clonally
propagated crop. Clonally propagated crops are thought to be difficult
to breed because, as a result of not requiring seed production, they
accumulate genetic alterations that are detrimental to breeding and
hence require
[[Page 89574]]
heterozygosity for vigorous growth (Brown, et al., 2017; Kardile, et
al., 2022). Recently much progress has been made in breeding inbred
diploid potato lines by overcoming self-incompatibility (Kardile, et
al., 2022) and purging deleterious alleles causing inbreeding
depression in homozygous lines (Zhang, et al., 2021). These
developments have led to the first potato elite inbred lines
established through selfing that were crossed to successfully exploit
heterosis in the F1 generation (Zhang, et al., 2021).
Similarly, in banana, another clonally propagated crop, low
fertility and seed viability, abnormal meiosis, and inbreeding
depression have been breeding challenges, but some progress has been
made in overcoming fertility problems and seed viability by screening
for fertile plants and using embryo rescue to improve seed germination
((Brown, et al., 2017; Batte, et al., 2019)). The insight gained in
overcoming inbreeding depression in potato will likely be used in other
clonal crops such as banana. We are witnessing conventional breeding
advancements that were once used nearly exclusively to improve easy to
breed crops, now being actively used in breeding programs for difficult
to breed crops.
Some crops that play key roles in nutrition security, sustainable
agriculture, biodiversity, and cultural traditions, have been
overlooked in agricultural crop development because they represent a
small percentage of total tonnage and acreage of production or belong
to resource poor nations. These crops may be difficult to breed because
genetic tools have yet to be developed. However, this situation could
change as advanced breeding tools become more affordable, due to the
steep decline in sequencing costs, and therefore more widely deployed
in all crops.
Commenters provided APHIS with examples demonstrating that many
more than four favorable alleles or QTL can be pyramided. In some
cases, modifications are made to more than one gene to create the
desired trait. In one example, (Ye, et al., 2008) noted that, in
theory, with marker assisted selection coupled with gene pyramiding and
double haploid practices, ``a plant having as many as twenty target
markers can be obtained at an almost perfect certainty in about three
rounds of selection.'' APHIS found several examples in rice where 10 to
11 favorable alleles or QTLs were successfully pyramided (Das, et al.,
2018; Dixit, et al., 2020; Sandhu, et al., 2021; Yadav, et al., 2021).
In one of the cases, the group initially pyramided 15 alleles and QTLs,
with at least some in a heterozygous (non-fixed) condition but lost
some in later generations that they might have retained had they chosen
to use double haploid technology to fix the alleles and QTLs of
interest. We found cases for pyramiding eight alleles or QTLs in tomato
(Hanson, et al., 2016), eight and perhaps more in wheat (Tyagi, et al.,
2014; Rahman, et al., 2020), seven in canola (Wang, et al., 2023b), six
in potato (Rogozina, et al., 2021), five in apple (Baumgartner, et al.,
2015), five in tobacco (Lewis, et al., 2020), five in soybean (Diers,
et al., 2023), five in grape (H[aacute]dl[iacute]k, et al., 2024), four
in coffee (de Almeida, et al., 2021; Saavedra, et al., 2023), and three
in poplar (Lv, et al., 2021). In many cases, these pyramids were fixed
in the homozygous state, while in other species that are typically
vegetatively propagated, some were present in the heterozygous state.
For the potato and grape examples, the papers describe cases where
breeder collections were screened with markers for resistance genes and
individuals in the collection, representing historical crosses, were
found to have pyramids of resistance genes. The other examples
represent cases where the pyramids were specifically bred de novo to
combine target genes in the population.
Given the breeding advances that have been made in many crops, the
number of modifications that can be made in any crop is not static.
Periodic updates to the modifications plants can contain and qualify
for exemption, like this one, will remain necessary moving forward. In
general, the greater the number of favorable alleles or QTLs to be
pyramided in a crop, the greater the number of plants that need to be
screened to obtain the desired plant. Various techniques, such as
second filial (F2) enrichment, are used to reduce the numbers of plants
required, but the numbers of plants required nonetheless rise
exponentially with the number of alleles or QTLs to be pyramided
(Bonnett, et al., 2005; Wang, et al., 2023a). The extent of pyramiding
that is possible also depends on whether the alleles or QTLs are all
present in elite lines, such that little or no backcrossing may be
required to remove deleterious alleles, or whether the alleles and QTLs
are being introgressed from multiple different non-elite lines and wild
relatives, requiring extensive backcrossing. Taking these factors and
the noted differences between species into consideration, in the final
notice we are establishing the number of allowable modifications based
on a number that is readily achievable in crops with advanced breeding
systems and extending this number to all crops as we see evidence of
breeding advances being widely deployed. As we described earlier, for
rice at least 10 modifications have already been achieved multiple
times (Das, et al., 2018; Dixit, et al., 2020; Rahman, et al., 2020;
Sandhu, et al., 2021; Yadav, et al., 2021). Given the rapid advances in
plant breeding this number of modifications will quickly, if not
already, become out of date. Therefore, in this final notice, AM2 will
allow up to 12 modifications made simultaneously or sequentially.
Setting the limit at 12 modifications also enables an even number of
modifications in diploids, triploids, tetraploids, hexaploids, and
octaploids. In terms of counting modifications, both a modification to
a single allele and a pair of functionally equivalent modifications to
a pair of alleles on homologous chromosomes will count as one
modification. Thus, where all alleles of a given locus are modified,
the maximum number of modified loci is 12 in diploids, 6 in
tetraploids, 4 in hexaploids, and 3 in octoploids. Triploids and
pentaploid modifications will be counted as tetraploids and hexaploids,
respectively. In polyploids, if only one allele is modified in the case
of a dominant mutation, the loci modified can exceed 6, 4, and 3 in
tetraploids, hexaploids, and octoploids, respectively. In terms of
counting, there are at least three cases where multiple DNA breaks or
edits can be made and ``counted'' as a single modification:
1. When two guide RNAs are used to cut out a single contiguous
portion of a gene or to otherwise make a single deletion of any size.
2. When multiple indels are created near the target site or at any
other unintended sites with near homology to the target site with one
indel being functional while the other indels have no additional
effect.
