Journal Pre-proof

Recent Advances to Enhance Nutritional Quality of Rice

Sundus Zafar, Xu Jianlong

PII: S1672-6308(23)00084-7
DOI: https://doi.org/10.1016/j.rsci.2023.05.004
Reference: RSCI 428

To appear in: Rice Science

Received Date: 10 December 2022

Accepted Date: 25 May 2023

Please cite this article as: S. Zafar, X. Jianlong, Recent Advances to Enhance Nutritional Quality of Rice,
Rice Science, https://doi.org/10.1016/j.rsci.2023.05.004.

This is a PDF file of an article that has undergone enhancements after acceptance, such as the addition
of a cover page and metadata, and formatting for readability, but it is not yet the definitive version of
record. This version will undergo additional copyediting, typesetting and review before it is published
in its final form, but we are providing this version to give early visibility of the article. Please note that,
during the production process, errors may be discovered which could affect the content, and all legal
disclaimers that apply to the journal pertain.

Copyright © 2023, China National Rice Research Institute. Hosting by Elsevier B.V.
 Available online at www.sciencedirect.com

ScienceDirect
Rice Science

Review

Recent Advances to Enhance Nutritional Quality of Rice

Sundus ZAFAR1, 2, XU Jianlong2, 3

(1School of Life Sciences, Jiangsu University, Zhenjiang 212013, China; 2Shenzhen Branch, Guangdong Laboratory for Lingnan
Modern Agriculture, Agricultural Genomics Institute at Shenzhen, Chinese Academy of Agricultural Sciences, Shenzhen 518120,
China; 3The National Key Facility for Crop Gene Resources and Genetic Improvement, Institute of Crop Sciences, Chinese Academy
of Agricultural Sciences, Beijing 100081, China)

f
oo
Abstract: The nutritional quality of rice is a major concern, along with the need to enhance productivity to
feed the continuously growing population. Therefore, there is a requirement to breed high-yield rice
varieties with improved nutritional quality that can help combat malnutrition, which affects global health.

r
Undoubtedly, breeding approaches have played a significant role in increasing rice yield while enhancing

-p
its nutritional content, thereby meeting the demands for this crop. In addition to traditional breeding
techniques, other recent approaches, such as genetic engineering, gene editing, omics methods, and
re
agronomic practices, must also be employed to meet the nutritional needs of the current population. In
this review, we offer detailed information on the development of nutritionally improved rice varieties
through the enhancement of protein content, micro- and macro-nutrients, vitamins, and oil quality using
lP

genetic engineering approaches. We also identify QTLs associated with amino acids, proteins, and
micro-nutrients in rice. Furthermore, omics approaches provide a range of tools and techniques for
effectively exploring resources and understanding the molecular mechanisms involved in trait
na

development. Omics branches, including transcriptomics, proteomics, ionomics, and metabolomics, are
efficiently utilized for improving rice nutrition. Therefore, by utilizing the information obtained from these
techniques and incorporating all of these recent approaches, we can effectively modify the rice genome,
ur

directly enhancing the nutritional value of rice varieties. This will help address the challenges of
malnutrition in the years to come.
Jo

Key words: rice; nutritional quality; quantitative trait locus; genetic engineering; omics; gene editing

Nutrition is a fundamental prerequisite for all living as these staples, while being the primary source of
beings to perform essential functions like growth, calories, fall short in providing essential nutrients.
development, and production. Rice serves as a primary Consequently, these countries face health issues related
food source for more than three billion people to nutrient deficiencies, such as anemia (iron deficiency),
worldwide, providing 20% of the world’s nutritional night blindness (vitamin A deficiency), growth retardation,
energy supply. In recent years, agriculture has and poor mental health (Oakley et al, 2004; Darnton-
predominantly focused on producing nutritionally Hill et al, 2005). A significant portion of staple crop
improved varieties of staple crops such as rice, wheat, research is dedicated to exploring various methods and
maize, and legumes to fulfill human nutritional techniques to enhance crop nutrition, encompassing both
requirements (Ricachenevsky et al, 2019). Developing micro- and macro- nutrients. Rice is a noteworthy
and underdeveloped countries have increasingly source of protein containing eight essential amino
emphasized the need to boost crop yield and accessibility, acids in balanced proportions, contributing to healthy

Received: 10 December 2022; Accepted: 25 May 2023

Corresponding author: XU Jianlong ([email protected])
Copyright © 2023, China National Rice Research Institute. Hosting by Elsevier B V
This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/)
Peer review under responsibility of China National Rice Research Institute
http://dx.doi.org/
 Rice Science

hair, skin, heart and lung health, and a well-functioning Table 1. Comparison of nutrient content in different staple crops.
nervous system (Han et al, 2015). However, rice typically
Nutrient Rice Maize Wheat Soybean Potato
contains only about 7% protein, which is less than
Protein (g/kg) 71.0 98.0 137.0 130.0 20.0
what is found in pulses and wheat (Chattopadhyay et al, Carbohydrate (g/kg) 800.0 600.0 710.0 110.0 170.0
2019). Despite being a major component of cereals, Fat (g/kg) 22.0 11.8 24.7 68.0 1.0
relatively little effort has been invested in increasing Iron (mg/kg) 43.1 30.0 35.2 35.5 7.8
Zinc (mg/kg) 20.0 30.0 41.6 2.9 9.9
its protein content. Nevertheless, elevating protein levels Selenium (μg/kg) 6.0 151.0 894.0 15.0 3.0
in rice can help alleviate malnutrition in countries Beta-carotene (mg/kg) 0.0 3.7 0.2 0.0 0.0
where rice serves as a main staple crop, meeting daily Riboflavin (mg/kg) 0.5 0.6 1.2 1.8 0.3
calorie requirements but lacking diverse sources of Thiamin (mg/kg) 5.8 2.0 4.2 4.4 0.8
Vitamin E (mg/kg) 1.1 0.7 0.0 0.1 0.0
essential nutrients. Recent attention has also been Lysine (g/16 g N) 3.80 2.50 2.30 6.20 6.30
given to enhancing the micronutrient content in rice Threonine (g/16 g N) 3.60 2.80 3.20 3.99 4.10
grains through agronomic biofortification (Prom-u-thai Tryptophan (g/16 g N) 1.10 0.60 1.00 1.05 1.70
Data are from FAO (http://www.fao.org/3/t0567e/T0567E0d.htm).
and Rerkasem, 2020; Utasee et al, 2022), apart from
other approaches like conventional breeding, marker-

f
into the nutritional aspect of rice.

oo
assisted breeding, genetic engineering, and omics
techniques aimed at improving the nutritional quality Rice contains a variety of vital nutrients, including
of rice. In this review, our focus is on rice quality, calcium (Ca), iron (Fe), zinc (Zn), potassium (K),

r
particularly its nutritional value and significance, and phosphorus (P), and sodium (Na), as well as vitamins
we also discuss various strategies for enhancing the
nutritional quality of rice. -p
and minerals crucial for human health (Birla et al,
2017). However, when compared with other crops
re
such as maize, wheat, and legumes, rice tends to have
Rice quality characteristics and nutritional lower mineral and vitamin B contents (Mahender et al,
lP

aspects 2016). A comparison of the nutrient contents of

different staple crops is provided in Table 1. Brown
The traits and parameters used to assess rice grain
rice is considered more nutritious than white rice
quality differ across countries. However, there are
na

because the brown layer contains protein, fatty acids,

four main quality traits commonly used for evaluation:
vitamins, minerals, and antioxidants (Schramm et al,
physical appearance, milling, cooking, and nutritional
2007). Rice bran, in particular, contains about 80%
ur

qualities (Yu et al, 2008). Achieving a meaningful

fatty acids. Rice oil contains unsaturated fatty acids
increase in the economic and nutritional values of rice
such as oleic acid (C18:1) and α-linolenic acid
Jo

hinges on a comprehensive understanding of the factors

(C18:3), which are essential for maintaining cell
governing the production of different rice varieties
membranes and the proper functioning of the nervous
(Prom-u-thai and Rerkasem, 2020). Therefore, rice
system (Swanson et al, 2012). Despite its superior
quality depends upon numerous factors, including
nutritional value, consumers often prefer white rice
variety, harvesting conditions, environmental factors
over brown rice due to its inferior texture (Mahender
such as light, temperature, humidity, and postharvest
et al, 2016). However, the polishing processenhances
management (Fig. 1). In this discussion, we will delve
the rice texture and eating and
cooking quality but reduces the
nutrient content of rice (Wu et al,
2013). A comparison of the
nutritional value of brown and white
rice is provided in Table 2.
Rice can be categorized into two
types based on its color: non-pigmented
rice (accounting for approximately
85% of rice) and pigmented rice
(about 15%), including varieties like
red, brown, black, and purple rice
(Wijaya et al, 2017). Pigmented rice,
Fig. 1. Factors affect grain quality of rice. such as black and purple rice,
Sundus ZAFAR and XU Jianlong. Recent Advances to Enhance Nutritional Quality of Rice

