Bicol University • College of Science • 1st Sem, AY: 2021 - 2022 | Technical Elective: BIOTECHNOLOGY

IN A CUP OF RICE:
THE GOLDEN BENEFITS OF GENETICALLY
MODIFIED GRAINS
Andrie Benzon Jr., Arzhel Jhon Consulta, La Arnie Gata, Karl
Monreal, Earl Gerard Nipas, & Jose Joseph Ongonion

Globally known as the most essential staple human food,

rice (Oryza sativa) and its co-products are among the
nourishment a normal table would always have. However,
rice endosperm has shown a lack of many micronutrients
or phytonutrients vital in human development and
nourishment. Ingo Potrykus and Peter Beyer developed
and engineered Golden Rice from ordinary rice in the
1990s to aid public health. Marci Baranski (2013)
stated that golden rice's genome contains a
designed multi-gene biochemical pathway. This
process creates beta-carotene, a chemical that
when digested by humans produces vitamin A.
Golden Rice takes its title from beta-
carotene, which gives it its golden hue.
The endosperm—the starchy and largest
component of the rice seed, which is normally
an off-white color—does not express beta-
carotene in normal rice (Oryza sativa) (Baranski,
2013). Beta-carotene is a yellow-orange chemical
that belongs to the carotenoid’s family of compounds
and is among one of the hundreds of species produced
IN A CUP OF RICE: THE GOLDEN BENEFITS naturally by plants.
OF GENETICALLY MODIFIED GRAINS
As carotenoids are precursors to chemicals needed in
SCAN THE CODE the metabolic reaction pathway, they are hence necessary
to read the research journals nutrients for humans. These produced beta-carotenes
used in the submitted summary are then converted into provitamin A and Vitamin A which
are necessary for the regeneration of retinal and retinoic
acid. The lack of beta-carotene acquisition and other
Bicol University • College of Science • 1st micronutrient deficiencies has often led people to be at
Sem, AY: 2021 - 2022 | Technical Elective: risk of blindness and many other diseases.
BIOTECHNOLOGY Human micronutrient deficiencies are caused by a
lack of iron, zinc, and vitamin A, which leads to a high
Earl Gerard Nipas, summary on “Genetic prevalence of morbidity and mortality. Because Asian
Manipulation for Improved Nutritional Quality countries rely on rice as their primary food supply, vitamin
in Rice” • Andrie Benzon Jr., summary on A deficiency is unavoidable, impacting youngsters and
“Compositional Analysis of Genetically Engineered pregnant women the most. Although rice grains contain
GR2E “Golden Rice” in Comparison to That of other essential elements, they lack provitamin A, even in
Conventional Rice” • Karl Monreal, summary on their unrefined form. Improved maize crops with inserted
“Improving nutritional quality of rice for human features are integrated into rice grains have biological
health” • Arzhel Jhon Consulta & La Arnie implications on the product’s nutrient content. Golden rice
Gata, summary on “Rice Biofortification: High will be used to create new types of rice that do not have
Iron, Zinc, and Vitamin-A to Fight against ‘Hidden nutritional attributes that have been purposely modified.
Hunger’” • Jose Joseph Ongonion, summary The improvement of nutritional quality of food crops
editor, layout & graphics, conclusion. through metabolic engineering, in this case, rice, has
widely known as Biofortification. Rice biofortification via
genetic engineering, based GENETIC MANIPULATION...
on transgenic and RNAi-me- transgenic rice. The level of other rice plants with 230- fold higher level of tryptophan was gen- Agronomic biofortification by application of optimized
diated silencing of antinutrient amino acids was also increased. erated, by harboring OASA2 with S126F/L530D, Y367A/L530D, fertilizers for nutrient enhancement in rice. Zinc was applied
pathways led to the develop- Over accumulation of lysine-rich and Y367A mutations. Mutant gene Mtr1 (mtr1-D) encodes a through the soil and foliar methods increasing the maximum
binding protein in transgenic rice prephenate dehydratase with a point mutation in the protein yield of high zinc rice grains. Furthermore, high zinc accumulation
ment of some rice varieties
rice resulted in high lysine regulatory section, altering phenylalanine biosynthetic enzyme resulted in higher iron accumulation and loss of phytic acid.
that showed a 6.3-fold germplasm increasing the when introduced in rice callus, thus accumulates phenylalanine Nitrogen fertilizer enhanced GPC, Zn, and Fe content. Variation
increase in iron, a 2.0- lysine content of the grains and tryptophan. in location alters the absorption of micro and macronutrients in
fold increase in zinc, Threonine content To successfully breed a rice variety with higher absolute rice by applying nitrogen fertilizers (Garg et al 2018, Graham et al
and 37 μg/g total is related to the lysine mineral content, IRRI and the University of Adelaide documented 2007, Saha et al 2017, Chandel et al 2010).
carotenoid in biofor- content in terms of the zinc and iron content of several rice varieties. There are Agronomic biofortification by application of optimized
tified rice gain. The regulation. By combining three possible ways of attaining higher bioavailability; raise the fertilizers for nutrient enhancement in rice. Zinc was applied
genetic manipulation endogenous rice genes percentage of bioavailability, raise the nutrient concentration in through the soil and foliar methods increasing the maximum
with lysine (K)/threonine the grain, and a mixture of the two. Major mineral contents (Fe, yield of high zinc rice grains. Furthermore, high zinc accumulation
of rice has been ben- (T) motif (TKTKK) coding Zn, Cu, Mn) was higher in traditional rice varieties compared to resulted in higher iron accumulation and loss of phytic acid.
eficial not only in a cu- stretches, two genes were modern yield variety (Graham et al., 1999). Nitrogen fertilizer enhanced GPC, Zn, and Fe content. Variation
mulative increase in the synthesized [TKTKK1 and Agronomic traits and genes responsible for micronutrient in location alters the absorption of micro and macronutrients in
micronutrient content TKTKK2) with 73.1% lysine content in rice are studied and manipulated by QTL mapping for rice by applying nitrogen fertilizers (Garg et al 2018, Graham et al
but as well as to the pur- and 83.5% threonine and future breeding programs. Brown rice had been mapped with five 2007, Saha et al 2017, Chandel et al 2010).
