Advancements in multi-omics for nutraceutical enhancement and traits improvement in buckwheat

Abstract

Buckwheat (Fagopyrum spp.) is a typical pseudocereal, valued for its extensive nutraceutical potential as well as its centuries-old cultivation. Tartary buckwheat and common buckwheat have been used globally and become well-known nutritious foods due to their high quantities of: proteins, flavonoids, and minerals. Moreover, its increasing demand makes it critical to improve nutraceutical, traits and yield. In this review, bioactive compounds accumulated in buckwheat were comprehensively evaluated according to their chemical structure, properties, and physiological function. Biosynthetic pathways of flavonoids, phenolic acids, and fagopyrin were methodically summarized, with the regulation of flavonoid biosynthesis. Although there are classic synthesis pathways presented in the previous research, the metabolic flow of how these certain compounds are being synthesized in buckwheat still remains uncovered. The functional genes involved in the biosynthesis of flavonols, stress response, and plant development were identified based on multi-omics research. Furthermore, it delves into the applications of multi-omics in improving buckwheat's agronomic traits, including: yield, nutritional content, stress resilience, and bioactive compounds biosynthesis. While pangenomics combined with other omics to mine elite genes, the regulatory network and mechanism of specific agronomic traits and biosynthetic of bioactive components, and developing a more efficient genetic transformation system for genetic engineering require further investigation for the execution of breeding designs aimed at enhancing desirable traits in buckwheat. This critical review will provide a comprehensive understanding of multi-omics for nutraceutical enhancement and traits improvement in buckwheat.

Full text

Full Terms & Conditions of access and use can be found at
https://www.tandfonline.com/action/journalInformation?journalCode=ibty20
Critical Reviews in Biotechnology
ISSN: (Print) (Online) Journal homepage: www.tandfonline.com/journals/ibty20
Advancements in multi-omics for nutraceutical
enhancement and traits improvement in
buckwheat
Yingjie Song, Chunlin Long, Ying Wang, Yuxing An & Yinglin Lu
To cite this article: Yingjie Song, Chunlin Long, Ying Wang, Yuxing An & Yinglin Lu (19 Aug
2024): Advancements in multi-omics for nutraceutical enhancement and traits improvement in
buckwheat, Critical Reviews in Biotechnology, DOI: 10.1080/07388551.2024.2373282
To link to this article: https://doi.org/10.1080/07388551.2024.2373282
© 2024 The Author(s). Published by Informa
UK Limited, trading as Taylor & Francis
Group.
View supplementary material
Published online: 19 Aug 2024.
Submit your article to this journal
View related articles
View Crossmark data

REVIEW ARTICLE
CRITICAL REVIEWS IN BIOTECHNOLOGY
Advancements in multi-omics for nutraceutical enhancement and traits
improvement in buckwheat
Yingjie Songa, Chunlin Longb, Ying Wangc, Yuxing Ana, and Yinglin Lua
aInstitute of Nanfan and Seed Industry, Guangdong Academy of Sciences, Guangzhou, P.R. China; bCollege of Life and Environmental
Sciences, Minzu University of China, Beijing, China; cKey Laboratory of South China Agricultural Plant Molecular Analysis and Genetic
Improvement & Guangdong Provincial Key Laboratory of Applied Botany, South China Botanical Garden, Chinese Academy of Sciences,
Guangzhou, China
ABSTRACT
Buckwheat (Fagopyrum spp.) is a typical pseudocereal, valued for its extensive nutraceutical
potential as well as its centuries-old cultivation. Tartary buckwheat and common buckwheat have
been used globally and become well-known nutritious foods due to their high quantities of:
proteins, flavonoids, and minerals. Moreover, its increasing demand makes it critical to improve
nutraceutical, traits and yield. In this review, bioactive compounds accumulated in buckwheat
were comprehensively evaluated according to their chemical structure, properties, and physiological
function. Biosynthetic pathways of flavonoids, phenolic acids, and fagopyrin were methodically
summarized, with the regulation of flavonoid biosynthesis. Although there are classic synthesis
pathways presented in the previous research, the metabolic flow of how these certain compounds
are being synthesized in buckwheat still remains uncovered. The functional genes involved in the
biosynthesis of flavonols, stress response, and plant development were identified based on
multi-omics research. Furthermore, it delves into the applications of multi-omics in improving
buckwheat’s agronomic traits, including: yield, nutritional content, stress resilience, and bioactive
compounds biosynthesis. While pangenomics combined with other omics to mine elite genes, the
regulatory network and mechanism of specific agronomic traits and biosynthetic of bioactive
components, and developing a more efficient genetic transformation system for genetic
engineering require further investigation for the execution of breeding designs aimed at enhancing
desirable traits in buckwheat. This critical review will provide a comprehensive understanding of
multi-omics for nutraceutical enhancement and traits improvement in buckwheat.
GRAPHIC ABSTRACT
© 2024 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.
CONTACT Yingjie Song yjsong0517@163.com Institute of Nanfan and Seed Industry, Guangdong Academy of Sciences, Guangzhou 510220, P.R.
China.
Supplemental data for this article can be accessed online at https://doi.org/10.1080/07388551.2024.2373282.
https://doi.org/10.1080/07388551.2024.2373282
This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivatives License (http://creativecommons.org/licenses/
by-nc-nd/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited, and is not altered, transformed,
or built upon in any way. The terms on which this article has been published allow the posting of the Accepted Manuscript in a repository by the author(s) or with their
consent.
ARTICLE HISTORY
Received 8 February 2024
Revised 10 April 2024
Accepted 31 May 2024
KEYWORDS
Buckwheat; multi-omics;
agronomic traits;
bioactive components;
biosynthesis; nutraceutical

2 Y. SONG ETAL.
HIGHLIGHTS
• Buckwheat (Fagopyrum spp.) is considered as promising and sustainable nutrient crop for
abundant flavonoids, phenolic acids and fagopyrum production with impressive biosynthetic
capacity.
• The chemical structure, properties, physiological function, and biosynthesis pathways of these
bioactive components are summarized.
• The comprehensive information of multi-omics including genome, transcriptome, proteome,
and metabolism for buckwheat nutraceutical traits improvement has been concluded.
• The pangenomics combined with other omics to mine elite genes, and regulatory network and
mechanism of specific agronomic traits and biosynthetic of bioactive components are explored.
1. Introduction
In the last two years, with: climate change, the increas-
ing global population and the COVID-19 pandemic
impact, numbers of undernourished and hungry people
have increased to 821 million and 720 million, respec-
tively [1, 2]. The intensification of conflict, economic
shocks and climate extremes, with the high cost of
nutritious foods, will continue to challenge food nutri-
tion and security [3]. Agrifood systems are transformed
and provide low-cost nutritious foods and affordable
healthy diets. In addition, incorporating ancient
pseudocereals with unparalleled nutritive value in mod-
ern food systems could also combat hunger [4].
Buckwheat (Fagopyrum spp.) is a typical pseudocereal
valued for its extensive nutraceutical potential (such as:
protein, dietary fiber, slowly edible starch, polyphenols
and bioactive compounds) as well as its centuries-old
cultivation [5]. These biologically active compounds
have heightened interest in the health advantages of
buckwheat, particularly its hypoglycemic tendency. The
global buckwheat production in 2021 is 1.88 million
tonnes, and Russia and China are the top buckwheat
producers with 0.92 million and 0.50 tonnes, respec-
tively (https://www.fao.org/faostat). Out of the twenty-six
species of genus Fagopyrum that have been recorded,
F. esculentum (common buckwheat) and F. tataricum
(Tartary buckwheat) are the greatest two cultivated spe-
cies [6]. F. tataricum and F. cymosum are mostly grown
in the highlands of China and the Himalayas, but com-
mon buckwheat is found all over the world [7].
The groats and flour of common buckwheat and
Tartary buckwheat have been used globally and
become well-known nutritious foods due to their high
quantities of proteins, flavonoids, and minerals. There
are numerous studies reporting the potential health
advantages from ingesting buckwheat, which can take
the form of: food, dietary supplements, home cures, or
prescription medications [8]. These buckwheats were
shown to have a wide range of bioactivities, including:
anti-tumor [9], anti-oxidant [10], hepatoprotective [11],
anti-inflammatory [12], anti-allergic [13], anti-hyperglycemic
[14], anti-fatigue [15] and anti-bacterial properties [16]
(Figure 1). In southwest China, common buckwheat
and Tartary buckwheat were also traditionally used in
Figure 1. Three buckwheat including Fagopyrum esculentum, Fagopyrum tataricum, and Fagoprum cymosum, with the bioac-
tive compound including avonoids, phenolics, triterpenoids and sterols, and their benets to human health. Created with
BioRender.com.

CRITICAL REVIEWS IN BIOTECHNOLOGY 3
folk medicine for various medicinal purposes by the Yi
people [17, 18]. Tartary buckwheat is widely used as
an: antioxidant, anticancer, hypoglycemic, and hypolip-
idemic agent based on traditional folk medicine [19–21].
The rhizome of F. cymosum was also used in traditional
medicine to eliminate heat and toxic materials, and to
treat: dysmenorrhea, cancer, lumbago, snakebite, and
traumatic injuries [22] (Figure 1). Based on these func-
tions, buckwheat has received increasing attention in
recent decades, especially in common and Tartary
buckwheat, with significant improvements to be a
commercially viable crop.
Over the past few decades, a number of promising
omics technologies have emerged as a result of the
development of next-generation sequencing (NGS)
technologies and the ability to regularly acquire high
quality whole genome sequences. Through: changes in
DNA, transcript levels, proteins, metabolites, and min-
eral nutrients against the backdrop of environmental
and physiological stress response, these omics-base
approaches (including: genomics, transcriptome,
metabolome, proteome, ionomics and phenomics)
have demonstrated their value for examining the
genetic and molecular basis of crop development [23].
The combination of these various omics techniques
may help to clarify how genes function and interact
with one another under physiological and environmen-
tal stress [24]. Various economically significant crops,
including: wheat, millet, soybean and rice, have been
studied to identify and decipher key elements of: stress
responses, senescence, and yields, using reliable tech-
niques and comprehensive multi-omics approaches
[25–28]. In addition, as a powerful and indispensable
integrated technique, multi-omics has greatly facili-
tated the discovery and synthesis pathway of second-
ary metabolites [29], and is essential for finding
undiscovered plant signaling molecules. For instance,
based on the integrated approach of omics tools, the
canadine synthase enzyme involved in berberine bio-
synthesis pathway in Coptis chinensis Franch was dis-
covered [30]. Above all: improved genetic development,
agricultural production, crop breeding knowledge,
nutraceutical improvement, crop resilience to physio-
logical and environmental stress, may be possible with
the help of multi-omics methods [24].
Research from many groups regarding the
F.esculentum and F. tataricum reference genome was
published with ∼1.2 Gbp and ∼0.48 Gbp genome size
respectively [31, 32], beginning a new phase in the
study of omics in buckwheat. Recent research of the
application of multi-omics was conducted in buck-
wheat research. In this review, our aim is: to highlight
recent progress in the biosynthesis pathway of
bioactive compounds, especially flavonoids; to high-
light the potential of multi-omics for nutraceutical
improvement and enhanced stress tolerance; and to
identify nutrition-related, yield-related and stress
tolerance-related genes to improve buckwheat yield.
We hypothesize that integrating all omics technologies
could serve as a foundation for: enhancing molecular
breeding, yields of buckwheat, and enhancing resil-
ience to physiological and environmental stress.
2. The major bioactive components and
biosynthesis pathway analysis in Fagopyrum
2.1. Bioactive components
Buckwheat is a nutritious pseudocereal that contains a
variety of bioactive chemicals, such as: bioactive flavo-
noids, fagopyrins, fagopyritols, d-fagomine, and phenolic
acids. Buckwheat grains bioactive ingredient boosts
their healing ability against illnesses. Flavonoids, pheno-
lics, fagopyritols, triterpenoids, steroids, and fatty acids
are the principal groups of bioactive chemicals identi-
fied from three Fagopyrum buckwheat species (Figure 2,
chemical structure). Flavonoids and phenolic compounds
were considered to be the major active components.
2.1.1. Flavonoids
The main flavonoids in three buckwheat mainly include:
rutin, orientin, isoorientin, quercetin, kaempferol and
their derivative based on phenylbenopyrone structure
[33, 34]. Thirty-two species of flavonoids have been
identified in buckwheat (Figure 2), with 15 species found
in F. tataricum, 16 species in F. esculentum, and 15 spe-
cies in F. cymosum. The flavonoid present in buckwheat
contribute to its antioxidant, anti-inflammatory, and car-
diovascular health-promoting properties. Rutin is the
most abundant and well-studied flavonoid in buck-
wheat. It is predominantly concentrated in the outer
layers of buckwheat seeds, as well as in the leaves and
flowers [35]. Rutin is a potent antioxidant and has been
associated with improving blood vessel function, reduc-
ing the risk of blood clots, and lowering LDL cholesterol
levels [36]. Its cardiovascular benefits make it a standout
flavonoid in buckwheat. The content of rutin in F. tatar-
icum was higher than that in F. cymosum and F. esculen-
tum with about 14.00 mg/g of dry weight in seeds, and
110 mg/100g of fresh weight in sprouts [37, 38].
Quercetin is another prominent flavonoid present in
buckwheat. It has well-documented antioxidant and
anti-inflammatory properties. Quercetin is known for
its ability to help regulate is important to note that
cooking or processing buckwheat can affect the quer-
cetin content.

4 Y. SONG ETAL.
Kaempferol, though present in smaller quantities
compared to rutin and quercetin, is a valuable flavo-
noid with antioxidant and anti-inflammatory proper-
ties. It is found in various plant parts, including the
leaves and flowers of buckwheat. Kaempferol has been
studied for its potential to reduce the risk of chronic
diseases and promote overall well-being. In general,
the kaempferol content in buckwheat can range from
approximately 0.1 to 2 mg/100 g of raw buckwheat
groats [39]. It’s essential to consider that the kaemp-
ferol content might be higher in other parts of the
buckwheat plant, such as the leaves and flowers, com-
pared to the seeds [38, 40].
Additionally, based on HPLC-MS and NMR, four flavonse,
including: vitexin, orientin, isoorientin, and isovitexin, have
been identified in F. tataricum and F. esculentum [41, 42]
(Figure 2). The content of these four flavonse in F. tataricum
is lower than that in F. esculentum [43]. Luteolin, catechin,
hesperidin, rhamnetin, 3-methylquercetin and
3,5-dimethylquercetin were only determined in F. cymosum
[44, 45]. Quercetin-3-O-glucoside, myricetin, kaempferol-
3-O-sophoroside, kaempferol-3-O-glucoside-7-O-glucoside,
catechin-7-O-glucoside and aromadendrin-3-O-D-galacto-
side were just identified in F.esculentum [46, 47]. In F. tatari-
cum 5,7,3′,4′-tetramethylquercetine-3-O-rutinoside, quercetin-3-O-
rutinoside-7-O-galac toside, kaempferol-3-O-glucoside, kaempferol-
3-O-galactoside, kaempferol-3-O-rutinoside, cyanidin
3-O-glucoside, cyanidin 3-O-rutinoside, four proanthocyani-
dins (procyanidin A2, A3, B2 and B3), and some specific
flavonoids, including fagopyrin A to F were identified [38,
39, 48, 49].
2.1.2. Phenolics acids
Phenolic acids are a diverse group of phytochemicals
found in various plant-based foods, and they are
known for their potential health benefits. And two
main groups, including hydroxybenzoic acids (gallic
acid, protocatechuic acid, p-hydroxybenzoic acid, syrin-
gic acid, vanillic acid) and hydroxycinnamic acids (caf-
feic acid, ferulic acid, p-coumaric acid, chlorogenic acid)
have been determined in buckwheat (Figure 2) [41,
50]. In addition, some new phenylpropanoid glyco-
sides, namely tatarisides A to tatarisides G, have been
detected in F. tataricum [9]. Just protocatechuic acid
and resveratrol were detected in F.esculentum [44]. The
content of phenolics acids in buckwheat can vary
depending on factors such as the growing conditions
and processing methods. Raw buckwheat groats can
contain anywhere from approximately 50 to
400 mg/100g of total phenolic compounds [51].
2.1.3. Cyclitol
Fagopyritols are a group of soluble carbohydrates
found in F. esculentum, namely fagopyritol A1 to A3,
and B1 to B3 [52, 53]. These compounds are often
referred to as "anti-nutrients" because they can inter-
fere with the absorption of minerals, particularly
Figure 2. The bioactive components identied in buckwheat including avonoids, phenolics, tannins, cyclitol, triterpenoids, ste-
roids, and fatty acids (Dierent color areas represent dierent types of bioactive compounds).