Table 2. Comparison of nutritional value in brown and white rice quality of various crops. Similarly, omics approaches
(one medium cup of white rice or brown rice).
provide various tools and techniques for exploring
Nutrient Brown rice White rice genetic resources and understanding the genetic
Carbohydrate (g) 51.70 53.20 mechanisms involved in trait development. Therefore,
Protein (g) 5.54 4.43
Fat (g) 1.96 0.39
incorporating omics approaches is profitable for
Calories (Kcal) 248.00 242.00 attaining desired enhancements in rice varieties. Thus,
Iron (mg) 1.10 2.80 developing these superior varieties that can be used
Zinc (mg) 1.40 0.80
Magnesium (mg) 78.80 24.20
further in breeding is the basis of biofortification (Brar
Copper (mg) 0.20 0.10 et al, 2012). The major strategies are shown in Fig. 2.
Phosphorus (mg) 208.00 68.80
Manganese (mg) 2.00 0.70 Genetic engineering approach for nutritious
Thiamine (mg) 0.40 0.30 rice
Folate (mg) 18.20 108.00
Niacin (mg) 5.20 3.40 Genetic engineering is considered an effective and
efficient method for enhancing the nutritional quality
contains secondary metabolites like anthocyanin,

f
traits of plants (Dias and Ortiz, 2012). Numerous

oo
flavonoids, and phenolics, which contribute to their efforts have also been made to enhance the nutritional
status as functional foods with associated health value of rice.
benefits (Fitri et al, 2021; Fongfon et al, 2021). Red

r
rice is rich in Fe and Zn, which are crucial for Protein and amino acids
hemoglobin production and enzymatic processes,
respectively (Boue et al, 2016). Additionally, black -p
Rice protein is considered nutritionally incomplete
re
due to a deficiency in lysine and other essential amino
and red rice have been found to inhibit the proliferation acids. Constitutive expression of maize lysine
of breast cancer (Ghasemzadeh et al, 2018). dihydrodipicolinate synthase (DHPS) leads to an
lP

Need to improve nutritional quality of rice increase in free lysine (2.5-fold) in transgenic rice
(Lee et al, 2001). Long et al (2013) reported an
na

Though rice is a good source of micro- and effective way to enhance lysine concentration by
macro-nutrients, recent climate change has reduced expressing aspartate kinase (AK) and DHPS, and by
the nutrient content of rice and other crops (Smith and down-regulating LKR/SDH (lysine ketoglutaric acid
ur

Myers, 2019). Rice grown with elevated levels of CO2 reductase/saccharopine dehydropine dehydrogenase)
has mainly decreased vitamins, Fe, Zn, and protein expression through RNAi (RNA interference), which
Jo

contents compared with rice grown with a normal increases lysine concentration in rice grains up to
concentration of CO2 (Smith and Myers, 2019). 60-fold. Yang et al (2016) targeted bacterial AK and
Though about half of the world acquires 25% of its DHPS in the intragenic region of the rice genome.
calories from rice, it is still estimated that an increase Two pyramid transgenic lines (high free lysine; HFL1
in proteins or micronutrients could fulfill the nutritional
demands of rice-dependent people (FAO, 2018).
Hence, there is a need to enhance micro- and macro-
nutrients in rice and develop varieties with high
nutritional quality using different approaches.
Strategies to enhance nutritional quality of rice
Genetic engineering is a quick and cost-effective
method to enhance nutrients in crops. Genetic engineering
of rice is a potential option as desirable plant parts can
be obtained using tissue-specific promoters and gene
stacking or transformation and co-transformation of
the same and different plasmids that allow multiple
traits to be transferred (Naqvi et al, 2009, 2010; Farré
et al, 2014). Furthermore, genome editing has become
an essential genomic tool for improving the nutritional Fig. 2. Different approaches to enhance nutritional quality of rice.
 Rice Science

and HFL2) were obtained with 25-fold higher lysine Table 3. Genetic engineering approaches to incorporate nutritional
concentration in the seeds than the wild type (WT) genes into rice varieties.

plants. In another approach, Liu et al (2016) over- Nutrient Gene Expression level Reference
expressed lysine-rich protein (LRP) gene in rice from Lysine DHPS 2.5-fold Lee et al, 2001
Psophocarpus tetragonolobus using an endosperm- AK + DHPS 60-fold Long et al, 2013
RLRH1 and 35-fold Wong et al, 2015
specific promoter and obtained transgenic rice with RLRH2
30% lysine higher than the control plants. Further, two AK + DHPS 25-fold Yang et al, 2016
lysine-rich histone proteins (RLRH1 and RLRH2) LRP 30-fold Liu et al, 2016
β-Carotene psy+ crtI + lcy Ye et al, 2000
were overexpressed in rice, which showed a 35% Maize Psy 23-fold Paine et al, 2005
increase in lysine concentration, along with balanced Folate Pterin + 10-fold Storozhenko et al,
amino acids as instructed by the WHO dietary Aminobenzoate 2007
Fe SoyferH1 2-fold Goto et al, 1999
standard (Wong et al, 2015).
OsNAS3 2.9-fold Lee and An, 2009
OsNAS2 4.2-fold Johnson, 2011
Manipulation of micronutrients OsVIT1 and 1.4-fold Bashir et al, 2013
OsVIT2
Micronutrient deficiency is a primary global health

f
Fe and Zn OsYSL2 4-fold Fe; Ishimaru et al, 2010;

oo
concern. It negatively affects an individual’s physical 4-fold Zn Masuda et al, 2013
and mental development, productivity, and intellectual OsNAS1, OsNAS2 2-fold Fe; Johnson et al, 2011
and OsNAS3 2-fold Zn
growth (Siddiqui et al, 2020). The inadequacy of

r
Osfer2 2.09-fold Fe; Paul et al, 2012
essential micronutrients in the human diet is termed

-p
1.37-fold Zn
‘hidden hunger’. Due to hidden hunger and micronutrient α-Linolenic OsFAD3 23.8- to 27.9-fold
Liu et al, 2012b
malnutrition, more than 24 000 people die worldwide acid
re
DHPS, Dihydrodipicolinate synthase; AK, Aspartate kinase; LRP,
every day (Fiaz et al, 2019). The main micronutrients, Lysine-rich protein; RLRH, Lysine-rich histone protein; Fe, Iron; Zn,
such as vitamin A, zinc, and iron deficiencies, are the Zinc.
lP

most common and cause severe health consequences

(Parashar et al, 2023). increase β-carotene concentration by 63-fold in rice
endosperm.
na

β-Carotene
Anthocyanins and folate
Rice cannot produce β-carotene, the precursor of Anthocyanins have high antioxidant activities and are
ur

vitamin A, which is essential for retinal function and beneficial to human health. Zhu et al (2017) reported
can prevent night blindness. Ye et al (2000) reported the engineering of eight anthocyanin-related genes
Jo

the invention of golden rice (GR), known as the (two maize genes and six coleus genes) with an
prototype of GR1. They acquired the psy (phytoene endosperm-specific promoter, producing a novel
synthase) gene from daffodil and two other genes, crtI biofortified germplasm called ‘Purple Endosperm
(phytoene desaturase) and lcy (lycopene cyclase), Rice’ that contains a high concentration of anthocyanins
from Erwinia uredovora. However, psy from daffodils (about 1 mg/g). The human body cannot synthesize
has limitations in synthesizing higher levels of folate (vitamin B9); thus, a folate deficiency can cause
carotenoids. Paine et al (2005) replaced the daffodil neural tube defects, increasing the risk of cardiovascular
psy with its homolog from maize, enhancing the and coronary diseases (Iyer and Tomar, 2009).
nutritional value of GR by increasing β-carotene Storozhenko et al (2007) enhanced the folate level in
content by 23-fold, with up to approximately 37 µg/g rice by overexpressing two genes from Arabidopsis
in rice endosperm. Compared with the original GR, a thaliana with a rice endosperm-specific promoter. The
psy gene from maize enhances carotenoid storage, transgenic rice contains approximately 100 times more
termed ‘Golden Rice 2’. Subsequently, numerous folate than WT plants. Dong et al (2014a) evaluated
efforts have been made to develop new versions of the folate biosynthesis pathway in japonica rice
GR (Ha et al, 2010; Bai et al, 2016; Jeong et al, 2017). (Kitaake) by overexpressing two genes from A.
Tian et al (2019) expressed psy from maize (ZmPsy) thaliana coding for GTP cyclohydrolase I and amino
and Crtl from Pantoea ananatis (PaCrtI) along with deoxychorismate synthase, which increase folate
tHMG1, encoding truncated HMGR from Saccharomyces content in rice seeds by 3.2 times.
cerevisiae. They reported that these chemically
synthesized genes (tHMG1, ZmPsy1, and PaCrtI) Fe and Zn
Sundus ZAFAR and XU Jianlong. Recent Advances to Enhance Nutritional Quality of Rice

Fe and Zn are the most dominant deficient micronutrients and linoleic acid (C18:2, 37%)] that are substandard
affecting 2 billion people globally and causing 0.8 for rice storage (Tiwari et al, 2016). The shelf life of
million annual deaths (Ritchie and Roser, 2017). On RBO reduces due to the existence of a double bond in
average, polished rice comprises approximately 2 µg/g C18:2 (linoleic acid), resulting in a waste of
of Fe and 16 µg/g of Zn. Goto et al (1999) attained a 60%–70% of RBO (Chaiyasit et al, 2007). Thus, fatty
2-fold increase in Fe content in brown rice compared acid desaturase 2 (FAD2) is suppressed by RNAi,
with WT by overexpressing the soybean ferritin gene reducing linoleic acid and enhancing oleic acid in
(SoyferH1). Subsequently, research on Fe and Zn many plants, such as Arabidopsis (Lei et al, 2014) and
biofortification in rice was conducted using single- rice (Mikami et al, 2015). Liu et al (2012a) conducted
gene approaches like introducing rice ferritin gene and transgenic analyses of three annotated FAD genes and
soybean ferritin gene into rice (Lucca et al, 2001; confirmed that OsFAD3 converted C18:2 to C18:3
Vasconcelos et al, 2003). Many approaches for Fe (α-linolenic acid, ALA). Similarly, other researchers
biofortification also lead to an increase in Zn content reported a 23.8- to 27.9-fold increase in ALA content
because most components related with Fe uptake and by introducing the FAD3 gene into rice (Liu et al,
translocation are also related with Zn biofortification 2012a; Yin et al, 2014). However, Cheah et al (2013)

f
oo
except for a few Fe-specific transporters like barley only attained a slight increase in ALA content in
yellow stripe 1 (HvYS1) (Banakar et al, 2017). Paul et al transgenic rice (japonica) by overexpressing the
(2012) overexpressed rice ferritin (Osfer2), which FAD3 gene. ALA has gained massive attention due to

r
increases Fe content by 2.09-fold and Zn content by its use in producing health-stimulating products, and
1.37-fold in transgenic rice. Furthermore, the
transformation of the rice nicotianamine synthases -p its potential to reduce the risk of cardiovascular
diseases (Saravanan et al, 2010; Swanson et al, 2012).
re
(OsNAS3-D1) gene increases Fe content by 2.9-fold,
Zn content by 2.2-fold, and Cu content by 1.7-fold Lowering phytic acid (PA)
lP

compared with controls at the seedling stage (Lee et al, PA serves as the primary storage form of phosphorus
2012). To improve micronutrient content in staple (P) in cereal grains. It carries a negative charge and
crops, an initiative was taken by HarvestPlus in forms chelates with cations such as Zn, Ca, P, and Fe,
na

association with International Rice Research Institute resulting in the creation of insoluble salts referred to
(IRRI) to reduce micronutrient deficiency. In 2013, as phytate or phytin (Raboy, 2003). These insoluble
ur

they released a Zn biofortified rice variety in Bangladesh salts inhibit the absorption of vital nutrients in the
that can meet 60% of the daily Zn requirement human intestine, ultimately leading to micronutrient
Jo