pose that it has served – were introduced into the rice QTL in different chromosomes for iron and zinc concentration. Adopting biotechnological or breeding methods in optimizing
helping people to combat genome. Increasing lysine, 14 QTL in unpolished rice grain through genome-wide mapping the nutritional quality of rice. QTLs play crucial roles in developing
threonine, total amino acids, and from the cross of Madhukar x Swarna variety. SNP and SSR required genotypes. Molecular studies on grain amino acid
micronutrient deficiency. crude protein. markers were used to enhance zinc and iron content through and protein content, glycemic index value, vitamins, minerals,
Due to its tendency to produce Synthesis of tryptophan in plants MAS. and their transporters, phytic acid, phenolic, and flavonoid
sufficient amounts, increasing is feedback regulated at the anthrani- Double-haploid (DH) mapping populations of rice reported compounds, zinc, and iron content created potentials.
the levels of some essential late synthase step. Insensitivity of an- higher grain zinc in rice. Candidate genes with rice grain zinc Although, it is believed that some genes in transgenic methods
amino acids in crop plants has thranilate synthase to feedback inhibition (OsNRAMP, OsNAS, OsZIP, OsYSL, OsFER, and OsZIFL family) are not safe in the environment. Continuous studies are still in
piqued researchers’ interest. enhances tryptophan accumulation. A mu- were identified in association with the QTL. Korean Japonica progress focusing on the nutrient, environmental benefits, and
Genetic research and crop genetic tation breeding approach was used by gene cultivar DH population was used to identify the QTL effect on risks on human health, environmental assessment, and public
targeting mediated mutagenesis of OASA2-an grain iron and zinc content (Jeong et al. 2020, Anuradha et al. accessibility to standardize the impact of the new varieties.
engineering have aided in enrichment
a-subunit of anthranilate synthase. Transgenic 2012, Agarwal et al. 2014, Swamy et al., 2018).
by increasing the amino acid and protein
content, particularly lysine and tryptophan. Because
genetic breeding procedures resulted in high quantities
of lysine and tryptophan, the plant grew unnaturally, and
these features are not highly functional in a seed-specific
manner. However, using a genetic engineering technique, Improving Nutritional Quality of Rice
success in seed-specific features of interest was
achieved. The features can be implanted into a variety of for Human Health
plant species and genotypes. Mingchao Zhao et al., 2019 | summary by Karl Monreal
Polished rice and brown rice are biosynthetic pathway which cannot endosperms. Jeong et al. (2017)
provitamin A-free which is mainly be found in rice. GR contained only synthesized a new gene version
carotenoids (β-carotene) since 1.6 μg/gm of carotenoids which for PAC: stPAC. However, it did not
β-carotene synthetic pathway is was inadequate to supply enough surpass β-carotene concentration
Genetic absent in the rice endosperm. In
plants, β-carotene biosynthetic
vitamin A for a child (Ye, et al., 2000).
Several studies were conducted to
of GR2. Some studies showed that
higher β-carotene accumulation
Manipulation pathway, as shown in Fig. 1, consists
of five enzymes namely phytoene
improve GR. Syngenta scientists
further developed GR into GR2
may be achieved by increasing the
upstream flux from either MVA or
for Improved Nutritional synthase (PSY), phytoene desaturase which contained significantly higher MEP pathways. Bai et al. (2016)
Quality in Rice (PDS), ζ-carotene desaturase (ZDS), β-carotene content by combining used maize Psy gene (ZmPsy1) and
Priyanka Das et al., 2020 carotene cis–trans-isomerase the Psy gene from maize with CRTI the Pantoea CrtI (PaCrtI) gene and
summary by Earl Gerard Nipas (CRTISO), lycopene-β cyclase from GR. The main rate-limiting combined it with Arabidopsis thaliana
(LCYB) to produce β-carotene from step for β-carotene synthesis in GR genes AtDXS or AtOR to increase
Since lysine production in rice is minimal, researchers are GGPP while rice can only produce depends on the type of Psy (Paine supply of isoprenoid precursors
focusing their efforts on figuring out how to regulate lysine the precursor of carotenoids et al. 2005). In a study conducted upstream of 2-C-methyl-D-erythritol
geranylgeranyl diphosphate by Ha et al. (2010), GR expressing 4-phosphate (MEP) pathway. It
metabolism and use it to boost the amount of free lysine in (GGPP). This led to the biofortified constructs combined with Korea resulted to higher concentration
the seeds by employing proteins with high lysine content. rice called Golden Rice (GR). GR pepper (Capsicum annuum) Psy and of β-carotene in the endosperms
The seed lysine content of rice was increased by using a is a transgenic rice that possess Pantoea CrtI in a single bicistronic of rice plants. Tian et al. (2019)
recombinant tRNA-lysine to inject lysine into alternative Phytoene synthase (Psy) from gene, using the 2A sequence from studied boosting the upstream flux
codons during translation. Targeting bacterial aspartate daffodil (Narcissus pseudonarcissus) the foot-and-mouth disease virus of cytoplasmic mevalonate (MVA)
kinase and dihydrodipicolinate synthase into the intragenic and Carotene desaturase (CRTI) from resulted to PAC (Psy-2A-CrtI) and PIC pathway which involved combining
region of the rice genome was also employed. the bacterial phytoene desaturase (Psy-IRES-CrtI). The two bicistronic ZmPSY1 + PaCRTI with tHMG1,
The hybrid of the transgenic rice lines also revealed Pantoea ananatis (formerly, Erwinia genes were introduced into rice under which encodes a truncated HMGR
increased lysine content, level of lysine increased up to 35% uredovora). Psy and CRTI replaces the control of endosperm-specific from Saccharomyces cerevisiae.
(Wong et al., 2015). The introduction of recombinant AmA1 four plant enzymes (PDS, ZISO, promoters however resulted to limited It resulted to higher β-carotene
increases the level of total protein content (1.06-12.87%) of ZDS, and CRTISO) in the β-carotene accumulation of carotenoids in their concentration.
Bicol University • College of Science • 1st Sem, AY: 2021 - 2022 | Technical Elective: BIOTECHNOLOGY 02
Bicol University • College of Science • 1st Sem, AY: 2021 - 2022 | Technical Elective: BIOTECHNOLOGY 03