CRITICAL REVIEWS IN BIOTECHNOLOGY 5
calcium, iron, and zinc, in the digestive tract.
Fagopyritols are specifically known for their ability to
chelate or bind to these minerals, reducing their bio-
availability [54]. Moreover, many traditional culinary
practices involve the soaking, fermenting, or cooking
of buckwheat, which can help reduce the levels of
fagopyritols and improve mineral bioavailability.
2.1.4. Triterpenoids
Triterpenoids are a diverse class of naturally occurring
compounds found in various plants, particularly in
medicinal herbs and certain vegetables, with
anti-inflammatory and antioxidant effects [55]. Several
triterpenoids have been detected in buckwheat, includ-
ing: ursolic acid, α-thujene, α-terpineol, α-amyrin,
β-amyrin, lupeol and olsolic acid in F. tataricum [40,
56], olean-12-en-3-ol and urs-12-en-3-ol in F. esculen-
tum [57], glutinone and glutinol in F. cymosum [44].
2.1.5. Steroids
Steroids are a class of organic compounds with a char-
acteristic structure, consisting of four interconnected
carbon rings. They are found in both animals and
plants and play various essential roles in biology [58].
A total of 10 steroids have been determined in buck-
wheat (Figure 2), including: 6-hydroxystigmasta-4,22-dien-3-
one, 23S-methylcholesterol, stigmast-5-en-3-ol, stigmast-5,24-
dien-3-ol, trans-stigmast-5,22-dien-3-ol in F. esculentum
[57], and β-sitosterol, β-sitosterol-palmitate, daucosterol,
peroxidize-ergosterol, stigmsat-4-en-3,6-dione in F. tatari-
cum [40].
2.1.6. Other compounds
In addition, buckwheat also contains several substances
with noteworthy bioactivities. For instance, some aroma
compounds, including: phenylacetaldehyde, decanal, hex-
anal, salicylaldehyde, 2-methoxy-4-vinylphenol, 2,5-di methy l-4-
hydroxy-3(2H)-furanone, 2-nonenal and 2,4-decadienal
were isolated in F. esculentum [59]. Ten fatty acids,
including: palmitic acid (PA), palmitoleic acid (PLA),
stearic acid(SA), oleic acid (OA), linoleic acid (LA),
α-linoleic acid (LNA), arachidonic acid (AA), eicosapen-
taenoic acid (EPA), and docosahexaenoic acid (DHA)
were detected in seeds of F. tataricum [60], and three
new fatty acids (4,7-dihydroxy-3,7-dimethyl-octa-2(E),5(
E)-dienoic acid, 6,7-dihydroxy-3,7-dimethhy-octa-2(Z),4(
E)-dienoic acid, and 6,7-dihydroxy-3,7-dimethyl-oc-
ta-2(E),4(E)-dienoic acid) in F. esculentum hulls[61].
Tropane alkaloids its derivatives determined in three
buckwheat [33, 62]. In addition, these three buck-
wheats also possess bioactive amines, such as N-trans-
feruloyltyramine [41].
2.2. Pharmacological bioactivities and applications
Multiple in vitro and in vivo studies have demonstrated
that the bioactive compounds of three buckwheats
possess versatile bioactivities, acting as: anti-tumor
[63–65], anti-inflammatory [41, 66], hyperglycemic [67],
hepatoprotective [68, 69], anti-oxidant [51, 63, 70, 71],
anti-hypertension [72], anti-obesity [67, 73] and
anti-diabetic [74, 75] (Figure 3). Some reviews have
summarized the mechanisms of action attributed to
bioactive compounds [33, 43].
Based on versatile bioactivities, buckwheat has a
wide range of applications, making it a versatile and
valuable agricultural crop, such as: culinary use, animal
use, buckwheat honey, cosmetics, and beverage indus-
try. According to market survey, some healthy foods,
including: buckwheat flour, buckwheat wine, buck-
wheat tea, buckwheat noodle, buckwheat pastry, and
buckwheat cake have been authorized by the State
Administration for Market Regulation of China [17].
Also the products of buckwheat tea, bread, and noo-
dles have widely consumed in: China, Japan, India,
Korea, Nepal, Canada, USA and Europe countries [41].
2.3. Biosynthetic pathways and transcriptional
regulation of bioactive compounds
2.3.1. Flavonoids
Flavonoids are produced through a multi-step flavo-
noid biosynthesis pathway, a subset of the phenylpro-
panoid biosynthesis pathway, which exhibits a degree
of conservation across various plant taxa. However, the
specific enzymes involved and their genetic encoding
may exhibit diversity. Notably, numerous enzymes
within the buckwheat flavonoid biosynthetic pathway
have not been characterized, and the dynamics of
metabolite flux through this pathway remain to be elu-
cidated. With the advancement of multi-omics technol-
ogies, an increasing number of functional genes are
being identified.
The biosynthesis of buckwheat flavonoid com-
pounds originates from the phenylpropanoid meta-
bolic pathway (specifically the shikimate branch
pathway). Phenylalanine is an important precursor for
the synthesis of flavonoid compounds. Under the cata-
lytic action of phenylalanine ammonia-lyase (PAL), phe-
nylalanine is converted into cinnamic acid. Cinnamic
acid is catalyzed by cinnamic acid 4-hydroxylase (C4H)
to form p-coumaric acid. Subsequently, under the
action of 4-coumaroyl-CoA ligase (4CL), p-coumaric
acid is converted into coumaroyl-CoA. Coumaroyl-CoA,
together with malonyl-CoA derived from the tricarbox-
ylic acid cycle, serves as the initial substrates for

6 Y. SONG ETAL.
flavonoid biosynthesis. Under the catalysis of chalcone
synthase (CHS), these substrates lead to the formation
of tetrahydroxychalcone (naringenin chalcone). CHS is
the first key rate-limiting enzyme in the flavonoid bio-
synthesis pathway. Tetrahydroxychalcone is catalyzed
by chalcone isomerase (CHI) to form naringenin.
Naringenin is an intermediate in the anthocyanin bio-
synthesis pathway, initiating multiple branches of the
entire anthocyanin biosynthetic process. This leads to
the synthesis of various flavonoid compounds such as
flavones, flavonols, or isoflavones, as well as anthocy-
anin substances (Figure 4A) [76]. Additionally, in legu-
minous plants and others, coumaroyl-CoA and
malonyl-CoA are catalyzed jointly by CHS (chalcone
synthase) and chalcone reductase (CHR) to form trihy-
droxychalcone. Trihydroxychalcone, under the catalysis
of CHI (chalcone isomerase), is converted into liquiriti-
genin, which then undergoes transformation into
isoflavone compounds under the action of isoflavone
synthase (IFS). This process results in the formation of
isoflavones [77]. Existing research has identified the
presence of isoflavone compounds in both common
and Tartary buckwheat, and has identified genes
related to isoflavone synthesis in buckwheat (Figure
4A) [48, 78]. However, the isoflavone synthesis branch
pathway in buckwheat requires further evidence for
confirmation.
Quercetin Synthesis: The fundamental flavonoid
scaffold is subject to additional modifications, notably
hydroxylation and glycosylation, culminating in the
generation of quercetin. This transformation is facili-
tated by enzymes such as flavonol synthase (FLS).
Rutin Formation via Glycosylation: In the terminal
phase, quercetin undergoes glycosylation, leading to
the production of rutin. This process encompasses the
attachment of a rhamnose sugar molecule to
Figure 3. Pharmacological bioactivities and applications of bioactive compounds in buckwheat, including anti-tumor,
anti-inammatory, hyperglycemic, hepatoprotective, anti-oxidant, anti-hypertension, anti-obesity and anti-diabetic. Created with
BioRender.com.

CRITICAL REVIEWS IN BIOTECHNOLOGY 7
Figure 4A. A Flavonoids biosynthesis pathway including rutin, anthocyanins, proanthocyanidin in buckwheat. PAL: Phenylalanine
Ammonia-Lyase; C4H: Cinnamate 4-Hydroxylase; 4CL: 4-Coumarate Coenzyme A Ligase; CHS: Chalcone Synthase; CHI: Chalcone
Isomerase; F3H: Flavanone 3-Hydroxylase; F3′H: Flavonoid 3′-Hydroxylase; F3′5′H: Flavonoid 3′,5′-Hydroxylase; DFR: Dihydroavonol
4-Reductase; FLS: Flavonol Synthase; LAR: Leucoanthocyanidin Reductase; LDOX/ANS: Leucoanthocyanidin Dioxygenase/
Anthocyanidin Synthase; UFGT: UDP-Glucose Flavonoid 3-O-Glucosyltransferase; RT: Rhamnosyltransferase; 3GT: 3-Glucosyltransferase.
Figure 4B. Phenolic acids biosynthesis pathway including gallic acid, 4-hydroxybenzoate, caeic acid, ferulic acid, sinapic acid and chlo-
rogenic acid in buckwheat. PAL: Phenylalanine Ammonia-Lyase; C4H: Cinnamate 4-Hydroxylase; 4CL: 4-Coumarate Coenzyme A Ligase.

8 Y. SONG ETAL.
quercetin. A specific UDP-glycosyltransferase enzyme
typically mediates this glycosylation step (Figure 4A).
Among the cloned and identified genes associated
with rutin synthesis in buckwheat, the following have
been noted: F3H (Flavnone-3-hydroxylase), FLS (Flavnol
synthase), F3′H (Flavonoid-3′-hydroxylase), F3′5′H (Flavonoid-3′5′-
hydroxylase), 3GT (Flavonoid 3-O-glucosyltransferase), RT
(3-O-Rhamnosyltransferase) [32, 79, 80].
Proanthocyanidins (PAs) and anthocyanins commence
their biosynthetic pathways at a convergent point, initi-
ated by the action of dihydroflavonol 4-reductase (DFR),
which leads to the production of leucoanthocyanidins
(Figure 4A). These intermediates may either be trans-
formed into (+)-catechins through the enzymatic activity
of leucoanthocyanidin reductase (LAR) or converted into
anthocyanidins via anthocyanidin synthase (ANS), and
subsequently to (−)-epicatechins by anthocyanidin
reductase (ANR). The biosynthesis of PAs involves the
polymerization of either or both (+)-catechins and
(−)-epicatechins. Among the cloned and identified genes
associated with rutin synthesis in buckwheat, the genes
have been noted, including: DFR (Dihydroflavonol reduc-
tase), ANS (Anthocyanin synthase), UFGT(glucose-flavonoid
glucosyl transferase), LAR(leucoanthocyanidin reductase)
[81–83].
2.3.2. Phenolic acids
The biosynthesis pathway of phenolic acids in buck-
wheat is part of the larger phenylpropanoid pathway,
which is a key route for the production of a wide
range of secondary metabolites in plants. Phenolic
acids, such as ferulic acid, caffeic acid, and p-coumaric
acid, are important components of this pathway (Figure
4B). The pathway starts with phenylalanine, and con-
verts into cinnamic acid catalyzed by PAL. Cinnamic
acid is then hydroxylated to form p-coumaric acid. The
C4H typically catalyzes this reaction. To Caffeic Acid:
p-Coumaric acid can be further hydroxylated by the
C3H to form caffeic acid. To Ferulic Acid: Alternatively,
p-coumaric acid can be methoxylated by the enzyme
p-coumarate O-methyltransferase (COMT) to produce
ferulic acid [84–86]. While some key enzymes and steps
have been identified, the entire biosynthetic pathway
of phenolic acids in buckwheat may not be fully eluci-
dated, such as the biosynthesis of chlorogenic acid,
and require comprehensive mapping. There is a need
for more in-depth research on the genetic and molec-
ular factors that regulate the biosynthesis of phenolic
acids in buckwheat, including the identification and
characterization of more genes and transcription fac-
tors involved in this pathway.
Figure 4C. Fagopyrin, hypericin and pseudohypericin biosynthesis pathway in buckwheat. CHS: Chalcone Synthase; Hyp-1:
Hypericum perforatum.

