Artigo 01 - Crop Biotechnology and The Future of Food

Review Article
https://doi.org/10.1038/s43016-020-0072-3
Crop biotechnology and the future of food

Michael A. Steinwand1 and Pamela C. Ronald 1,2,3 ✉
The global population continues to rise, as does the likelihood of reduced yields of major food crops due to the changing
climate, thus making the development of genetically improved, stress-resilient crops a research priority. The convergence
of low-cost genome sequencing with improved computational power and high-throughput molecular phenotyping technolo-
gies has accelerated the identification of genes underlying important agronomic traits relevant to food production and quality.
Here, we discuss the evolution of plant improvement, and how researchers leverage genomic analyses and revolutionary
new plant breeding technologies like site-directed nucleases to enhance food crop traits through agricultural biotechnology.
Deployment of these products from the laboratory to the field remains hindered by biological and regulatory bottlenecks that
require further development.
T
he production of food via agriculture began approximately for characteristics including upright vegetative structure, uniform
10,000 years ago, and its history is punctuated by strides in flowering, seed retention on the plant for easier harvest, and reduc-
progress both technological and biological. During the rela- tions in seed dormancy and toxic chemicals in edible tissues13. This
tively recent Green Revolution of the 1960s, international research suite of traits, termed the ‘domestication syndrome’, typically ren-
investment in agricultural improvement of the cereal grains wheat, ders domesticated crops poorly adapted to growth outside of human
rice and maize resulted in new high-yielding varieties that saw cultivation. Through iterative selective breeding, specific versions of
widespread cultivation and gave greater food security in many parts genes (alleles) underlying these traits and their surrounding DNA
of the world1. The subsequent rise in molecular genetic tools has regions became uniformly present across the domesticated breed-
ushered in the era of genomic breeding, wherein molecular breeding population14. (Box 1 provides a glossary of key terms.) Examples
ing and genetic engineering have gained prominence2. include the teosinte branched 1 allele that is a major contributor to
By 2050, the global population is predicted to reach 9.7 billion. modern-day maize plant architecture15 and the Bitter fruit allele
Meeting this higher food demand, if consumption practices and responsible for reducing cucumber fruit bitterness16. Continued
food waste do not change, requires estimated food production selection of these domesticated species further optimized agro-
increases of 25 to 100 per cent3,4. At the same time, crop yield is stag- nomic traits, and geographic dispersal established locally adapted
nating in many parts of the world5, and climate change threatens the landrace cultivars17. For some species, further breeding led to glob-
worldwide agricultural system6,7 with yields and nutritional content ally adapted elite cultivars produced by seed companies and research
predicted to decline for major crops8–10. Additionally, crop patho- institutions. While substantial research has been invested in pro-
gen and insect pest ranges are shifting into new territory towards ducing new varieties of select crops (for example, maize, wheat and
the global poles11. These challenges to sustained food security will rice), so-called orphan crops—often staple foods like cassava, millet
require multiple solutions encompassing social, technological and and sweet potato for people in developing areas of the world—have
economic change. One part of the solution is intrinsic improvement received less international genomic breeding focus18.
of cultivated crops2. Genetic diversity is crucial for continued crop improvement.
Increasing genomic and phenotypic information is becoming Many crops possess reduced genetic diversity relative to their
available as technology improves and costs decrease. New plant wild ancestors, though generally this effect is more prevalent in
breeding techniques reduce the time necessary to enhance agro- annual plants (for example, soybean and wheat) than in long-lived
nomic traits relative to conventional breeding. These techniques perennial species that extensively outcross and are often clonally
have potential to deliver improvements such as greater abiotic and propagated19. Domesticated species may harbour elevated numbers
biotic stress tolerance to minimize yield losses, and the improve- of deleterious mutations (see ref. 20 and references within). Novel
ment of food nutrition and quality. Underutilized and regionally genetic diversity can be introduced via induced mutagenesis using
important crops, often adapted to grow on marginal lands, can be irradiation (for example, X-rays and gamma rays) or chemical
further improved and grown more widely to diversify the global treatment, causing DNA breakages, deletions and/or base changes
diet. We discuss how the application of new biomolecular and that result in new alleles; a subset will have effects on agronomic
mechanistic tools provides greater understanding of the genetics and quality traits21. This process creates random genome-wide
and physiology underlying crop plant performance, and we address mutations, necessitating a multigenerational screening and selec-
how those new tools are being applied to innovations in food tion pipeline to identify high-performing mutant individuals. The
production and quality. global FAO/IAEA Mutant Variety Database (http://mvd.iaea.org)
documents over 3,300 mutant varieties representing over 230 plant
Evolution of plant breeding in the computational era species that have been officially released since 1950. Examples
Humans have manipulated plant genomes for millennia, long before include mutant alleles for the semi-dwarfing trait key in Green
the modern understanding of DNA underlying heritable genetics12. Revolution varieties of wheat, maize and rice22, and seedless
Early domestication of wild annual species occurred via selection mandarin citrus (PAU Kinnow-1)23. Landrace varieties and related
Department of Plant Pathology, University of California, Davis, CA, USA. 2The UC Davis Genome Center, University of California, Davis, CA, USA.
1
Center on Food Security and the Environment, Stanford University, Stanford, CA, USA. ✉e-mail: [email protected]
3
Nature Food | VOL 1 | May 2020 | 273–283 | www.nature.com/natfood 273

Review Article NaTure FooD
Box 1 | Glossary of terminology
Agrobacterium tumefaciens. A naturally occurring plant patho- Nanomaterials. A broad class of chemically and structurally diverse
genic bacterium that delivers DNA into host cells as part of its synthetic materials with at least one dimension between 1 and 100
infection process. Modified lab strains deliver DNA of interest nanometres, such as gold particles or carbon nanofilaments, which
into susceptible plant cells, where it is randomly integrated into can be used as delivery systems for biological molecules into plant
the host genome. This property was exploited to develop mutant and animal cells. Nanomaterials can be designed for cell and
populations in model systems for functional gene analysis, which organelle specificity, making them applicable to the agricultural
has provided the basis of much fundamental plant research over and biomedical fields.
the past decades.
Omics. The generic term for study of large-scale data of a
Allele. Alternative forms of a gene at a chromosomal location that biological class, such as the total complement of genes or chemical
arise through mutation. Different alleles may vary by small DNA metabolites present in an organism. Detection technologies
changes, encode multiple forms of a protein or possess structural vary by type of molecular component being assessed, may
changes that change when and how a gene is active. The diversity quantitate total or relative abundance, or provide other specific
of alleles for a trait in a population is often greater than those information.
present in any single individual; breeding is the directed accumu
lation of desired alleles from a population into an elite variety. Orphan crops. Also termed underutilized crops, underinvested
crops, or crops for the future. Orphan crops encompass all food
Clonal propagation. Asexual propagation that retains specific types: cereals, root and tubers, legumes, and vegetables. Although
combinations of alleles present in the parent plant. This process they provide nutrition to large numbers of people, these crops
can be used for rapid generation of additional crop plants like have historically received little attention by the international
those that reproduce infrequently or are sterile (for example, research community, and thus genomic data and molecular tools
bananas), or in grafting to fuse plants with desired traits to a remain limited.
genetically dissimilar rootstock.
Particle bombardment. Also termed biolistics or gene gun,
Epigenetics. Heritable chemical markers on DNA that influences this delivery system uses helium under high pressure to propel
how genes are read, causing activation or repression of gene microparticles bonded with DNA, RNA and/or protein into cells.
function. These markers are enzymatically added or removed, and These particles break through cell walls and penetrate to the
typically vary based on cell identity and response to external cues. interior, where the biomolecules dissociate and perform biological
functions.
Genomic selection. An accelerated breeding approach that
utilizes models trained on genomic and phenotypic data to Selective breeding. The reiterative process of assessing genetically
computationally predict breeding values for candidate lines based heterogeneous organisms for desired traits and combining
on their genotype, to improve varieties. these advantageous characteristics to improve cultivars. Genetic
diversity can originate from multiple sources, including DNA
Haploid/diploid. The number of complete genome copies an recombination, sexual outcrossing or induced mutations.
organism possesses is termed ploidy. Gamete cells like pollen
and ovules typically are haploid, containing just one copy of each Site-directed nuclease. A protein enzyme that cleaves DNA at
chromosome, one genome copy. Adult animals and many plant locations specified by chemical interactions between cellular
species are typically a fusion of two gametes and possess one pair of DNA and protein molecules (for example, zinc-finger nuclease
each chromosome, termed diploid. Additional copies of genomes or TALEN) or RNA (for example, CRISPR-Cas) sequences.
can arise through whole genome duplication or hybridization. For Canonical nucleases break both strands of double-stranded DNA,
example, bread wheat is hexaploid (six genomic copies), while while modified enzyme variants can cleave only one strand, or
strawberries are octoploid. perform other biochemical functions.
Hybrid vigour. The improved growth and yield displayed by specific Transgenic. Plants that have integrated genetic material artificially
hybrid offspring relative to either parent line. This performance is introduced from a foreign source are classified as transgenic. This
due to a unique combination of alleles created by crossbreeding transgene DNA is commonly from related plant species, bacteria
specific genetically distinct inbred parents and becomes variable in or synthetic origin, and includes regulatory sequences that can
subsequent generations due to further segregation of these alleles. specify cell- or tissue-specific gene expression.
wild populations also serve as sources to introduce diversity, such Precise genetic-marker-assisted breeding was used to introduce
as desired stress-adaptive traits, using genetic markers strongly the Sub1A-1 gene into several farmer-preferred rice cultivars
linked to or underlying the desired phenotype to guide breeding throughout South and Southeast Asia29, while selecting against
selection24,25. For example, the landrace rice cultivar Flood Resistant introduction of undesired genes of the donor parent, accruing
(FR) 13A possesses the ability to survive in complete water sub- benefits to generationally disadvantaged farmers30.
mergence for multiple weeks, but lacks the desired high yield and
high-quality grain of other commercial cultivars26. Severe flooding Using omics technology to augment traditional breeding
and prolonged, complete submergence of rice plants results in crop programmes. Recent technological advances have increased the
and yield losses for most commercial varieties. Crossbreeding gen- ubiquity of molecular ‘-omics’ studies in plant science, a term
erated an FR13A-derived rice population, and subsequent genetic denoting the total complement of a biological unit like genes
mapping identified the gene underlying its submergence tolerance (genome) or proteins (proteome). With modern high-throughput
as Sub1A-1, a gene absent from intolerant varieties26–28 (Fig. 1b). genome sequencing technology more accessible at lowering costs
274 Nature Food | VOL 1 | May 2020 | 273–283 | www.nature.com/natfood