3. A gene in the gene pool is inserted into the genome or an
existing gene is edited several times to correspond to a gene in the
gene pool.
As noted previously, in this final notice, the proposed AM4 is
renumbered as AM2 and is revised as follows: ``Plants with up to 12
modifications, made simultaneously or sequentially, are exempt from
regulation if each modification individually qualifies the plant for
exemption and occurs in a different gene.''
With respect to this final version of AM2, we wish to clarify that
the phrase ``individually qualifies the plant for exemption'' refers to
the modifications described at Sec. 340.1(b) that qualify
[[Page 89575]]
plants for exemption and does not include the exemptions described in
Sec. 340.1(c). We also wish to note that when AM2 is used in
combination with AM1, we are restricting the use of repair templates to
create modifications across subgenomes. As noted above, we expect that
ordered mutant libraries could be used to identify similar but not
identical deletions across subgenomes in allopolyploid species. We have
not yet identified any literature demonstrating that identical indel or
deletion modifications can be achieved across subgenomes using
conventional breeding methods. For this reason, we are restricting the
application of AM2 in combination with AM1, when a repair template is
used, to allow modification to one pair of homologous chromosomes. If
new literature emerges demonstrating an identical indel or deletion
modification can be achieved across subgenomes using conventional
breeding methods, we will reconsider this restriction.
Comment: Several commenters asked APHIS to clarify whether AM5
applies to plants that have been cleared through the regulatory status
review or petition process. Another concern raised was that AM5 would
change a voluntary consultation process into a mandatory process with
the requirement that the exemption only applied to plants that are
``produced, grown, and observed consistent with conventional breeding
methods.'' Another commenter suggested removing the requirement for a
plant to be produced, grown, and observed consistent with conventional
breeding methods because it is not clear what APHIS meant. Some
commenters noted that APHIS could restrict hypothetical, successively
modified plants from AM5 by stating in associated guidance that plants
that are merely hypothetical in nature would not be eligible for
subsequent hypothetical modifications because they have not yet been
produced, grown, and observed consistent with conventional breeding
methods for the appropriate plant species.
Response: APHIS acknowledges that plants that are not subject to
part 340, because they have undergone the petition process, the
regulatory status review process, or meet the criteria for regulatory
exemption, may be modified in accordance with the exemption. Therefore,
it is no longer necessary to use proposed AM5 to describe this
allowance. APHIS wishes to clarify that an exempt plant can only
contain a single modification to a particular gene. For example, this
means that once a modification has been made to a particular gene and
that plant is not subject to part 340, plants with successive
modifications to the same gene will not qualify for exemption because
such modifications are not achievable through conventional breeding.
APHIS agrees with the commenters who suggested that APHIS should no
longer consider hypothetical modifications for confirmation requests.
APHIS is concerned that allowing large numbers of hypothetical
modifications will overburden APHIS with confirmation requests for
plants that have little or no value because the plants may not be
viable, may not have the intended phenotype, or have a different
genotype than originally requested.
Response to General Comments on the Proposed Modifications
Comment: Pay special attention to the massive lawsuits resulting
from the human health impacts of glyphosate, which would not have
happened if glyphosate-resistant genetically modified organisms (GMOs)
had not been released into the environment.
Response: While it is true that glyphosate has been the subject of
litigation, APHIS does not agree with the commenter that glyphosate use
on glyphosate resistant (GR) crops has been the primary subject of the
litigation. Glyphosate is widely used in the residential lawn and
garden market business segment. When glyphosate is used in the lawn and
garden markets, glyphosate is not sprayed on GR crops. According to
Werner Baumann, CEO of Bayer AG, more than 90 percent of the Roundup
litigation claims Bayer has faced in recent years have come from the
U.S. residential lawn and garden market business segment that do not
involve the application of glyphosate onto GR crops (Brooks, 2021).
Comment: Absent case-specific government oversight, testing, and
approval of individual GMO products, how would ``voluntary'' testing by
manufacturers protect Americans from potentially negative health
effects of consuming products engineered under such broad exemptions?
Response: The modifications (AM1 and AM2) described in this final
notice pertain to products that otherwise could be produced by
conventional breeding. Although conventional breeding is not risk free,
the risks associated with it are manageable by accepted standards
(National Research Council, 1989). The health effects of products that
qualify for exemption are not expected to be different than the risks
posed by conventionally bred crops and likewise manageable by accepted
standards.
Comment: What level of documentation and data transparency would be
required of GMO producers who might exploit the proposed exemptions?
Response: The developers of crops that qualify for exemption have
no requirements to submit documentation to APHIS. If they wish
confirmation from APHIS that their particular crop meets the criteria
for exemption, the developer can request a confirmation request.
Information needed for a confirmation request is detailed in a guide
found on APHIS' Biotechnology Regulatory Services website (<a href="https://www.aphis.usda.gov/sites/default/files/requesting-confirmation-of-exemption.pdf">https://www.aphis.usda.gov/sites/default/files/requesting-confirmation-of-exemption.pdf</a>). Again, however, we wish to reiterate that this final
notice describes modifications pertaining to products that could
otherwise have been developed through conventional breeding. This
limitation on the scope of the modifications that plants can contain
and qualify for exemption precludes the sort of abuse envisioned by the
commenters.
Comment: Would third-party testing be required before releasing
food products produced using the proposed modifications and exempt from
regulation?
Response: Oversight of all food products including those produced
using plants that qualify for exemption is conducted by the U.S. Food
and Drug Administration (FDA). FDA recently released guidance for
industry on foods derived from plants produced using genome editing
(U.S. Food and Drug Administration, 2024). FDA explained in the New
Plant Variety (NPV) policy that the regulatory status of a food,
irrespective of the method by which it is developed, is dependent upon
objective characteristics of the food and the intended use of the food
(or its components) (57 FR 22984 at 22984).\2\ Please see the FDA's
guidance for more information (U.S. Food and Drug Administration,
2024).
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\2\ May 29, 1992 (57 FR 22984-23005; Docket No. 92N-0139).
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Comment: One commenter suggested that USDA conduct public trials to
establish the modifications are safe before finalizing the exemptions.