CRITICAL REVIEWS IN BIOTECHNOLOGY 9
2.3.3. Fagopyrin
Fagopyrin, a type of naturally occurring polyphenolic
compound found in buckwheat, particularly Tartary
buckwheat, belongs to the group of naphthodian-
thrones, which are chemically similar to hypericin
found in St. John’s Wort [87]. The biosynthesis pathway
of fagopyrin in buckwheat, while not fully elucidated,
is believed to be part of the shikimate and polyketide
pathways. The biosynthetic process of fagopyrin initi-
ates within the shikimate pathway, a predominant bio-
synthetic conduit for aromatic amino acids, transforming
simple carbohydrate precursors into shikimic acid. The
subsequent conversion of shikimic acid into chorismic
acid marks a critical juncture, where one divergent
pathway culminates in the generation of protocate-
chuic acid, an essential precursor for a variety of phe-
nolic compounds. Concurrently, the polyketide
synthesis pathway, characterized by the sequential
condensation of acetyl-CoA units, plays a pivotal role
in constructing the naphthodianthrone structure, fun-
damental to fagopyrin. The synthesis of naphthodian-
throne compounds is orchestrated by specific enzymes
through a series of intricate reactions, encompassing
cyclization and oxidative modifications. The terminal
phase of fagopyrin biosynthesis entails the intricate
coupling of the naphthodianthrone structure with
additional molecular fragments, potentially originat-
ing from the shikimate pathway, thus culminating in
the distinctive molecular architecture of fagopyrin
(Figure 4C).
2.4. Regulation of buckwheat avonoid
biosynthesis
Current research on the biosynthesis of buckwheat fla-
vonoids has extensively explored the key enzyme genes
involved in the pathway. The primary steps of flavonoid
biosynthesis in buckwheat are relatively well under-
stood. Research on regulatory genes, however, is in a
phase of rapid development. Studies focusing on the
regulation of flavonoid biosynthesis in buckwheat are
predominantly concentrated on the transcription factor
MYB. MYB achieves regulatory control over the tran-
scription of target genes either by directly binding to
the promoter regions of these genes or through inter-
actions with other proteins, such as WD40 and bHLH.
The regulatory effects exerted by MYB are characterized
by pronounced temporal and spatial specificity. To date,
twenty MYB transcription factors have been identified
in buckwheat. Among these, the known gene sequences
of sixteen MYB transcription factors belong to the
R2R3-MYB subfamily. Transcription factors primarily
associated with buckwheat flavonoid biosynthesis
include: FtMYB1, FtMYB2, FtMYB3, FtMYB7, FtMYB9,
FtMYB11, FtMYB13, FtMYB14, FtMYB15, FtMYB16,
FtMYB116, and FtMYB123L. These factors are distributed
across various tissues and organs in buckwheat. During
specific stages of growth and development, they regu-
late the concentration of different flavonoid compounds
in various tissues and organs by inducing or inhibiting
the expression of key enzyme genes in flavonoid syn-
thesis. This regulation activates or deactivates specific
branch pathways of flavonoid biosynthesis [88–91].
MYB can directly regulate specific flavonoid biosyn-
thesis pathways by directly interacting with the key
enzyme genes involved in flavonoid synthesis. The
FtMYB11, FtMYB13, FtMYB14, FtMYB15, and FtMYB16 in
buckwheat exhibit a tissue-specific expression in vari-
ous tissues and are regulated by jasmonic acid. Among
these, FtMYB13, FtMYB14, and FtMYB15 are subject to
jasmonic acid-induced proteolytic degradation at the
protein level, directly inhibiting the expression of the
FtPAL gene, thereby reducing flavonoid accumulation.
Additionally, FtSAD2 and FtJAZ1, which interact syner-
gistically with: FtMYB11, FtMYB13, FtMYB14, and
FtMYB15, significantly enhance the repressor activity of
FtMYBs [92, 93]. In addition, MYB regulate the tran-
scription of target genes through interactions with
other proteins. Simultaneously, they are modulated by
inducers, such as jasmonic acid, abscisic acid, and
environmental factors such as light exposure/UV radi-
ation. Abscisic acid, drought sensitivity, and jasmonic
acid signaling inhibitors can synergistically interact
with MYB to inhibit flavonoid biosynthesis in buck-
wheat. For example, the FtMYB8 mRNA is predomi-
nantly distributed in the roots of buckwheat during
the true leaf stage and flowering stages. Overexpression
of the FtMYB8 gene can inhibit the accumulation of
anthocyanins/proanthocyanidins. The FtMYB8 modu-
lates the content variations of anthocyanins/proantho-
cyanins in the roots and flowers of buckwheat, thereby
influencing the distribution of trichomes on the floral
buds of buckwheat [94].
Additionally, research on the relationship between
MYB and the relative expression levels of multiple key
enzyme genes involved in flavonoid synthesis has been
conducted. The FtMYB7 exhibits its highest relative
expression levels during the true leaf and budding
stages. The relative expression patterns of: Ft4CL, FtCHS,
FtF3H, FtUFGT, and FtMYB7, are positively correlated
with the fluctuations in rutin content [95]. However, the
situation concerning the expression correlation between
key enzyme genes: PAL, CHI, FLS and FtMYB1, FtMYB2
and FtMYB3, is relatively complex. There is a lack of
strong regularity in the transcription of key enzyme
genes in relation to the expression patterns of these

10 Y. SONG ETAL.
transcription factors. Therefore, it is conjectured that
certain key enzyme genes in flavonoid synthesis may
be regulated exclusively by specific transcription fac-
tors, precisely initiating one or more branch pathways
for the synthesis of flavonoid compounds. This charac-
teristic cannot be adequately reflected solely by
changes in the total flavonoid content. Research on the
regulation of buckwheat flavonoid biosynthesis is still
in its nascent stages. Consequently, elucidating the key
genes and regulatory factors within the buckwheat fla-
vonoid biosynthetic pathway can lay the foundation for
further revealing the molecular mechanisms underlying
the biosynthesis of flavonoid compounds in buckwheat.
3 Applications of multi-omics technologies for
nutraceutical improvement
3.1. Genomics research
Genomics research in buckwheat, encompassing both
common buckwheat and Tartary buckwheat, has been
progressing significantly, unlocking insights into this
crop’s unique traits, evolutionary history, and potential
for crop improvement.
The genome of common buckwheat is relatively
large and complex. It is characterized by a high level
of repetitive DNA, which comprises a significant por-
tion of the genome. The exact size can vary among
cultivars, but it is generally approximately 1.2 to 1.5 Gb
[96–98]. The chromosome-scale genome assembly
helps in understanding the evolution of the buckwheat
genome and the origins of the cultivated crop. The
expansion of several gene families in common buck-
wheat is associated with its wider distribution com-
pared to Tartary buckwheat. Notably, copy number
variations in genes involved in flavonoid metabolism
correlate with differences in rutin content between
common and Tartary buckwheat [99]. The expansion of
several gene families in common buckwheat, such as
FhFAR genes, is associated with its broader distribution
compared to Tartary buckwheat. Genome-wide associ-
ation analyses identified genes like Fh05G014970 as
potential major regulators of the flowering period, a
crucial trait for crop yield. Another gene, Fh06G015130,
is crucial for flavor-associated flavonoids [99].
Comprehensive genomic variation based on whole-genome
resequencing of multiple accessions revealed genetic
variation associated with environmental adaptability
and floral development. However, a comprehensive
understanding of the genetic diversity in common
buckwheat remains limited, especially concerning
genetic variations across different geographical regions
and environmental conditions.
The Tartary buckwheat genome, sized at 489.3 Mb,
was assembled using a combination of whole-genome
shotgun sequencing, single-molecule real-time long
reads, and sequence tags of a large DNA insert fosmid
library, Hi-C sequencing data, and BioNano genome
maps [32]. This enabled the construction of a compre-
hensive database of genome variation, facilitated pop-
ulation structure and domestication studies. In addition,
an extensive database encompassing genomic variation
was established through whole-genome resequencing of
510 germplasms. This endeavor uncovered evidence of
two distinct domestication events, occurring inde-
pendently in southwestern and northern China. The
genetic diversity elucidated by this study significantly
contributes to the phenotypic diversity observed in
contemporary Tartary buckwheat cultivars [100].
Genome-wide association studies identified several
candidate genes, such as FtUFGT3 and FtAP2YT1, which
are significantly correlated with flavonoid accumula-
tion and grain weight, respectively. Also, the AP2/ERF
gene family which regulates flower and fruit develop-
ment was preliminarily identified based on
genome-wide investigation [101]. Additionally, quanti-
tative trait loci (QTLs) associated with 1000-grain
weight (TGW) and genes governing hull type were
mapped across various environmental conditions,
underscoring their influence in multiple contexts. Also,
it was found that nine QTLs for TGW were detected
and distributed on four loci on chromosome 1 and 4.
The genes responsible for hull type were localized to
chromosome 1, situated between markers Block330
and Block331. This genomic region is in close proxim-
ity to the major locus that influences TGW, indicating
a potential linkage or interaction between these
genetic elements [102]. The Tartary buckwheat
genome also provided insights into genes predicted
to be involved in: rutin biosynthesis and regulation,
aluminum stress resistance, and drought and cold
stress responses [32, 103–106]. Thus, genomic studies
could be used to mine elite genes for crop
improvement.
The F. cymosum complete chloroplast genome is
159,919 bp long, with a typical quadripartite structure,
including inverted repeat regions, a large single copy
region, and a small single copy region. It encodes 131
genes, comprising 80 protein-coding genes, 28 tRNA
genes, and 4 rRNA genes. The genome structure and
gene order are typical of angiosperm complete chloro-
plast genomes [107, 108]. The genome of F. cymosum
exhibits a size exceeding twice that of its closely
related counterpart, Tartary buckwheat, which pos-
sesses a genome of approximately 0.48 Gb [32]. Despite
this substantial difference in genome size, the gene

CRITICAL REVIEWS IN BIOTECHNOLOGY 11
count in golden buckwheat (38,919 genes) is compara-
ble to that in Tartary buckwheat (33,366 genes). This
observation collectively suggests that the expansion in
genome size of F. cymosum, relative to Tartary buck-
wheat, predominantly results from the large-scale
amplification of repetitive sequences [108].
Predominantly, golden buckwheat ecotypes with an
erect growth habit are observed at lower altitudes,
whereas decumbent ecotypes are more prevalent at
higher altitudes [109]. Genomic comparisons between
these ecotypes have led to the identification of candi-
date genes that may be instrumental in the pheno-
typic divergence of these two ecotypes. Based on
genomic research, a notable alteration was identified
in the erect ecotype: the predicted start codon of a
CRF gene was absent, which could potentially lead to
either a loss of gene expression or the production of a
mutant protein. Such a genetic modification might
inhibit the erect ecotype’s ability to thrive in colder,
high-altitude environments, thereby confining its distri-
bution to the warmer, low-altitude regions [108, 109].
The utilization of a resequencing panel, encompassing
a wide range of genetic variation, significantly enhances
the capacity to investigate the population structure of
golden buckwheat.
Genome-Wide Association Studies (GWAS) in buck-
wheat, particularly focusing on Tartary buckwheat,
have led to significant insights into the genetic basis
of important agronomic traits. The resequencing of
510 Tartary buckwheat germplasms from around the
world, including 483 landraces and 27 wild acces-
sions, generated a massive dataset of genomic varia-
tions. GWAS was performed and identified genetic
factors associated with the yield and growth period,
including the FtAP2YT1 gene, which influences
1000-grain weight. In addition, GWAS analysis was
also used to identify genes correlated with the con-
tent of flavonols, such as: quercetin, rutin, and
kaempferol-3-O-rutinoside. A significant association
with kaempferol-3-O-rutinoside content was identi-
fied, leading to the identification of the FtUFGT3
gene, which plays a role in flavonoid metabolism
[100]. Zarger et al. provided insights into the genetic
architecture of seed metabolome in buckwheat based
on metabolic-GWAS, and identified 27 SNPs/QTLs
linked to 18 metabolites [110]. Naik et al. found a
total of 71 significant quantitative trait loci revealed
the genomic regions associated with major
yield-attributing traits in buckwheat with GWAS anal-
ysis, and 71 significant marker-trait associations across
eight chromosomes were identified [111]. Yang et al.
revealed FtCOMT1 reinstated S lignin biosynthesis
based on GWAS detection, which could enhance
dehulling efficiency in buckwheat [112]. In common
buckwheat, GWAS examining multi-year agronomic
traits and flavonoid content have identified
Fh05G014970 as a putative principal regulator of
the flowering period, a critical agronomic trait influ-
encing the yield of outcrossing crops. Furthermore,
Fh06G015130 has been pinpointed as a vital gene
associated with the flavor-related flavonoids. Notably,
our research revealed that gene translocation and
sequence variation in FhS-ELF3 play a significant role
in the homomorphic self-compatibility observed in
common buckwheat [99]. These findings highlight the
power of GWAS in uncovering the genetic underpin-
nings of critical traits in buckwheat, offering a path-
way for targeted breeding and improvement of this
important crop.
3.2. Transcriptomics research
The progress in transcriptomic research in buckwheat
has been notable in recent years, particularly focusing
on its qualities as a functional food source and its
environmental benefits (Table S1).
3.2.1. Stress response studies
Transcriptomic analysis has been instrumental in study-
ing buckwheat’s response to environmental stresses,
such as: cadmium, cold stress, drought stress, salt
stress, nitrogen stress, Pb stress and Al stress. Studies
reveal that Tartary buckwheat can tolerate high con-
centrations of cadmium, suggesting its potential use in
phytoremediation, and found Cd stress triggered the
induction of nine pivotal genes, which play roles in an
array of biological processes, including: binding of
metal ions, calcium signal transduction, cell wall orga-
nization, and antioxidant activities [113]. Different
buckwheat landrace different responses cold stress in
proanthocyanidin and rutin synthesis, suggesting a
high rutin content to enhance cold tolerance with GTR,
F3’H and FLS upregulation [104]. Differential regulation
of genes involved in oxidoreductase activity in cotyle-
don and root, ABA (Abscisic Acid)-dependent and
ABA-independent pathway, transcription factors, includ-
ing: NAC, bZIP, MYB and WRKY families were take part
in the response to drought stress, and gene involved
in phenylpropanoid synthesis overrepresented to
response drought stress [114–116]. Also, small secreted
peptides (SSPs) may regulate the adaptability of buck-
wheat under low nitrogen stress by modulating the
expression of the genes involved in N transport and
assimilation and IAA (Indole-3-Acetic Acid) signaling,
53 low nitrogen-responsive RLKs encoding genes were

12 Y. SONG ETAL.
identified and they were predicted as potential SSP
receptors, which was revealed based on transcriptomic
research [117]. The UV-B stress treatment of hairy roots
resulted in a striking increase of rutin and quercetin
production, demonstrated that the UV-B radiation was
an effective elicitor that dramatically changed in the
transcript abundance of: FtPAL, FtCHI, FtCHS, FtF3H, and
FtFLS [118]. In addition, FtTCP15 and FtTCP18 were also
revealed the potential function in response to UV
stress in buckwheat [119]. Above all, transcriptomic
research: plays a crucial role in understanding plant
stress responses, is vital for unraveling the complex
molecular mechanisms underlying plant stress
responses, and paving the way for the development of
crops that are better equipped to withstand environ-
mental challenges.
3.2.2. Medicinal and nutritional metabolites
Transcriptomic research plays a significant role in
understanding and enhancing the production of
medicinal and nutritional metabolites in plants. This
research area focuses on how genes are expressed in
relation to the biosynthesis of these valuable com-
pounds. Transcriptomics helps in identifying the genes
and enzymes involved in the biosynthetic pathways of
various metabolites. Hou et al. researched flavonoid
metabolism in different tissues in buckwheat, and
revealed the roadmap of the rutin synthesis pathway
[120]. Transcriptomic studies reveal the complex regu-
latory networks that control the synthesis of medicinal
and nutritional compounds. Key enzymes such as: PAL,
CHS, CHI, and FLS were differentially expressed, influ-
encing the total flavonoid through transcriptomic
research in the buckwheat flower [121]. The key role of
FtPinG0009153900.01 in flavonoid biosynthesis, particu-
larly that regulating flavanones and flavonols, was
found [79], which can guide breeding and genetic
engineering efforts to improve both the quantity and
quality of flavonoids. Overall, transcriptomic research is
crucial for advancing our understanding and manipula-
tion of the biosynthetic pathways of medicinal and
nutritional metabolites in plants, leading to improved
health benefits and therapeutic applications.
3.2.3. Improving crop agronomic traits
Transcriptomic studies help identify genes and gene
networks responsible for important agronomic traits
such as yield and quality. By understanding which
genes are active during the different stages of plant
growth or under stress conditions, researchers can
identify key genetic targets for crop improvement.
Jiang etal. researched seeds during filling stages based
on transcriptomics, and found Phytohormone: ABA,
AUX, ET, BR and CTK, and related TFs which could sub-
stantially regulate seed development [120]. Fang et al.
found AP2 and bZIP transcription factors, BR-signal and
ABA were considered to be important regulators of
seed size based on transcriptomic analysis [122].
In summary, transcriptomic research is instrumental
in advancing our understanding of plant biology,
which directly translates into the development of
improved crop varieties with: enhanced yields, better
stress tolerance, improved nutritional quality, and dis-
ease resistance. This is increasingly important for ensur-
ing global food security in the face of a growing
population and changing climate conditions.
3.3. Proteomics research
Recent advancements in proteomics research related
to buckwheat have contributed significantly to the
understanding of its growth, nutrient enrichment, and
stress responses. A key study using iTRAQ-based quan-
titative proteomic analysis on buckwheat sprouts
treated with slightly acidic electrolyzed water (SAEW)
revealed changes in proteins associated with: energy
metabolism, oxidative stress, and flavonoid biosynthe-
sis, which demonstrated how the SAEW treatment can
induce flavonoid enrichment in buckwheat sprouts by
modulating enzymes involved in the phenylpropanoid
biosynthesis pathway [123]. In addition, FeALS1.1 and
FeALS1.2 were identified based on proteomic analysis
in common buckwheat, which is involved in the inter-
nal detoxification of Al in the roots and leaves, respec-
tively [124]. Wang et al. provide a comprehensive
picture of the seed’s proteome, and this analysis was
pivotal in understanding the functional role of these
proteins in seed germination, nutrient composition,
and metabolism of Tartary buckwheat seeds [125].
Investigation in proteomic changes in flowers and
leaves of two common buckwheat with varying heat
tolerance, found that a high temperature up-regulated
the expression of 182 proteins, with the proteomic
response varying between the accessions and their
organs [126]. Proteomic research also used to evaluate
nutritional properties. The comparative proteomic
study of wheat, oat, and buckwheat genotypes showed
that buckwheat has the best nutritional quality based
on its high coefficient of nutritional quality and con-
tent of essential amino acids [127].
The proteomics research in buckwheat has signifi-
cantly advanced our understanding of: the seed’s nutri-
tional properties, stress responses, allergenic potential,
and germination mechanisms. These findings are vital
for enhancing buckwheat’s agricultural and nutritional