NaTure FooD Review Article
a b c
d e
WT mult 8
Fig. 1 | Examples of genetically altered agricultural species. a, Inflorescence with fruits from maize progenitor teosinte (left) and primitive domesticated
maize (right). b, Submergence tolerance is conferred by the presence of the Sub1a-1 gene (right) compared with control rice (left) after two weeks of
submergence, followed by two weeks of recovery30. c,d, Eggplant variety Uttara showing injury from eggplant fruit and shoot borer (c) and lack of injury in
Bt eggplant variety131 (d). e, Wild tomato S. pimpinellifolium flower WT morphology (left) and after Cas9 targeted mutagenesis of the MULTIFLORA locus
(right) during de novo domestication113. Scale bars, 2 cm. Figure reproduced with permission from John Doebley (a); ref. 30, Laboratory of Pamela Ronald
(b); ref. 131, Cold Spring Harbor Laboratory Press (c,d); ref. 113, Springer Nature America, Inc (e).
(US$1,000–10,000s), the depth of crop variety and breadth of Computational correlative association studies synthesize the
species genomic data is quickly expanding. For example, collabora- information contained in these agronomic, proteomic, transcrip-
tive sequencing efforts have produced whole genome sequences for tomic and/or metabolomic datasets to find genes with large and
3,000 rice varieties from 89 countries31, and the African Orphan small effects in crop phenotype to guide manipulation of complex
Crops Consortium aims to sequence 101 genomes of impor- traits42 (Fig. 2). Recent research for tomato flavour improvement
tant native crops, including finger millet (Eleusine coracana) and exemplifies this application. Flavour is a complex combination of
Bambara groundnut (Vigna subterranea)32. To capture the diver- the taste of numerous sugars, acids and bitter chemical metabolites
sity of specific gene families within a large group, genomic DNA and the smell of volatile aromatic compounds, governed by numer-
samples can be preferentially enriched prior to sequencing. This ous genetic determinants that make it a difficult breeding goal43.
method has been used to define genetic variation in disease resis- Indeed, historic selection for a gene underlying pale green, evenly
tance gene repertoires in Solanaceae and Triticeae (RenSeq33,34), and ripening tomato fruits also had a deleterious effect on total sugar
gluten gene families in bread wheat (GlutEnSeq35). content44; breeding for increased fruit size or pink colouration also
Robotics and other technologies are speeding acquisition of altered its chemical profile45. Multiple studies paired quantification
data on macrophysiology of crop plants36, while an assortment of of a select number of tomato metabolites (~20–80) with genetic
high-throughput detection technologies can be employed to pro- markers in wild and cultivated tomato fruits to identify genes
duce physiological data on RNA, protein, metabolites and other underlying specific potential flavour determinants46–48. Consumer
molecular metrics shaped by the plant’s genomics and its environ- flavour preference surveys were paired with analysis of 398 mod-
ment. Network-based analyses using a broad array of transcrip- ern, heirloom and wild accessions to identify which compounds
tomic data can identify major regulatory hubs underlying adaptive and genetic markers were correlated with positive taste preference49.
responses to environmental stress (for example, refs. 37–39). Plant Work by Zhu et al. expanded the metabolite panel to quantify 980
proteomic analysis may detect context-dependent protein abun- distinct fruit chemical metabolites in 610 wild and commercial
dance and the presence of transient chemical modifications that red-fruited tomato accessions and paired that information with
alter their biological function40. Recently, a proteomic survey of 13 genetic sequence and gene expression network analysis, describing
species across the green plant lineage has been reported, identifying how domestication and breeding have altered tomato metabolites45.
known and novel conserved multi-protein complexes and protein Most recently, the first meta-analysis of previous association studies
interactions; genes encoding proteins in these complexes likely play in tomato46,47,49 compiled data from 775 tomato accessions and iden-
a role in important agronomic traits41. tified 305 genes tied to sugars, acids and flavour-related volatiles50.

Wild progenitors, landrace cultivars

and diversity of species
Crop performance phenotyping
Transcriptomics
Gene introgression from Computational Conventional transgene

related/wild species via Genomics analysis Proteomics introduction from plant or
conventional breeding other sources
Metabolomics
O
N N
O N N
Marker-assisted Improvement via

breeding site-directed nucleases
Genomic selection
breeding
Fig. 2 | Omics data can be used to inform marker-assisted breeding, genomic selection and site-directed nuclease targets. Plant population diversity
can be characterized for agronomic traits through field phenotyping and using omics technologies that capture genome sequence (genomics) and
RNA (transcriptomics), protein (proteomics) and metabolite abundance (metabolomics) under a number of conditions. Computational algorithms and
correlation analysis of these different data sets can uncover linkages between genes and macro and molecular phenotypes and identify novel genes with
potential agronomic value. These data can be used to generate markers for use in marker-assisted selection breeding, to produce predictive models for
use in genomic selection or guide introgression of genes from wild sources without the use of biotechnology (blue text). Use of omics data and phenotypic
analysis can also identify gene candidates for manipulation via site-directed nucleases (including targeted mutagenesis and targeted gene insertion)
and inform conventional introduction of genes or multi-gene clusters for creation of stable transgenic lines using biotechnology tools (green text).
Credit: Scott Camazine / Alamy Stock Photo (protein structure)
The genetic locations identified in these studies represent potential non-homologous end joining, in which broken DNA strands are
targets for manipulation, either through conventional breeding or reattached, often creating small deletions or insertions of DNA
biotechnological approaches. bases on either end of the break, and homology directed repair,
Additionally, data from genome sequencing, gene association in which the gap is bridged using a template that shares sequence
studies and phenotypic data can be used to generate and train with the DNA region surrounding the break54. These repair pro-
machine-learning predictive models for use in genomic selec- cesses can be exploited to enable the introduction of several genetic
tion plant breeding (reviewed in ref. 51). This accelerated process changes of different classifications, including: mutation of gene
uses a trained computational model to select lines based on their function through random error-prone DNA repair via end joining
genetic markers without repeated phenotyping during breeding (classified as site-directed nuclease application 1 (SDN1), see Fig. 3);
cycles. Such models can be repeatedly improved with data from allele replacement through repair mechanisms using supplied
high-throughput phenotyping technologies in both research and short DNA fragments as a template (SDN2); or targeted insertion
field settings, utilizing stress-phenotyping via imaging sensors to of DNA at the cut site using supplied long template DNA (SDN3).
detect disease, drought and nutrient deficiency, among others52. In non-reproductive plant cells, non-homologous end joining
repair occurs at high efficiency, while homology directed repair
Gene-editing tools enable customization of germplasm occurs with low efficiency55. Introduced DNA breakages can also
The discovery and application of targetable site-directed nuclease stimulate targeted recombination between homologous chromo-
(SDN) enzymes ushered in a new era of plant mutagenesis breeding, somes. Cas9-induced cuts were used to trigger recombination
giving researchers powerful tools for precision manipulation of crop in tomato somatic cells to alter fruit colour56, suggesting the poten-
genomes (broadly termed ‘gene editing’) to leverage the wealth of tial application of the technology to break inheritance linkages
genomic data. Among these, RNA-guided CRISPR-Cas technology between physically close genes or combine favourable alleles onto
has become a dominant tool since 2013, when gene editing capac- one chromosome.
ity was demonstrated in plant cells. The gene editing toolkit has Improvement of cultivar performance and product quality can
bloomed in the short period that has followed, with Cas enzyme be achieved via deactivation of genes with undesirable effects, or
variants discovered or developed that enable A/T to G/C DNA modification of regulatory regions governing when and how genes
base-pair swapping, direct gene repression or activation, directed are expressed57. Targeted mutagenesis without inserting foreign
DNA methylation and RNA targeting. For specific details of current DNA at the target site (SDN1) accounts for roughly 90 per cent of
CRISPR technology, Cas variants and gene-editing methods, read- SDN usage thus far in published agricultural research58 due to its
ers are referred to a recent review53. relative ease and can be viewed as an evolution of mutation breed-
ing previously discussed. Cas cutting specificity is determined by
Application of site-directed nucleases to molecular breeding an associated guide RNA molecule that can bind to the target DNA
of food crops. Site-directed nucleases, also called sequence- sequence. Supplying the enzyme with multiple guide RNAs allows
specific nucleases, create a targeted breakage in DNA. Two distinct for simultaneous changes at multiple locations across chromosomes
mechanisms within the plant cell exist to repair this damage: within a single cell59 or allows for large deletions of DNA60 when