Response: We disagree. The modifications described in this final
notice only pertain to plants with modifications that could otherwise
be achieved through conventional breeding. Conventionally bred crops
have a history of safe use. Public field trials of crops with
modifications eligible for exemption would not be expected to reveal
otherwise because
[[Page 89576]]
the use of genetic engineering, in and of itself, does not present an
increased plant pest risk (National Research Council, 1987; National
Research Council, 1989; National Academies of Sciences Engineering and
Medicine, 2016).
Comment: The proposed modifications sidestep National Environmental
Protection Act (NEPA) review, transparency, and public participation.
Response: We disagree with this comment. The exemption at Sec.
340.1(b) excludes from the scope of regulation at part 340, modified
plants that could have been created through conventional breeding to
ensure that plants with similar characteristics are treated similarly
from a regulatory perspective. APHIS assessed this exemption in the
Programmatic Environmental Impact Statement (PEIS) prepared to support
the 2020 revisions to part 340, which included a thorough, detailed,
and transparent review, and invited public comment on, the description
of why modified plants described at Sec. 340.1(b) fall outside of
APHIS's authority under the regulations. APHIS explained that modified
plants that qualify for exemption under Sec. 340.1(b), are no
different, as a class, and in terms of plant pest risk, from comparable
plants that are made through conventional breeding, which, likewise, do
not come before APHIS. In May 2020, when APHIS adopted the revised part
340, APHIS expressly stated in the final rule that it would continue to
update the modifications that plants can contain and qualify for
exemption to further clarify the types of modified plants that do not
fall within the scope of regulation. As described in the PEIS, where,
as here, modified plants are not within APHIS's scope of regulation or
jurisdictional authority, a NEPA analysis is not required. It is also
worth noting that the modifications described in this final notice
would have also fallen outside the scope of the legacy regulations
previously codified at part 340, because plants with such modifications
would not have met the definition of a ``regulated article.''
Sec. Sec. 340.0, 340.1 (2019). Many developers provide transparency by
voluntarily submitting confirmation requests to APHIS. When APHIS
confirms a modified plant meets the criteria for exemption from
regulation, APHIS posts on its website the incoming submission and our
response, redacted to protect Confidential Business Information, as
appropriate.
Comment: The modifications may increase the amount of genome edited
crops in the food supply and lead to an increase in commingling of
genome edited crops with crops that are not produced with genetic
engineering or genome editing including organic crops. Crops created
using genome editing may not be disclosed as bioengineered. For these
two reasons, consumers wishing to purchase food made without this
technology may have more limited consumer choice.
Response: Again, it is worth noting that the modifications
described in this final notice would have also fallen outside the scope
of the legacy regulations previously codified at 7 CFR part 340,
because plants with such modifications would not have meet definition
of a ``regulated article.'' Sec. Sec. 340.0, 340.1 (2019). With that
said, genome edited crops that meet the criteria for exemption from
part 340 are currently not permitted to be used in organic production
(National Organic Standards Board, 2019). Inadvertent commingling of
crops exempted from part 340 would not result in loss of organic
certification to the organic producer, however. Although commingling is
possible, if it were to occur, we expect it to occur at a low
frequency.
As we noted in the PEIS associated with the 2020 revisions to part
340, on average 1 to 3 percent of non-GE farmers have reported
commodity rejection by suppliers due to the presence of GE crop
material, and the number of organic farms reporting economic losses
from the presence of GE material was 0.7 percent in 2010 (U.S.
Department of Agriculture Animal and Plant Health Inspection Service,
2020a). In the PEIS, we also noted that we expected innovation in the
agricultural biotechnology to increase under revised part 340, and
there could be seen a wider variety of modified crop plants in
commercial production. If development and adoption by growers of new
varieties of modified crop plants does occur, there may be an increase
in the potential for incidents of unintended presence of modified crop
material in non-modified crops or crop products. This would primarily
be due to the possibility that there would be more modified crop
varieties in production and therefore more non-modified crop types that
could potentially have commingling issues with the corresponding GE
crops. An increase in development and adoption of new varieties of
modified crops would entail maintaining segregation of modified crop
products from a wider variety of non-modified and identity-preserved
cropping systems along supply chains.
Though the likelihood of commingling could increase, there are
incentives to keep it low. Identity preserved systems are in place to
guard against commingled products entering the marketplace and non-
modified producers have economic incentives to keep it low.
Furthermore, most modified plants exempt from Sec. 340.1(b) are not
immediately commercialized as they may still be subject to regulation
by FDA and U.S. Environmental Protection Agency (EPA), as appropriate.
From our experience with the Am I Regulated Program (AIR) under the
legacy regulations, there were roughly 80 cases of plants that
completed the AIR process, but only three of the modified plants were
or are being grown in the United States for commercial purposes (High
Oleic Acid soybean, waxy corn, and a reduced pungency mustard green).
Additionally, it has been our experience that many developers whose
products meet the criteria for exemption nonetheless ask for
confirmation letters because the letters help them market their
products domestically and overseas. These letters are posted on the
APHIS website and are available to the public. Organic and other
growers of non-modified crops have this resource to become aware of new
genome edited crops. Conversations between neighbors and other
voluntary interactions are another way for an organic grower to learn
whether their neighbors are growing GE crops, and if so, to take steps
to minimize commingling.
Comment: Some commenters expressed concern about off target and
unintended effects.
Response: APHIS considers some off-target and unintended effects.
For example, APHIS considers the unintended retention of exogenous DNA
inserted as part of the modification process to be an unintended
modification (e.g., DNA encoding genome modification machinery such as
the Cas9 protein). APHIS also considers modifications to DNA sequences
that are highly similar to the target sequence as unintended
modifications (e.g., sequences found in multigene families that have
the same or highly similar sequences as the intended target,
pseudogenes, or other conserved sequences), as those sequences would
likely be modified at frequencies exceeding low-similarity promiscuous
binding. Except for Sec. 340.1(b)(3) and AM2 involving Sec.
340.1(b)(3) type modifications (i.e., modifications that allow for the
insertion of a gene from a plant's gene pool), the modified plant must
be free of any DNA that was deliberately inserted as part of the
modification process, including vector sequences, and requests to
confirm a plant's exempt status should include
[[Page 89577]]
scientific methodology describing the design or verification steps
taken to anticipate, reduce, and monitor for off-target modifications
to highly similar sequences. For Sec. 340.1(b)(3) and AM2 involving
Sec. 340.1(b)(3) type modifications, only DNA from within the gene
pool may be retained in the plant.