CRITICAL REVIEWS IN BIOTECHNOLOGY 13
value and for developing strategies to utilize it more
effectively in food and health industries. To date, pro-
teomics researches in buckwheat remain limited. Future
research should also focus on the detection of func-
tional protein.
3.4. Metabolomics research
Metabolomics research in buckwheat has seen consid-
erable progress, focusing on the crop’s nutritional and
medicinal properties, particularly its rich flavonoid con-
tent. 784 metabolites, including a large group of: flavo-
noids, organic acids, and amino acids were identified
based on metabolomics analysis, which revealed the
nutrition profile of different common buckwheat culti-
vars [128]. Metabolomics analysis is also employed to
nutrition evaluation between buckwheat and other
crops, and reveal nutritional diversity of buckwheat
[129]. Much metabolomics research focuses on buck-
wheat because of its high and diverse flavonoid con-
tent. These studies have led to well understood
biosynthesis and regulatory mechanisms for flavonoids
[78, 82, 130]. The metabolites vary often in different
tissues of buckwheat. The rutin and chlorogenic acid in
cotyledons were higher than that in seeds and radicles
during germination [131].
Metabolomics research in response to environmen-
tal stress. Based on metabolomic analysis, Song et al.
revealed the phenylalanine pathway’s role in cold tol-
erance and flavonoid synthesis in Tartary buckwheat
[104]. Du et al. found 1798 metabolites enrichment in:
galactose metabolism, glycerol metabolism, phenylpro-
pane biosynthesis, glutathione metabolism, starch, and
sucrose metabolism, and resistance to cadmium [132].
Gene-to-metabolite network analysis in buckwheat
salinity stress was conducted based on metabolomics
and transcriptomics, and found changes in secondary
metabolite biosynthesis, including phenylpropanoid
and flavonoid biosynthesis [133]. Metabolomics
research employed in buckwheat response to environ-
mental stress, which are the small molecule end prod-
ucts of cellular processes, contributes significantly to
understanding how buckwheat, a valuable crop known
for its nutritional qualities and adaptability to stress.
The field of Weighted Gene Co-expression Network
Analysis (WGCNA) has made significant strides in the
study of buckwheat biology, encompassing a diverse
range of research aspects. Notably, Meng et al. con-
ducted a comprehensive study involving comparative
physiological, transcriptomic, and WGCNA analyses to
elucidate key genes and regulatory pathways that con-
fer drought tolerance in Tartary buckwheat [134]. This
study highlighted pronounced disparities in drought
responses between drought-tolerant and drought-susceptible
genotypes, pinpointing central drought-resistant genes and
delineating their functional roles in Tartary buckwheat.
In another vein, Huang et al. integrated RNA-seq
analyses with WGCNA on buckwheat varieties with
varying starch contents [135]. This approach facilitated
the identification of differentially expressed genes
(DEGs) and the determination of trait-specific modules
directly correlated with starch-related characteristics.
Furthermore, Li et al. employed WGCNA to investigate
the mechanism underlying the easy dehulling of
rice-Tartary buckwheat [136]. This research identified
pivotal modules and genes associated with hull forma-
tion, revealing variations in the expression of genes
responsible for secondary cell wall biosynthesis, which
significantly influence the hull’s physical attributes.
Additionally, WGCNA has been instrumental in uncov-
ering core genes activated by cadmium stress in buck-
wheat, as demonstrated by Ye, etal. [113]. These genes
play critical roles in various biological processes, includ-
ing: metal ion binding, signal transduction, and antiox-
idant activities, shedding light on the plant’s adaptive
mechanisms in the face of heavy metal stress.
Metabolic Genome-Wide Association Studies
(mGWAS) in buckwheat has been an evolving field,
blending advanced genetic analysis with metabolom-
ics to understand the genetic basis of metabolite
variation. A study by Zargar et al. utilized high-
throughput metabolomic analysis to identify 24
metabolites in the seed endosperm of 130 diverse
buckwheat genotypes [110]. The genotyping-by-se-
quencing (GBS) of these genotypes revealed a vast
number of SNPs (3,728,028). The Genome Association
and Prediction Integrated Tool (GAPIT) was instru-
mental in identifying 27 SNPs/QTLs linked to 18 dif-
ferent metabolites, providing valuable insights into
several metabolic and biosynthetic pathways in buck-
wheat [110]. This study highlights the significant
strides made in understanding the genetic basis of
metabolite variation in buckwheat through mGWAS.
Additionally, most mGWAS studies might focus on a
limited set of metabolites, potentially overlooking
others that could be important for specific traits like
disease resistance or nutritional quality, and further
research to confirm the relevance of identified mark-
ers and the complexity of incorporating these mark-
ers into breeding strategies.
4. Functional genes
Buckwheat, a highly nutritious pseudocereal, pos-
sesses various functional genes that contribute to its
valuable agricultural and nutritional qualities. The

14 Y. SONG ETAL.
potential functions of these functional genes have
been identified, including: biosynthesis of flavonols
genes, stress response genes, and plant development
genes (Table 1).
4.1. Biosynthesis of avonols genes
A substantial number of genes implicated in the bio-
synthesis of flavonols have been isolated and
characterized from buckwheat. Within the UFGT
(UDP-glucose: flavonoid 3-O-glucosyltransferase) gene
family, several members, including: FtUFGT8, FtUFGT15,
and FtUFGT41, have been identified. These genes are
known to play a regulatory role in the biosynthesis of
anthocyanins in Tartary buckwheat [139]. Furthermore,
within the R2R3-MYB family, genes, such as: FtMYB43,
FtMYB4R, FtMYB3, FtF3′H1, and FtWD40 have been
identified as regulators of anthocyanin biosynthesis
Table 1. Functional gene identied in buckwheat.
Category Gene Name Role in buckwheat Reference
MADS FtMADS Regulate the easy dehulling of Tartary buckwheat fruit [137]
FaesPl_1, FaesPl_2 Regulating lament development [138]
UFGT FtUFGT8, FtUFGT15, FtUFGT41 Regulating anthocyanin synthesis [139]
UGT FtUGT73BE5 Glucosylate avonol [140]
FeCGTa, FeCGTb C-glucosyltransferases responsible for the biosynthesis of
C-glucosylavones
[141]
miRNAs fes-miR156j-5p Regulating seed size [142]
fta-miR167c-5p, FtPinG0002560000.01 Regulating seed development [143]
GRAS FtGRAS22 Relating-fruit development [144]
Hsf FtHsf Relating-fruit development [145]
NAC FtNAC70 Resistance to salt and drought, involved in the development of owers [146]
FtNAC43, FtNAC46, FtNAC58 Play a role in the roots and leaves development [147]
FtNAC4,7 Response to salt, drought, abscisic acid, and salicylic acid [148]
ABC FeALS1.1, FeALS1.2 Resistance to Al [124]
CHS FtCHS Responded to Cd and drought stress [149]
CHI FtCHI Response to the hormone and environment factors, and promote the
avonoid synthesis
[150]
F3′HFtF3′H1 Involve in anthocyanin and avonol metabolism [151]
FLS FtFLS1 Play a key role in rutin biosynthesis [152]
DFR FeDFR1a Contribute to substrate specicity [153]
AAT FtAAP12, FtCAT7 Relating-grain development and response to abiotic stress [154]
TIFY FtTIFY1, FtJAZ7 Relating ABA-mediated germination and stress responses [106]
HMA FtP1bA1 Response to Cd stress [155]
AP2/ERF FtAP2/ERF Regulate ower and fruit development [101]
R2R3-MYB FtMYB Response to abiotic stresses [156]
FtMYB3 Negatively regulated anthocyanin and PA biosynthesis [157]
FtMYB43 Regulation of anthocyanin or proanthocyanidin biosynthesis [158]
FtMYB4R1 Regulation of anthocyanin or proanthocyanidin biosynthesis [159]
FtMYBF1 Regulate avonol synthesis [80]
FtMYB30 Enhancing drought/salt tolerance [160]
FtMYB1, FtMYB2 Enhance the accumulation of proanthocyanidins [89]
WRKY
FtWRKY6,74,31 Response to salt and drought treatments [161]
FtWRKY29 Response to low-P-induced stress [162]
GBSSI FtGBSSI Relating-starch synthesis [163]
ARF FtARF Respond positively to auxin during fruit development [164]
FtPinG0001427400.01,
FtPinG0001428600.01,
FtPinG0002492200.01
Regulate the easily-shelled [165]
LOX FtLOX1, FtLOX4-7 Respond to light [166]
SPL FtSPLs Regulate ower and fruit development [167]
NF-Y FtNF-Ys Relating-fruit development [168]
TH FtTH Related to the growth and development of Tartary buckwheat [169]
GST FeGST1 segregated with stem color [170]
MT FeMT3 Respond to heavy metal [171]
Dof FtDofs Regulate growth and development of buckwheat and response to
abiotic and biotic stresses
[172]
TCP FtTCP15,18 cis-elements in response to abiotic stress and conserved nature in
evolution
[119]
HDAC FtHDAC Relating-avonoid synthesis and cold response [105]
MAPK FtMAPK1, FtMAPK2, FtMAPK3, FtMAPK12 Regulate the expression of other transcription factors and participate in
the abiotic stress response
[173]
AP3 FaesAP3 Regulating the development of stamens [174]
ATG FtATG8a, FtATG8f Response to drought and salt stresses [175]
CBF/DREB FeDREB1 Enhanced the drought and freezing tolerance [176]
AG FaesAG Regulate the ower development [177]
PI FaesPI Involved only in stamen development [178]
WD40 FtWD40 Regulate the anthocyanin biosynthesis [179]
GH FtGH1 Hydrolyse rutin to quercetin [180]

CRITICAL REVIEWS IN BIOTECHNOLOGY 15
[151, 158, 159, 179]. Notably, FtMYB3 specifically has
been characterized as a negative regulator of both
anthocyanin and proanthocyanidin (PA) biosynthetic
pathways [157]. Within the flavonol metabolic pathway,
genes such as: FtF3′H1, FtCHI, FtGH1 and FtHDAC, have
been identified as regulators of flavonol synthesis [105,
150, 151, 180]. Additionally, the gene FtUGT73BE5 in
Tartary buckwheat, along with FeCG Ta and FeCGTb in
common buckwheat, have been associated with the
glucosylation of flavonols [140, 141]. Despite the iden-
tification of numerous metabolic genes in buckwheat,
the functional roles of the majority of these genes
remain largely undefined. Consequently, it is impera-
tive to employ genetic modification methodologies to
elucidate the specific functions of these genes in buck-
wheat. Furthermore, among these three buckwheat,
functional gene exploration in golden buckwheat has
been comparatively limited. Therefore, there is a neces-
sity for further investigation into the functional genes
involved in flavonols synthesis present in golden
buckwheat.
4.2. Stress response genes
Buckwheat, characterized as sessile organisms, have
developed intricate mechanisms to adapt to a diverse
array of environmental stressors. These include condi-
tions, such as: drought, mechanical damage, soil salin-
ity and acidification stress, and extreme temperature
variations, encompassing both high and low tempera-
tures. Some key functional genes which were reported
could response abiotic stress, including FtAAP12 and
FtCAT7 belonging to AAT [154], FtTIFY1 and FtJAZ7
belonging to TIFY [106], FtMYB belong to R2R3-MYB
[156], FtDofs [172], FtTCP15 and 18 [119], FtMAPK1,
FtMAPK2, FtMAPK3 and FtMAPK12 belong to MAPK
[173]. In addition, some genes response to specific
stress were also identified. FtNAC4,7 and 70 belonging
to NAC, FtWRKY6,74 and 31 belong to WRKY, FtATG8a
and FtATG8f belonging ATG, FtMYB30, FtCHS, were
response to drought and salt stress in Tartary and
common buckwheat [146, 148, 149, 157, 161, 175].
While FeDREB1 in common could enhance the drought
and freezing tolerance [176]. Another reported
response to cold stress gene was FtHDAC [105].
Furthermore, some genes response to heavy metal
were also identified, including FeALS1.1 and FeALS1.2
(Al), FtCHS and FtP1bA1 (Cd), and FeMT3 (heavy metal)
[113, 124, 149, 171]. Plant growth is markedly influ-
enced by environmental factors. The identification
and characterization of stress-specific genes represent
a potent strategy to augment plant resistance,
utilizing advanced biotechnological approaches.
However, current efforts in the identification and
characterization of genes associated with pest and
disease resistance are notably lacking, necessitating
further research in this area.
4.3. Plant development genes
Agronomic traits, being critical indicators that affect
crop yield and quality, have led to the discovery of
several functional genes influencing the agronomic
characteristics of buckwheat. In this section, we review
functional genes related to buckwheat plant develop-
ment to help crop improvement (Figure 5).
The FaesAG belonging to AG gene family and
Fh05G014970, were reported regulate the flower devel-
opment in common buckwheat [99, 152]. In Tartary
buckwheat the FtSPLs belonging to the SPL gene fam-
ily, FtAP2 belonging to AP2/ERF, and FtNAC70 were
identified which were involved in the development of
flowers [101, 144, 146]. In addition, the development
of flowers in common buckwheat was also detected,
and found FaesPI and FaesAP3 which majorly regulate
the development of stamens [174, 176]. The rate and
success of fruit development can be influenced by var-
ious factors, including: environmental conditions (like
temperature and light), the nutritional status, and
genetic factors. The functional genes involved in fruit
development exploration was currently a strong focus.
Some genes were identified with fruit development,
including: FtHsF, FtAAP12, FtCAT7, FtAP2, FtARF, FtSPLs,
and FtNF-Ys [101, 145, 154, 167, 168, 181]. Additionally,
a miRNA named fta-miR167c-5p was identified in
Tartary buckwheat, which could regulate seed develop-
ment [143]. All the aforementioned functional genes
related to fruit and seed development are specific to
Tartary buckwheat. Correspondingly, there is a need for
further exploration of similar genes within common
and golden buckwheat. Seed size, as a crucial agro-
nomic trait in buckwheat, is a significant factor in
enhancing yield. One miRNA fes-miR156j-5p was identi-
fied, which regulates the seed size [142]. The trait of
easy dehulling primarily affects the convenience of
harvesting and seed processing. FtMADS and a further
three genes belonging to ARF gene family, including:
FtPinG0001427400.01, FtPinG0001428600.01 and
FtPinG0002492200.01, were identified, which regulates
those easily-shelled [137, 165]. In the detection of a
functional gene regulating stem development, only
FeGST1 was identified, which segregated with stem
color [170]. In the NAC gene family, the function of
FtNAC43, FtNAC46, and FtNAC58 were identified, which