SDN-induced break isolated, naturally occurring phenotypes and creating new ones that
Genome sequence
can be incorporated into breeding programmes. In Brassica napus
(canola), leaf and seed number65 and seed yield per plant66 were
SDN1
No template provided
increased by mutating development and plant architecture genes.
or
Targeted mutagenesis or
In cereal crops such as rice, genes restraining grain size and seed
via random repair number67–69 are promising targets. New alleles for yield genes Grain
number 1a and DENSE AND ERECT PANICLES 1 were created
that each had superior performance relative to naturally occurring
SDN2 + high-yield alleles68 in field trials, and combining alleles generated
Template with small DNA difference
flanked by homologous sequence
in elite rice lines had additive effects on grain size and grain num-
Targeted mutagenesis via HDR ber67. Altering the response to plant growth regulating hormones
also increased plant size and flower number, resulting in 30 per cent
greater grain yield in field trials70.
SDN3 + Plant diseases and pests, including fungi, bacteria, oomyce-
Template of large DNA tes and nematodes, reduce global yield of major crops by an esti-
flanked with homologous sequence
Targeted insertion via HDR mated 17 to 30 per cent71, with higher losses found in food-insecure
regions. One method to engineer enhanced disease resistance
Fig. 3 | Classification of possible products produced via site-directed is to remove plant genes that facilitate disease susceptibility,
nuclease technology. Three classifications for application of site-directed either because they suppress plant immune responses or they are
nucleases (SDN) are established based on European Union working group required by the plant pathogen for its growth and proliferation72.
guidelines186, resulting in end products possibly governed by differing Such susceptibility genes have been identified widely in crop spe-
regulations. Recovery of plants with targeted mutations that result from cies in relation to many disease-causing organisms of agronomic
only the random non-homologous end joining repair mechanism of the importance (reviewed in ref. 73) and are often conserved between
cell with no donor template are classified as SDN1—the most commonly species. For example, breeders have used a naturally occurring
applied SDN approach in plants to date. Such edits are largely insertions or mutant allele of the MILDEW RESISTANCE LOCUS O (MLO) gene
deletions that occur surrounding the double stranded DNA break. Plants to confer heritable broad-spectrum immunity against powdery
classified as SDN2 or SDN3 both utilize a supplied repair template and mildew races in susceptible barley cultivars for decades; research-
the homology directed repair (HDR) process but vary in degree of the ers used SDNs to edit the corresponding MLO genes in tomato74
introduced change. SDN2 is typically classified by templates that induce and wheat75 to generate similar broad resistance against the pow-
small alterations (<20 base pairs), while SDN3 templates can include one dery mildew species infecting these crops. The first application of
or multiple genes flanked by homologous regions and result in either CRISPR technology in the cocoa tree Theobroma cacao deactivated
gene replacement or targeted insertion of foreign DNA at DNA cut a conserved immune suppression gene to hinder development of
sites. Two cut sites can be cleaved simultaneously to excise larger DNA Phytophthora pathogen infection76. Editing of susceptibility gene
fragments to achieve complete gene deletion (SDN1) or to effect gene promoter regions needed by bacterial leaf blight and citrus canker
replacement via SDN3. pathogens significantly reduced disease symptoms in rice77,78 and
orange79. Broad viral resistance in cucumber was deployed through
editing of the cucumber eIF4E gene necessary for viral spread80,
narrowly targeted to one location. This multiplexing allows the while viral infection symptoms were attenuated in eIF4E-edited
stacking of multiple altered traits in a single generation59 and can cassava81. Because susceptibility genes may possess necessary func-
streamline trait development in species like wheat and potato, which tion in the host plant in the absence of disease, not all may be
contain complex genomes with many copies of redundant genes acceptable targets for modification in this manner.
(summarized in ref. 61). Because these mutations do not directly Targeted mutagenesis has also been used to enhance the quality
incorporate foreign genetic material, it is possible to produce of food, particularly related to removing undesired compounds
transgene-free plants carrying these edited traits that are virtually and increasing nutritional value. For example, many plant food
indistinguishable from their conventionally produced counterparts, products contain allergens, often seed storage proteins, perceived
and in some countries these products are exempt from transgenic by the human immune system in a small segment of the popula-
regulation62. Application of this technology can accelerate innova- tion82. The main therapeutic strategy for those with a food allergy
tion in both annual crops and perennial woody crops. For exam- is avoidance of these inciting compounds. Gene-editing tools allow
ple, gene editing techniques can reduce the time for coffee cultivar removal of genes encoding these allergens in food products, poten-
improvement from thirty years in a traditional breeding scheme to tially creating a more hypoallergenic product. Upon completion of
as few as six63. Here, we highlight the diversity of potential agricul- the hexaploid bread wheat genome, genes encoding gluten proteins
tural and food trait improvement with select examples. linked to wheat intolerance (for example, coeliac disease and baker’s
Breeding to increase crop yield performance remains a high asthma) were identified and mapped across all chromosomes83; the
priority. Yield is a complex trait representing the cumulative output large number and distribution of these genes has likely hindered
of plant architecture, nutrient acquisition, and resource allocation breeding efforts to produce low-allergenic wheat. Gluten is a poly-
limited by disease, pests and abiotic stress encountered throughout mer largely composed of two classes of protein: glutenins and α-, γ-
the growing season per area of land. Developing new allelic variation and ω-gliadins. Gene editing successfully disrupted ~30 α-gliadin
in genes modulating plant developmental pathways, among others, genes in bread wheat84, resulting in up to an 85 per cent reduction
holds potential for improving crop productivity64 and land usage, in detectable immunoreactive gluten in seed grains. Unlike previous
and is particularly important for genomic regions where domestica- efforts to reduce gluten proteins through RNA silencing methods
tion selectively reduced the allele diversity of the breeding popula- that require a permanent transgene in the wheat85, transgene-free
tion. Rodríguez-Leal and colleagues demonstrated this approach in lines were isolated with these heritable α-gliadin mutations. Genes
tomato, targeting DNA controlling when and how developmental encoding major allergens from apple86, peanut87, rice88 and soy89
genes underlying fruit size, flower number, or plant shoot archi are potential gene editing targets to ameliorate food allergy symp-
tecture are active60. The resulting edited populations possessed toms. Likewise, manipulation of innate metabolism genes can
new alleles with a spectrum of plant changes, reiterating previously alter the nutritional profile of the food. The tomato antioxidant

compound lycopene, responsible for the fruit’s red colour, and the closer to future use as a winter cover crop and source of oilseed
health-promoting compound γ-aminobutyric acid were highly for food114.
enriched90–92 by manipulating biosynthesis genes to favour accumu- As costs further decline and genomic data and resources accrue,
lation of these compounds. In lettuce, removal of regulatory DNA gene editing can facilitate rapid improvement and greater adoption
regions enhanced production of vitamin C in the leaf threefold93. of orphan crops and landrace cultivars115. Lemmon and co-workers
In sorghum, deletion of multiple α-kafirin gene regions encoding generated whole genome sequence, gene expression data, and gene
seed storage proteins enhanced protein digestibility and increased editing protocols for the groundcherry Physalis grisea with the goal
overall essential amino acid lysine and total protein content of the of transforming this fruit into a mass-produced berry crop among
grain94. Soybean seed oil genes were altered to increase fourfold the the rank of strawberry and raspberry. Weedy, sprawling ground-
percentage of the polyunsaturated fat oleic acid95, and oleic acid lev- cherry plant growth was compacted, and fruit size and number
els could also be modulated by different mutation combinations of increased116,117. Genome editing protocols have also proved suc-
redundant genes in Camelina sativa96. cessful in cassava, an orphan crop grown for uses in both food and
Gene editing has also been applied to traits that prolong the post- manufacturing. The cassava tuber starch profile was altered through
harvest shelf life in certain food crops, potentially reducing food modification of amylose synthesis genes, conferring desirable cook-
waste along the supply chain97. Transgenic silencing of polyphenol ing and food processing properties118. Careful elimination of genes
oxidase genes reduces potato tuber browning after processing98, responsible for toxic compound formation could also enhance
and this trait was subsequently commercialized after application to nutritional quality and processing in cassava119 and grass pea
transgenic non-browning Arctic apples99. Recently, targeted muta- (Lathyrus sativus)120. As genomic information and biotech methods
genesis was used to create varieties of non-browning, transgene-free on wild and orphan species continues to develop, editing of known
white button mushrooms100, potatoes101 and romaine lettuce102,103. domestication genes111,121 will further expand crop diversity and
Development of ripe tomatoes that remain firm longer has been a food options.
long-sought goal briefly realized in the Flavr Savr tomato, in which
transgenic gene silencing reduced activity of cell-wall modify- Insertion of transgenes and synthetic gene clusters
ing enzyme that contributes to fruit softening104. Recent manipu- The process of plant transformation, in which genetic material is
lation of a different gene has yielded firmer ripe fruits105 with introduced and integrated into the heritable plant genome, has been
greater resistance to fungal rot106 that retain flavour-contributing a fundamental tool in the development of cultivated genetically
metabolite profiles107. engineered food, fibre and biofuel crops. Trait design can introduce
single or multiple genes to affect a phenotype, or introduce DNA
Creating multi-generational hybrid vigour. Agriculture of the sequences that cause silencing of endogenous genes. Inclusion of
past century has widely exploited the genetic phenomenon of regulatory DNA elements can produce selective effects that are diffi-
hybrid vigour, or heterosis, to produce high-performing, genetically cult to achieve via targeted mutagenesis. For example, seed-specific
uniform F1 hybrid crops that possess variable yield in subsequent gene silencing of widely expressed gossypol synthesis genes in cot-
generations due to genes segregating during sexual reproduction. ton was used to engineer seed with minimal levels of the toxic com-
Mutagenesis of three genes creates the genotype Mitosis instead of pound, making them fit for human consumption while preserving
Meiosis (MiMe) in diploid rice, which results in clonal diploid gam- gossypol content in vegetative parts as a necessary chemical deter-
etes instead of haploid gametes due to the doubling of the genome rent against pathogens and insect pests122. Transgenic products have
without two rounds of segregation and cell division108. When historically received higher scrutiny from government regulatory
MiMe is paired with the pollen mutation matrilineal that prevents bodies, vastly increasing costs and time to develop and commercial-
transfer of pollen-derived DNA to the embryo, self-fertilization in ize transgenics compared with conventional varieties123,124. In 2018,
F1 hybrids produced clonal seeds that retain the F1 diploid geno- 26 countries cultivated 191.7 million hectares of genetically engi-
type109. Another study in rice achieved a similar result by combining neered crops, with the United States, Brazil, Argentina, Canada and
the MiMe genotype with transgenic cell-specific expression of the India collectively representing 91 per cent of the global transgenic
embryogenic gene Baby Boom 1, which stimulates embryo develop- crop area125.
ment without fertilization110. However, this application remains in One of the most prevalent engineered traits is insect resistance
the early phases and further optimization is necessary before com- conferred by genes originating from the soil bacterium Bacillus
mercial adoption, as clonal seed production was reported to occur thuringiensis (Bt). The Bt trait has been applied to many crops,
at low rates ranging from 5 to 29 per cent. including maize, soybean, cotton and eggplant126, and the cumula-
tive use of Bt maize, soybean and cotton crops has resulted in 37 per
Increase the diversity of cultivated species through rapid domes- cent less global pesticide use127 and potential insect pest-suppressive
tication. Another promising approach to expand the food palate benefits to nearby non-Bt crops128. In India, Bt cotton pesticide
is through de novo domestication, in which domestication application was reduced by 50 per cent and, accordingly, reduced
alleles are introduced to wild species possessing desired stress acute pesticide poisonings were seen in cotton growers129. The Bt
tolerance or adaptation to marginal lands, compressing thousands trait in maize has also led to increased consumer safety through
of years of human mutagenesis and selective breeding to a few reduced grain contamination with mycotoxins produced by fungal
plant generations111. This strategy has been demonstrated in the infections that can follow insect damage130. In Bangladesh, where
wild tomato relative Solanum pimpinellifolium, as simultane- introduction of four varieties of Bt eggplant (Fig. 1c,d) in 2014
ous editing via CRISPR of four112 and six113 known domestication marked the first genetically engineered food crop released in a
genes was sufficient to remodel shoot and flower morphology developing country and first Bt vegetable, net returns for farmers
(Fig. 1e) and increase the size and number of fruits to be similar increased sixfold in part due to a 61% reduction in pesticide costs131.
to cultivated tomato Solanum lycopersicum within a single genera- The breadth of phenotypic change possible with transgenesis
tion. De novo domesticated fruits contained 500 per cent higher is far greater than can be achieved through gene editing alone in
levels of lycopene than a commercial tomato variety113 and retained part due to the wide genetic diversity that can be utilized, includ-
disease resistance and salinity tolerance traits of the wild parent112. ing across species with innate reproductive barriers (for example,
Similarly, recent stacking of mutations modifying flowering time, ref. 132). Assembling a repertoire of disease-resistance genes from
seed retention, removal of undesired chemicals and enrichment of numerous accessions or species, as well as rationally designed novel
oleic acids in seeds brings the weed pennycress (Thlaspi arvense L.) resistance genes133, could provide durable, broad-based resistance