(Goldstein, 2018). To date, the transformation of two deficiencies (Perera et al, 2018). Rice bran contains
genes (OsNAS2 and SferH1) into rice produced the approximately 8.7% PA, prompting numerous efforts
most promising rice variety (NASFer-274) with high to reduce its PA levels to enhance the absorption rates
Fe and Zn contents. NASFer-274 polished rice of Fe and Zn (Gupta et al, 2015). In this context,
contains Fe (15 μg/g) and Zn (45.7 μg/g), both of research has been conducted to downregulate the
which are above the target levels of Fe and Zn set by OsINO1 gene expression, thereby decreasing PA levels,
HarvestPlus. Additionally, no harmful effects related utilizing the Olesin18 (Ole18) promoters (Kuwano et al,
to heavy metal accumulation in NASFer-274 grains 2009). Since the animal system lacks the phytase
were observed (Trijatmiko et al, 2016). Wu et al (2019) enzyme, it cannot digest phytate. Consequently, a low
attained Fe (12.67 μg/g) and Zn (45.60 μg/g) contents phytate rice variety has been recently developed by
in Nipponbare, and Fe (13.65 μg/g) and Zn (48.18 overexpressing the appA gene from E. coli, resulting
μg/g) contents in IR64 using the intracellular Fe in a 4-fold increase in inorganic P (Pi) levels, along
transportation gene. Both of these varieties reached with higher levels of Zn and Fe (Bhattacharya et al,
the recommended Fe and Zn contents by HarvestPlus. 2019).
High nutritional oil Undoubtedly, genetic engineering represents a
potent tool for increasing the micronutrient content in
Rice bran oil (RBO) is extracted from rice bran and is rice grains. For example, successful developments
commercially available as food-grade vegetable oil. include rice varieties fortified with lysine, β-carotene,
Rice contains different concentrations of fatty acids anthocyanin, folate, Zn, and Fe. However, despite the
[palmitic acid (C16:0, 18%), oleic acid (C18:1, 36%), scientific achievements, genetically engineered plants
 Rice Science

face a significant challenge in terms of public acceptance to increased β-carotene accumulation in rice endosperm
and biosafety. To address concerns related to public (Yang et al, 2017).
safety and the potential escape of transgenes into the
environment, various methods have been implemented, Amylose content (AC)
albeit at an increased production cost for transgenic Cereals with high AC represent a good source of
plants (Ronald, 2014). resistant starch (Regina et al, 2006), and foods with
Gene editing for nutritious rice high resistant starch are nutritious, potentially reducing
the risk of various diseases (Jiang et al, 2010). The
Over the past decade, genetic engineering has undergone expression levels of two isoforms of SBE (starch
a significant transformation thanks to the CRISPR/Cas9 branching enzyme) genes (SBEI and SBEIIb) are
system, which has emerged as a groundbreaking tool simultaneously inhibited through RNAi, resulting in
for genome editing. CRISPR/Cas9 offers an effective an increase in AC from 27.2% to 64.8%, and resistant
approach that allows for the manipulation of genes at starch from 0% to 14.6% in transgenic rice grains
multiple genomic positions (Wang et al, 2018). Genome compared with WT plants (Zhu et al, 2012). Later,

f
editing has also been employed to enhance the CRISPR/Cas9 was employed for directed mutagenesis

oo
nutritional quality of rice, encompassing alterations in of SBEI and SBEIIb in rice. Despite sbeI-edited plants,
oil composition and other biofortification initiatives. the sbeII-edited plants exhibit significantly enhanced
amylose and resistant starch contents, increasing by

r
Oleic acid content 25.0% and 9.8%, respectively (Sun et al, 2017).
Augmenting the C18:1 content in rice at the expense
-p
Furthermore, Guo et al (2020) used CRISPR/Cas9 to
induce site-specific mutations at the SBEIIb locus in
re
of C18:2 through genetic manipulation of fatty acid
profiles can elevate the nutritional value of rice, thereby japonica rice and observed a 1.8-fold increase in AC
enhancing its health benefits for consumers. Zaplin et al and 6% increase in resistant starch content. Additionally,
lP

(2013) identified four FAD2 genes in rice, namely nutritional quality analysis showed that soluble sugar
OsFAD2-1, OsFAD2-2, OsFAD2-3, and OsFAD2-4. and lipid contents, mainly sucrose and unsaturated
fatty acids, increased significantly in the mutant lines.
na

They elucidated the pivotal role of OsFAD2-1 in

converting C18:1 to C18:2 in rice seeds and successfully Recently, Biswas et al (2022) adopted CRISPR/Cas9
generated rice varieties with elevated C18:1 content to simultaneously target the four SBE genes in rice
ur

through RNAi. Tiwari et al (2016) explored the using the tRNA (endogenous transfer RNA) processing
function of OsFAD2-1 in rice development via RNAi system to express the sgRNA (single-guided RNA)
Jo

and observed its impact on several key genes in the targeting the genes. SBE-edited lines exhibit a 15%
lipid biosynthesis pathway. Abe et al (2018) employed increase in resistant starch content compared with WT
CRISPR/Cas9-mediated targeted mutagenesis to disrupt plants. All these findings pave the way for gene editing
OsFAD2-1, resulting in homozygous OsFAD2-1 to enhance the nutritional quality of rice crops.
knockout rice plants with a 2-fold increase in C18:1 Methods such as CRISPR/Cas9 circumvent regulatory
content and an absence of detectable C18:2. This mechanisms associated with genetically modified crops
advancement improves the fatty acid composition of addressing biosafety concerns related to transgenic
rice bran oil. crops. Currently, very few nutrients have been targeted
in rice using CRISPR/Cas9, but in the near future, the
β-Carotene majority of nutrients should also be fortified. However,
Biofortification of β-carotene is a crucial target for the lack of extensive knowledge about the biosynthetic
rice improvement. CRISPR/Cas9 was used to target pathways of essential nutrients, along with the related
the Osor gene in cauliflower, which primarily accumulates genes and transcription factors involved, represents
β-carotene in cauliflower. The Osor gene, an ortholog some of the main challenges for further improving the
of the Orange (Or) gene in cauliflower was targeted in nutritional quality of rice.
rice using CRISPR/Cas9. This genome editing resulted Omics approaches for nutritious rice
in enhanced β-carotene accumulation in rice (Endo et al,
2019). Furthermore, five rice carotenoid genes The omics approach is considered a sustainable, safe,
(OsCYP97A4, OsDSM2, OsCCD4a, OsCCD4b, and and efficient method for crop improvement. Researchers
OsCCD7) were knocked out by CRISPR/Cas9, leading can detect and measure vairous biological molecules
Sundus ZAFAR and XU Jianlong. Recent Advances to Enhance Nutritional Quality of Rice

like DNA, RNA, proteins, ions, and metabolites in patterns of phenolic compounds, anthocyanin, and
living systems using omics approaches such as genetics, antioxidants in two high-yielding rice lines [KDML105
genomics, metabolomics, proteomics and ionomics and Mali Daeng (MD)], finding that the concentrations
(Deshmukh et al, 2010; Chaudhary et al, 2015). of these compounds are higher in red rice (MD) than
in white rice (KDML105) (Maksup et al, 2018).
Transcriptomics approach Proteomics methods also assess changes in gene
Transcriptomics is the study of RNA expression expression in transgenics due to translation activities
profiles inside the cell addressing both coding and or food nutritional quality. Ramli and Md Zin (2015)
non-coding RNA sequences. Many techniques such as analyzed the expression levels of seed storage proteins
RNA sequencing and microarrays have been developed and their association with the nutritional quality of
to study gene expression in various crops under rice varieties. They also investigated the glycomic and
different conditions (Pandit et al, 2018). Initial efforts proteomic contents of chalky rice grains under high-
to study whole transcriptomes were initiated in the temperature stress, revealing that starch degradation,
1990s (Lowe et al, 2017). Using RNA-Seq, whole- rather than starch synthesis, is involved in rice
chalkiness (Kaneko et al, 2016). Thus, proteome analysis,

f
genome transcription profiles of indica and japonica

oo
subspecies were obtained to understand the complexity combined with crop genetics, provides insights into
of the rice transcriptome. The analysis of large-scale protein content and its genes in agronomical parts of
RNA-Seq has improved rice genome annotation by plants under variable conditions.

r
discovering 1 584 novel peptides and 101 new loci
matched by novel peptides (Ren et al, 2019). However,
to date, less efforts have been made in exploring the -p Metabolomics approach
Metabolomics is a qualitative and quantitative study of
re
nutritional quality of rice using the transcriptomics small molecules in a biological system. Metabolomics
approach. analyses have shown the differences in bioactive
lP

compounds in various cooked and uncooked rice

Ionomics approach
varieties. One study identified about 3 097 compounds
and single nucleotide polymorphisms (SNPs) for
na