APHIS does not consider modifications occurring at sites without
similarity to the target region, as these are associated with
spontaneous or other types of background mutation that occur naturally
in plants and do not raise plant pest risk concerns in conventional
breeding programs. APHIS does not believe it is necessary to regulate
such modifications of genome editing in plants because (1) the mutation
rate from genome editing at sites without similarity to the target
region is low relative to the background mutation rate that occurs in
conventional breeding, and (2) whatever changes do occur are likely to
be segregated away from the target mutation during the breeding
process. Comprehensive CRISPR/Cas off-target analysis on a genome-wide
scale has been performed in rice, maize, tomato, and Arabidopsis (Feng,
et al., 2014; Peterson, et al., 2016; Nekrasov, et al., 2017; Feng, et
al., 2018; Tang, et al., 2018; Lee, et al., 2019). In these cases where
the frequency of mutation at sites without similarity to the target
region was measured in CRISPR/Cas expressing lines and their progeny,
the authors concluded that the rate of mutation was below the level of
background mutation induced during seed multiplication or tissue
culture (Hahn and Nekrasov, 2019). Although there can be variation in
mutation rates due to the nature of the technique used and the
biological system to which it is applied, the mutation rates in such
conventional breeding techniques as chemical and irradiation-based
mutagenesis dwarf the rate associated with genome editing methods.
Due to the nature of plant breeding--in which populations are
created and evaluated, and individual plants are selected for the
intended modifications--untargeted modifications (or untargeted
mutations) are likely to be lost unless they are genetically linked to
the targeted modification that is introduced. For these reasons, APHIS
does not consider untargeted modifications (untargeted mutations) when
determining eligibility for an exemption. This is also consistent with
APHIS' approach regarding conventional breeding techniques.
APHIS believes that similar products should have similar regulatory
requirements. Crops made by conventional breeding are not reviewed for
spontaneous and/or background mutations.
Comment: There should be no exemptions. There needs to be
comprehensive safety testing and long-term environmental monitoring for
all GE crops.
Response: This comment is outside the scope of this notice, and,
for reasons discussed in the final rule (U.S. Department of Agriculture
Animal and Plant Health Inspection Service, 2020c), we disagree with
the commenter.
Comment: USDA does not and cannot demonstrate that GE plants thus
exempted would not pose increased plant pest or noxious weed risks.
Plants that are exempt are more disease susceptible, e.g. Nicotiana
attenuata, low lignin plants.
Response: Consistent with the provisions in Sec. 340.1(b)(4), the
modifications that APHIS has described are not based on plant pest risk
per se but, instead, are based on whether the modified plant could have
been achieved through conventional breeding. Plants produced through
conventional breeding are not risk free; rather, their risks are at an
acceptable level that has historically not merited regulation. Plants
with additional modifications listed in this final notice are not
expected to have any greater risk than those having a history of safe
use.
Comment: USDA has placed limitations on the modifications and these
limitations are not based on plant pest risk.
Response: As described in the regulations, the modifications
described in this final notice are based on modifications that could be
achieved through conventional breeding. For each modification, APHIS
has identified literature and publicly available information indicating
proof of concept that the additional modifications are achievable
through conventional breeding.
Comment: Modifications should be inclusive of the current state of
scientific knowledge and not just the literature record because the
literature does not capture the full range of modifications that are
achievable through conventional breeding.
Response: Consistent with the provision at Sec. 340.1(b)(4), APHIS
has developed the modifications based on available literature and
public information (including the comments we received in response to
the proposal) describing modifications achievable through conventional
breeding.
Comment: The modifications should broaden the origin boundaries for
insertions to include any sequences in the kingdom Plantae versus
sexual compatibility.
Response: We acknowledge that examples of horizontal gene transfer
have occurred in plants on an evolutionary time scale. Our review of
the literature indicates these types of insertions do not routinely
occur during the conventional plant breeding process. At this time, we
will not broaden the modifications to allow insertions from any species
within the kingdom Plantae.
Comment: USDA should broadly exempt all gene edited products.
Response: The exemption at Sec. 340.1(b) is for DNA modifications
that could be achieved through conventional plant breeding. Based on
the available literature and public information, some types of gene
editing can accomplish modifications beyond what can currently be
achieved through conventional breeding. Although products with these
types of edits are not currently exempt from regulation, most non-
exempt plants have a pathway for commercialization through the
regulatory status review process to evaluate the plant pest risk of
those products.
Comment: A commenter advised APHIS to conduct regular and frequent
review of regulations to stay relevant in light of new scientific
developments.
Response: APHIS agrees and in fact does so. APHIS also reminds
stakeholders that under Sec. 340.1(b)(4), they can help APHIS ensure
the regulations are current by informing APHIS of new scientific
developments that demonstrate that additional modifications are
possible through conventional breeding.
Response To Specific Comments on the Proposed Modifications
Comment: APHIS should also consider the de-regulation of cis
genetically engineered crops, made by targeted insertion or CRISPR
transposition systems (emerging tools to be utilized in crops).
Response: Plants with targeted insertions qualify for the exemption
listed at Sec. 340.1(b)(3) if the inserted sequence is found within
the plant's gene pool. CRISPR transposition systems can be used to make
cisgenic modifications to plants that qualify for exemption provided
the CRISPR tools (or any foreign DNA) are segregated away from the
final product.
Comment: APHIS should provide guidance for when a plant contains a
modification meets more than one of the criteria for exemption.
Response: The commenter has presented an example where two cuts
[[Page 89578]]
are made to a single locus, a deletion that would qualify under AM1 and
a targeted insertion that would qualify under Sec. 340.1(b)(3). In
cases where a plant has been edited in a manner that meets the
description of more than one of the modifications listed under Sec.
340.1(b), developers can claim either type of modification as the basis
for their confirmation request.