against plant pathogens. Applications of synthetic biology, the cre- Additionally, the transformation method employed can trigger dif-
ation of modular DNA components and rationale design of genetic fering governmental regulations and approval timelines for end
circuits to confer novel traits, can accelerate improvement of funda- products; for example, products produced in the United States using
mental plant biological processes (reviewed in ref. 134). For example, Agrobacterium fall under United States Department of Agriculture
the enzyme RuBisCo is responsible for atmospheric carbon fixa- (USDA) oversight152,153 and cannot be released until deregulated or
tion in photosynthesis but can also utilize atmospheric oxygen in non-regulated status is granted.
a process called photorespiration to produce glycolate, which can In plants, Agrobacterium transformation is restricted to suscep-
inhibit photosynthesis. Photorespiration rate increases with tem- tible host plant species. Bombardment can carry both DNA and/or
perature, thus reducing photosynthetic efficiency, and ultimately other biomolecules such as gene-editing protein/RNA complexes154,
sacrificing growth of the plant. Introducing alternative biosynthetic but the projectiles damage the impacted tissues. In both cases, pro-
pathways using bacterial or plant enzymes to convert glycolate into duction of suitable starting material and recovery of transformed
a usable form with introduction of three to five genes was sufficient plants can be time and labour intensive, typically extending several
to increase photosynthetic efficiency and vegetative biomass in field months, and unintended changes to the genome are likely155,156. A key
trials in tobacco135 and laboratory tests in the seed oil crop Camelina research goal going forward is expanding the scope of transformable
sativa136. Bacterial-derived traits that allow rice and cotton to species and increasing transformation efficiency. Recent protocols
metabolize phosphite in addition to phosphate137,138 permits phos- couple transformation with expression of growth and developmen-
phite’s dual use as both weed control agent and fertilizer. Extending tal regulator genes Baby Boom and Wuschel to markedly improve
symbiotic nitrogen fixation capability (reviewed in ref. 139) and pro- transformation rates in the monocots maize, sorghum, sugarcane
duction of nutritional and biopharmaceutical compounds are also and rice157, and refinement of this system shortened the timeframe
likely routes to further augment agricultural productivity and prod- for recovery of transformed plants to as little as four weeks158. The
uct value. Examples of biofortification of foods with new nutrients technique was successfully applied to vegetative leaf tissue for the
not found in conventional varieties include the enhanced accumula- first time in maize, reducing the time needed to grow adult plants
tion of the vitamin A precursor beta-carotene in edible tissue of rice to the flowering stage for source tissues. Similarly, developmental
(Golden Rice140,141) and Cavendish bananas (Banana21142) and the regulators were recently used to induce de novo transgenic shoots
carotenoid antioxidant astaxanthin in rice grains143. from vegetative tissues of tobacco, tomato, grape and potato159.
Use of site-directed nucleases for targeted gene insertion Bypassing tissue culture methods for producing modified
or replacement. Commonly used methods of Agrobacterium plants. The use of tissue culture to generate germplasm can yield
tumefaciens-mediated and DNA-coated particle bombardment unintended genomic changes affecting fitness or yield of the end
transformation introduce transgenic sequences randomly in the product, necessitating additional screening of individual lines.
crop genome with potential for endogenous gene disruption or This is a long-known phenomenon named somaclonal variation160.
misexpression of neighbouring genes through trans- or cis-gene Whole genome sequencing revealed an increase in DNA deletions
regulation. Another potential drawback of random DNA inser- and variations in cultured tissues of rice161, cotton162 and potato163.
tion is that, when multiple transgenes are used, these transgenes Epigenetic marks widespread in the genome can be lost or retained,
will likely integrate into different chromosomes and each will with these changes persisting for multiple generations after regen-
segregate independently, complicating downstream breeding. To eration from tissue culture164–166. In some cases, these plants have
address these potential shortcomings, researchers are investigating altered gene expression profiles that resemble their source tissue167
the use of SDNs to precisely introduce DNA elements into the plant and display aberrant interactions with beneficial and pathogenic
genome at targeted DNA breakages via the plant’s DNA homology microorganisms. While sufficient screening and field testing can
directed repair pathway. Multiple introduced genes can be incor- identify properly functioning individuals, the development of new
porated at a single genomic safe harbour, a defined chromosomal methods bypassing or limiting tissue culture is desirable151.
region with minimal positional effects to maximize efficacy of the Newly developed nanomaterials enable passive delivery of bio-
transgene without perturbing necessary cell functions144. Ainley molecules such as DNA, RNA and proteins into cells for, among
et al. developed a DNA ‘landing pad’ that when integrated into the other applications, integration into, or gene editing of, host
genome allows sequential stacking of modular transgenes through genomes. Physical and chemical properties of these materials such
defined sequence-specific nuclease cut sites. Its use in maize dem- as size and electric charge can be tailored to allow uptake across
onstrated co-segregation of inserted traits in the subsequent gen- plant cell walls and membranes of both plant and animal cells (see
eration145. Despite these advances, site-specific integration of DNA ref. 168 and references within), with application as simple as a foliar
via SDNs remains challenging in plants, as homology-directed spray. Carbon nanomaterials have been demonstrated to passively
repair is not always active during the plant cell life cycle, and it is traverse the plant cell wall to deliver DNA into leaf cells of arugula,
difficult to deliver sufficient target DNA near the DNA breakage wheat, cotton and the tobacco relative Nicotiana benthamiana169,
for high-efficiency insertion146. Efforts to address this include bias- and selectively into arugula, spinach and tobacco leaf cell chloro-
ing the cell’s repair machinery to favour homology-directed repair plasts170. Expression of the delivered genes was short lived (less than
(for example, ref. 147) and employing plant viral replication machin- 10 days) as the DNA did not integrate into the host genome169; such
ery to increase the amount of donor DNA, the latter of which a system could be useful for CRISPR gene-editing in which integra-
has enhanced targeted insertion efficiency in tobacco148, tomato149 tion is not necessary for function.
and wheat150. Gametic tissues in crop plants that propagate via sexual repro-
duction are being targeted to bypass tissue culture regeneration.
New methods to expand plant transformation technology Zhao et al. used a magnetic field to deliver transgene-carrying
Generating transformed plants is often a major bottleneck for crop nanoparticles through apertures where the cell wall is thin and
improvement151. Biological characteristics, such as recalcitrance more permeable in cotton, pepper and pumpkin pollen, produc-
to foreign DNA uptake and genomic integration or capacity for ing viable transgenic pollen171. Manual fertilization of flowers
regeneration into whole plants after transformation, limit genetic using this pollen produced seeds with stable, heritable transgene
engineering to amenable plant cultivars. The rigid plant cell wall integration. Transgenic pollen can also serve as the delivery vector
presents a specific challenge to plant transformation protocols for gene-editing protein/RNA complexes. Using methods deve
relative to those of mammalian cells, which lack such a barrier. loped previously172,173, a specific type of maize pollen carrying the