Ionomics is the study of mineral nutrients and the

composition of trace elements in diverse physiological genes regulating metabolic pathways of nutritional
environments and developmental phases of plants. It is significance (Heuberger et al, 2010). This study
ur

an efficient omics approach that can efficiently identify investigated the effect of SNPs on rice nutritional
the genes and gene networks, regulating the ionome compounds like vitamin E and phenolics, as well as
Jo

using various techniques such as X-ray crystallography, changes in the metabolome of cooked rice. Likewise,
inductively coupled plasma mass spectrometry, and metabolomics profiling of normal and giant embryo
neutron activation analysis (Huang and Salt, 2016). rice was performed, suggesting that giant embryo has
Furthermore, a genome-wide association study (GWAS) better quality due to its higher bioactive compounds
was conducted on 17 mineral elements in grains from (Zhao et al, 2019). However, the main chanllenge in
529 rice accessions for ionome analysis (Yang et al, metabolomics is extracting and interpreting vast
2018), providing insights into changes in mineral amounts of data in a biological context.
compositions and micronutrients across rice accessions.
Nutrigenomics study
Previously, a study was also performed to identify rice
germplasm with increased mineral content, nutritional Nutrigenomics is a study that investigates the effect of
quality and trace elements of 1 763 rice accessions for nutrient intake on human health. Its primary goal is to
whole grains (Pinson et al, 2015). Such efforts are enhance food quality by improving macro- and micro-
essential to explore genetic resources and understand nutrients in cereals, fruits, and vegetables or by integrating
ionomic dynamics in rice tissue. bioactive compounds into crop plants (Tran and
Kumar, 2016). Imam and Ismail (2015) investigated
Proteomics study
the nutrigenomic effects of bioactive compounds and
Proteomics mainly explores the nutritional quality of their functional properties to identify their roles in
rice by understanding the expression of bioactive healthy diets. However, despite being a promising
compounds. Sarkar et al (2015) performed comparative avenue for personalized disease prevention, nutrigenomics
proteomics to evaluate the differences in the expression faces limitations related to individual’s dietary habits,
 Rice Science

as well as cost and time-consuming protocols (Castle challenging to improve these quality traits through
and Ries, 2009). Although the significance of conventional breeding methods. The development of
nutrigenomics is widely recognized, only a few efforts genomic tools, such as molecular markers, offers
have been made towards integrating it into the crop practical ways to enhance the efficacy of plant breeding
improvement programs. for transferring these quantitatively inherited traits.
Omics approaches have revealed the molecular The application of marker-assisted selection has become
mechanisms of grain quality, contributing to the possible for identifying precise genomic regions/QTLs
enhancement of nutritional quality. However, the controlling nutritional traits by adopting molecular
genetic architecture underlying rice nutritional quality markers (Choudhary et al, 2007).
traits such as chalkiness and nutritional quality is
complex to dissect. Currently, rational design for these QTLs for micro-nutrients
traits seems complicated, and the practical implementation Many genetic mapping populations have been developed
of these designs still needs improvement. Furthermore, to dissect Fe- and Zn-related traits. Numerous new
this approach is expensive and requires additional QTLs for Fe and Zn contents have been mapped using
support from bioinformatics platforms.

f
SSR (simple sequence repeat) markers in double-

oo
Breeding techniques for nutritious rice haploid and BIL (backcross inbred line) populations
(Dixit et al, 2019; Islam et al, 2020; Jeong et al, 2020;
Biofortification of crops through breeding involves Pradhan et al, 2020) as summarized in Table 4.

r
identifying genetic resources with high nutrient content Stangoulis et al (2007) mapped two QTLs for Zn and
from existing germplasm. However, quality traits are
polygenic and quantitatively controlled, making it -p three QTLs for Fe on chromosomes 1, 2, 8, and 12.
Garcia-Oliveira et al (2009) identified one main effect
re
lP

Table 4. QTLs associated to different nutritional traits in rice.

Trait Chromosome QTL PVE (%) Marker Population Reference

na

Zn 1, 12 qZn.1, qZn.12 15, 13 SSR DHs Stangoulis et al, 2007

Zn 8 qZn8-1 11‒19 SSR (RM152) ILs Garcia-Oliveira et al, 2009
Zn 2, 9, 10 qGZn9, qGZn9a, qGZn9b 15‒21 BRILs Ishikawa et al, 2017
Zn 1,12 qZn.1, qZn.12 13‒15 RM34‒RM237, RM235‒RM17 DHs James, 2007
ur

Zn 6, 8 qZn.6, qZn.8 RZ398‒RM204, RM25‒R1629 RILs Lu et al, 2008

Zn 2, 5, 9, 11 qZn2.1, qZn5.1, qZn9.1, qZn11.1 8.6‒27.7 DHs Descalsota-Empleo et al,
Jo

2019
Zn 3, 7, 12 qZn3, qZn7.3, qZn12.2 29‒71 RM7‒RM517, RM501‒OsZip, RILs Anuradha et al, 2012
RM260‒RM7102
Fe 2, 12 qFe.2, qFe.12 14‒18 RM53‒RM300 DHs James, 2007
Fe 2, 8, 12 qFe.2, qFe.8, qFe.12 17, 18, 14 Stangoulis et al, 2007
Fe 1, 11 qFe.1, qFe.11 RM259‒RM243, RZ536‒TEL3 RILs Lu et al, 2008
Fe 5, 6 qFC-5, qFC-6 RG480‒RM274, RM190‒RZ516 RILs Yu et al, 2009
Fe 1, 5, 7, 12 qFe1.1, qFe5.1, qFe7.1, qFe12.2, 29‒71 RM243‒RM48, RM574‒RM122, RILs Anuradha et al, 2012
qFe12.1 RM234‒RM248, M260‒RM7102,
RM17‒RM260
MAC-Ca 1, 5 qCa1-1, qCa5-1 RM6480, RM598 ILs Garcia-Oliveira et al, 2009
MAC-Mg 6, 10, 11 qMg.6, qMg.10, qMg.11 OSR21, RM467, RM332 LT/TL-RILs Zhang et al, 2014
LT-RILs
LT/TL-RILs
MAC-P 1 qP.1 RM3411 LT/TL-RILs Zhang et al, 2014
MIC-Mn 7 qMn.7 RM214 LT/TL-RILs Zhang et al, 2014
Folate 2, 1 qQTF-3-1, qQTF-3-2, qQTF-3-3 7. 8, RIL, BIL Dong et al, 2014b
11.1‒15.8,
25.3
GPC 6, 7 qPc.6, qPc.7 13.0, 6 C952-Wx, R1245‒RM234 RILs Tan et al, 2001
GPC 6 qPc.6 4.8‒15.0 DHs Aluko et al, 2004
GPC 1 qPC-1 10.0‒15.4 CSSL Yang et al, 2015
PC 3 qPC-3 RM251‒RM282 RILs Yu et al, 2009
PC 1 qPr1 RM493‒RM562 RILs Zhong et al, 2011
GPC 1, 2, 7 qGPC1.1, qSGPC2.1, qSGPC7.1 13, 14, 7.8 Chattopadhyay et al, 2019
Zn, Zinc; Fe, Iron; MAC, MIC, GPC, PC, PVE, DH, IL, BRIL, RIL, LT, TL, CSSL
Sundus ZAFAR and XU Jianlong. Recent Advances to Enhance Nutritional Quality of Rice

QTL for Zn, flanked by RM152 on chromosome 8. protein content, which are important for the biosynthesis
Norton et al (2010) also identified Fe (qFe-1) and of some amino acids/proteins in rice grains (Sun et al,
other QTLs related to elements like Zn, Fe, Mn and 2019). This study offers a new approach to phenotype
Ca. Dong et al (2014b) identified three QTLs for folate important biochemical traits for determining rice
(qQTF-3-1, qQTF-3-2, and qQTF-3). Kumar et al (2014) quality that could be used in genetic studies.
used an F4 population (579 individuals) and detected
one Zn QTL and five Fe QTLs. These detected QTLs QTLs for PA
can significantly enhance the efficiency of breeding Significant reductions in nutrient element bioavailability
programs by improving Zn and Fe contents in rice. have been observed due to PA present in the form of
Swamy et al (2018) identified eight QTLs for Zn and phytate (Gemede, 2014). Liu et al (2005) identified a
one QTL for Fe on chromosomes 2, 3, 4, 6, 8, 11, and broad range of PA contents, ranging from 0.685%–
12, and also detected candidate genes near Zn-related 1.030% by assaying 72 rice varieties. They also observed
genes (OsZIP and OsNRAMP) that will help in the major effects of varieties, environments, and their
breeding using markers to improve Zn content in rice. interactions on PA. While Liu et al (2007) reported

f
genetic variation for PA, Stangoulis et al (2007) detected
QTLs for proteins and amino acids

oo
two QTLs for grain phytate from an IR64 × Azucena
Rice grain protein content (GPC) significantly affects double haploid population. Mn, Fe, and Zn contents
the nutritional value and taste of cooked rice. About showed different genetic regulation due to their location

r
80 QTLs for GPC have been identified and mapped on on other chromosomes compared with the phytate
all 12 rice chromosomes. Aluko et al (2004) detected
four QTLs for GPC, with one of them, named Pro6, -p position. Thus, there is an opportunity to discover
segregants containing high Fe, Zn, and Mn and low
re
being related to the Wx gene that affects rice quality PA content. The QTLs detected in these studies can be
(Table 4). Yang et al (2015) identified seven QTLs in valuable in improving rice nutritional quality.
lP