Rice reproduction was started following a successful pollen continuous search in the gene pool of rice varieties and other Despite the fact
Biofortification:
culture of those plants. As a result, efficient homozygous GR
plants were produced.
organisms for candidate genes useful for rice biofortification
projects to fight against hidden hunger. that there are still
High Iron, Zinc, & Vitamin-A to
Fight against “Hidden Hunger”
As the techniques are directly beneficial to rice consumers
of affected nations, biofortified rice developers, policymakers, a lot of work to be
HIDDEN HUNGER
Shuvobrata Majumder et al., 2019
stakeholders, and philanthropists should focus on policies
such as the PPP model (public-private partnership) in Agri- done, biofortified
crops like Golden
summary by Arzhel Jhon Consulta
and La Arnie Gata
Biotech research, ‘freedom to operate (FTO) biofortified Despite the gradual and continuous progress of
rice varieties developed by private companies, area- industrial processes, particularly in food commerce which
BIOFORTIFICATION
specific production, better storage facilities, international
rice distribution policies, and developing awareness on the
helped humanity to lower health-related concerns, “Hidden
Hunger” or commonly known as micronutrient deficiencies Rice could be
The metabolic engineering of Golden Rice was
nutritional value of other locally available foods.
Rice biofortification for folic acid (or folate), thiamin,
still possess huge risks as one out of every three persons
especially in countries of Africa, South Asia, and Latin worth their weight
in gold when it
initially done in japonica rice cultivar (Taipei-309) by the riboflavin, niacin, pantothenic acid, pyridoxine, biotin, America suffers from insufficiencies of thereof.
introduction of phytoene synthase (PSY) and lycopene vitamin-B12, ascorbic acid, vitamin-D, and vitamin-E are in This occurs when people do not get the vitamins
comes to disease
beta-cyclase (LYC) both from Narcissus pseudonarcissus steady progress. Zinc assists in the synthesis of vitamin-A1 and minerals they need for growth and development, and
and both were under the control of the endosperm-specific (retinol) binding protein and increases lymphatic absorption the food they eat does not meet their nutritional needs.
prevention. After
glutelin promoter together with the sequence coding for a and transport of vitamin-A. Less fortunate people are disproportionately harmed. Iron,
bacterial Erwinia uredovora phytoene desaturase (CRTI) Similarly, vitamin-A also affects zinc lymphatic absorption zinc, and vitamin A deficiency are the most often reported
all, it is nice to have
via Agrobacterium-mediated transformation. and transport via regulating zinc-dependent proteins. micronutrient deficiencies among people worldwide. Because
Subsequently, this was achieved in Indica rice cultivars. Descalsota et al. recently searched through the genome-wide these countries consume a lot of rice, a technique called
what we previously
Various approaches have been made in the development association studies (GWAS) using a multi-parent advanced biofortification was created to address the hidden hunger
of Golden Rice; this includes the diploid/double haploid generation inter-cross (MAGIC) plus rice population to identify problem by improving rice quality using genetic engineering
lack in just a cup
homozygosity approach. This approach first developed QTLs (quantitative trait locus) and SNP (single-nucleotide and molecular sequencing.
the GR plant through molecular breeding and then further polymorphism) markers for biofortification. There is a
of golden rice
GOLD VS. WHITE IN A CUP OF RICE: THE GOLDEN BENEFITS
OF GENETICALLY MODIFIED GRAINS