nuclease Cas9 protein fertilizes the egg cell, transferring gene-editing References
complexes that mutate the egg cell genome. Due to defects in the 1. Pingali, P. L. Green revolution: impacts, limits, and the path ahead.
pollen, none of the genetic material it carries is permanently incor- Proc. Natl Acad. Sci. USA 109, 12302–12308 (2012).
2. Wallace, J. G., Rodgers-Melnick, E. & Buckler, E. S. On the road to breeding
porated to the resulting haploid offspring, which are edited but 4.0: unraveling the good, the bad, and the boring of crop quantitative
transgene-free. The gene-editing complex could be passed from genomics. Annu. Rev. Genet. 52, 421–444 (2018).
corn pollen to corn ovule, as well as corn pollen to wheat ovule. The 3. Hunter, M. C., Smith, R. G., Schipanski, M. E., Atwood, L. W. &
mechanisms underpinning this haploid-inducing pollen appear Mortensen, D. A. Agriculture in 2050: recalibrating targets for sustainable
intensification. Bioscience 67, 386–391 (2017).
conserved across monocot and dicot species172, suggesting this
4. Tilman, D., Balzer, C., Hill, J. & Befort, B. L. Global food demand and the
method may have broad applicability, including genotypes recalci- sustainable intensification of agriculture. Proc. Natl Acad. Sci. USA 108,
trant to transformation themselves. However, as the development 20260–20264 (2011).
of the technologies described in this section remain active areas of 5. Ray, D. K., Ramankutty, N., Mueller, N. D., West, P. C. & Foley, J. A. Recent
research, further testing by the research community at large will be patterns of crop yield growth and stagnation. Nat. Commun. 3, 1293 (2012).
6. Tietjen, B. et al. Climate change-induced vegetation shifts lead to more
necessary to determine reproducibility, efficiency and ease of use ecological droughts despite projected rainfall increases in many global
for particular crop species. temperate drylands. Glob. Chang. Biol. 23, 2743–2754 (2017).
7. Hirabayashi, Y. et al. Global flood risk underclimate change. Nat. Clim.
Further considerations Chang. 3, 816–821 (2013).
The convergence of omics technology presents an unprecedented 8. Rosenzweig, C. et al. Assessing agricultural risks of climate change in the
21st century in a global gridded crop model intercomparison. Proc. Natl
opportunity to identify genes controlling agronomically valuable Acad. Sci. USA 111, 3268–3273 (2014).
traits and speed the development of genetically improved cultivars. 9. Zhao, C. et al. Plausible rice yield losses under future climate warming. Nat.
Still, the lag time between fundamental research advances and com- Plants 3, 16202 (2017).
mercialization is often lengthy174. For example, the process for com- 10. Myers, S. S. et al. Increasing CO2 threatens human nutrition. Nature 510,
mercialization of transgenic varieties is affected by a diverse array of 139–142 (2014).
11. Bebber, D. P., Ramotowski, M. A. T. & Gurr, S. J. Crop pests and pathogens
political and socioeconomic concerns and can span decades124, mak- move polewards in a warming world. Nat. Clim. Chang. 3, 985–988 (2013).
ing it difficult to address urgent agricultural needs. Consequently, in 12. Meyer, R. S. & Purugganan, M. D. Evolution of crop species: genetics of
many parts of the world, breeders and farmers do not have access to domestication and diversification. Nat. Rev. Genet. 14, 840–852 (2013).
transgenic technologies. For example, while farmers in Bangladesh 13. Olsen, K. M. & Wendel, J. F. A bountiful harvest: genomic insights into
continue to adopt and cultivate Bt eggplant, the varieties remain crop domestication phenotypes. Annu. Rev. Plant Biol. 64, 47–70 (2013).
14. Hufford, M. B., Berny Mier y Teran, J. C. & Gepts, P. Crop biodiversity: an
prohibited in neighbouring India despite scientific support and unfinished magnum opus of nature. Annu. Rev. Plant Biol. 70, 727–751 (2019).
similar farmer need131. Similarly, organic farmers do not have access 15. Studer, A., Zhao, Q., Ross-Ibarra, J. & Doebley, J. Identification of a
to Bt and other transgenic varieties because methods that employ functional transposon insertion in the maize domestication gene tb1.
genetic engineering techniques are excluded from use in certified Nat. Genet. 43, 1160–1163 (2011).
organic production175, although other types of genetic alteration 16. Shang, Y. et al. Biosynthesis, regulation, and domestication of bitterness in
cucumber. Science 346, 1084–1088 (2014).
such as chemical and radiation mutagenesis are permitted176. 17. Casañas, F., Simó, J., Casals, J. & Prohens, J. Toward an evolved concept of
The application of gene editing in agriculture has been met landrace. Front. Plant Sci. 8, 145 (2017).
with mixed support among the general public177, and regulation 18. Varshney, R. K. et al. Can genomics boost productivity of orphan crops?
of transgene-free, gene-edited SDN1 products diverge across the Nat. Biotechnol. 30, 1172–1176 (2012).
19. Gaut, B. S., Díez, C. M. & Morrell, P. L. Genomics and the contrasting
globe (discussed in ref. 62). For example, in the European Union,
dynamics of annual and perennial domestication. Trends Genet. 31,
the products of gene editing are regulated as “genetically modified” 709–719 (2015).
according to a 2018 decision by the Court of Justice of the European 20. Moyers, B. T., Morrell, P. L. & McKay, J. K. Genetic costs of domestication
Union178, a designation that complicates European scientific field tri- and improvement. J. Hered. 109, 103–116 (2018).
als for SDN1/2 improved plants179 and restricts farmer adoption178. 21. Oladosu, Y. et al. Principle and application of plant mutagenesis in crop
improvement: a review. Biotechnol. Biotechnol. Equip. 30, 1–16 (2016).
By contrast, the USDA has no plans to regulate SDN1 crops as long 22. Peng, J. et al. ‘Green revolution’ genes encode mutant gibberellin response
as these crops are not plant pests or developed using plant pests180. modulators. Nature 400, 256–261 (1999).
This decision potentially saves years or even decades in bring- 23. Rattanpal, H. S., Singh, G. & Gupta, M. Studies on mutation breeding in
ing new varieties to US farmers181. The USDA National Organic mandarin variety Kinnow. Curr. Sci. 116, 483–487 (2019).
Program (which develops the rules and regulations for the produc- 24. Dwivedi, S. L. et al. Landrace germplasm for improving yield and abiotic
stress adaptation. Trends Plant Sci. 21, 31–42 (2016).
tion, handling, labelling and enforcement of all USDA organic prod- 25. Mickelbart, M. V., Hasegawa, P. M. & Bailey-Serres, J. Genetic mechanisms
ucts) has not yet ruled on the use of SDN1 technologies in certified of abiotic stress tolerance that translate to crop yield stability. Nat. Rev.
organic production and there is disagreement on how to proceed. Genet. 16, 237–251 (2015).
For instance, whereas the National Organic Program Standards 26. Xu, K. & Mackill, D. J. A major locus for submergence tolerance mapped
Board (a federal advisory board made up of 15 volunteers from the on rice chromosome 9. Mol. Breed. 2, 219–224 (1996).
27. Xu, K., Xu, X., Ronald, P. C. & Mackill, D. J. A high-resolution linkage map
organic community) has recommended that diverse SDN technolo- of the vicinity of the rice submergence tolerance locus Sub1. Mol. Gen.
gies be disallowed; researchers in the US and Europe have called for Genet. 263, 681–689 (2000).
the allowance of such technologies in organic farming systems182–184. 28. Xu, K. et al. Sub1A is an ethylene-response-factor-like gene that confers
These examples reflect the need for ongoing engagement of the submergence tolerance to rice. Nature 442, 705–708 (2006).
scientific community with diverse stakeholders, including consum- 29. Ismail, A. M., Singh, U. S., Singh, S., Dar, M. H. & Mackill, D. J. The
contribution of submergence-tolerant (Sub1) rice varieties to food security
ers and politicians, on the challenges faced by farmers and the use in flood-prone rainfed lowland areas in Asia. Field Crop. Res. 152,
of plant biotechnologies to address these challenges. As described 83–93 (2013).
by communications scientists Scheufele and Krause, the increas- 30. Emerick, K. & Ronald, P. C. Sub1 rice: engineering rice for climate change.
ingly polarized political environments and fundamental changes in Cold Spring Harb. Perspect. Biol. https://doi.org/10.1101/cshperspect.
how information is shared by media and audiences have given new a034637 (2019).
31. Li, Z. et al. The 3,000 rice genomes project. Gigascience 3, 7 (2014).
urgency to the problem of the disconnect between public opinion in 32. Hendre, P. S. et al. African Orphan Crops Consortium (AOCC): status of
the United States and the scientific consensus on scientific topics185. developing genomic resources for African orphan crops. Planta 250,
989–1003 (2019).
Received: 22 October 2019; Accepted: 1 April 2020; 33. Jupe, F. et al. Resistance gene enrichment sequencing (RenSeq) enables
Published online: 19 May 2020 reannotation of the NB-LRR gene family from sequenced plant genomes