three locations using chromosome segment substitution In the recent years, significant advancements have
lines (CSSL), and only qPC-1 was consistently found been made in genetic studies on Zn and Fe contents,
in all three locations. Chattopadhyay et al (2019) used vitamins and minerals, protein and amino acids, and
na

a 40 K Affymetrix custom SNP array and detected QTLs linked to these traits. However, further research
three QTLs for GPC (qGPC1.1, qSGPC2.1, and is needed on processing and curative properties. The
ur

qSGPC7.1). Several QTLs for protein content have availability of gene-based markers and other advanced
been identified in rice, including qPro.1, qPC1.2, and tools will assist breeders in selecting specific gene
Jo

qPC1 on chromosome 1, and qAAC7.1, qPC6.2, and alleles that play a critical role in the nutritional traits
qPro-2 on chromosomes 7, 6, and 2, respectively of rice. Although breeding methods can be resource-
(Kinoshita et al, 2017; Islam et al, 2020; Jang et al, intensive, the benefits from this approach have proven
2020). Numerous studies have used linkage methods to make it cost-effective. Thus, effective steps such as
to improve rice amino acid composition by providing increased financial assistance, training of plant breeders
genetic information from different mapping populations. and technicians, improved infrastructure, and the
Yoo (2017) mapped six main effect QTLs located on availability of raw materials should be taken to facilitate
chromosome 3, contributing to 10.2%–12.4% of the adoption of breeding methods worldwide.
phenotypic variations for six amino acids. The QTL
Agronomic biofortification
cluster (qAla3, qVal3, qPhe3, qIle3, and qLeu3) was
associated with five amino acids. These results are Apart from breeding approaches and transgenic methods,
valuable for marker-assisted breeding programs and agronomic biofortification is a technique based on
the identification of candidate genes that enhance applying fertilizers to enhance the nutritional quality
amino acid content in rice. It is also possible to of rice. Generally, plants absorb micro-nutrients from
phenotype the biochemical attributes of rice using the soil, and applying fertilizers in the soil and foliar
hyperspectral techniques and this data for GWAS. The methods can address the deficiency of micro-nutrients
Normalized difference spectral index (NDSI) has a in plants (Garg et al, 2018). An agronomic practice
high correlation with protein content. Based on was conducted in five countries to enhance the contents
GWAS analysis, NDSI identified all the 43 genes and of Zn, iodine (I), Fe, and selenium (Se) in rice grains
annotated 3 pathways that are precisely the same as through foliar application. On average, brown rice
 Rice Science

shows an increase in Zn (21.4–28.1 mg/kg) and I ACKNOWLEDGEMENTS

(11.0–204.0 mg/kg) contents by foliar application of Zn
This study was funded by the Hainan Yazhou Bay Seed Lab
and I. In contrast, a 26.8 mg/kg increase in Zn content Project, China (Grant No. B21HJ0216), the Key Research and
and an 181.0 mg/kg increase in I were reported with Development Project of Hainan Province, China (Grant No.
the cocktail application (Zn + I + Fe + Se) application ZDYF2021XDNY128), and the Agricultural Science and
(Prom-u-thai et al, 2020). Furthermore, high Zn Technology Innovation Program and the Cooperation and
accumulation also increases Fe accumulation and Innovation Mission, China (Grant No. CAAS-ZDXT202001).
reduces PA content (Saha et al, 2017). Different
REFERENCES
locations can also alter the levels of micro- and
macro-nutrients in rice by application of fertilizers Abe K, Araki E, Suzuki Y, Toki S, Saika H. 2018. Production of
(Chandel et al, 2010). high oleic/low linoleic rice by genome editing. Plant Physiol
Agronomic biofortification of crops with Fe, Se, Biochem, 131: 58–62.
and Zn using organic fertilizers holds remarkable Aluko G, Martinez C, Tohme J, Castano C, Bergman C, Oard J H.
2004. QTL mapping of grain quality traits from the interspecific
potential in addressing nutrient deficiencies. However,
cross Oryza sativa × O. glaberrima. Theor Appl Genet, 109(3):
the success of agronomic biofortification depends on

f
630–639.

oo
various factors, such as field experiments, the influence Anuradha K, Agarwal S, Rao Y V, Rao K V, Viraktamath B C,
of soil properties on micro-nutrients, and the responses Sarla N. 2012. Mapping QTLs and candidate genes for iron and
of crops or different varieties of the same crop on Zn, zinc concentrations in unpolished rice of Madhukar × Swarna

r
Se, and Fe fertilization. These factors still need to be RILs. Gene, 508(2): 233–240.
explored in detail to achieve desired results.
-p
Bai C, Capell T, Berman J, Medina V, Sandmann G, Christou P,
Zhu C F. 2016. Bottlenecks in carotenoid biosynthesis and
re
Conclusion and future prospects accumulation in rice endosperm are influenced by the precursor-
product balance. Plant Biotechnol J, 14(1): 195–205.
Enhancing the nutritional quality of crops is a
lP

Banakar R, Alvarez Fernández Á, Abadía J, Capell T, Christou P.

promising and cost-effective agricultural strategy to 2017. The expression of heterologous Fe(III) phytosiderophore
improve the nutritional status of undernourished transporter HvYS1 in rice increases Fe uptake, translocation and
populations worldwide. However, crop biofortification seed loading and excludes heavy metals by selective Fe transport.
na

presents significant challenges. Therefore, there is a Plant Biotechnol J, 15(4): 423–432.

Bashir K, Takahashi R, Akhtar S, Ishimaru Y, Nakanishi H,
need to combine recent technologies such as plant
Nishizawa N K. 2013. The knockdown of OsVIT2 and MIT
ur

breeding, genetic engineering, omics techniques, and affects iron localization in rice seed. Rice, 6(1): 31.
agronomic approaches. Breeding approaches have Bhattacharya S, Sengupta S, Karmakar A, Sarkar S N, Gangopadhyay
Jo

been widely used due to their high success rate and G, Datta K, Datta S K. 2019. Genetically engineered rice with
acceptance. While transgenic plants have also shown appA gene enhanced phosphorus and minerals. J Plant Biochem
promise, they face challenges such as cost, public Biotechnol, 28(4): 470–482.
acceptance, and lengthy approval processes in various Birla D S, Malik K, Sainger M, Chaudhary D, Jaiwal R, Jaiwal P K.
2017. Progress and challenges in improving the nutritional
countries. Despite these challenges, biofortified crops
quality of rice (Oryza sativa L.). Crit Rev Food Sci Nutr, 57(11):
offer great potential to combat micronutrient malnutrition,
2455–2481.
especially in developing countries. In recent years, Biswas S, Ibarra O, Shaphek M, Molina-Risco M, Faion-Molina M,
genome editing research has made significant progress Bellinatti-Della Gracia M, Thomson M J, Septiningsih E M.
in enriching food crops with higher nutritional value, 2022. Increasing the level of resistant starch in ‘Presidio’ rice
offering a bright future in addressing malnutrition through multiplex CRISPR-Cas9 gene editing of starch branching
challenges. Additionally, the integration of omics tools, enzyme genes. Plant Genome, 16(2): e20225.
including transcriptomics, metabolomics, proteomics, Boue S M, Daigle K W, Chen M H, Cao H P, Heiman M L. 2016.
Antidiabetic potential of purple and red rice (Oryza sativa L.)
and ionomics, is pivotal in developing highly nutritious
bran extracts. J Agric Food Chem, 64(26): 5345–5353.
rice varieties. However, there is a widening gap Brar D S, Virk P S, Grewal D, Slamet-Loedin I, Fitzgerald M,
between the development of omics resources and their Khush G S. 2012. Breeding rice varieties with improved grain
effective utilization, requiring more efforts to bridge and nutritional quality. Qual Assur Saf Crop, 4(3): 137.
this gap. The recent advancements in genetic engineering, Castle D, Ries N. 2009. Nutrition and Genomics: Issues of Ethics,
agronomic biofortification, and omics approaches can Law, Regulation and Communication. San Diego, CA, USA:
complement breeding programs and contribute to Academic Press.
improving the nutritional quality of rice. Chaiyasit W, Elias R J, McClements D J, Decker E A. 2007. Role
of physical structures in bulk oils on lipid oxidation. Crit Rev
Sundus ZAFAR and XU Jianlong. Recent Advances to Enhance Nutritional Quality of Rice

Food Sci Nutr, 47(3): 299–317. Ali F. 2019. Applications of the CRISPR/Cas9 system for rice
Chandel G, Banerjee S, See S, Meena R, Sharma D J, Verulkar S B. grain quality improvement: Perspectives and opportunities. Int J
2010. Effects of different nitrogen fertilizer levels and native soil Mol Sci, 20(4): 888.
properties on rice grain Fe, Zn and protein contents. Rice Sci, Fitri I G S, Nurhasanah, Handoyo T. 2021. Genetic and phytochemical
17(3): 213–227. analysis of Indonesian black rice cultivars. J Crop Sci Biotechnol,
Chattopadhyay K, Behera L, Bagchi T B, Sardar S S, Moharana N, 24(5): 567–578.
Patra N R, Chakraborti M, Das A, Marndi B C, Sarkar A, Fongfon S, Pusadee T, Prom-u-thai C, Rerkasem B, Jamjod S.
Ngangkham U, Chakraborty K, Bose L K, Sarkar S, Ray S, 2021. Diversity of purple rice (Oryza sativa L.) landraces in
Sharma S. 2019. Detection of stable QTLs for grain protein northern Thailand. Agronomy, 11(10): 2029.
content in rice (Oryza sativa L.) employing high throughput Garcia-Oliveira A L, Tan L B, Fu Y C, Sun C Q. 2009. Genetic
phenotyping and genotyping platforms. Sci Rep, 9: 3196. identification of quantitative trait loci for contents of mineral
Chaudhary J, Patil G B, Sonah H, Deshmukh R K, Vuong T D, nutrients in rice grain. J Integr Plant Biol, 51(1): 84–92.
Valliyodan B, Nguyen H T. 2015. Expanding omics resources Garg M, Sharma N, Sharma S, Kapoor P, Kumar A, Chunduri V,
for improvement of soybean seed composition traits. Front Plant Arora P. 2018. Biofortified crops generated by breeding, agronomy,
Sci, 6: 1021. and transgenic approaches are improving lives of millions of
Cheah T W, Ismail I, Sidek N M, Wagiran A, Abdullah R. 2013. people around the world. Front Nutr, 5: 12.
Biosynthesis of very long polyunsaturated Omega-3 and Gemede H M. 2014. Potential health benefits and adverse effects