With the new AM2, there will be cases where a plant may have
modifications of multiple types listed under paragraph 340.1(b). For
example, a developer might make an indel modification to one gene and a
single nucleotide substitution to a second gene. In that case the
developer should claim AM2 for the multiple modifications and specify
the type of each modification made in the plant. APHIS will provide
additional examples on its website for greater clarity. It will be fact
specific based on the specific nature of the plant. We invite
developers to consult with us to determine the appropriate path.
Comment: Commenters raised the point that the notice did not
address triploid crops such as watermelon, banana, and plantain and
aneuploids such as peppermint and complex auto/allopolyploids such as
sweet potato. A commenter also pointed out that for many species the
distinction between auto and allopolyploids is not always
straightforward. For example, homologous recombination, one of the
distinguishing characteristics of autopolyploid is thought to occur to
varying degrees in allopolyploids.
Response: As we are no longer making a distinction between
autopolyploids and allopolyploids in the modifications described in
this final notice, these points are now moot.
Comment: A comment was made that the term ``loci'' is not precise
when applied to allopolyploids because it implies a positional
relationship remains intact in evolution and positional relationships
between homoeologs could have changed during speciation prior to
polyploidization.
Response: We agree with the commenter. It can be difficult to tell
whether a gene in one subgenome directly corresponds to a similar gene
on another subgenome. Confusion can result because gene families may
have arisen due to gene duplication prior to the hybridization event
that resulted in the speciation, and after speciation genetic
rearrangements may have altered positional information (Adams and
Wendel, 2005; Soltis, et al., 2014). Furthermore, after speciation gene
inactivation may have reduced the number of gene family members on one
subgenome relative to another further confounding the evolutionary
relationships between genes (Adams and Wendel, 2005; Soltis, et al.,
2014). We wish to clarify that our meaning for genetic locus in
allopolyploids pertains to a single pair of alleles in each subgenome
at a fixed location and need not reflect positional relationships
across other subgenomes.
Comment: Commenters requested clarification as to when an external
template may be used.
Response: An external repair template may be used to generate a
modification and the plant will qualify for an exemption when creating:
1. An indel without insertion of DNA or a single contiguous
deletion of any size provided the final product does not retain foreign
DNA (AM1). When combined with AM2, application of AM1/AM2 is restricted
in creating exact modifications across subgenomes. For indels or
deletions that require exact modifications for the desired outcome, the
exemption allows modification to one pair of homologous chromosomes. If
an external template is used to make an indel or deletion that need not
be specific, such as for gene inactivation, the restriction of AM1/AM2
to one pair of homologous chromosomes does not apply;
2. A single base pair (nucleotide) substitution (Sec.
340.1(b)(2)); and
3. Insertion based on sequences within the gene pool (Sec.
340.1(b)(3)).
When an external repair template is used to make a targeted
insertion representing a sequence outside the gene pool, the plant
would not qualify for exemption.
Comment: The proposed modifications are at odds with international
regulations especially on the number of edits allowed and with respect
to ploidy. The USDA should consider evaluations undertaken by expert
agencies in other geographies such as Argentina, Brazil, Canada, and
the European Union.
Response: In response to these comments, APHIS has reviewed the
frameworks for other international and domestic regulatory agencies
that oversee products of biotechnology. Globally, regulatory frameworks
for biotechnology leverage different authorities and definitions, and
subsequently have different approaches to regulation. One approach uses
the definition of a ``living modified organism'' from the Cartagena
Protocol on Biosafety (Secretariat of the Convention on Biological
Diversity, 2000) to determine what biotechnology products fall under a
regulatory scope. This approach is now used by many countries,
including Argentina. Beginning in 2015, and continuing with updates
through 2021, Argentina has maintained a regulatory framework \3\ for
new breeding technologies, including genome editing (Lema, 2020). In
Argentina, all modified plants require evaluation to determine whether
or not they are considered a GMO under Argentina law. Under the
``Argentina Model,'' products developed using genome editing are not
considered genetically modified organisms unless they contain a ``new
combination of genetic material,'' which it defines as ``change
produced in the genome of the organism by the incorporation, in a
stable and joint manner, of ONE (1) or more genes or nucleic acid
sequences that are part of a defined genetic construction.'' Regardless
of the outcome of this analysis, Argentina may impose monitoring
requirements on any plant product based on its characteristics and/or
novelty. Countries that have adopted approaches that are similar to the
Argentina Model, include Chile, Brazil, Paraguay, Uruguay, Colombia,
Guatemala, Honduras, Japan, the Philippines, and Israel.
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\3\ <a href="https://www.argentina.gob.ar/normativa/nacional/resoluci%C3%B3n-21-2021-346839/texto">https://www.argentina.gob.ar/normativa/nacional/resoluci%C3%B3n-21-2021-346839/texto</a>.
---------------------------------------------------------------------------
Other countries have also recently considered how to regulate the
products of genome editing within their existing regulatory frameworks.
For example, in 2023, the Canadian Food Inspection Agency updated their
guidance to clarify that genome edited crops do not present novel risks
and, like certain other crops grown in Canada, do not require review
unless the crop has an herbicide resistance trait or has both a novel
trait and a potential to have significant environmental impacts
(Government of Canada, 2023b; Government of Canada, 2023a). The United
Kingdom also finalized a ``Genetic Technology Act'' \4\ in 2023 to
establish new regulatory and marketing standards for plants and animals
that are ``precision bred'' and remove such products from regulation as
genetically modified organisms. Under this law, a modified plant is
``precision bred'' if ``(a) any feature of its genome results from the
application of modern biotechnology, (b) every feature of its genome
that results from the application of modern biotechnology is stable,
(c) every feature of its genome that results from the application of
modern biotechnology could have resulted from traditional processes,
whether or not in conjunction with selection techniques, alone, and (d)
its genome does not
[[Page 89579]]
contain any feature that results from the application of any artificial
modification technique other than modern biotechnology produced through
precision breeding techniques, so long as they could have resulted from
traditional processes.'' (emphasis added).
---------------------------------------------------------------------------
\4\ Genetic Technology (Precision Breeding) Act 2023
(<a href="http://legislation.gov">legislation.gov</a>.uk).