16 Y. SONG ETAL.
play a role in roots and leaves development [147].
These functional genes are crucial for enhancing our
understanding of plant biology and for the develop-
ment of improved buckwheat varieties. However, sev-
eral areas require further research. For instance, the
molecular mechanisms by which functional genes reg-
ulate agronomic traits need more in-depth investigation.
Additionally, there is a need to identify and characterize
functional miRNAs and to explore functional genes that
regulate the plant height in buckwheat.
5. Integration of multi-omics for crop
improvement
5.1. Multi-omics strategies for improving bioactive
compound biosynthesis
The application of multi-omics approaches are to
enhance the biosynthesis of bioactive compounds in
buckwheat which necessitates a holistic and integra-
tive methodology. This strategy aims to elucidate and
optimize the synthesis of critical compounds, including
flavonoids, phenolic acids, and various antioxidants
(Figure 6).
At the genomic level, key genes implicated in the
biosynthesis of secondary metabolites, such as flavo-
noids and phenolic acids, are identified through com-
prehensive sequencing of the buckwheat genome.
Transcriptomic analyses elucidate the expression
dynamics of these genes throughout the growth and
developmental phases of buckwheat, with a particular
emphasis on their response to environmental stressors,
including variations in light and temperature [32, 104].
For instance, to pinpoint the genes responsible for rutin
hydrolysis, a collection of 200 genetic varieties from
Tartary buckwheat was extensively analyzed using rese-
quencing technology at a depth of 30-fold coverage.
Integrating the data on intermediate rutin metabolism
substances with the genomic resequencing results, a
comprehensive metabolite-genome association study
was carried out, and successfully highlighted the FtGH1
glycosyl hydrolase gene as capable of converting rutin
Figure 5. Regulatory genes involve in root, stem, ower, and seed development in buckwheat.

CRITICAL REVIEWS IN BIOTECHNOLOGY 17
into quercetin [180]. Additionally, combining: phyloge-
netic, transcriptomic, metabolomic, and comparative
genomic analyses, identified a high expression of cate-
chin biosynthesis-related genes in flower and seed in
common buckwheat and high expression of rutin
biosynthesis-related genes in seed in Tartary buckwheat
as being important for the differences in flavonoid type,
and identified a candidate key rutin-degrading enzyme
gene (Ft8.2377) [98]. Proteomic investigations further
delineate the expression and functionality of enzymes
and regulatory proteins integral to these metabolic
pathways. Metabolomic analysis, in turn, provides an
intricate profile of the diverse metabolites present in
buckwheat, with a specific focus on the types and con-
centrations of targeted secondary metabolites. The
amalgamation and scrutiny of these data streams facil-
itate a thorough understanding of the molecular mech-
anisms governing the synthesis and accumulation of
Figure 6. Application of multi-omics (genome, epigenome, transcriptome, proteome and metabolome) in traits improvement and
bioactive compounds biosynthesis.

18 Y. SONG ETAL.
secondary metabolites in buckwheat. This knowledge
underpins strategies for genetic enhancement or culti-
vation management aimed at augmenting the yield of
these health-promoting compounds.
Based on multi-omics studies, researchers are able
to pinpoint crucial genetic and metabolic determinants
influencing the synthesis of secondary metabolites and
their interplay. For instance, transcriptomic and pro-
teomic data can reveal which biosynthetic pathways
are activated or repressed under particular environ-
mental conditions [123]. Furthermore, metabolic engi-
neering techniques may be utilized to manipulate
gene expression in buckwheat, thereby boosting the
efficiency of certain secondary metabolic pathways
and increasing the production of specific compounds.
These multi-omics revelations offer precise molecular
targets and methodologies for enhancing the content
of health-beneficial compounds in buckwheat. Such
advancements contribute significantly to the cultiva-
tion of more nutritious and medicinally valuable buck-
wheat varieties.
5.2. Multi-omics used in agronomy traits
improvement
With the advent of multi-omics technologies and com-
putational methodologies, integrative omics approaches
have been increasingly adopted in crop science
research, which used to improve the agronomy traits
of buckwheat (Figure 6). The combination of genomic
information with advanced phenotyping methodolo-
gies offers valuable insights into complex traits, which
can be pivotal for the enhancement of crop species.
The integration GWAS and metabolite profiling meth-
odologies have emerged as a robust technique for
unraveling the biochemical and genetic mechanisms
underlying various traits in: buckwheat, rice, maize,
and tomato [99, 110, 182–184]. Based on GWAS analy-
sis, Fh06G015130 was identified, which is a crucial gene
underlying flavor-associated flavonoids in common
buckwheat [99]. In addition, the integration of omics
approaches has facilitated the development of abiotic
stress-tolerant phenotypes in crops. Also, functional
genomics and mutagenomics have been employed to
identify many mutants exhibiting specific variations in
growth, development, and stress tolerance across a
variety of crops, which has been widely used in rice,
wheat, and maize. Also the application of QTL map-
ping combined with the analysis of agronomic traits
has been instrumental in identifying numerous QTLs in
staples.
Genomic analysis provides comprehensive insights
into the buckwheat genome, aiding researchers in
identifying genes associated with key agronomic traits,
such as those related to: yield, seed size, and pest and
disease resistance. Transcriptomic analysis reveals the
expression patterns of these genes across different
growth stages and environmental conditions, thereby
understanding how genes regulate agronomic traits at
the physiological level. Proteomics delves deeper into
the functionality of these gene products, particularly
proteins playing crucial roles in signaling pathways and
metabolic processes. Metabolomics offers a comprehen-
sive view of the metabolites in buckwheat, especially
those directly related to agronomic traits, such as
metabolites affecting seed nutritional components. By
integrating and analyzing these multi-omics data, com-
plex biological networks underlying the agronomic traits
of buckwheat could be mapped. The integrative
approach not only elucidates the molecular mechanisms
behind various agronomic traits but also guides precise
genetic improvement strategies, such as developing
new buckwheat varieties with higher yields, enhanced
nutritional values, and improved resistance to pests and
diseases through molecular breeding techniques. Thus,
multi-omics technologies provide an efficient and accu-
rate pathway for the improvement of agronomic traits
in buckwheat, aiding in meeting the diverse needs of
agricultural production and consumer demand.
Above all, in buckwheat research, particularly in
nutraceutical enhancement and traits improvement,
several limitations persist. These include incomplete
elucidation of metabolic pathways for key bioactive
compounds, unverified functionality of many identified
genes, a lack of in-depth understanding of regulatory
networks, and inefficiencies in current genetic transfor-
mation systems. These gaps highlight the need for
comprehensive research efforts to fully understand and
exploit buckwheat’s potential.
6. Conclusion and perspective
Due to its high nutritive value, buckwheat has
garnered considerable interest among researchers and
breeders. An array of biological processes in buckwheat
is currently subject to extensive research. Within this
context, omics methodologies are recognized as potent
tools, capable of yielding an extensive dataset encom-
passing: gene sequences, gene expression patterns,
protein expression profiles, and the spectrum of metab-
olites present in buckwheat. Consequently, compre-
hensive investigations into the molecular mechanisms
driving various biological processes have now become
feasible and reliable. This includes studies focused on
the accumulation of bioactive components and agron-
omy traits improvement. In recent years, significant

CRITICAL REVIEWS IN BIOTECHNOLOGY 19
advancements have been achieved in the field of
buckwheat research, such as the synergistic integration
of GWAS with metabolomics, transcriptomics, and pro-
teomics has emerged as a promising approach for elu-
cidating biochemical processes and enhancing
understanding of abiotic stress tolerance in buckwheat.
Research has demonstrated the efficacy of combining
multi-omics approaches in: identifying potential candi-
date genes, elucidating biosynthesis pathways, nutra-
ceutical improvement, and traits improvement.
Through significant advancements in enhancing our
comprehension of buckwheat, there still exist notable
limitations in the field. Pangenomics refers to the com-
prehensive genomic composition of a species, encom-
passing a set of core and dispensable genes. It
contributes to crop improvement, including: under-
standing genetic diversity, nutritional improvement, cli-
mate adaptation, and speeding up crop improvement.
Also pangenomics can be integrated with other omics
approaches for a more holistic understanding of how
genetic variation influences phenotypic traits. However,
the application of pangenomics has not yet been
employed in buckwheat research. Genetic engineering
has been widely used to elucidate gene function and
for crop improvement. Although Agrobacterium-mediated
transformation has been successful in hairy roots of
buckwheat, the efficiency is notably low. Additionally,
many functional genes related biosynthesis of flavonols,
stress response, and plant development, have been
identified based on multi-omics, but the functionalities
of the majority of these genes remain unverified through
empirical functional investigation, and the regulatory
mechanisms governing them are yet to be elucidated.
To facilitate the advancement of research pertaining
to buckwheat, future endeavors should concentrate on
the following areas of work: (1) Although, some genes
have been identified based on the multi-omics research
in buckwheat, pangenomics should be used to identify
elite genes in buckwheat landraces and wild species
for traits improvement. In particular, the combination
research between pangenomics and other omics.
(2) The biosynthetic pathways of bioactive compo-
nents, along with the regulatory networks and mecha-
nisms of specific agronomic traits, necessitate more
in-depth investigation. (3) Developing a more efficient
genetic transformation system, in conjunction with
multi-omics techniques and genetic engineering tech-
nologies, is imperative for the execution of breeding
designs aimed at enhancing desirable traits in buck-
wheat. (4) Employing multi-omics technologies to
investigate the nutrient utilization mechanisms in
buckwheat, with the goal of cultivating plants that
exhibit high efficiency in nutrient utilization.
Author contributions
S. Y. conceived and supervised the study, and wrote the
manuscript. H. W., L.C., W.Y., A.X., and L.Y. revised the manu-
script. All the authors read and approved the nal manuscript.
Disclosure statement
No potential competing interest was reported by the
author(s).
Funding
This work was nancially supported by the National Nature
Science Foundation of China (31761143001 and 31870316).
the Ministry of Ecology and Environment (2019HB2096001006),
the Key Laboratory of Ethnomedicine (Minzu University of
China) of Ministry of Education of China (KLEM-ZZ201904
and KLEM-ZZ201906), and the Minzu University of China
(Collaborative Innovation Center for Ethnic Minority Development
and yldxxk201819).
Data availability
The data that support the ndings of this study are availabil-
ity from the corresponding author upon reasonable request.
References
[1] FAO I, UNICEF, WEP, WHO. The State of Food Security
and Nutrition in the World 2021: transforming food
systems for food security, improved nutrition and af-
fordable healthy diets for all. Rome, Italy: FAO; 2021.
[2] FAO I, UNICEF, WEP, WHO. The State of Food Security
and Nutrition in the World 2022: help eliminate hunger
and malnutrition. Rome, Italy: FAO; 2022.
[3] FAO I, UNICEF, WEP, WHO. The State of Food Security
and Nutrition in the World 2021: repurposing food and
agricultural policies to make healthy diets more aord-
able. Rome, Italy: FAO; 2022.
[4] Boukid F, Folloni S, Sforza S, et al. Current trends in
ancient grains-based foodstus: insights into nutrition-
al aspects and technological applications. Compr Rev
Food Sci Food Saf. 2018;17:123–136. doi: 10.1111/1541-
4337.12315.
[5] Jha R, Zhang K, He Y, et al. Global nutritional challeng-
es and opportunities: buckwheat, a potential bridge
between nutrient deciency and food security. Trend
Food Sci Technol. 2024;145:104365. doi: 10.1016/j.
tifs.2024.104365.
[6] Joshi DC, Chaudhari GV, Sood S, et al. Revisiting the ver-
satile buckwheat: reinvigorating genetic gains through
integrated breeding and genomics approach. Planta.
2019;250:783–801. doi: 10.1007/s00425-018-03080-4.
[7] Ohsako T, Ohnishi O. Intra- and interspecic phylogeny
of wild Fagopyrum (Polygonaceae) species based on
nucleotide sequences of noncoding regions in chloro-
plast DNA. Am J Bot. 2000;87:573–582. doi: 10.2307/
2656601.