f
oo
Omega-6 fatty acids in transgenic japonica rice (Oryza sativa associated with phytate in foods. Food Sci Qual Manage, 27: 45.
L.). Aust J of Crop Sci, 7(9): 1227–1234. Ghasemzadeh A, Karbalaii M T, Jaafar H Z E, Rahmat A. 2018.
Choudhary K, Choudhary O P, Shekhawat N S. 2007. Marker Phytochemical constituents, antioxidant activity, and antiproliferative

r
assisted selection: A novel approach for crop improvement. properties of black, red, and brown rice bran. Chem Central J,
Am-Eurasian J Agron, 1(2): 26–30.
Darnton-Hill I, Webb P, Harvey P W, Hunt J M, Dalmiya N, Chopra
M, Ball M J, Bloem M W, de Benoist B. 2005. Micronutrient -p 12(1): 17.
Goldstein P. 2018. HarvestPlus talks zinc rice with farmers in
southeastern Bangladesh. [2019-04-14]. https://www.harvestplus.
re
deficiencies and gender: Social and economic costs. Am J Clin org/knowledgemarket/in-the-news/harvestplus-talks-zinc-rice-fa
Nutr, 81(5): 1198S–1205S. rmerssoutheasternbangladesh.
lP

Descalsota-Empleo G I, Amparado A, Inabangan-Asilo M A, Tesoro Goto F, Yoshihara T, Shigemoto N, Toki S, Takaiwa F. 1999. Iron
F, Stangoulis J, Reinke R, Swamy B P M. 2019. Genetic fortification of rice seed by the soybean ferritin gene. Nat
mapping of QTL for agronomic traits and grain mineral elements Biotechnol, 17(3): 282–286.
na

in rice. Crop J, 7(4): 560–572. Guo D S, Ling X T, Zhou X G, Li X, Wang J Y, Qiu S, Yang Y W,
Deshmukh R, Singh A, Jain N, Anand S, Gacche R, Singh A, Gaikwad Zhang B L. 2020. Evaluation of the quality of a high-resistant
K, Sharma T, Mohapatra T, Singh N. 2010. Identification of starch and low-glutelin rice (Oryza sativa L.) generated through
ur

candidate genes for grain number in rice (Oryza sativa L.). Funct CRISPR/Cas9-mediated targeted mutagenesis. J Agric Food
Integr Genom, 10(3): 339–347. Chem, 68(36): 9733–9742.
Jo

Dias J S, Ortiz R. 2012. Transgenic vegetable breeding for nutritional Gupta R K, Gangoliya S S, Singh N K. 2015. Reduction of phytic
quality and health benefits. Food Nutr Sci, 3(9): 1209–1219. acid and enhancement of bioavailable micronutrients in food
Dixit S, Singh U M, Abbai R, Ram T, Singh V K, Paul A, Virk P S, grains. J Food Sci Technol, 52(2): 676–684.
Kumar A. 2019. Identification of genomic region(s) responsible Ha S H, Liang Y S, Jung H, Ahn M J, Suh S C, Kweon S J, Kim D
for high iron and zinc content in rice. Sci Rep, 9: 8136. H, Kim Y M, Kim J K. 2010. Application of two bicistronic
Dong W, Cheng Z J, Lei C L, Wang X L, Wang J L, Wang J, Wu F systems involving 2A and IRES sequences to the biosynthesis of
Q, Zhang X, Guo X P, Zhai H Q, Wan J M. 2014a. Overexpression carotenoids in rice endosperm. Plant Biotechnol J, 8(8): 928–938.
of folate biosynthesis genes in rice (Oryza sativa L.) and Han S W, Chee K M, Cho S J. 2015. Nutritional quality of rice
evaluation of their impact on seed folate content. Plant Foods bran protein in comparison to animal and vegetable protein.
Hum Nutr, 69(4): 379–385. Food Chem, 172: 766–769.
Dong W, Cheng Z J, Xu J L, Zheng T Q, Wang X L, Zhang H Z, Heuberger A L, Lewis M R, Chen M H, Brick M A, Leach J E,
Wang J, Wan J M. 2014b. Identification of QTLs underlying Ryan E P. 2010. Metabolomic and functional genomic analyses
folate content in milled rice. J Integr Agric, 13(9): 1827–1834. reveal varietal differences in bioactive compounds of cooked
Endo A, Saika H, Takemura M, Misawa N, Toki S. 2019. A novel rice. PLoS One, 5(9): e12915.
approach to carotenoid accumulation in rice callus by mimicking Huang X Y, Salt D E. 2016. Plant ionomics: From elemental
the cauliflower Orange mutation via genome editing. Rice, 12(1): profiling to environmental adaptation. Mol Plant, 9(6): 787–797.
81. Imam M, Ismail M. 2015. An overview on germinated brown rice
FAO. 2018. World food and agriculture: Statistical pocketbook and its nutrigenomic implications. In: Bagchi D, Swaroop A,
2018. Rome, Italy: Food and Agriculture Organization. Bagchi M. Genomics, Proteomics and Metabolomics in Nutraceuticals
FarréG, Blancquaert D, Capell T, van der Straeten D, Christou P, and Functional Foods. 2nd edn. Hoboken, NJ, USA: Wiley:
Zhu C F. 2014. Engineering complex metabolic pathways in 504–517.
plants. Annu Rev Plant Biol, 65: 187–223. Ishikawa R, Iwata M, Taniko K, Monden G, Miyazaki N, Orn C,
Fiaz S, Ahmad S, Noor M, Wang X K, Younas A, Riaz A, Riaz A, Tsujimura Y, Yoshida S, Ma J F, Ishii T. 2017. Detection of
 Rice Science

quantitative trait loci controlling grain zinc concentration using 2001. Constitutive and seed-specific expression of a maize lysine-
Australian wild rice, Oryza meridionalis, a potential genetic feedback-insensitive dihydrodipicolinate synthase gene leads to
resource for biofortification of rice. PLoS One, 12(10): e0187224. increased free lysine levels in rice seeds. Mol Breed, 8(1):
Ishimaru Y, Masuda H, Bashir K, Inoue H, Tsukamoto T, Takahashi 75–84.
M, Nakanishi H, Aoki N, Hirose T, Ohsugi R, Nishizawa N K. Lei Y, Lu L, Liu H Y, Li S, Xing F, Chen L L. 2014. CRISPR-P: A
2010. Rice metal-nicotianamine transporter, OsYSL2, is required web tool for synthetic single-guide RNA design of CRISPR-
for the long-distance transport of iron and manganese. Plant J, system in plants. Mol Plant, 7(9): 1494–1496.
62(3): 379–390. Liu H L, Yin Z J, Xiao L, Xu Y N, Qu L Q. 2012a. Identification
Islam M Z, Arifuzzaman M, Banik S, Hossain M A, Ferdous J, and evaluation of ω-3 fatty acid desaturase genes for
Khalequzzaman M, Pittendrigh B R, Tomita M, Ali M P. 2020. hyperfortifying α-linolenic acid in transgenic rice seed. J Exp
Mapping QTLs underpin nutrition components in aromatic rice Bot, 63(8): 3279–3287.
germplasm. PLoS One, 15(6): e0234395. Liu J, Hua W, Yang H L, Zhan G M, Li R J, Deng L B, Wang X F,
Iyer R, Tomar S K. 2009. Folate: A functional food constituent. J Liu G H, Wang H Z. 2012b. The BnGRF2 gene (GRF2-like gene
Food Sci, 74(9): R114–R122. from Brassica napus) enhances seed oil production through
Jang S, Han J H, Lee Y K, Shin N H, Kang Y J, Kim C K, Chin J regulating cell number and plant photosynthesis. J Exp Bot,
H. 2020. Mapping and validation of QTLs for the amino acid 63(10): 3727–3740.
and total protein content in brown rice. Front Genet, 11: 240. Liu Q L, Xu X H, Ren X L, Fu H W, Wu D X, Shu Q Y. 2007.

f
oo
Jeong O Y, Lee J H, Jeong E G, Chun A, Bombay M, Banzon Generation and characterization of low phytic acid germplasm in
Ancheta M, Ahn S N. 2020. Analysis of QTL responsible for rice (Oryza sativa L.). Theor Appl Genet, 114(5): 803–814.
grain iron and zinc content in doubled haploid lines of rice Liu X, Zhang C C, Wang X R, Liu Q Q, Yuan D Y, Pan G, Sun S

r
(Oryza sativa) derived from an intra-japonica cross. Plant Breed, S M, Tu J M. 2016. Development of high-lysine rice via
139(2): 344–355.
Jeong Y S, Ku H K, Kim J K, You M K, Lim S H, Kim J K, Ha S
H. 2017. Effect of codon optimization on the enhancement of the -pendosperm-specific expression of a foreign LYSINE RICH
PROTEIN gene. BMC Plant Biol, 16(1): 147.
Liu Z H, Cheng F M, Zhang G P. 2005. Grain phytic acid content
re
β-carotene contents in rice endosperm. Plant Biotechnol Rep, in japonica rice as affected by cultivar and environment and its
11(3): 171–179. relation to protein content. Food Chem, 89(1): 49–52.
lP