---------------------------------------------------------------------------
In February 2024, the European Parliament voted in favor of
proposed legislation \5\ that would consider plants produced through
``New Genomic Techniques'' (NGT) (like genome editing) as conventional
equivalents if such plants could also occur naturally or be produced by
conventional breeding. Under the proposal, an NGT plant ``is considered
equivalent to conventional plants when it differs from the recipient/
parental plant by no more than 20 genetic modifications'' of various
types (European Commission, 2023b). These include targeted
modifications are similar to those APHIS has identified in Sec.
340.1(b)(1) through (3) and in AM1 and AM2 (small insertions, deletions
of any length, nucleotide substitutions, and insertions or
substitutions of DNA present in the gene pool of the plant). The
proposal, which has not yet reached consensus agreement among EU
members, includes a mandatory verification that a plant meets the NGT
criteria. Most recently, on July 11, 2024, the European Food Safety
Authority (EFSA) published an opinion (European Food Safety Authority
Panel on Genetically Modified Organisms, et al., 2024) on the
definitions and scientific justification of the NGT proposal in
response to an analysis by the French Agency for Food, Environmental
and Occupational Health & Safety. EFSA concluded that ``it is
scientifically justified to consider [certain NGT plants identified in
the proposal] as equivalent to conventionally bred plants.'' As a next
step, the Council of the European Union will begin negotiations with
member states about the specifics of the legislation--that is to say,
this law is not yet final.
---------------------------------------------------------------------------
\5\ <a href="https://food.ec.europa.eu/document/download/c03805a6-4dcc-42ce-959c-e4d609010fa3_en?filename=gmo_biotech_ngt_proposal_2023-411_en.pdf">https://food.ec.europa.eu/document/download/c03805a6-4dcc-42ce-959c-e4d609010fa3_en?filename=gmo_biotech_ngt_proposal_2023-411_en.pdf</a>.
---------------------------------------------------------------------------
Changes in regulatory approaches involving products of genome
editing are also being made in southeast Asia. Most recently, in July
of 2024, Thailand revised its regulations to allow for the
certification and subsequent release into the environment of
``organisms developed from gene editing technology,'' defined as
``organisms that have been genetically improved in a manner similar to
mutation or hybridization, where the final product contains genetic
material from donor organisms that can naturally crossbreed with the
recipient organisms.'' In August 2024, the Singapore Food Agency (SFA)
published its framework for genome edited crops (Singapore Food Agency,
2024). SFA will regulate crops that contain foreign DNA, which includes
crops with DNA that could not have been inserted naturally or been
introduced into the crop using conventional breeding techniques. In
cases where the developer determines their crop contains foreign DNA,
SFA requires the crop to undergo a pre-market safety assessment. For
crops with modifications made through genome editing that do not
involve the retention of foreign DNA, developers are encouraged (but
not required) to notify SFA in cases where they determine their crop
does not contain foreign DNA.
Within the United States, in May 2023, the EPA issued a final rule
exempting a class of plant-incorporated protectants (PIPs) created
using genetic engineering from registration requirements under the
Federal Insecticide, Fungicide, and Rodenticide Act (FIFRA), and from
the food or feed residue tolerance requirements under the Federal Food,
Drug, and Cosmetic Act (FFDCA) (U.S. Environmental Protection Agency,
2023). The final rule exempts PIPs from FIFRA registration and FFDCA
tolerance requirements in cases where they both pose no greater risk
than PIPs that EPA has already concluded meet safety requirements, and
when they could have otherwise been created through conventional
breeding, as follows: PIPs in which genetic engineering has been used
to insert or modify a gene to match a gene found in a sexually
compatible plant; and, loss-of-function PIPs in which the genetically
engineered modification reduces or eliminates the activity of a gene,
which then helps make the plant resistant to pests. EPA's PIP exemption
does not limit the number of modifications developers can make using
genetic engineering provided the resulting PIPs meet the criteria for
exemption. More recently, on February 22, 2024, FDA issued updated
guidance related to the handling of NPV to affirm that ``the regulatory
status of foods derived from plant varieties produced using genome
editing will, like that of food from other plant varieties, be based on
the objective characteristics of the food and the intended use of the
food (or its components)'' (U.S. Food and Drug Administration, 2024).
Although there are some differences in specific details, in
general, we see countries around the world adopting a similar approach
as we have for the movement of plants developed using new genome
editing techniques: If a modified plant could have been developed using
conventional breeding, the level of regulatory oversight will more
closely align with a conventionally developed product. In 2020, when
APHIS first adopted the exemption for plants with modifications
achievable through conventional breeding, APHIS explained:
``There are many biological and practical factors that affect a
plant breeder's ability to develop a new crop variety by introducing
genetic variation and intentionally selecting for desired traits. These
include the number of targeted loci and type of desired genetic
changes, the genetic distance between the desired changes, generation
time, breeding system (sexual or asexual), ploidy type and level and
genomic complexity, resource availability (time, money, labor, and
genomic resources), extent of domestication, and other factors. These
factors, and thus the extent of intentionally selected genetic
variation that can be introduced, vary widely among plant species.
Moreover, new plant breeding techniques can make possible more complex
combinations of genetic modifications than can practically be achieved
through conventional breeding methods.
Initially, the exemptions will apply only to plants containing a
single targeted modification in one of the categories listed. APHIS
anticipates scientific information and/or experience may, over time,
allow APHIS to list additional modifications that plants can contain
and still be exempted from the regulations so that the regulatory
system stays up to date and keeps pace with advances in scientific
knowledge, evidence, and experience. This may include multiple
simultaneous genomic changes.'' (U.S. Department of Agriculture Animal
and Plant Health Inspection Service, 2020c).
As discussed above, APHIS has received numerous comments and
supporting literature and has conducted our own extensive literature
review indicating that 12 modifications are within the scope of
conventional breeding for diploids and polyploids. Based on this new
information, we have eliminated most restrictions on the modification
of allopolyploids, eliminated the restrictions with regard to GOF
modifications, and increased to 12 the number of modifications that can
be made simultaneously or sequentially in plants that qualify for
exemption. As such, the modifications described in this final notice
bring APHIS' treatment of plants with modifications that are
[[Page 89580]]
achievable through conventional breeding into greater alignment with
other countries that have adopted regulatory approaches that consider
most genome edited plants as conventional equivalents, including those
that allow multiple modifications and modifications in ploidy plants.