20 Y. SONG ETAL.
[8] Kreft M. Buckwheat phenolic metabolites in health and
disease. Nutr Res Rev. 2016;29:30–39. doi: 10.1017/
S0954422415000190.
[9] Zheng CJ, Hu CL, Ma XQ, et al. Cytotoxic phenylpro-
panoid glycosides from Fagopyrum tataricum (L.)
Gaertn. Food Chem. 2012;132:433–438. doi: 10.1016/j.
foodchem.2011.11.017.
[10] Jiang P, Burczynski F, Campbell C, et al. Rutin and a-
vonoid contents in three buckwheat species Fagopyrum
esculentum, F-tataricum, and F-homotropicum and their
protective eects against lipid peroxidation. Food Res
Int. 2007;40:356–364. doi: 10.1016/j.foodres.2006.10.009.
[11] Choi I, Seog H, Park Y, etal. Suppressive eects of ger-
minated buckwheat on development of fatty liver in
mice fed with high-fat diet. Phytomedicine.
2007;14:563–567. doi: 10.1016/j.phymed.2007.05.002.
[12] Ishii S, Katsumura T, Shiozuka C, et al. Anti-inammatory
eect of buckwheat sprouts in lipopolysaccharide-activated
human colon cancer cells and mice. Biosci Biotechnol
Biochem. 2008;72:3148–3157. doi: 10.1271/bbb.80324.
[13] Kim CD, Lee WK, No KO, et al. Anti-allergic action of
buckwheat (Fagopyrum esculentum Moench) grain ex-
tract. Int Immunopharmacol. 2003;3:129–136. doi:
10.1016/s1567-5769(02)00261-8.
[14] Neeraj Vasudeva N, Sharma S. Chemical composition of
Fagopyrum esceulentum moench seed through GC-MS.
Int J Pharm Sci Res. 2019;10:2392–2396.
[15] Jin HM, Wei P. Anti-fatigue properties of tartary buck-
wheat extracts in mice. Int J Mol Sci. 2011;12:4770–
4780. doi: 10.3390/ijms12084770.
[16] Singh S, Dubey V, Meena A, et al. Rutin restricts hydro-
gen peroxide-induced alterations by up-regulating the
redox-system: an in vitro, in vivo and in silico study.
Eur J Pharmacol. 2018;835:115–125. doi: 10.1016/j.ej-
phar.2018.07.055.
[17] Song Y, Dong Y, Wang J, et al. Tartary buckwheat
(Fagopyrum tataricum Gaertn.) landraces cultivated by
Yi people in Liangshan, China. Genet Resour Crop Evol.
2020;67:745–761. doi: 10.1007/s10722-019-00852-z.
[18] He Y, Zhang K, Shi Y, et al. Genomic insight into the
origin, domestication, dispersal, diversication and hu-
man selection of Tartary buckwheat. Genome Biol.
2024;25:61. doi: 10.1186/s13059-024-03203-z.
[19] Guo X, Zhu KX, Zhang H, et al. Purication and charac-
terization of the antitumor protein from chinese Tartary
buckwheat (Fagopyrum tataricum Gaertn.) water-soluble
extracts. J Agric Food Chem. 2007;55:6958–6961. doi:
10.1021/jf071032+.
[20] Liu CL, Chen YS, Yang JH, et al. Antioxidant activity of
Tartary (Fagopyrum tataricum (L.) Gaertn.) and common
(Fagopyrum esculentum moench) buckwheat sprouts. J
Agric Food Chem. 2008;56:173–178. doi: 10.1021/
jf072347s.
[21] Tomotake H, Yamamoto N, Yanaka N, et al. High pro-
tein buckwheat our suppresses hypercholesterolemia
in rats and gallstone formation in mice by hypercholes-
terolemic diet and body fat in rats because of its low
protein digestibility. Nutrition. 2006;22:166–173. doi:
10.1016/j.nut.2005.01.012.
[22] Chan PK. Inhibition of tumor growth in vitro by the ex-
tract of Fagopyrum cymosum (fago-c). Life Sci.
2003;72:1851–1858. doi: 10.1016/s0024-3205(03)00013-4.
[23] Muthamilarasan M, Singh NK, Prasad M. Multi-omics
approaches for strategic improvement of stress toler-
ance in underutilized crop species: a climate change
perspective. Adv Genet. Vol 103 2019;103:1–38. doi:
10.1016/bs.adgen.2019.01.001.
[24] Yang YD, Saand MA, Huang LY, et al. Applications of
multi-omics technologies for crop improvement. Front
Plant Sci. 2021;12:563953. doi: 10.3389/fpls.2021.563953.
[25] Zhou M, Zheng SG. Multi-omics uncover the mecha-
nism of wheat under heavy metal stress. Int J Mol Sci.
2022;23:15968. doi: 10.3390/ijms232415968.
[26] Li X, Gao J, Song J, et al. Multi-omics analyses of 398
foxtail millet accessions reveal genomic regions associ-
ated with domestication, metabolite traits, and
anti-inammatory eects. Mol Plant. 2022;15:1367–
1383. doi: 10.1016/j.molp.2022.07.003.
[27] Cao P, Zhao Y, Wu FJ, et al. Multi-omics techniques for
soybean molecular breeding. Int J Mol Sci. 2022;23:4994.
doi: 10.3390/ijms23094994.
[28] Muthuramalingam P, Jeyasri R, Rakkammal K, et al.
Multi-omics and integrative approach towards under-
standing salinity tolerance in rice: a review. Biology
(Basel). 2022;11:1022. doi: 10.3390/biology11071022.
[29] Luo F, Yu ZJ, Zhou Q, et al. Multi-omics-based discov-
ery of plant signaling molecules. Metabolites.
2022;12:76. doi: 10.3390/metabo12010076.
[30] Liu Y, Wang B, Shu S, et al. Analysis of the Coptis chin-
ensis genome reveals the diversication of
protoberberine-type alkaloids. Nat Commun.
2021;12:3276. doi: 10.1038/s41467-021-23611-0.
[31] Yasui Y, Hirakawa H, Ueno M, et al. Assembly of the
draft genome of buckwheat and its applications in
identifying agronomically useful genes. DNA Res.
2016;23:215–224. doi: 10.1093/dnares/dsw012.
[32] Zhang L, Li X, Ma B, et al. The Tartary buckwheat ge-
nome provides insights into rutin biosynthesis and abi-
otic stress tolerance. Mol Plant. 2017;10:1224–1237.
doi: 10.1016/j.molp.2017.08.013.
[33] Jing R, Li HQ, Hu CL, etal. Phytochemical and pharma-
cological proles of three fagopyrum buckwheats. Int J
Mol Sci. 2016;17:589. doi: 10.3390/ijms17040589.
[34] Zhang ZL, Zhou ML, Tang Y, et al. Bioactive compounds
in functional buckwheat food. Food Res Int.
2012;49:389–395. doi: 10.1016/j.foodres.2012.07.035.
[35] Ahmed A, Khalid N, Ahmad A, et al. Phytochemicals and
biofunctional properties of buckwheat: a review. J Agric
Sci. 2014;152:349–369. doi: 10.1017/S0021859613000166.
[36] Rahmani S, Naraki K, Roohbakhsh A, et al. Nutrition: the
protective eects of rutin on the liver, kidneys, and heart
by counteracting organ toxicity caused by synthetic and
natural compounds. Food Sci & Nutrition. 2023;11:39–56.
[37] Zhu F. Chemical composition and health eects of
Tartary buckwheat. Food Chem. 2016;203:231–245. doi:
10.1016/j.foodchem.2016.02.050.
[38] Joshi DC, Zhang KX, Wang CL, etal. Strategic enhancement
of genetic gain for nutraceutical development in buck-
wheat: a genomics-driven perspective. Biotechnol Adv.
2020;39:107479. doi: 10.1016/j.biotechadv.2019.107479.
[39] Borovaya SA, Klykov AG. Some aspects of avonoid
biosynthesis and accumulation in buckwheat plants.
Plant Biotechnol Rep. 2020;14:213–225. doi: 10.1007/
s11816-020-00614-9.

CRITICAL REVIEWS IN BIOTECHNOLOGY 21
[40] Lee JM, Lee S, Lee KH, et al. Identication of triter-
penoids and avonoids from the seeds of tartary buck-
wheat. Nat Prod Sci. 2013;19:137–144.
[41] Huda MN, Lu S, Jahan T, et al. Treasure from garden:
bioactive compounds of buckwheat. Food Chem.
2021;335:127653. doi: 10.1016/j.foodchem.2020.127653.
[42] Kim S-J, Zaidul ISM, Suzuki T, et al. Comparison of phe-
nolic compositions between common and Tartary
buckwheat (Fagopyrum) sprouts. Food Chem.
2008;110:814–820. doi: 10.1016/j.foodchem.2008.02.050.
[43] Zou L, Wu D, Ren G, et al. Bioactive compounds, health
benets, and industrial applications of Tartary buck-
wheat (Fagopyrum tataricum). Crit Rev Food Sci Nutr.
2023;63:657–673. doi: 10.1080/10408398.2021.1952161.
[44] Shao M, Yang Y, Gao H, etal. Studies on the chemical
constituents of Fagopyrum dibotrys (D.Don) Hara. J.
Shenyang Pharmaceut Univ. 2005;22:100–102. 160.
[45] Zhang J, Kuang Y, Liu L, et al. Chemical constituents
from root tubers of Fagopyrum dibotrys. Chin. Trad.
Herbal Drug. 2016;47:722–725.
[46] Watanabe M. Catechins as antioxidants from buck-
wheat (Fagopyrum esculentum Moench) groats. J Agric
Food Chem. 1998;46:839–845. doi: 10.1021/jf9707546.
[47] Watanabe M, Ohshita Y, Tsushida T. Antioxidant com-
pounds from buckwheat (Fagopyrum esculentum
Moench) hulls. J Agric Food Chem. 1997;45:1039–1044.
doi: 10.1021/jf9605557.
[48] Li J, Yang P, Yang Q, et al. Analysis of avonoid metabo-
lites in buckwheat leaves using UPLC-ESI-MS/MS.
Molecules. 2019;24:1310. doi: 10.3390/molecules24071310.
[49] Ren Q, Wu C, Ren Y, et al. Characterization and identica-
tion of the chemical constituents from Tartary buckwheat
(Fagopyrum tataricum Gaertn) by high performance liquid
chromatography/photodiode array detector/linear ion trap
FTICR hybrid mass spectrometry. Food Chem.
2013;136:1377–1389. doi: 10.1016/j.foodchem.2012.09.052.
[50] Liu Y, Cai C, Yao Y, et al. Alteration of phenolic proles
and antioxidant capacities of common buckwheat and
Tartary buckwheat produced in China upon thermal
processing. J Sci Food Agric. 2019;99:5565–5576. doi:
10.1002/jsfa.9825.
[51] Chen Y, Qin L, Wen A, et al. Three-solvent extracting
method comprehensively evaluates phenolics prole
and antioxidant activities of Tartary buckwheat. J Food
Process Preserv. 2021;45:15020. doi: 10.1111/jfpp.15020.
[52] Obendorf RL, Steadman KJ, Fuller DJ, et al. Molecular
structure of fagopyritol Al (O-α-D-galactopyranosyl-
(1→3)-D-chiro-inositol) by NMR. Carbohydr Res.
2000;328:623–627. doi: 10.1016/s0008-6215(00)00133-6.
[53] Horbowicz M, Obendorf RL. Fagopyritol accumulation
and germination of buckwheat seeds matured at 15,
22, and 30 °C. Crop Sci. 2005;45:1264–1270. doi:
10.2135/cropsci2004.0431.
[54] Wu W, Wang L, Qiu J, et al. The analysis of fagopyritols
from Tartary buckwheat and their anti-diabetic eects
in KK-Ay type 2 diabetic mice and HepG2 cells. J Funct
Foods. 2018;50:137–146. doi: 10.1016/j.j.2018.09.032.
[55] Mäki-Opas I, Hämäläinen M, Moilanen LJ, et al.
Pyrazine-fused triterpenoids block the TRPA1 Ion chan-
nel in Vitro and inhibit TRPA1-mediated acute inam-
mation in Vivo. ACS Chem Neurosci. 2019;10:2848–
2857. doi: 10.1021/acschemneuro.9b00083.
[56] Lv L, Xia Y, Zou D, et al. Fagopyrum tataricum (L.)
Gaertn.: a review on its traditional uses, phytochemical
and pharmacology. FSTR. 2017;23:1–7. doi: 10.3136/
fstr.23.1.
[57] Fan Z, Song Q, Qiang G, et al. GC/MS analysis of fatty
acid in fagopyrum esculentum seed oil. Food Sci.
2004;25:267–269.
[58] Agarwal DS, Sakhuja R, Beteck RM, et al. Steroid-triazole
conjugates: A brief overview of synthesis and their ap-
plication as anticancer agents. Steroids. 2023;197:109258.
doi: 10.1016/j.steroids.2023.109258.
[59] Janeš D, Kantar D, Kreft S, et al. Identication of buck-
wheat (Fagopyrum esculentum Moench) aroma com-
pounds with GC-MS. Food Chem. 2009;112:120–124.
doi: 10.1016/j.foodchem.2008.05.048.
[60] Lv QY, Wang JR, Sun PY, et al. Evaluation of the
Composition and accumulation pattern of fatty acids in
Tartary buckwheat seed at the germplasm level.
Agronomy-Basel. 2022;12:2447. doi: 10.3390/agrono-
my12102447.
[61] Cho JY, Moon JH, Kim HK, et al. Isolation and structur-
al elucidation of antimicrobial compounds from buck-
wheat hull. J Microbiol Biotechnol. 2006;16:538–542.
[62] Caligiani A, Palla G, Bonzanini F, et al. A validated
GC-MS method for the detection of tropane alkaloids
in buckwheat (Fagopyron esculentum L.) fruits, ours
and commercial foods. Food Chem. 2011;127:204–209.
doi: 10.1016/j.foodchem.2010.11.141.
[63] Sedem Dzah C, Duan Y, Zhang H, et al. Ultrasound-,
subcritical water- and ultrasound assisted subcritical
water-derived Tartary buckwheat polyphenols show su-
perior antioxidant activity and cytotoxicity in human
liver carcinoma cells. Food Res Int. 2020;137:109598.
doi: 10.1016/j.foodres.2020.109598.
[64] Zhou X-L, Chen Z-D, Zhou Y-M, et al. The eect of tartary
buckwheat avonoids in inhibiting the proliferation of
MGC80-3 cells during seed germination. Molecules.
2019;24:3092. doi: 10.3390/molecules24173092.
[65] Li W, Zhang X, He X, et al. Eects of steam explosion
pretreatment on the composition and biological activ-
ities of Tartary buckwheat bran phenolics. Food Funct.
2020;11:4648–4658. doi: 10.1039/d0fo00493f.
[66] Hwang D, Kang M-J, Kang C-W, etal. Kaempferol-3-O-β-
rutinoside suppresses the inammatory responses in
lipopolysaccharide-stimulated RAW264.7 cells via the
NF-κB and MAPK pathways. Int J Mol Med.
2019;44:2321–2328. doi: 10.3892/ijmm.2019.4381.
[67] Zhou Y, Zhao S, Jiang Y, et al. Regulatory function of
buckwheat-resistant starch supplementation on lipid pro-
le and gut microbiota in mice fed with a high-fat diet. J
Food Sci. 2019;84:2674–2681. doi: 10.1111/1750-3841.14747.
[68] Yang Q, Luo C, Zhang X, et al. Tartary buckwheat ex-
tract alleviates alcohol-induced acute and chronic liver
injuries through the inhibition of oxidative stress and
mitochondrial cell death pathway. Am J Transl Res.
2020;12:70–89.
[69] Lee S, Lee J, Lee H, et al. Relative protective activities
of quercetin, quercetin-3-glucoside, and rutin in
alcohol-induced liver injury. J Food Biochem.
2019;43:e13002. doi: 10.1111/jfbc.13002.
[70] Luo X, Fei Y, Xu Q, et al. Isolation and identication of anti-
oxidant peptides from Tartary buckwheat albumin

