Movement of Organisms Modified or Produced Through Genetic Engineering; Notice of Additional Modifications Exempt Plants Can Contain, 89569-89585 [2024-26232]

Agencies

• DEPARTMENT OF AGRICULTURE
• Animal and Plant Health Inspection Service

[Federal Register Volume 89, Number 219 (Wednesday, November 13, 2024)]
[Notices]
[Pages 89569-89585]
From the Federal Register Online via the Government Publishing Office [www.gpo.gov]
[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; [email protected]; (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 
https://www.regulations.gov/docket/APHIS-2018-0034.
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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):
     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
     The genetic modification is a targeted single base pair 
substitution; or
     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 https://www.aphis.usda/gov/biotech-exemptions.
    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

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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 (https://plants.ensembl.org/). 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 (https://www.aphis.usda.gov/sites/default/files/requesting-confirmation-of-exemption.pdf). 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).
---------------------------------------------------------------------------

    \2\ May 29, 1992 (57 FR 22984-23005; Docket No. 92N-0139).
---------------------------------------------------------------------------

    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\ https://www.argentina.gob.ar/normativa/nacional/resoluci%C3%B3n-21-2021-346839/texto.
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    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 
(legislation.gov.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\ https://food.ec.europa.eu/document/download/c03805a6-4dcc-42ce-959c-e4d609010fa3_en?filename=gmo_biotech_ngt_proposal_2023-411_en.pdf.
---------------------------------------------------------------------------

    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