Compositional Analysis of Genetically Engineered GR2E

Andrie Benzon Jr., Arzhel Jhon Consulta,
La Arnie Gata, Karl Monreal, Earl Gerard Nipas,

“Golden Rice” in Comparison to That of Conventional Rice

& Jose Joseph Ongonion • October 2020

Mallikarjuna Swammy et al., 2019 | summary by Andrie Benzon Jr. The genetic modification of rice into its golden form
have had probe and proven to contain more nutritional
Vitamin A deficiency (VAD) has been found as a significant grain. Total dietary fiber was determined by gelatinization with value. Many countries supplement their meals with
these vitamins to enhance micronutrients ingestion. The
factor that has a detrimental influence on the health and heat-stable -amylase, followed by enzymatic digestion with
production of genetically modified organisms is another
economic condition of people in low-income South Asian protease and amyloglucosidase to remove protein and starch,
technique for increasing vitamin intake and preventing
countries, and it is more common in poorer areas where there respectively. Inductively coupled plasma atomic emission sickness, particularly in developing countries (GMOs).
is economic suffering. Low dietary intake of preformed vitamin spectroscopy was used to examine minerals. Undernutrition is a deadly crisis in the underdeveloped
A and its precursors is frequently linked to VAD, according to It was found out that there were no statistically significant countries, claiming the lives of millions of children every
the World Health Organization (2009). differences in any of the evaluated parameters when year. Genetic improvement may not be a perfect answer
In the compositional analysis conducted by Mallikarjuna proximates, fiber, and minerals in grain (paddy), straw, and to these countries' poverty problems, but it has the ability
Swammy et al. (2019), four sites of experimental set-ups were bran samples obtained from golden rice and control rice to substantially lower ailment tolls. Despite the fact that
conducted during the growing seasons of rice grains in the were compared. Amino acids characterizations yield also no there are still a lot of work to be done, biofortified crops
Philippines. Grain and straw samples were taken from at least statistical difference between the concentrations of its content. like Golden Rice could be worth their weight in gold when
150 developed rice plants in each plot, ready for analysis and However, several fatty acids found in trace levels in both it comes to disease prevention. After all, it is nice to have
processing. In triplicate quantities of paddy rice, straw, and Golden rice and control rice grain samples were found to be what we previously lack in just a cup of golden rice.
bran from the collected samples, nutrients and antinutrients below the limit of detection and were not reported. In each
were examined. Bio-Analytical services were provided by case, provitamin A carotenoids concentrations in samples
several methodologies from various chemical associations for produced from control rice were below the LOQ. Rice has a few IN A CUP OF RICE: THE GOLDEN
the characterization. chemicals that aren’t good for human or animal nutrition, but BENEFITS OF GENETICALLY
During the study, the percentage of moisture in the samples they’re rarely found in high enough amounts to be dangerous. MODIFIED GRAINS
was determined by gravimetric weight loss after drying in a
forced-air oven. An automated Kjeldahl approach was used
16 Antinutritional substances such as phytic acid and trypsin
inhibitors are found in a variety of food crops, including rice. SCAN THE CODE
to view the references used in the
to determine crude protein in rice straw, bran, and paddy rice summary article