22 Y. SONG ETAL.
(Fagopyrum tataricum Gaertn.) and their antioxidant activi-
ties. J Food Sci. 2020;85:611–617. doi: 10.1111/1750-
3841.15004.
[71] Zhou Y, Jiang Y, Shi R, et al. Structural and antioxidant
analysis of Tartary buckwheat (Fagopyrum tartaricum
Gaertn.) 13S globulin. J Sci Food Agric. 2020;100:1220–
1229. doi: 10.1002/jsfa.10133.
[72] Hou Z, Hu Y, Yang X, et al. Antihypertensive eects of
Tartary buckwheat avonoids by improvement of vascu-
lar insulin sensitivity in spontaneously hypertensive rats.
Food Funct. 2017;8:4217–4228. doi: 10.1039/c7fo00975e.
[73] Kim S-Y, Lee M-S, Chang E, etal. Tartary buckwheat ex-
tract attenuated the obesity-induced inammation and
increased muscle PGC-1a/SIRT1 expression in high fat
diet-induced obese rats. Nutrients. 2019;11:654. doi:
10.3390/nu11030654.
[74] Zhu F. Buckwheat proteins and peptides: biological func-
tions and food applications. Trend Food Sci Technol.
2021;110:155–167. doi: 10.1016/j.tifs.2021.01.081.
[75] Wu W, Li Z, Qin F, et al. Anti-diabetic eects of the sol-
uble dietary ber from Tartary buckwheat bran in dia-
betic mice and their potential mechanisms. Food Nutr
Res. 2021;65:4998. doi: 10.29219/fnr.v65.4998.
[76] Li X, Park NI, Kim YB, et al. Accumulation of avonoids
and expression of avonoid biosynthetic genes in
Tartary and rice-Tartary buckwheat. Process Biochem.
2012;47:2306–2310. doi: 10.1016/j.procbio.2012.09.009.
[77] Cui Y, Li W, Chang W, et al. Dierential expression anal-
ysis of genes associated with isoavone synthesis in
soybean (Glycine max Merr.). J. Plant Genetic Res.
2016;17:719–725.
[78] Li J, Hossain MS, Ma H, et al. Comparative metabolom-
ics reveals dierences in avonoid metabolites among
dierent coloured buckwheat owers. J Food Compos
Anal. 2020;85:103335. doi: 10.1016/j.jfca.2019.103335.
[79] Li H, Lv Q, Ma C, et al. Metabolite proling and tran-
scriptome analyses provide insights into the avonoid
biosynthesis in the developing seed of tartary buck-
wheat (Fagopyrum tataricum). J Agric Food Chem.
2019;67:11262–11276. doi: 10.1021/acs.jafc.9b03135.
[80] Matsui K, Oshima Y, Mitsuda N, et al. Buckwheat R2R3
MYB transcription factor FeMYBF1 regulates avonol
biosynthesis. Plant Sci. 2018;274:466–475. doi: 10.1016/j.
plantsci.2018.06.025.
[81] Matsui K, Hisano T, Yasui Y, et al. Isolation and charac-
terization of genes encoding leucoanthocyanidin re-
ductase (FeLAR) and anthocyanidin reductase (FeANR)
in buckwheat (Fagopyrum esculentum). J Plant Physiol.
2016;205:41–47. doi: 10.1016/j.jplph.2016.08.010.
[82] Huang J, Wang L, Tang B, et al. Integrated transcrip-
tomics and widely targeted metabolomics analyses
provide insights into avonoid biosynthesis in the rhi-
zomes of golden buckwheat (Fagopyrum cymosum).
Front Plant Sci. 2022;13:803472. doi: 10.3389/
fpls.2022.803472.
[83] Li X, Thwe AA, Park CH, et al. Ethephon-induced phenyl-
propanoid accumulation and related gene expression in
Tartary buckwheat (Fagopyrum tataricum (L.) Gaertn.)hairy
root. Biotechnol Biotechnol Equip. 2017;31:304–311. doi:
10.1080/13102818.2017.1282835.
[84] Park CH, Park YE, Yeo HJ, et al. Eect of light and dark
on the phenolic compound accumulation in tartary
buckwheat hairy roots overexpressing ZmLC. Int J Mol
Sci. 2021;22:4702. doi: 10.3390/ijms22094702.
[85] Thwe AA, Kim JK, Li X, et al. Metabolomic analysis and
phenylpropanoid biosynthesis in hairy root culture of
tartary buckwheat cultivars. PLoS One. 2013;8:e65349.
doi: 10.1371/journal.pone.0065349.
[86] Claude S-J, Park S, Park S-J. Transcriptome-wide identi-
cation and quantication of caeoylquinic acid bio-
synthesis pathway and prediction of its putative BAHDs
gene complex in A. spathulifolius. Int J Mol Sci.
2021;22:6333. doi: 10.3390/ijms22126333.
[87] Sytar O, Brestic M, Rai M. Possible ways of fagopyrin bio-
synthesis and production in buckwheat plants. Fitoterapia.
2013;84:72–79. doi: 10.1016/j.tote.2012.10.008.
[88] Li X, Sathasivam R, Park NI, et al. Enhancement of
phenylpropanoid accumulation in Tartary buckwheat
hairy roots by overexpression of MYB transcription fac-
tors. Ind Crops Prod. 2020;156:112887. doi: 10.1016/j.
indcrop.2020.112887.
[89] Bai Y-C, Li C-L, Zhang J-W, et al. Characterization of
two Tartary buckwheat R2R3-MYB transcription factors
and their regulation of proanthocyanidin biosynthesis.
Physiol Plant. 2014;152:431–440. doi: 10.1111/ppl.12199.
[90] Zhao H, Wu X, Bai Y, et al. Gene expression analysis of
key enzymes and MYB transcription factors in avo-
noid biosynthesis pathway during germination of
Fagopyrum tataricum. J. Agric. Biotechnol. 2012;20:121–
128.
[91] Zhao H, Yao P, Zhao J, et al. A Novel R2R3-MYB tran-
scription factor FtMYB22 negatively regulates salt and
drought stress through ABA-dependent pathway. Int J
Mol Sci. 2022;23:14549. doi: 10.3390/ijms232314549.
[92] Zhou M, Sun Z, Ding M, et al. FtSAD2 and FtJAZ1 reg-
ulate activity of the FtMYB11 transcription repressor of
the phenylpropanoid pathway in Fagopyrum tataricum.
New Phytol. 2017;216:814–828. doi: 10.1111/nph.14692.
[93] Zhang K, Logacheva MD, Meng Y, et al.
Jasmonate-responsive MYB factors spatially repress ru-
tin biosynthesis in Fagopyrum tataricum. J Exp Bot.
2018;69:1955–1966. doi: 10.1093/jxb/ery032.
[94] Huang Y, Wu Q, Wang S, et al. FtMYB8 from Tartary
buckwheat inhibits both anthocyanin/Proanthocyanidin
accumulation and marginal Trichome initiation. BMC
Plant Biol. 2019;19:263. doi: 10.1186/s12870-019-1876-x.
[95] Sun Z, Hou S, Linghu B, et al. Correlation analysis on
rutin accumulation and gene expression of rutin syn-
thetic enzymes and MYBs in the whole developmental
stage of Fagopyrum tataricum. Sci. Agric. Sin.
2017;50:3473–3481.
[96] Fawcett JA, Takeshima R, Kikuchi S, et al. Genome se-
quencing reveals the genetic architecture of heterosty-
ly and domestication history of common buckwheat.
Nat Plants. 2023;9:1236–1251. +. doi: 10.1038/
s41477-023-01474-1.
[97] Penin AA, Kasianov AS, Klepikova AV, et al.
High-resolution transcriptome atlas and improved ge-
nome assembly of common buckwheat, Fagopyrum
esculentum. Front Plant Sci. 2021;12:612382. doi:
10.3389/fpls.2021.612382.
[98] He Q, Ma D, Li W, et al. High-quality Fagopyrum esculen-
tum genome provides insights into the avonoid accumu-
lation among dierent tissues and self-incompatibility.

CRITICAL REVIEWS IN BIOTECHNOLOGY 23
J Integr Plant Biol. 2023;65:1423–1441. doi: 10.1111/
jipb.13459.
[99] Zhang K, He Y, Lu X, etal. Comparative and population
genomics of buckwheat species reveal key determi-
nants of avor and fertility. Mol Plant. 2023;16:1427–
1444. doi: 10.1016/j.molp.2023.08.013.
[100] Zhang K, He M, Fan Y, et al. Resequencing of global
Tartary buckwheat accessions reveals multiple domes-
tication events and key loci associated with agronomic
traits. Genome Biol. 2021;22:23. doi: 10.1186/
s13059-020-02217-7.
[101] Liu M, Sun W, Ma Z, et al. Genome-wide investigation
of the AP2/ERF gene family in Tartary buckwheat
(Fagopyum Tataricum). BMC Plant Biol. 2019;19:84. doi:
10.1186/s12870-019-1681-6.
[102] Shi T-X, Li R-Y, Zheng R, et al. Mapping QTLs for
1000-grain weight and genes controlling hull type us-
ing SNP marker in Tartary buckwheat (Fagopyrum tatar-
icum). BMC Genomics. 2021;22:142. doi: 10.1186/
s12864-021-07449-w.
[103] Wang A, Liu Y, Li Q, et al. FlbZIP12 gene enhances
drought tolerance via modulating avonoid biosynthe-
sis in Fagopyrum leptopodum. Front Plant Sci.
2023;14:1279468. doi: 10.3389/fpls.2023.1279468.
[104] Song Y, Feng J, Liu D, et al. Dierent phenylalanine
pathway responses to cold stress based on metabolo-
mics and transcriptomics in Tartary buckwheat landra-
ces. J Agric Food Chem. 2022;70:687–698. doi: 10.1021/
acs.jafc.1c06915.
[105] Hou Y, Lu Q, Su J, et al. Genome-wide analysis of the HDAC
gene family and its functional characterization at low tem-
peratures in Tartary buckwheat (Fagopyrum tataricum). Int J
Mol Sci. 2022;23:7622. doi: 10.3390/ijms23147622.
[106] Zhao Z, Meng G, Zamin I, et al. Genome-wide identi-
cation and functional analysis of the TIFY family genes
in response to abiotic stresses and hormone treat-
ments in Tartary buckwheat (Fagopyrum tataricum). Int
J Mol Sci. 2023;24:10916. doi: 10.3390/ijms241310916.
[107] Wang X, Zhou T, Bai G, etal. Complete chloroplast genome
sequence of Fagopyrum dibotrys: genome features, com-
parative analysis and phylogenetic relationships. Sci Rep.
2018;8:12379. doi: 10.1038/s41598-018-30398-6.
[108] He M, He Y, Zhang K, et al. Comparison of buckwheat
genomes reveals the genetic basis of metabolomic di-
vergence and ecotype dierentiation. New Phytol.
2022;235:1927–1943. doi: 10.1111/nph.18306.
[109] Cheng C, Zhang K, Tang Y, et al. Investigation on wild
Fagopyrum cymosum resources in Yunnan and analysis
of genetic diversity. J. Plant Genetic Res. 2019;20:1438–
1446.
[110] Zargar SM, Manzoor M, Bhat B, et al. Metabolic-GWAS
provides insights into genetic architecture of seed me-
tabolome in buckwheat. BMC Plant Biol. 2023;23:373.
doi: 10.1186/s12870-023-04381-x.
[111] Naik S, Sudan J, Urwat U, et al. Genome-wide SNP discov-
ery and genotyping delineates potential QTLs underlying
major yield-attributing traits in buckwheat. Plant Genome.
2024;17:e20427. doi: 10.1002/tpg2.20427.
[112] Yang W, Duan H, Yu K, et al. Integrative dissection of
lignin composition in tartary buckwheat seed hulls for
enhanced dehulling eciency. Adv Sci. 2024;11:2400916.
doi: 10.1002/advs.202400916.
[113] Ye X, Li Q, Liu C, et al. Transcriptomic, cytological, and
physiological analyses reveal the potential regulatory
mechanism in Tartary buckwheat under cadmium
stress. Front Plant Sci. 2022;13:1004802. doi: 10.3389/
fpls.2022.1004802.
[114] Huang J, Chen Q, Rong Y, et al. Transcriptome analysis
revealed gene regulatory network involved in
PEG-induced drought stress in Tartary buckwheat
(Fagopyrum Tararicum). PeerJ. 2021;9:e11136. doi:
10.7717/peerj.11136.
[115] Wu Q, Zhao G, Bai X, et al. Characterization of the tran-
scriptional proles in common buckwheat (Fagopyrum
esculentum) under PEG-mediated drought stress.
Electron J Biotechnol. 2019;39:42–51. doi: 10.1016/j.
ejbt.2019.03.005.
[116] Hou Z, Yin J, Lu Y, et al. Transcriptomic analysis reveals
the temporal and spatial changes in physiological pro-
cess and gene expression in common buckwheat
(Fagopyrum esculentum Moench) grown under drought
stress. Agronomy-Basel. 2019;9:569. doi: 10.3390/agron-
omy9100569.
[117] Liu C, Xiang D, Wu Q, etal. Dynamic transcriptome and
co-expression analysis suggest the potential roles of
small secreted peptides from Tartary buckwheat
(Fagopyrum tataricum) in low nitrogen stress response.
Plant Sci. 2021;313:111091. doi: 10.1016/j.plants-
ci.2021.111091.
[118] Huang X, Yao JW, Zhao YY, et al. Ecient rutin and
quercetin biosynthesis through avonoids-related gene
expression in Fagopyrum tataricum Gaertn. Hairy root
cultures with UV-B irradiation. Front Plant Sci. 2016;7:63.
doi: 10.3389/fpls.2016.00063.
[119] Yang MF, He GD, Hou QD, etal. Systematic analysis and
expression proles of TCP gene family in Tartary buck-
wheat (Fagopyrum tataricum (L.) Gaertn.) revealed the
potential function of FtTCP15 and FtTCP18 in response
to abiotic stress. BMC Genomics. 2022;23:415. doi:
10.1186/s12864-022-08618-1.
[120] Jiang L, Liu C, Fan Y, et al. Dynamic transcriptome anal-
ysis suggests the key genes regulating seed develop-
ment and lling in Tartary buckwheat (Fagopyrum ta-
taricum Garetn.). Front Genet. 2022;13:990412. doi:
10.3389/fgene.2022.990412.
[121] Yao H, Li C, Zhao H, et al. Deep sequencing of the
transcriptome reveals distinct avonoid metabolism
features of black Tartary buckwheat (Fagopyrum tatari-
cum Garetn.). Progress Biophys Mol. Biology.
2017;124:49–60.
[122] Fang X, Zhang Y, Zhang Y, etal. De novo transcriptome
assembly and identication of genes related to seed
size in common buckwheat (Fagopyrum esculentum
M.). Breed Sci. 2019;69:487–497. doi: 10.1270/
jsbbs.18194.
[123] Rao H, Wang Q, Han X, et al. Quantitative proteomics
reveals the mechanism of slightly acidic electrolyzed
water-induced buckwheat sprouts growth and avo-
noids enrichment. Food Res Int. 2021;148:110634. doi:
10.1016/j.foodres.2021.110634.
[124] Lei GJ, Yokosho K, Yamaji N, etal. Functional character-
ization of two half-size ABC transporter genes in
aluminium-accumulating buckwheat. New Phytol.
2017;215:1080–1089. doi: 10.1111/nph.14648.