Jiang H X, Lio J, Blanco M, Campbell M, Jane J L. 2010. Long X H, Liu Q Q, Chan M L, Wang Q, Sun S S M. 2013.
Resistant-starch formation in high-amylose maize starch during Metabolic engineering and profiling of rice with increased lysine.
kernel development. J Agric Food Chem, 58(13): 8043–8047. Plant Biotechnol J, 11(4): 490–501.
na

Johnson A A T, Kyriacou B, Callahan D L, Carruthers L, Stangoulis Lowe R, Shirley N, Bleackley M, Dolan S, Shafee T. 2017.
J, Lombi E, Tester M. 2011. Constitutive overexpression of the Transcriptomics technologies. PLoS Comput Biol, 13(5): e1005457.
OsNAS gene family reveals single-gene strategies for effective Lu K Y, Li L Z, Zheng X F, Zhang Z H, Mou T M, Hu Z L. 2008.
ur

iron- and zinc-biofortification of rice endosperm. PLoS One, Quantitative trait loci controlling Cu, Ca, Zn, Mn and Fe content
6(9): e24476. in rice grains. J Genet, 87(3): 305–310.
Jo

Kaneko K, Sasaki M, Kuribayashi N, Suzuki H, Sasuga Y, Shiraya Lucca P, Hurrell R, Potrykus I. 2001. Genetic engineering
T, Inomata T, Itoh K, Baslam M, Mitsui T. 2016. Proteomic and approaches to improve the bioavailability and the level of iron in
glycomic characterization of rice chalky grains produced under rice grains. Theor Appl Genet, 102(2): 392–397.
moderate and high-temperature conditions in field system. Rice, Mahender A, Anandan A, Pradhan S K, Pandit E. 2016. Rice grain
9(1): 26. nutritional traits and their enhancement using relevant genes and
Kinoshita N, Kato M, Koyasaki K, Kawashima T, Nishimura T, QTLs through advanced approaches. SpringerPlus, 5(1): 2086.
Hirayama Y, Takamure I, Sato T, Kato K. 2017. Identification of Maksup S, Pongpakpian S, Roytrakul S, Cha-Um S, Supaibulwatana
quantitative trait loci for rice grain quality and yield-related traits K. 2018. Comparative proteomics and protein profile related to
in two closely related Oryza sativa L. subsp. japonica cultivars phenolic compounds and antioxidant activity in germinated
grown near the northernmost limit for rice paddy cultivation. Oryza sativa ‘KDML105’ and Thai brown rice ‘Mali Daeng’ for
Breed Sci, 67(3): 191–206. better nutritional value. J Sci Food Agric, 98(2): 566–573.
Kuwano M, Mimura T, Takaiwa F, Yoshida K T. 2009. Generation Masuda H, Kobayashi T, Ishimaru Y, Takahashi M, Aung M S,
of stable ‘low phytic acid’ transgenic rice through antisense Nakanishi H, Mori S, Nishizawa N K. 2013. Iron-biofortification
repression of the 1D-myo-inositol 3-phosphate synthase gene in rice by the introduction of three barley genes participated in
(RINO1) using the 18-kDa oleosin promoter. Plant Biotechnol J, mugineic acid biosynthesis with soybean ferritin gene. Front
7(1): 96–105. Plant Sci, 4: 132.
Lee S, An G. 2009. Over-expression of OsIRT1 leads to increased Mikami M, Toki S, Endo M. 2015. Comparison of CRISPR/Cas9
iron and zinc accumulations in rice. Plant Cell Environ, 32(4): expression constructs for efficient targeted mutagenesis in rice.
408–416. Plant Mol Biol, 88(6): 561–572.
Lee S, Kim Y S, Jeon U S, Kim Y K, Schjoerring J K, An G. 2012. Naqvi S, Zhu C F, Farre G, Ramessar K, Bassie L, Breitenbach J,
Activation of rice nicotianamine synthase 2 (OsNAS2) enhances Perez Conesa D, Ros G, Sandmann G, Capell T, Christou P.
iron availability for biofortification. Mol Cells, 33(3): 269–275. 2009. Transgenic multivitamin corn through biofortification of
Lee S I, Kim H U, Lee Y H, Suh S C, Lim Y P, Lee H Y, Kim H I. endosperm with three vitamins representing three distinct
Sundus ZAFAR and XU Jianlong. Recent Advances to Enhance Nutritional Quality of Rice

metabolic pathways. Proc Natl Acad Sci USA, 106(19): 7762–7767. of large-bowel health in rats. Proc Natl Acad Sci USA, 103(10):
Naqvi S, FarréG, Sanahuja G, Capell T, Zhu C F, Christou P. 2010. 3546–3551.
When more is better: Multigene engineering in plants. Trends Ren Z, Qi D, Pugh N, Li K, Wen B, Zhou R, Xu S H, Liu S Q,
Plant Sci, 15(1): 48–56. Jones A R. 2019. Improvements to the rice genome annotation
Norton G J, Deacon C M, Xiong L Z, Huang S Y, Meharg A A, through large-scale analysis of RNA-seq and proteomics data
Price A H. 2010. Genetic mapping of the rice ionome in leaves sets. Mol Cell Proteom, 18(1): 86–98.
and grain: Identification of QTLs for 17 elements including Ricachenevsky F K, Vasconcelos M W, Shou H X, Johnson A A T,
arsenic, cadmium, iron and selenium. Plant Soil, 329(1): 139–153. Sperotto R A. 2019. Editorial: Improving the nutritional content
Oakley Jr G P, Bell K N, Weber M B. 2004. Recommendations for and quality of crops: Promises, achievements, and future challenges.
accelerating global action to prevent folic acid-preventable birth Front Plant Sci, 10: 738.
defects and other folate-deficiency diseases: Meeting of experts Ritchie H, Roser M. 2017. Micronutrient Deficiencies. Geneva,
on preventing folic acid-preventable neural tube defects. Birth Swiss: World Health Organization (WHO).
Defects Res A Clin Mol Teratol, 70(11): 835–837. Saha S, Chakraborty M, Padhan D, Saha B, Murmu S, Batabyal K,
Paine J A, Shipton C A, Chaggar S, Howells R M, Kennedy M J, Seth A, Hazra G C, Mandal B, Bell R W. 2017. Agronomic
Vernon G, Wright S Y, Hinchliffe E, Adams J L, Silverstone A biofortification of zinc in rice: Influence of cultivars and zinc
L, Drake R. 2005. Improving the nutritional value of Golden application methods on grain yield and zinc bioavailability.
Rice through increased pro-vitamin A content. Nat Biotechnol, Field Crops Res, 210: 52–60.

f
oo
23(4): 482–487. Saravanan P, Davidson N C, Schmidt E B, Calder P C. 2010.
Pandit A A, Shah R A, Husaini A M. 2018. Transcriptomics: A Cardiovascular effects of marine omega-3 fatty acids. Lancet,
time-efficient tool with wide applications in crop and animal 376: 540–550.

r
biotechnology. J Pharmacogn Phytochem, 7: 1701–1704. Sarkar A, Singh A A, Agrawal S B, Ahmad A, Rai S. P. 2015.
Parashar R, Afzal S, Mishra M, Singh N K. 2023. Improving
biofortification success rates and productivity through zinc
nanocomposites in rice (Oryza sativa L.). Environ Sci Pollut Res, -pCultivar specific variations in antioxidative defense system,
genome and proteome of two tropical rice cultivars against ambient
and elevated ozone. Ecotoxicol Environ Saf, 115: 101–111.
re
30(15): 44223–44233. Schramm R, Abadie A, Hua N, Xu Z M, Lima M. 2007.
Paul S, Ali N, Gayen D, Datta S K, Datta K. 2012. Molecular Fractionation of the rice bran layer and quantification of vitamin
lP

breeding of Osfer2 gene to increase iron nutrition in rice grain. E, oryzanol, protein, and rice bran saccharide. J Biol Eng, 1(1): 9.
GM Crops Food, 3(4): 310–316. Siddiqui F, Salam R A, Lassi Z S, Das J K. 2020. The intertwined
Perera I, Seneweera S, Hirotsu N. 2018. Manipulating the phytic relationship between malnutrition and poverty. Front Public
na

acid content of rice grain toward improving micronutrient Health, 8: 453.

bioavailability. Rice, 11: 4. Smith M R, Myers S S. 2019. Global health implications of
Pinson S R M, Tarpley L, Yan W G, Yeater K, Lahner B, Yakubova nutrient changes in rice under high atmospheric carbon dioxide.
ur

E, Huang X Y, Zhang M, Guerinot M L, Salt D E. 2015. Worldwide GeoHealth, 3(7): 190–200.

genetic diversity for mineral element concentrations in rice grain. Stangoulis J C R, Huynh B L, Welch R M, Choi E Y, Graham R D.
Jo