Comment: A commenter noted that several modifications might be made
to the same genetic locus if successive rounds of mutagenesis were
used. Thus, it seems unnecessary to limit targeted base pair
substitutions to one base pair in Sec. 340.1(b)(2).
Response: APHIS is not aware, and the commenter did not provide an
example of this type of modification made by conventional breeding.
Until we have more concrete proof of concept, APHIS will limit targeted
modifications to a single modification per gene. This limitation
applies to successive modifications made to a plant that qualifies for
exemption under Sec. 340.1(b).
Comment: A commenter noted that a certain number of nucleotides can
always be present in a plant's genome simply by chance. In the European
Union's proposal for the regulation of NGT, insertions or substitutions
of up to twenty nucleotides are considered to be exempted from the GMO
regulations, irrespective if they result in GOF or LOF. A similar
sentiment was expressed in the comment that sequences of smaller sizes
from outside the breeder's gene pool should be exempted.
Response: As noted above, the European Union proposal is not yet
final and remains under negotiation within the European Union. As part
of considering this proposal, the European Commission has made
available a document entitled, ``Potential criteria to determine
whether a plant obtained by targeted mutagenesis or cisgenesis could
also occur naturally or be produced by conventional breeding
techniques,'' which includes a disclaimer indicating this ``draft has
not been adopted or endorsed by the European Commission (European
Commission, 2023a). Any views expressed are the preliminary views of
the Commission services and may not in any circumstances be regarded as
stating an official position of the Commission.'' Although we are not
revising the modifications to incorporate this suggestion at this time,
we will continue to follow developments in the European Union as they
are finalized. With that said, we wish to note that within this final
notice, in AM1, we allow insertions that occur in the absence of a
repair template. This repair could result in a sequence not within the
gene pool and there is no restriction on the size of the repair
(insertion).
Comment: One commenter asked for clarification as to whether, in
proposed AM4 and AM5, heterozygosity refers to genomic rather than
allelic.
Response: In the proposed modifications, the heterozygosity
referred to allelic. However, the modifications described in this final
notice no longer make distinctions between allopolyploids and
autopolyploids, so this point in now moot.
Comment: One commenter noted that the observation mandate in AM5
unfairly penalizes crops with excessively long breeding cycles such as
trees or berries, and research groups with limited access to field
trials such as small universities.
Response: Moving forward, we will only consider confirmation
requests for actual plants with up to 12 modifications. Our standard
for the exemption is based on a conventional breeding standard and
crops with long breeding cycles are also at a similar disadvantage
compared to short cycle crops under conventional breeding. The
regulatory status review process provides another pathway to
commercialization that may be more advantageous for long cycle crops
that require more than 12 simultaneous modifications.
Comment: There is ongoing litigation on the revisions to 7 CFR part
340. New modifications should not be finalized prior to judicial ruling
on the ongoing litigation.
Response: We disagree with this comment.
In May 2020, when APHIS issued the final rule outlining the updates
to 7 CFR part 340, APHIS anticipated scientific information and/or
experience would, over time, allow APHIS to list additional
modifications that plants can contain and be exempted from the
regulations so that the regulatory system stays up to date and keeps
pace with advances in scientific knowledge, evidence, and experience.
To ensure the regulations do not apply to plants that are equivalent to
those that could be developed through conventional breeding, the May
2020 final rule established a regulatory process for continuously
identifying and updating modifications that are achievable through
conventional breeding and, thus, exempt from regulation (85 FR 29791-
29796; Sec. 340.1(b)). To this end, Sec. 340.1(b)(4) provides that
the Administrator may propose to exempt plants with additional
modifications, based on what could be achieved through conventional
breeding through a notice published in the Federal Register.
As of August 2, 2024, APHIS has issued 96 responses confirming the
exempt status of modified plants, reviewed 70 other modified plants
through the regulatory status review process, and continued to gather
information and literature about what can be achieved through
conventional breeding methods. For example, as discussed more fully
above, since APHIS initially adopted its exemption 4 years ago,
advances in conventional breeding methods have enabled the steady
introgression of desired genes, alleles, and QTLs in several crops
(Krishna, et al., 2023; Abdul Aziz and Masmoudi, 2024). Genomic
assisted breeding, genetic mapping and studies, high through-put
genotyping, speed breeding, multi-parent advance generation inter-
crosses, and pyramid breeding strategies have advanced quickly and are
now affordable for many crop types (Krishna, et al., 2023; Abdul Aziz
and Masmoudi, 2024), and new methods are consistently emerging to
improve and accelerate breeding methods for difficult to breed crops,
particularly in crops with a complex autopolyploid genome or with
predominant asexual reproduction (Chen, et al., 2021). It is important
that APHIS update its list of modifications plants can contain and
qualify for exemption from regulations to ensure its regulations
reflect these advances in science and technology and remain rooted in
the best science.
Indeed, since July 2021, APHIS has followed the established
regulatory processes to identify modifications that plants can contain
without being subject to part 340 (86 FR 37988 (July 19, 2021); 88 FR
78285). In late July 2021, plaintiffs filed a lawsuit in the United
States District Court for the Northern District of California to
challenge APHIS' May 2020 final rule.\6\ During the pendency of this
litigation, countries around the globe have updated their biotechnology
policies and regulations related to new plant breeding techniques (or
plants with modifications achievable through conventional breeding). As
described in greater detail above, many of these countries, including
the United Kingdom, the Philippines, Singapore, and Thailand, treat
genome edited plants (including polyploid plants) that are free of
exogenous DNA as conventional plants irrespective of the number of
modifications made to the plants. In contrast, because APHIS was an
early
[[Page 89581]]
leader in establishing a regulatory exemption for plants with
modifications that are achievable through conventional breeding, APHIS
initially limited developers to a single modification of the type
described in Sec. 340.1(b)(1) through (3)--a narrower standard for
conventional equivalence compared to both international regulatory
frameworks and scientific literature describing what can be
accomplished today through conventional breeding methods. To ensure the
United States maintains its position as a global leader in agricultural
biotechnology regulation and that its regulatory system and list of
modifications exempt plants can contain is current and accurately
reflects what can be achieved through conventional breeding methods, it
is essential that APHIS issue this final notice updating the types of
modifications plants can contain and qualify for exemption from
regulation.