24 Y. SONG ETAL.
[125] Wang J, Xiao J, Liu X, et al. Analysis of Tartary buck-
wheat (Fagopyrum tataricum) seed proteome using of-
ine two-dimensional liquid chromatography and tan-
dem mass spectrometry. J Food Biochem.
2019;43:e12863. doi: 10.1111/jfbc.12863.
[126] Kopeć P, Hornyák M, Pastuszak J, et al. Changes in the
ower and leaf proteome of common buckwheat
(Fagopyrum esculentum Moench) under high temperature.
Int J Mol Sci. 2021;22:22. doi: 10.3390/ijms22052678.
[127] Colgrave ML, Goswami H, Byrne K, et al. Proteomic
proling of 16 cereal grains and the application of tar-
geted proteomics to detect wheat contamination. J
Proteome Res. 2015;14:2659–2668. doi: 10.1021/acs.
jproteome.5b00187.
[128] Deng J, Dong F, Wu C, et al. Comparative metabolom-
ics analysis between red- and white-owered common
buckwheat cultivars. Phyton-Int. J. Experim. Botany.
2021;90:859–870. doi: 10.32604/phyton.2021.014625.
[129] Zhao Y, Zhai G, Li X, et al. Metabolomics reveals nutri-
tional diversity among six coarse cereals and antioxi-
dant activity analysis of grain sorghum and sweet sor-
ghum. Antioxidants. 2022;11:1984. doi: 10.3390/
antiox11101984.
[130] Kim YB, Park S-Y, Thwe AA, et al. Metabolomic analysis
and dierential expression of anthocyanin biosynthetic
genes in white- and red-owered buckwheat cultivars
(Fagopyrum esculentum). J Agric Food Chem.
2013;61:10525–10533. doi: 10.1021/jf402258f.
[131] Ling A, Li X, Hu X, et al. Dynamic changes in polyphe-
nol compounds, antioxidant activity, and PAL gene ex-
pression in dierent tissues of buckwheat during ger-
mination. J Sci Food Agric. 2018;98:5723–5730. doi:
10.1002/jsfa.9119.
[132] Du D, Xiong H, Xu C, et al. Nutrient metabolism path-
ways analysis and key candidate genes identication
corresponding to cadmium stress in buckwheat
through multiomics analysis. Genes (Basel).
2023;14:1462. doi: 10.3390/genes14071462.
[133] Ma W, Kim JK, Jia C, et al. Comparative transcriptome
and metabolic proling analysis of buckwheat
(Fagopyrum tataricum (L.) Gaertn.) under salinity stress.
Metabolites. 2019;9:225. doi: 10.3390/metabo9100225.
[134] Meng H-L, Sun P-Y, Wang J-R, et al. Comparative phys-
iological, transcriptomic, and WGCNA analyses reveal
the key genes and regulatory pathways associated
with drought tolerance in Tartary buckwheat. Front
Plant Sci. 2022;13:985088. doi: 10.3389/fpls.2022.985088.
[135] Huang J, Tang B, Ren R, et al. Understanding the po-
tential gene regulatory network of starch biosynthesis
in Tartary buckwheat by RNA-Seq. Int J Mol Sci.
2022;23:15774. doi: 10.3390/ijms232415774.
[136] Li H-Y, Wu C-X, Lv Q-Y, etal. Comparative cellular, phys-
iological and transcriptome analyses reveal the poten-
tial easy dehulling mechanism of rice-Tartary buck-
wheat (Fagopyrum Tararicum). BMC Plant Biol.
2020;20:505. doi: 10.1186/s12870-020-02715-7.
[137] Liu MY, Fu QK, Ma ZT, etal. Genome-wide investigation
of the MADS gene family and dehulling genes in
Tartary buckwheat (Fagopyrum tataricum). Planta.
2019;249:1301–1318. doi: 10.1007/s00425-019-03089-3.
[138] You W, Chen XJ, Zeng LT, et al. Characterization of
PISTILLATA-like genes and their promoters from the di-
styly Fagopyrum esculentum. Plants-Basel. 2022;11:1047.
doi: 10.3390/plants11081047.
[139] Yao P, Deng R, Huang Y, etal. Diverse biological eects
of glycosyltransferase genes from Tartary buckwheat.
BMC Plant Biol. 2019;19:339. doi: 10.1186/
s12870-019-1955-z.
[140] Yin Q, Han X, Han Z, et al. Genome-wide analyses re-
veals a glucosyltransferase involved in rutin and emo-
din glucoside biosynthesis in Tartary buckwheat. Food
Chem. 2020;318:126478. doi: 10.1016/j.foodchem.2020.
126478.
[141] Nagatomo Y, Usui S, Ito T, etal. Purication, molecular
cloning and functional characterization of avonoid
C-glucosyltransferases from Fagopyrum esculentum M.
(buckwheat) cotyledon. Plant J. 2014;80:437–448. doi:
10.1111/tpj.12645.
[142] Li HY, Sun XQ, Ma C, et al. Identication and character-
ization of micrornas during common buckwheat
(Fagopyrum esculentum) seed development. Int J Agric
Biol. 2020;24:1115–1124.
[143] Li HY, Meng HL, Sun XQ, et al. Integrated microRNA
and transcriptome proling reveal key miRNA-mRNA
interaction pairs associated with seed development in
Tartary buckwheat (Fagopyrum tataricum). BMC Plant
Biol. 2021;21:132. doi: 10.1186/s12870-021-02914-w.
[144] Liu MY, Huang L, Ma ZT, et al. Genome-wide identica-
tion, expression analysis and functional study of the
GRAS gene family in Tartary buckwheat (Fagopyrum
tataricum). BMC Plant Biol. 2019;19:342. doi: 10.1186/
s12870-019-1951-3.
[145] Liu MY, Huang Q, Sun WJ, et al. Genome-wide investi-
gation of the heat shock transcription factor (Hsf) gene
family in Tartary buckwheat (Fagopyrum tataricum).
BMC Genomics. 2019;20:871. doi: 10.1186/
s12864-019-6205-0.
[146] Huang J, Ren RR, Rong YP, et al. Identication, expres-
sion, and functional study of seven NAC transcription
factor genes involved in stress response in Tartary
buckwheat (Fagopyrum tataricum (L.) Gaertn.).
Agronomy-Basel. 2022;12:849. doi: 10.3390/agrono-
my12040849.
[147] Liu MY, Ma ZT, Sun WJ, et al. Genome-wide analysis of
the NAC transcription factor family in Tartary buck-
wheat (Fagopyrum tataricum). BMC Genomics.
2019;20:113. doi: 10.1186/s12864-019-5500-0.
[148] Deng RY, Zhao HX, Xiao YH, et al. Cloning, characteri-
zation, and expression analysis of eight stress-related
NAC genes in Tartary buckwheat. Crop Sci. 2019;59:266–
279. doi: 10.2135/cropsci2018.06.0368.
[149] Li L, Yan XY, Li J, et al. Physiological and FtCHS gene
expression responses to peg-simulated drought and
cadmium stresses in Tartary buckwheat seedlings. J
Plant Growth Regul. 2022;41:3518–3529. doi: 10.1007/
s00344-021-10530-z.
[150] Nan GH, Wu HL, Wu Q, et al. Cloning, identication,
and functional analysis of chalcone isomerase gene
and its promoter from Tartary buckwheat. Acta Physiol
Plant. 2022;44:78. doi: 10.1007/s11738-022-03410-w.
[151] Li CL, Yang K, Yang JJ, et al. Tartary buckwheat FtF3H1
as a metabolic branch switch to increase anthocyanin
content in transgenic plant. Front Plant Sci.
2022;13:1056857. doi: 10.3389/fpls.2022.959698.

CRITICAL REVIEWS IN BIOTECHNOLOGY 25
[152] Li CL, Bai YC, Li SJ, et al. Cloning, characterization, and
activity analysis of a avonol synthase gene FtFLS1 and
its association with avonoid content in Tartary buck-
wheat. J Agric Food Chem. 2012;60:5161–5168. doi:
10.1021/jf205192q.
[153] Katsu K, Suzuki R, Tsuchiya W, etal. A new buckwheat di-
hydroavonol 4-reductase (DFR), with a unique substrate
binding structure, has altered substrate specicity. BMC
Plant Biol. 2017;17:239. doi: 10.1186/s12870-017-1200-6.
[154] Yang Y, Wang XF, Zheng J, etal. Amino acid transport-
er (AAT) gene family in Tartary buckwheat (Fagopyrum
tataricum L. Gaertn.): characterization, expression anal-
ysis and functional prediction. Int J Biol Macromol.
2022;217:330–344. doi: 10.1016/j.ijbiomac.2022.07.059.
[155] Ye XL, Liu CY, Yan HL, etal. Genome-wide identication
and transcriptome analysis of the heavy metal-associated
(HMA) gene family in Tartary buckwheat and their reg-
ulatory roles under cadmium stress. Gene.
2022;847:146884. doi: 10.1016/j.gene.2022.146884.
[156] Gao F, Zhao HX, Yao HP, et al. Identication, isolation
and expression analysis of eight stress-related
R2R3-MYB genes in Tartary buckwheat (Fagopyrum ta-
taricum). Plant Cell Rep. 2016;35:1385–1396. doi:
10.1007/s00299-016-1971-5.
[157] Wang L, Deng RY, Bai YC, et al. Tartary buckwheat
R2R3-MYB gene FtMYB3 negatively regulates anthocy-
anin and proanthocyanin biosynthesis. Int J Mol Sci.
2022;23:2775. doi: 10.3390/ijms23052775.
[158] Deng J, Wang L, Damaris RN, et al. Genome-wide identi-
cation of R2R3-MYB transcription factor family in tartary
buckwheat (Fagopyrum tataricum) identies a member in-
volved in anthocyanin biosynthesis. Agronomy-Basel.
2023;13:2117. doi: 10.3390/agronomy13082117.
[159] Liu MY, Sun WJ, Ma ZT, et al. Integrated network anal-
yses identify MYB4R1 neofunctionalization in the UV-B
adaptation of Tartary buckwheat. Plant Commun.
2022;3:100414. doi: 10.1016/j.xplc.2022.100414.
[160] Wang S, Wu HL, Cao XX, et al. Tartary buckwheat
FtMYB30 transcription factor improves the salt/drought
tolerance of transgenic Arabidopsis in an
ABA-dependent manner. Physiol Plant. 2022;174:e13781.
doi: 10.1111/ppl.13781.
[161] He X, Li JJ, Chen Y, et al. Genome-wide analysis of the
WRKY gene family and its response to abiotic stress in
buckwheat (Fagopyrum tataricum). Open Life Sci.
2019;14:80–96. doi: 10.1515/biol-2019-0010.
[162] Wang S, Lv BB, Wang AH, etal. Cloning and functional
characterization of FtWRKY29, a low phosphorus
stress-inducible transcription factor in Tartary buck-
wheat. Plant Physiol Biochem. 2023;203:107997. doi:
10.1016/j.plaphy.2023.107997.
[163] Wang X, Feng B, Xu ZB, et al. Identication and char-
acterization of granule bound starch synthase I (GBSSI)
gene of Tartary buckwheat (Fagopyrum tataricum
Gaertn.). Gene. 2014;534:229–235. doi: 10.1016/j.
gene.2013.10.053.
[164] Liu M, Ma Z, Wang A, et al. Genome-Wide Investigation
of the Auxin Response Factor Gene Family in Tartary
Buckwheat (Fagopyrum tataricum). Int J Mol Sci.
2018;19:3526. doi: 10.3390/ijms19113526.
[165] Duan Y, Yin G, He R, et al. Identication of candidate
genes for easily-shelled traits in Tartary buckwheat
based on BSA-Seq and RNA-Seq methods. Euphytica.
2022;218:91. doi: 10.1007/s10681-022-03023-x.
[166] Ji QQ, Zhang TY, Zhang D, et al. Genome-wide identi-
cation and expression analysis of lipoxygenase genes
in Tartary buckwheat. Biotechnol Biotechnol Equip.
2020;34:273–286. doi: 10.1080/13102818.2020.1738956.
[167] Liu MY, Sun WJ, Ma ZT, et al. Genome-wide identica-
tion of the SPL gene family in Tartary Buckwheat
(Fagopyrum tataricum) and expression analysis during
fruit development stages. BMC Plant Biol. 2019;19:299.
doi: 10.1186/s12870-019-1916-6.
[168] Yan HL, Liu CY, Zhao JL, et al. Genome-wide analysis of
the NF-Y gene family and their roles in relation to fruit
development in Tartary buckwheat (Fagopyrum tatari-
cum). Int J Biol Macromol. 2021;190:487–498. doi:
10.1016/j.ijbiomac.2021.09.001.
[169] Ma ZT, Liu MY, Sun WJ, et al. Genome-wide identica-
tion and expression analysis of the trihelix transcrip-
tion factor family in Tartary buckwheat (Fagopyrum ta-
taricum). BMC Plant Biol. 2019;19:344. doi: 10.1186/
s12870-019-1957-x.
[170] Matsui K, Tomatsu T, Kinouchi S, et al. Identication of
a gene encoding glutathione S-transferase that is relat-
ed to anthocyanin accumulation in buckwheat
(Fagopyrum esculentum). J Plant Physiol. 2018;231:291–
296. doi: 10.1016/j.jplph.2018.10.004.
[171] Nikolić DB, Samardzić JT, Bratić AM, et al. Buckwheat
(Fagopyrum esculentum Moench) FeMT3 gene in heavy
metal stress: protective role of the protein and induc-
ibility of the promoter region under Cu2+ and
Cd2+treatments. J Agric Food Chem. 2010;58:3488–
3494. doi: 10.1021/jf904483a.
[172] Li J, Zhang YC, Xu L, et al. Genome-wide identication
of DNA binding with one nger (Dof) gene family in
tartary buckwheat (Fagopyrum tataricum) and analysis
of its expression pattern after exogenous hormone
stimulation. Biology (Basel). 2022;11:173. doi: 10.3390/
biology11020173.
[173] Yao YJ, Zhao HX, Sun L, et al. Genome-wide identica-
tion of MAPK gene family members in Fagopyrum ta-
taricum and their expression during development and
stress responses. BMC Genomics. 2022;23:96. doi:
10.1186/s12864-022-08293-2.
[174] Fang ZW, Qi R, Li XF, et al. Ectopic expression of
FaesAP3, a Fagopyrum esculentum (Polygonaceae) AP3
orthologous gene rescues stamen development in an
Arabidopsis ap3 mutant. Gene. 2014;550:200–206. doi:
10.1016/j.gene.2014.08.029.
[175] Fang Y, Wang S, Wu HL, et al. Genome-Wide
Identication of ATG Gene Family Members in
Fagopyrum tataricum and Their Expression during
Stress Responses. Int J Mol Sci. 2022;23:14845. doi:
10.3390/ijms232314845.
[176] Fang ZW, Zhang XH, Gao JF, et al. A Buckwheat
(Fagopyrum esculentum) DRE-Binding Transcription
Factor Gene, FeDREB1, Enhances Freezing and Drought
Tolerance of Transgenic Arabidopsis. Plant Mol Biol Rep.
2015;33:1510–1525. doi: 10.1007/s11105-015-0851-4.
[177] Li LY, Fang ZW, Li XF, etal. Isolation and Characterization
of the C-class MADS-box Gene from the Distylous
Pseudo-cereal Fagopyrum esculentum. J Plant Biol.
2017;60:189–198. doi: 10.1007/s12374-016-0488-3.

26 Y. SONG ETAL.
[178] Fang ZW, Li XP, Li XF, et al. FaesPI, a Fagopyrum escu-
lentum PISTILLATA ortholog, is involved only in stamen
development. J Plant Biol. 2015;58:102–109. doi:
10.1007/s12374-014-0390-9.
[179] Yao PF, Zhao HX, Luo XP, et al. Fagopyrum tataricum
FtWD40 Functions as a Positive Regulator of Anthocyanin
Biosynthesis in Transgenic Tobacco. J Plant Growth Regul.
2017;36:755–765. doi: 10.1007/s00344-017-9678-6.
[180] Lai D, Zhang K, He Y, et al. Multi‐omics identication of
a key glycosyl hydrolase gene FtGH1 involved in rutin
hydrolysis in Tartary buckwheat (Fagopyrum tataricum).
2023;22:1206–1223. doi: 10.1111/pbi.14259.
[181] Lu Q-H, Wang Y-Q, Song J-N, et al. Transcriptomic identi-
cation of salt-related genes and de novo assembly in com-
mon buckwheat (F. esculentum). Plant Physiol Biochem.
2018;127:299–309. doi: 10.1016/j.plaphy.2018.02.001.
[182] Yano K, Morinaka Y, Wang F, etal. GWAS with principal
component analysis identies a gene comprehensively
controlling rice architecture. Proc Natl Acad Sci U S A.
2019;116:21262–21267. doi: 10.1073/pnas.1904964116.
[183] Zhang X, Ren Z, Luo B, et al. Genetic architecture of
maize yield traits dissected by QTL mapping and GWAS
in maize. Crop J. 2022;10:436–446. doi: 10.1016/j.
cj.2021.07.008.
[184] Gai W, Yang F, Yuan L, et al. Multiple-model GWAS
identies optimal allelic combinations of quantitative
trait loci for malic acid in tomato. Hortic Res.
2023;10:uhad021. doi: 10.1093/hr/uhad021.