Crop Sci, 55(1): 294–311. 2007. Quantitative trait loci for phytate in rice grain and their
Pradhan S K, Pandit E, Pawar S, Naveenkumar R, Barik S R, relationship with grain micronutrient content. Euphytica, 154(3):
Mohanty S P, Nayak D K, Ghritlahre S K, Sanjiba Rao D, Reddy J 289–294.
N, Patnaik S S C. 2020. Linkage disequilibrium mapping for Storozhenko S, De Brouwer V, Volckaert M, Navarrete O, Blancquaert
grain Fe and Zn enhancing QTLs useful for nutrient dense rice D, Zhang G F, Lambert W, van der Straeten D. 2007. Folate
breeding. BMC Plant Biol, 20(1): 57. fortification of rice by metabolic engineering. Nat Biotechnol,
Prom-u-thai C, Rerkasem B. 2020. Rice quality improvement: A 25(11): 1277–1279.
review. Agron Sustain Dev, 40: 28. Sun D W, Cen H Y, Weng H Y, Wan L, Abdalla A, El-Manawy A
Prom-u-thai C, Rashid A, Ram H, Zou C Q, Guilherme L R G, I, Zhu Y M, Zhao N, Fu H W, Tang J, Li X L, Zheng H K, Shu
Corguinha A P B, Guo S W, Kaur C, Naeem A, Yamuangmorn Q Y, Liu F, He Y. 2019. Using hyperspectral analysis as a
S, Ashraf M Y, Sohu V S, Zhang Y Q, Dias Martins F A, Jumrus S, potential high throughput phenotyping tool in GWAS for protein
Tutus Y, Yazici M A, Cakmak I. 2020. Simultaneous biofortification content of rice quality. Plant Methods, 15: 54.
of rice with zinc, iodine, iron and selenium through foliar Sun Y W, Jiao G A, Liu Z P, Zhang X, Li J Y, Guo X P, Du W M,
treatment of a micronutrient cocktail in five countries. Front Du J L, Francis F, Zhao Y D, Xia L Q. 2017. Generation of high-
Plant Sci, 11: 589835. amylose rice through CRISPR/Cas9-mediated targeted mutagenesis
Raboy V. 2003. myo-Inositol-1,2,3,4,5,6-hexakisphosphate. Phytochemistry, of starch branching enzymes. Front Plant Sci, 8: 298.
64(6): 1033–1043. Swamy B P M, Descalsota G I L, Nha C T, Amparado A, Inabangan-
Ramli N S, Md Zin N H. 2015. Proteomic analysis of rice seed Asilo M A, Manito C, Tesoro F, Reinke R. 2018. Identification
storage proteins in relation to nutrient quality of three different of genomic regions associated with agronomic and biofortification
commercial rice types. J Teknol, 77(24): 7–11. traits in DH populations of rice. PLoS One, 13(8): e0201756.
Regina A, Bird A, Topping D, Bowden S, Freeman J, Barsby T, Swanson D, Block R, Mousa S A. 2012. Omega-3 fatty acids EPA
Kosar-Hashemi B, Li Z Y, Rahman S, Morell M. 2006. High- and DHA: Health benefits throughout life. Adv Nutr, 3(1): 1–7.
amylose wheat generated by RNA interference improves indices Tan Y F, Sun M, Xing Y Z, Hua J P, Sun X L, Zhang Q F, Corke H.
 Rice Science

2001. Mapping quantitative trait loci for milling quality, protein carotenoid catabolic genes in rice fails to boost carotenoid
content and color characteristics of rice using a recombinant accumulation, but reveals a mutation in strigolactone biosynthesis.
inbred line population derived from an elite rice hybrid. Theor Plant Cell Rep, 36(10): 1533–1545.
Appl Genet, 103(6): 1037–1045. Yang Y H, Guo M, Li R D, Shen L, Wang W, Liu M, Zhu Q, Hu Z,
Tian Y S, Wang B, Peng R H, Xu J, Li T, Fu X Y, Xiong A S, Gao He Q W, Xue Y, Tang S Z, Gu M H, Yan C J. 2015.
J J, Yao Q H. 2019. Enhancing carotenoid biosynthesis in rice Identification of quantitative trait loci responsible for rice grain
endosperm by metabolic engineering. Plant Biotechnol J, 17(5): protein content using chromosome segment substitution lines
849–851. and fine mapping of qPC-1 in rice (Oryza sativa L.). Mol Breed,
Tiwari G J, Liu Q, Shreshtha P, Li Z Y, Rahman S. 2016. RNAi- 35(6): 130.
mediated down-regulation of the expression of OsFAD2-1: Ye X D, Al-Babili S, Klöti A, Zhang J, Lucca P, Beyer P, Potrykus
Effect on lipid accumulation and expression of lipid biosynthetic I. 2000. Engineering the provitamin A (β-carotene) biosynthetic
genes in the rice grain. BMC Plant Biol, 16(1): 189. pathway into (carotenoid-free) rice endosperm. Science, 287:
Tran L S P, Kumar R. 2016. Plant quality improvement and 303–305.
nutrigenomics. Curr Genom, 17(3): 153–154. Yin Z J, Liu H L, Dong X B, Tian L H, Xiao L, Xu Y N, Qu L Q.
Trijatmiko K R, Dueñas C, Tsakirpaloglou N, Torrizo L, Arines F 2014. Increasing α-linolenic acid content in rice bran by embryo-
M, Adeva C, Balindong J, Oliva N, Sapasap M V, Borrero J, specific expression of ω3/Δ15-desaturase gene. Mol Breed, 33(4):
Rey J, Francisco P, Nelson A, Nakanishi H, Lombi E, Tako E, 987–996.

f
oo
Glahn R P, Stangoulis J, Chadha-Mohanty P, Johnson A A T, Yoo S C. 2017. Quantitative trait loci controlling the amino acid
Tohme J, Barry G, Slamet-Loedin I H. 2016. Biofortified indica content in rice (Oryza sativa L.). J Plant Biotechnol, 44(4):
rice attains iron and zinc nutrition dietary targets in the field. Sci 349–355.

r
Rep, 6: 19792. Yu T Q, Jiang W Z, Ham T H, Chu S H, Lestari F, Lee J H, Kim M
Utasee S, Jamjod S, Lordkaew S, Prom-U-Thai C. 2022. Improve
anthocyanin and zinc concentration in purple rice by nitrogen
and zinc fertilizer application. Rice Sci, 29(5): 435–450. -pK, Xu F R, Han L Z, Dai L Y, Koh H J. 2008. Comparison of
grain quality traits between japonica rice cultivars from Korea
and Yunnan Province of China. J Crop Sci Biotechnol, 11(2):
re
Vasconcelos M, Datta K, Oliva N, Khalekuzzaman M, Torrizo L, 135–140.
Krishnan S, Oliveira M, Goto F, Datta S K. 2003. Enhanced iron Yu Y H, Li G, Fan Y Y, Zhang K Q, Min J, Zhu Z W, Zhuang J Y.
lP

and zinc accumulation in transgenic rice with the ferritin gene. 2009. Genetic relationship between grain yield and the contents
Plant Sci, 164(3): 371–378. of protein and fat in a recombinant inbred population of rice. J
Wang M, Wang S B, Liang Z, Shi W M, Gao C X, Xia G M. 2018. Cereal Sci, 50(1): 121–125.
na

From genetic stock to genome editing: Gene exploitation in Zaplin E S, Liu Q, Li Z Y, Butardo V M, Blanchard C L, Rahman
wheat. Trends Biotechnol, 36(2): 160–172. S. 2013. Production of high oleic rice grains by suppressing the
Wijaya D N, Susanto F A, Purwestri Y A, Ismoyowati D, Nuringtyas expression of the OsFAD2-1 gene. Funct Plant Biol, 40(10):
ur

T R. 2017. NMR metabolite comparison of local pigmented rice 996–1004.

in Yogyakarta. Indones J Biotechnol, 22(2): 68–75. Zhang M, Pinson S R M, Tarpley L, Huang X Y, Lahner B, Yakubova
Jo

Wong H W, Liu Q, Sun S M. 2015. Biofortification of rice with E, Baxter I, Guerinot M L, Salt D E. 2014. Mapping and
lysine using endogenous histones. Plant Mol Biol, 87(3): 235–248. validation of quantitative trait loci associated with concentrations
Wu F F, Yang N, Touré A, Jin Z Y, Xu X M. 2013. Germinated of 16 elements in unmilled rice grain. Theor Appl Genet, 127(1):
brown rice and its role in human health. Crit Rev Food Sci Nutr, 137–165.
53(5): 451–463. Zhao G C, Zhang Y X, Sun S Y, Xie M X, Hu C Y, Shi Y Q, Shi J
Wu T Y, Gruissem W, Bhullar N K. 2019. Targeting intracellular X, Li J Y. 2019. Identification of the biochemical characteristics
transport combined with efficient uptake and storage significantly of developing giant embryo rice grains using non-targeted
increases grain iron and zinc levels in rice. Plant Biotechnol J, metabolomics. J Cereal Sci, 85: 70–76.
17(1): 9–20. Zhong M, Wang L Q, Yuan D J, Luo L J, Xu C G, He Y Q. 2011.
Yang M, Lu K, Zhao F J, Xie W B, Ramakrishna P, Wang G Y, Du Identification of QTL affecting protein and amino acid contents
Q Q, Liang L M, Sun C J, Zhao H, Zhang Z Y, Liu Z H, Tian J J, in rice. Rice Sci, 18(3): 187–195.
Huang X Y, Wang W S, Dong H X, Hu J T, Ming L C, Xing Y Zhu L J, Gu M H, Meng X L, Cheung S C K, Yu H X, Huang J,
Z, Wang G W, Xiao J H, Salt D E, Lian X M. 2018. Genome- Sun Y, Shi Y C, Liu Q Q. 2012. High-amylose rice improves
wide association studies reveal the genetic basis of ionomic indices of animal health in normal and diabetic rats. Plant
variation in rice. Plant Cell, 30(11): 2720–2740. Biotechnol J, 10(3): 353–362.
Yang Q Q, Zhang C Q, Chan M L, Zhao D S, Chen J Z, Wang Q, Zhu Q L, Yu S Z, Zeng D C, Liu H M, Wang H C, Yang Z F, Xie
Li Q F, Yu H X, Gu M H, Sun S S M, Liu Q Q. 2016. X R, Shen R X, Tan J T, Li H Y, Zhao X C, Zhang Q Y, Chen Y
Biofortification of rice with the essential amino acid lysine: L, Guo J X, Chen L T, Liu Y G. 2017. Development of ‘purple
Molecular characterization, nutritional evaluation, and field endosperm rice’ by engineering anthocyanin biosynthesis in the
performance. J Exp Bot, 67(14): 4285–4296. endosperm with a high-efficiency transgene stacking system.
Yang X Y, Chen L, He J X, Yu W C. 2017. Knocking out of Mol Plant, 10(7): 918–929.