and rapid mapping of resistance loci in segregating populations. Plant J. 76, 63. Breitler, J.-C. et al. CRISPR/Cas9-mediated efficient targeted mutagenesis
530–544 (2013). has the potential to accelerate the domestication of Coffea canephora. Plant
34. Steuernagel, B. et al. Rapid cloning of disease-resistance genes in plants using Cell Tiss. Org. Cult. 134, 383–394 (2018).
mutagenesis and sequence capture. Nat. Biotechnol. 34, 652–655 (2016). 64. Trevaskis, B. Developmental pathways are blueprints for designing
35. Jouanin, A. et al. Development of the GlutEnSeq capture system for successful crops. Front. Plant Sci. 9, 745 (2018).
sequencing gluten gene families in hexaploid bread wheat with deletions or 65. Yang, Y. et al. Precise editing of CLAVATA genes in Brassica napus L.
mutations induced by γ-irradiation or CRISPR/Cas9. J. Cereal Sci. 88, regulates multilocular silique development. Plant Biotechnol. J. 16,
157–166 (2019). 1322–1335 (2018).
36. Tardieu, F., Cabrera-Bosquet, L., Pridmore, T. & Bennett, M. Plant 66. Zheng, M. et al. Knockout of two BnaMAX1 homologs by CRISPR/
phenomics, from sensors to knowledge. Curr. Biol. 27, R770–R783 (2017). Cas9-targeted mutagenesis improves plant architecture and increases yield
37. Lee, T. et al. AraNet v2: An improved database of co-functional gene in rapeseed (Brassica napus L.). Plant Biotechnol. J. 18, 644–654 (2019).
networks for the study of Arabidopsis thaliana and 27 other nonmodel plant 67. Zhou, J. et al. Multiplex QTL editing of grain-related genes improves yield
species. Nucleic Acids Res. 43, D996–D1002 (2015). in elite rice varieties. Plant Cell Rep. 38, 475–485 (2019).
38. Brooks, M. D. et al. Network Walking charts transcriptional dynamics of 68. Huang, L. et al. Developing superior alleles of yield genes in rice
nitrogen signaling by integrating validated and predicted genome-wide by artificial mutagenesis using the CRISPR/Cas9 system. Crop J. 6,
interactions. Nat. Commun. 10, 1569 (2019). 475–481 (2018).
39. Lee, I. et al. Genetic dissection of the biotic stress response using a 69. Li, M. et al. Reassessment of the four yield-related genes Gn1a, DEP1, GS3,
genome-scale gene network for rice. Proc. Natl Acad. Sci. USA 108, and IPA1 in rice using a CRISPR/Cas9 system. Front. Plant Sci. 7, 377 (2016).
18548–18553 (2011). 70. Miao, C. et al. Mutations in a subfamily of abscisic acid receptor genes
40. Friso, G. & Van Wijk, K. J. Posttranslational protein modifications in plant promote rice growth and productivity. Proc. Natl Acad. Sci. USA 115,
metabolism. Plant Physiol. 169, 1469–1487 (2015). 6058–6063 (2018).
41. McWhite, C. D. et al. A pan-plant protein complex map reveals deep 71. Savary, S. et al. The global burden of pathogens and pests on major food
conservation and novel assemblies. Cell 181, 460–474 (2020). crops. Nat. Ecol. Evol. 3, 430–439 (2019).
42. Huang, X. & Han, B. Natural variations and genome-wide association 72. Pavan, S., Jacobsen, E., Visser, R. G. F. & Bai, Y. Loss of susceptibility
studies in crop plants. Annu. Rev. Plant Biol. 65, 531–551 (2014). as a novel breeding strategy for durable and broad-spectrum resistance.
43. Zhu, G., Gou, J., Klee, H. & Huang, S. Next-gen approaches to flavor-related Mol. Breed. 25, 1–12 (2010).
metabolism. Annu. Rev. Plant Biol. 70, 187–212 (2019). 73. van Schie, C. C. N. & Takken, F. L. W. Susceptibility genes 101: how to be a
44. Powell, A. L. T. et al. Uniform ripening encodes a Golden 2-like good host. Annu. Rev. Phytopathol. 52, 551–581 (2014).
transcription factor regulating tomato fruit chloroplast development. Science 74. Nekrasov, V. et al. Rapid generation of a transgene-free powdery mildew
336, 1711–1715 (2012). resistant tomato by genome deletion. Sci. Rep. 7, 482 (2017).
45. Zhu, G. et al. Rewiring of the fruit metabolome in tomato breeding. Cell 75. Wang, Y. et al. Simultaneous editing of three homoeoalleles in hexaploid
172, 249–261.e12 (2018). bread wheat confers heritable resistance to powdery mildew. Nat.
46. Bauchet, G. et al. Identification of major loci and genomic regions Biotechnol. 32, 947–951 (2014).
controlling acid and volatile content in tomato fruit: implications for flavor 76. Fister, A. S., Landherr, L., Maximova, S. N. & Guiltinan, M. J. Transient
improvement. New Phytol. 215, 624–641 (2017). expression of CRISPR/Cas9 machinery targeting TcNPR3 enhances defense
47. Sauvage, C. et al. Genome-wide association in tomato reveals 44 candidate response in Theobroma cacao. Front. Plant Sci. 9, 268 (2018).
loci for fruit metabolic traits. Plant Physiol. 165, 1120–1132 (2014). 77. Li, T., Liu, B., Spalding, M. H., Weeks, D. P. & Yang, B. High-efficiency
48. Zhang, J. et al. Genome-wide association mapping for tomato volatiles TALEN-based gene editing produces disease-resistant rice. Nat. Biotechnol.
positively contributing to tomato flavor. Front. Plant Sci. 6, 1042 (2015). 30, 390–392 (2012).
49. Tieman, D. et al. A chemical genetic roadmap to improved tomato flavor. 78. Oliva, R. et al. Broad-spectrum resistance to bacterial blight in rice using
Science 355, 391–394 (2017). genome editing. Nat. Biotechnol. 37, 1344–1350 (2019).
50. Zhao, J. et al. Meta-analysis of genome-wide association studies provides 79. Peng, A. et al. Engineering canker-resistant plants through CRISPR/
insights into genetic control of tomato flavor. Nat. Commun. 10, Cas9-targeted editing of the susceptibility gene CsLOB1 promoter in citrus.
1534 (2019). Plant Biotechnol. J. 15, 1509–1519 (2017).
51. Crossa, J. et al. Genomic selection in plant breeding: methods, models, and 80. Chandrasekaran, J. et al. Development of broad virus resistance in
perspectives. Trends Plant Sci. 22, 961–975 (2017). non-transgenic cucumber using CRISPR/Cas9 technology. Mol. Plant Pathol.
52. Singh, A., Ganapathysubramanian, B., Singh, A. K. & Sarkar, S. Machine 17, 1140–1153 (2016).
learning for high-throughput stress phenotyping in plants. Trends Plant Sci. 81. Gomez, M. A. et al. Simultaneous CRISPR/Cas9-mediated editing of
21, 110–124 (2016). cassava eIF4E isoforms nCBP-1 and nCBP-2 reduces cassava brown
53. Zhang, Y., Malzahn, A. A., Sretenovic, S. & Qi, Y. The emerging and streak disease symptom severity and incidence. Plant Biotechnol. J. 17,
uncultivated potential of CRISPR technology in plant science. Nat. Plants 5, 421–434 (2019).
778–794 (2019). 82. Breiteneder, H. & Radauer, C. A classification of plant food allergens.
54. Puchta, H. The repair of double-strand breaks in plants: mechanisms and J. Allergy Clin. Immunol. 113, 821–830 (2004).
consequences for genome evolution. J. Exp. Bot. 56, 1–14 (2005). 83. Juhász, A. et al. Genome mapping of seed-borne allergens and
55. Huang, T.-K. & Puchta, H. CRISPR/Cas-mediated gene targeting in plants: immunoresponsive proteins in wheat. Sci. Adv. 4, eaar8602 (2018).
finally a turn for the better for homologous recombination. Plant Cell Rep. 84. Sánchez-León, S. et al. Low-gluten, nontransgenic wheat engineered with
38, 443–453 (2019). CRISPR/Cas9. Plant Biotechnol. J. 16, 902–910 (2018).
56. Hayut, S. F., Bessudo, C. M. & Levy, A. A. Targeted recombination between 85. García-Molina, M., Giménez, M., Sánchez-León, S. & Barro, F. Gluten free
homologous chromosomes for precise breeding in tomato. Nat. Commun. 8, wheat: are we there? Nutrients 11, 487 (2019).
15605 (2017). 86. Dubois, A. E. J. et al. First successful reduction of clinical allergenicity of
57. Swinnen, G., Goossens, A. & Pauwels, L. Lessons from domestication: food by genetic modification: Mal d 1-silenced apples cause fewer allergy
targeting cis-regulatory elements for crop improvement. Trends Plant Sci. symptoms than the wild-type cultivar. Allergy Eur. J. Allergy Clin. Immunol.
21, 506–515 (2016). 70, 1406–1412 (2015).
58. Modrzejewski, D. et al. What is the available evidence for the range of 87. Dodo, H. W., Konan, K. N., Chen, F. C., Egnin, M. & Viquez, O. M.
applications of genome-editing as a new tool for plant trait modification Alleviating peanut allergy using genetic engineering: The silencing of the
and the potential occurrence of associated off-target effects: a systematic immunodominant allergen Ara h 2 leads to its significant reduction and a
map. Environ. Evid. 8, 27 (2019). decrease in peanut allergenicity. Plant Biotechnol. J. 6, 135–145 (2008).
59. Ma, X. et al. A robust CRISPR/Cas9 system for convenient, high-efficiency 88. Wakasa, Y., Hirano, K., Urisu, A., Matsuda, T. & Takaiwa, F. Generation of
multiplex genome editing in monocot and dicot plants. Mol. Plant 8, transgenic rice lines with reduced contents of multiple potential allergens
1274–1284 (2015). using a null mutant in combination with an RNA silencing method. Plant
60. Rodríguez-Leal, D., Lemmon, Z. H., Man, J., Bartlett, M. E. & Lippman, Z. Cell Physiol. 52, 2190–2199 (2011).
B. Engineering quantitative trait variation for crop improvement by genome 89. Herman, E. M., Helm, R. M., Jung, R. & Kinney, A. J. Genetic modification
editing. Cell 171, 470–480.e8 (2017). removes an immunodominant allergen from soybean. Plant Physiol. 132,
61. Weeks, D. P. Gene editing in polyploid crops: wheat, camelina, canola, 36–43 (2003).
potato, cotton, peanut, sugar cane, and citrus. Prog. Mol. Biol. Transl. Sci. 90. Li, X. et al. Lycopene is enriched in tomato fruit by CRISPR/Cas9-mediated
149, 65–80 (2017). multiplex genome editing. Front. Plant Sci. 9, 559 (2018).
62. Dederer, H.-G. & Hamburger, D. (eds) Regulation of Genome Editing in 91. Li, R. et al. Multiplexed CRISPR/Cas9-mediated metabolic engineering of
Plant Biotechnology. Regulation of Genome Editing in Plant Biotechnology γ-aminobutyric acid levels in Solanum lycopersicum. Plant Biotechnol. J. 16,
(Springer, 2019). 415–427 (2018).