---------------------------------------------------------------------------
\6\ National Family Farm Coalition, et al. v Vilsack, et al. No.
3:21-cv-05695.
---------------------------------------------------------------------------
Issuing this notice is also important to avoid differential
treatment for products produced through genetic engineering that are
otherwise equivalent to conventionally bred and/or developed products.
As discussed above, plants with modifications that are achievable
through conventional breeding that qualify for exemption, are no
different, as a class, and in terms of plant pest risk, from comparable
plants that are made through conventional breeding, which, likewise, do
not come before APHIS. Updating the list of modifications that plants
can contain and qualify for exemption will ensure that APHIS'
regulations do not impose unnecessary costs on modified plants that are
equivalent to those developed through conventional breeding, including
expenses associated with obtaining a permit, complying with permitting
conditions, and preparing submissions for regulatory status review
(i.e., the case-by-case method for determining whether a modified plant
is subject to part 340, described in Sec. 340.5).
To put these costs in perspective, developers with modified plants
that do not meet the criteria for regulatory exemption have the option
for obtaining a permit that authorizes the use of the modified plant
under conditions or submitting a regulatory status review request that
seeks a determination that the plant is not subject to part 340,
because it is unlikely to present an increased plant pest risk compared
to the non-modified version of the plant. To date, roughly 45 percent
of APHIS' regulatory status review submissions have involved plants
with modifications that would likely meet the criteria for exemption
described in this final notice. On average, APHIS has taken roughly 234
days to complete its evaluation of these modified plants and determine
they are not subject to regulation under part 340. Until now,
developers have incurred costs associated with regulatory uncertainty,
obtaining a permit and complying with associated conditions if they
wish to engage in regulated activities (which, could range in cost from
$13,000-$671,000, depending on a variety of factors) (U.S. Department
of Agriculture Animal and Plant Health Inspection Service, 2020b), and
preparing regulatory status review submissions for modified plants that
were intended to be exempt from regulation, while APHIS has expended
staff resources evaluating modified plants that were not intended to
fall within the scope of part 340, which has increased workloads, and,
in turn, drawn criticism for increased regulatory processing times and
calls for improvement (Bass and Kovak, 2024; Kovak and Bass, 2024; US
Congress Committee on Appropriations, 2024). Beyond this, if APHIS were
to continue imposing unnecessary regulatory costs on plants with
modifications achievable through conventional breeding, the United
States could face the risk of U.S. investors going to countries with
regulatory frameworks that already treat such modifications as
conventional equivalents, including global agricultural competitors
(Clayton Yeutter Institute Round Table Discussion, 2023), at a time
when the United States seeks to advance the U.S. bioeconomy and
biotechnology.
Along these lines, in September 2022, the President issued
Executive Order 14081, entitled ``Advancing Biotechnology and
Biomanufacturing Innovation for a Sustainable, Safe, and Secure
Bioeconomy,'' which directs regulatory agencies to improve the
efficiency of biotechnology regulations (Executive Office of the
President, 2022). Issuing this notice directly supports Section 8 of
this Executive Order, will aid the United States in maintaining its
position as a global leader in agricultural biotechnology, and will
help keep U.S. developers working in the United States on products that
help U.S. producers tackle climate, resource, and food security
challenges.
Lastly, it is important to note that the modified plants that are
described in this final notice and that are eligible for exemption
under Sec. 340.1(b) have never been subject to regulation under part
340--these modified plants were not intended to be within the scope of
the revised regulations (part 340 (2020)) and were not within the scope
of the legacy regulations (part 340 (2019)), and their conventionally
bred counterparts have not been subject to regulation. In fact, if the
May 2020 final rule that established the exemption for plants with
modifications achievable through conventional breeding were to be set
aside, it would mean that all the plants containing the modifications
described in this notice--and more--would still be outside the scope of
regulation.
The following table summarizes the modifications and their
applicability to polyploids:
Table 1--Summary of Modifications and Applicability to Polyploids
----------------------------------------------------------------------------------------------------------------
Notes Designation Modification
----------------------------------------------------------------------------------------------------------------
1 pair of homologous chromosomes....... Sec. 340.1(b)(1).................... The genetic modification is a
change resulting from cellular
repair of a targeted DNA break
in the absence of an
externally provided repair
template.
1 pair of homologous chromosomes....... Sec. 340.1(b)(2).................... The genetic modification is a
targeted single base pair
substitution.
1 pair of homologous chromosomes....... Sec. 340.1(b)(3).................... The genetic modification
introduces a gene known to
occur in the plant's gene pool
or makes changes in a targeted
sequence to correspond to a
known allele of such a gene or
to a known structural
variation present in the gene
pool.
1 pair of homologous chromosomes across 340.1(b)(4)(vi)(AM1).................. An indel or contiguous deletion
subgenomes without repair template and of any size, made at a
one pair of homologous chromosomes targeted location, with or
with repair template. without insertion of DNA if
generated without using a
repair template, or without
insertion of DNA if generated
using a repair template.
[[Page 89582]]
Allows up to 12 simultaneous 340.1(b)(4)(vi)(AM2).................. Plants with up to 12
(multiplex) or sequential modifications, made
modifications. simultaneously or
sequentially, are exempt from
regulation if each
modification individually
qualifies the plant for
exemption and occurs in a
different gene. Modifications
to either a single allele or
pair of alleles on homologous
chromosomes will count as one
modification. See website for
information on counting
modifications.
----------------------------------------------------------------------------------------------------------------
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Accordingly, pursuant to the process established under Sec.
340.1(b)(4), we are adopting the two additional modifications
articulated in this notice for the reasons set forth in our initial
notice and in this final notice.
Authority: 7 U.S.C. 7701-7772 and 7781-7786; 31 U.S.C. 9701; 7 CFR
2.22, 2.80, and 371.3.
Done in Washington, DC, this 6th day of November 2024.
Michael Watson,
Administrator, Animal and Plant Health Inspection Service.
[FR Doc. 2024-26232 Filed 11-12-24; 8:45 am]
BILLING CODE 3410-34-P
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