92. Nonaka, S., Arai, C., Takayama, M., Matsukura, C. & Ezura, H. Efficient 123. Bradford, K. J., Van Deynze, A., Gutterson, N., Parrott, W. & Strauss, S. H.
increase of γ-aminobutyric acid (GABA) content in tomato fruits by Regulating transgenic crops sensibly: lessons from plant breeding,
targeted mutagenesis. Sci. Rep. 7, 7057 (2017). biotechnology and genomics. Nat. Biotechnol. 23, 439–444 (2005).
93. Zhang, H. et al. Genome editing of upstream open reading frames enables 124. Napier, J. A., Haslam, R. P., Tsalavouta, M. & Sayanova, O. The challenges
translational control in plants. Nat. Biotechnol. 36, 894–898 (2018). of delivering genetically modified crops with nutritional enhancement traits.
94. Li, A. et al. Editing of an alpha-kafirin gene family increases digestibility Nat. Plants 5, 563–567 (2019).
and protein quality in sorghum. Plant Physiol. 177, 1425–1438 (2018). 125. Brief 54: Global Status of Commercialized Biotech/GM Crops in 2018
95. Haun, W. et al. Improved soybean oil quality by targeted mutagenesis (ISAAA, 2018).
of the fatty acid desaturase 2 gene family. Plant Biotechnol. J. 12, 126. GM Approval Database (ISAAA, accessed 1 October 2019);
934–940 (2014). http://www.isaaa.org/gmapprovaldatabase/
96. Morineau, C. et al. Selective gene dosage by CRISPR-Cas9 genome editing 127. Klümper, W. & Qaim, M. A meta-analysis of the impacts of genetically
in hexaploid Camelina sativa. Plant Biotechnol. J. 15, 729–739 (2017). modified crops. PLoS One 9, e111629 (2014).
97. Alexander, P. et al. Losses, inefficiencies and waste in the global food 128. Dively, G. P. et al. Regional pest suppression associated with widespread Bt
system. Agric. Syst. 153, 190–200 (2017). maize adoption benefits vegetable growers. Proc. Natl Acad. Sci. USA 115,
98. Bachem, C. W. B. et al. Antisense expression of polyphenol oxidase genes 3320–3325 (2018).
inhibits enzymatic browning in potato tubers. Nat. Biotechnol. 12, 129. Kouser, S. & Qaim, M. Impact of Bt cotton on pesticide poisoning
1101–1105 (1994). in smallholder agriculture: a panel data analysis. Ecol. Econ. 70,
99. Waltz, E. Nonbrowning GM apple cleared for market. Nature Biotechnology 2105–2113 (2011).
Blog http://blogs.nature.com/tradesecrets/2015/03/30/nonbrowning-gm- 130. Wu, F. Mycotoxin reduction in Bt corn: Potential economic, health, and
apple-cleared-for-market (2015). regulatory impacts. Transgenic Res. 15, 277–289 (2006).
100. Waltz, E. Gene-edited CRISPR mushroom escapes US regulation. Nature 131. Shelton, A. M. et al. Bt brinjal in Bangladesh: the first genetically
532, 293 (2016). engineered food crop in a developing country. Cold Spring Harb. Perspect.
101. Re: Confirmation that PPO _ KO Potato is not a Regulated Article (United Biol. https://doi.org/10.1101/cshperspect.a034678 (2019).
States Department of Agriculture, 2016); https://www.aphis.usda.gov/ 132. Kawashima, C. G. et al. A pigeonpea gene confers resistance to Asian
biotechnology/downloads/reg_loi/16-090-01_air_response_signed.pdf soybean rust in soybean. Nat. Biotechnol. 34, 661–665 (2016).
102. Re: Confirmation of the Regulatory Status of Genome edited Lactuca sativa 133. La Concepcion, J. D., Franceschetti, M., Terauchi, R., Kamoun, S. &
(Lettuce) (United States Department of Agriculture, 2019); https://www. Banfield, M. Protein engineering expands the effector recognition profile of
aphis.usda.gov/biotechnology/downloads/reg_loi/18-243-01_air_response_ a rice NLR immune receptor. eLife 8, e47713 (2019).
signed.pdf 134. Wurtzel, E. T. et al. Revolutionizing agriculture with synthetic biology.
103. Non-browning lettuce is making its way to the market. AG Daily Nat. Plants 5, 1207–1210 (2019).
https://www.agdaily.com/news/non-browning-lettuce-market/ 135. South, P. F., Cavanagh, A. P., Liu, H. W. & Ort, D. R. Synthetic glycolate
(3 June 2019). metabolism pathways stimulate crop growth and productivity in the field.
104. Bruening, G. & Lyons, J. M. The case of the FLAVR SAVR tomato. Calif. Science 363, eaat9077 (2019).
Agric. 54, 6–7 (2000). 136. Dalal, J. et al. A photorespiratory bypass increases plant growth and seed
105. Wang, D. et al. Characterisation of CRISPR mutants targeting genes yield in biofuel crop Camelina sativa. Biotechnol. Biofuels 8, 175 (2015).
modulating pectin degradation in ripening tomato. Plant Physiol. 179, 137. Manna, M., Achary, V. M. M., Islam, T., Agrawal, P. K. & Reddy, M. K. The
544–557 (2019). development of a phosphite-mediated fertilization and weed control system
106. Yang, L. et al. Silencing of SlPL, which encodes a pectate lyase in tomato, for rice. Sci. Rep. 6, 24942 (2016).
confers enhanced fruit firmness, prolonged shelf-life and reduced 138. Pandeya, D. et al. Selective fertilization with phosphite allows unhindered
susceptibility to grey mould. Plant Biotechnol. J. 15, 1544–1555 (2017). growth of cotton plants expressing the ptxD gene while suppressing weeds.
107. Uluisik, S. et al. Genetic improvement of tomato by targeted control of fruit Proc. Natl Acad. Sci. USA 115, E6946–E6955 (2018).
softening. Nat. Biotechnol. 34, 950–952 (2016). 139. Mus, F. et al. Symbiotic nitrogen fixation and the challenges to its extension
108. Mieulet, D. et al. Turning rice meiosis into mitosis. Cell Res. 26, to nonlegumes. Appl. Environ. Microbiol. 82, 3698–3710 (2016).
1242–1254 (2016). 140. Ye, X. & Beyer, P. Engineering the provitamin A (β-carotene) biosynthetic
109. Wang, C. et al. Clonal seeds from hybrid rice by simultaneous genome pathway into (carotenoid-free) rice endosperm. Science 287, 303–305 (2000).
engineering of meiosis and fertilization genes. Nat. Biotechnol. 37, 141. Paine, J. A. et al. Improving the nutritional value of Golden Rice through
283–286 (2019). increased pro-vitamin A content. Nat. Biotechnol. 23, 482–487 (2005).
110. Khanday, I., Skinner, D., Yang, B., Mercier, R. & Sundaresan, V. A 142. Paul, J.-Y. et al. Golden bananas in the field: elevated fruit pro-vitamin A
male-expressed rice embryogenic trigger redirected for asexual propagation from the expression of a single banana transgene. Plant Biotechnol. J. 15,
through seeds. Nature 565, 91–95 (2019). 520–532 (2017).
111. Østerberg, J. T. et al. Accelerating the domestication of new crops: feasibility 143. Zhu, Q. et al. From Golden Rice to aSTARice: bioengineering astaxanthin
and approaches. Trends Plant Sci. 22, 373–384 (2017). biosynthesis in rice endosperm. Mol. Plant 11, 1440–1448 (2018).
112. Li, T. et al. Domestication of wild tomato is accelerated by genome editing. 144. Dong, O. X. et al. Marker-free carotenoid-enriched rice generated through
Nat. Biotechnol. 36, 1160–1163 (2018). targeted gene insertion using CRISPR-Cas9. Nat. Commun. 11, 1178 (2020).
113. Zsögön, A. et al. De novo domestication of wild tomato using genome 145. Ainley, W. M. et al. Trait stacking via targeted genome editing. Plant
editing. Nat. Biotechnol. 36, 1211–1216 (2018). Biotechnol. J. 11, 1126–1134 (2013).
114. Chopra, R. et al. Identification and stacking of crucial traits required for the 146. Sun, Y., Li, J. & Xia, L. Precise genome modification via sequence-specific
domestication of pennycress. Nat. Food 1, 84–91 (2020). nucleases-mediated gene targeting for crop improvement. Front. Plant Sci.
115. Ribaut, J.-M. & Ragot, M. Modernising breeding for orphan crops: tools, 7, 1928 (2016).
methodologies, and beyond. Planta 250, 971–977 (2019). 147. Qi, Y. et al. Increasing frequencies of site-specific mutagenesis and gene
116. Lemmon, Z. H. et al. Rapid improvement of domestication traits in an targeting in Arabidopsis by manipulating DNA repair pathways. Genome Res.
orphan crop by genome editing. Nat. Plants 4, 766–770 (2018). 23, 547–554 (2013).
117. Kwon, C., Heo, J., Lemmon, Z. H., Capua, Y. & Hutton, S. F. Rapid adaptation 148. Baltes, N. J., Gil-Humanes, J., Cermak, T., Atkins, P. A. & Voytas, D. F.
of Solanaceae fruit crops for urban agriculture. Nat. Biotechnol. 38, DNA replicons for plant genome engineering. Plant Cell 26, 151–63 (2014).
182–188 (2020). 149. Dahan-Meir, T. et al. Efficient in planta gene targeting in tomato using
118. Bull, S. E. et al. Accelerated ex situ breeding of GBSS- and PTST1-edited geminiviral replicons and the CRISPR/Cas9 system. Plant J. 95, 5–16 (2018).
cassava for modified starch. Sci. Adv. 4, eaat6086 (2018). 150. Gil-Humanes, J. et al. High-efficiency gene targeting in hexaploid wheat
119. Jørgensen, K. et al. Cassava plants with a depleted cyanogenic glucoside using DNA replicons and CRISPR/Cas9. Plant J. 89, 1251–1262 (2017).
content in leaves and tubers. Distribution of cyanogenic glucosides, their 151. Altpeter, F. et al. Advancing crop transformation in the era of genome
site of synthesis and transport, and blockage of the biosynthesis by RNA editing. Plant Cell 28, 1510–1520 (2016).
interference technology. Plant Physiol. 139, 363–374 (2005). 152. Camacho, A., Van Deynze, A., Chi-Ham, C. & Bennett, A. B. Genetically
120. Tadele, Z. Orphan crops: their importance and the urgency of engineered crops that fly under the US regulatory radar. Nat. Biotechnol. 32,
improvement. Planta 250, 677–694 (2019). 1087–1091 (2014).
121. Fernie, A. R. & Yan, J. De novo domestication: an alternative route toward 153. McHughen, A. & Smyth, S. US regulatory system for genetically modified
new crops for the future. Mol. Plant 12, 615–631 (2019). [genetically modified organism (GMO), rDNA or transgenic] crop cultivars.
122. Sunilkumar, G., Campbell, L. A. M., Puckhaber, L., Stipanovic, R. D. & Plant Biotechnol. J. 6, 2–12 (2008).
Rathore, K. S. Engineering cottonseed for use in human nutrition by 154. Svitashev, S., Schwartz, C., Lenderts, B., Young, J. K. & Mark Cigan, A.
tissue-specific reduction of toxic gossypol. Proc. Natl Acad. Sci. USA 103, Genome editing in maize directed by CRISPR-Cas9 ribonucleoprotein
18054–18059 (2006). complexes. Nat. Commun. 7, 13274 (2016).

155. Liu, J. et al. Genome-scale sequence disruption following biolistic 175. National Organic Standards Board Materials/GMO Subcommittee Proposal
transformation in rice and maize. Plant Cell 31, 368–383 (2019). Excluded Methods Determinations (2019); https://www.ams.usda.gov/sites/
156. Jupe, F. et al. The complex architecture and epigenomic impact of plant default/files/media/MSExcludedMethodsProposaFall2019.pdf
T-DNA insertions. PLoS Genet. 15, e1007819 (2019). 176. National Organic Program Policy Memorandum 13 (USDA, 2013); https://
157. Lowe, K. et al. Morphogenic regulators Baby boom and Wuschel improve www.ams.usda.gov/sites/default/files/media/NOP-PM-13-1-CellFusion.pdf
monocot transformation. Plant Cell 28, 1998–2015 (2016). 177. Ishii, T. & Araki, M. Consumer acceptance of food crops developed by
158. Lowe, K. et al. Rapid genotype “independent” Zea mays L. (maize) genome editing. Plant Cell Rep. 35, 1507–1518 (2016).
transformation via direct somatic embryogenesis. In Vitro Cell. Dev. Biol. 178. Urnov, F. D., Ronald, P. C. & Carroll, D. A call for science-based review of
Plant 54, 240–252 (2018). the European court’s decision on gene-edited crops. Nat. Biotechnol. 36,
159. Maher, M. F. et al. Plant gene editing through de novo induction of 800–802 (2018).
meristems. Nat. Biotechnol. 38, 84–89 (2019). 179. Faure, J. D. & Napier, J. A. Europe’s first and last field trial of gene-edited
160. Larkin, P. J. & Scowcroft, W. R. Somaclonal variation — a novel source of plants? eLife 7, e42379 (2018).
variability from cell cultures for plant improvement. Theor. Appl. Genet. 60, 180. Secretary Perdue Issues USDA Statement on Plant Breeding Innovation
197–214 (1981). USDA Press Release https://www.usda.gov/media/press-releases/2018/03/28/
161. Tang, X. et al. A large-scale whole-genome sequencing analysis reveals secretary-perdue-issues-usda-statement-plant-breeding-innovation (2018).
highly specific genome editing by both Cas9 and Cpf1 (Cas12a) nucleases 181. Waltz, E. With a free pass, CRISPR-edited plants reach market in record
in rice. Genome Biol. 19, 84 (2018). time. Nat. Biotechnol. 36, 6–7 (2018).
162. Li, J. et al. Whole genome sequencing reveals rare off‐target mutations and 182. Gheysen, G., Custers, R., Gheysen, G. & Custers, R. Why organic farming
considerable inherent genetic or/and somaclonal variations in CRISPR / should embrace co-existence with cisgenic late blight–resistant potato.
Cas9‐edited cotton plants. Plant Biotechnol. J. 17, 858–868 (2019). Sustainability 9, 172 (2017).
163. Fossi, M., Amundson, K., Kuppu, S., Britt, A. & Comai, L. Regeneration of 183. Husaini, A. M. & Sohail, M. Time to redefine organic agriculture: can’t GM
Solanum tuberosum plants from protoplasts induces widespread genome crops be certified as organics? Front. Plant Sci. 9, 423 (2018).
instability. Plant Physiol. 180, 78–86 (2019). 184. Andersen, M. M. et al. Feasibility of new breeding techniques for organic
164. Han, Z. et al. Heritable epigenomic changes to the maize methylome farming. Trends Plant Sci. 20, 426–434 (2015).
resulting from tissue culture. Genetics 209, 983–995 (2018). 185. Scheufele, D. A. & Krause, N. M. Science audiences, misinformation, and
165. Lee, K. & Seo, P. J. Dynamic epigenetic changes during plant regeneration. fake news. Proc. Natl Acad. Sci. USA 116, 7662–7669 (2019).
Trends Plant Sci. 23, 235–247 (2018). 186. Hilscher, J., Bürstmayr, H. & Stoger, E. Targeted modification of plant
166. Stroud, H. et al. Plants regenerated from tissue culture contain stable genomes for precision crop breeding. Biotechnol. J. 12, 1600173 (2017).
epigenome changes in rice. eLife 2, e00354 (2013).
167. Wibowo, A. et al. Partial maintenance of organ-specific epigenetic marks Acknowledgements
during plant asexual reproduction leads to heritable phenotypic variation. We apologize to those authors whose research could not be cited due to space limits.
Proc. Natl Acad. Sci. USA 115, E9145–E9152 (2018). We thank H. Bartram for a careful reading of the manuscript. M.A.S. was supported
168. Cunningham, F. J., Goh, N. S., Demirer, G. S., Matos, J. L. & Landry, M. P. by the Corteva Agriscience Open Innovation programme grant entitled “Gene Editing
Nanoparticle-mediated delivery towards advancing plant genetic for Organic Agriculture.” P.C.R. was supported by grants from the US National Science
engineering. Trends Biotechnol. 36, 882–897 (2018). Foundation (award no. 1237975), the Crary Social Ecology Fund, the Foundation for
169. Demirer, G. S. et al. High aspect ratio nanomaterials enable delivery of Food and Agricultural Research (award no. 534683) and the National Institutes of
functional genetic material without DNA integration in mature plants. Health (GM122968).
Nat. Nanotechnol. 14, 456–464 (2019).
170. Kwak, S.-Y. et al. Chloroplast-selective gene delivery and expression in
planta using chitosan-complexed single-walled carbon nanotube carriers.
Competing interests
The authors declare no competing interests.
Nat. Nanotechnol. 14, 447–455 (2019).
171. Zhao, X. et al. Pollen magnetofection for genetic modification with
magnetic nanoparticles as gene carriers. Nat. Plants 3, 956–964 (2017). Additional information
172. Kelliher, T. et al. One-step genome editing of elite crop germplasm during Correspondence should be addressed to P.C.R.
haploid induction. Nat. Biotechnol. 37, 287–292 (2019). Reprints and permissions information is available at www.nature.com/reprints.
173. Wang, B. et al. Development of a haploid-inducer mediated genome editing
Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in
system for accelerating maize breeding. Mol. Plant 12, 597–602 (2019).
published maps and institutional affiliations.
174. Miller, J. K. & Bradford, K. J. The regulatory bottleneck for biotech specialty
crops. Nat. Biotechnol. 28, 1012–1014 (2010). © Springer Nature Limited 2020

Artigo 01 - Crop Biotechnology and The Future of Food

Uploaded by

Document Information

Original Description:

Original Title

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

Artigo 01 - Crop Biotechnology and The Future of Food

Uploaded by

Copyright:

Available Formats

Review Article

Crop biotechnology and the future of food

Nature Food | VOL 1 | May 2020 | 273–283 | www.nature.com/natfood 273

Box 1 | Glossary of terminology

274 Nature Food | VOL 1 | May 2020 | 273–283 | www.nature.com/natfood

Nature Food | VOL 1 | May 2020 | 273–283 | www.nature.com/natfood 275

Wild progenitors, landrace cultivars

Crop performance phenotyping

Gene introgression from Computational Conventional transgene

Marker-assisted Improvement via

276 Nature Food | VOL 1 | May 2020 | 273–283 | www.nature.com/natfood

Nature Food | VOL 1 | May 2020 | 273–283 | www.nature.com/natfood 277

278 Nature Food | VOL 1 | May 2020 | 273–283 | www.nature.com/natfood

Nature Food | VOL 1 | May 2020 | 273–283 | www.nature.com/natfood 279

280 Nature Food | VOL 1 | May 2020 | 273–283 | www.nature.com/natfood

Nature Food | VOL 1 | May 2020 | 273–283 | www.nature.com/natfood 281

282 Nature Food | VOL 1 | May 2020 | 273–283 | www.nature.com/natfood

Nature Food | VOL 1 | May 2020 | 273–283 | www.nature.com/natfood 283

You might also like