Phytonutritional Improvement of Crops, First Edition. Edited by Noureddine Benkeblia.
© 2017 John Wiley & Sons Ltd. Published 2017 by John Wiley & Sons Ltd.
‘Phytonutrients’ or ‘phytochemical’ are primary or secondary metabolites found in
plants and recognised to have nutritional quality attributes and potential health benefits.
From the etymological point of view, ‘phyto’ refers to the Greek word for plant, while
‘chemical’ refers to a substance in a form of matter that has constant chemical composi-
tion and characteristic properties, while ‘nutrient’ refers to a substance that provides
nourishment essential for the maintenance of life and for growth (Drewnowski and
Beside their protective roles in plants from diseases, disorders and other physiologi-
cal stresses (Agrawal 2007, Appel 1993, Close and McArthur 2002, Harborne 1991),
phytonutrients which include many other minor components in foods, particularly
plant‐derived foods, have been know from the ancient times to elicit biologic responses
in human and animal systems. These elicitors have been shown to reduce the risk of one
or more chronic diseases such as cancer, hypertension, diabetes and others (Beecher
1999, Dillard and German 2000, Percival 1997). Hence this new arisen interest in the
so‐called “healthy foods”, that is based on the intake of fruits, vegetables and many other
The use of plant foods to supress human health and nutrition needs, although ancient
in eastern cultures, has only raised attention for the last decades, and is becoming a
trend in Europe and America, especially since the 1990s (Kochian and Garvin 1999).
Moreover, IARC and WHO have recently made a press release classifying red meat and
processed meat as carcinogenic to humans and recommending to limit intake of meat.
Although a large fraction of consumers believe that phytonutrients are found only in
fruits and green leafy vegetables, many other plant‐based and well‐known crops also
contain these compounds. For example, whole grains, nuts, beans or tea also are plant‐
foods rich in phytonutrients. To date, more than 25,000 different phytonutrients have
been identified, and the most important classes are carotenoids, flavonoids, glucosi-
nolates, phytooestrogens, stillbens and sulphur‐containing compounds (Cassidy and
Strategies for Enhancing Phytonutrient Content
in Plant-Based Foods
Carla S. Santos1, Noureddine Benkeblia2 and Marta W. Vasconcelos1
1 CBQF‐Centro de Biotecnologia e Química Fina, Escola Superior de Biotecnologia, da Universidade Católica Portuguesa/Porto,
Rua Arquiteto Lobão Vital, Apartado 2511, EC Asprela, 4202‐401, Porto, Portugal
2 Department of Life Science, and the Biotechnology Centre, Faculty of Science and Technology|, The University of the West
Indies, Mona Campus, Kingston, Jamaica
Phytonutritional Improvement of Crops
Kay 2010). Many researchers report that c.a. 40,000 phytonutrients are found in plant
foods, and extensive literature reports that phytonutrients are recognised as “nutraceu-
ticals” (Rechkemmer 2001), having tremendous impact on the health care system and
therefore, having potential in providing medical health benefits (Gupta and Prakash
2014). Nonetheless, the effects of these phytochemicals depends more on their bioavail-
able dose rather than on the total dose ingested.
Because of the increased incidence of cancer and cardiovascular diseases, as well as
diabetes, one of the most pressing challenges for many governments is to promote a
healthy diet. Therefore, plant biotechnology programmes aiming at phytonutritional
improvement can make significant contributions to human health through the develop-
ment of phytonutrient‐rich plant‐based foods (Grusak and DellaPenna 1999, Kochian
and Garvin 1999, Martin 2013, Martin etal. 2011, Zhao 2007).
To achieve these goals, genetics and metabolic engineering of food crops is one the
ways to make crops improve their contents of specific phytonutrients. With the devel-
opment of omics technologies (Benkeblia 2012) for food science, genomic and prot-
eomic technologies have been used to identify these compounds, as well as genes and
proteins involved in their synthesis. Nowadays, a recent trend has put high‐throughput
tools as a valuable technique in the analysis of phytochemicals (Saito 2013). However,
the application of the data obtained with these approaches in an agricultural context
depends on further information on phenomics, which is the study of plant growth,
performance and composition with regards to the environment.
5.2 What are Phytonutrients?
Literally, a phytonutrient is a plant‐derived nutrient, which comprises proteins, lipids,
carbohydrates and essential minerals and vitamins. Besides these compounds, the term
phytonutrient or phytochemical has also been utilised to describe any organic or inor-
ganic compound in plant foods that is beneficial to human health or nutrition (Kochian
etal. 1999). Hence, the phytonutrient class also includes other secondary plant products
that present characteristics that contribute for health improvement. The phytochemi-
cals can be divided in three main groups, phenolic acids, flavonoids and lignans. The
ones with current preponderance in market and public health are polyphenols, terpe-
noids, resveratrol, flavonoids, isoflavonoids, carotenoids, lycopene, limonoids, phytos-
terols, phytooestrogens, glucosinolates, ω‐3 fatty acids, and anthocyanins (Gupta and
The applications of phytonutrients are extensive, and their efficacy has been tested in
different contexts. Generally, these compounds, especially phenolics (Mathew et al.
2015) and vitamin C, but also tocopherols and tocotrienols, as well as flavonoids and
carotenoids (Liang et al. 2014), have been associated with antioxidant properties.
Antioxidant activity can lower the risk of disease and slow biological ageing, by prevent-
ing chronic degenerative disease. For example, it can greatly improve cardiovascular
disease prevention by reducing cholesterol levels (Riccioni et al. 2012, Constans etal.
2015). In fact, consumption of nutrient‐rich plant foods has been associated with the
prevention of diabetes (Nunes etal. 2014) and hypertension (Rodriguez‐Casado 2016).
The plant‐based diet was shown to have a positive impact in the treatment of fibromy-
algia, a chronic fatigue syndrome, in an 8‐week pilot study (Lamb etal. 2011). Moreover,
5 Strategies forEnhancing Phytonutrient Content inPlant-Based Foods 205
quercetin, which is a flavonol compound also found in fruits and vegetables, has proven
to have an effect over atherosclerosis progression (Hung etal. 2015).
Also due to their antioxidant role, high‐intake of phytonutrients is vastly connoted
with cancer prevention. Early studies have suggested that flavonoids and flavones could
decrease cancer incidence, but with no statistical support (Ronco etal. 1999, Birt etal.
2001). Since then, and through the last decade (Birt 2006, Nakamura etal. 2009, Kang
etal. 2011), new studies have proven the beneficial effect of a combined chemopreven-
tion treatment with dietary phytochemicals intake. Nowadays, substantial proof has
been gathered and effective compounds in cancer treatment/prevention have been
identified (Cardeno etal. 2013, Chaithongyot etal. 2015, Li etal. 2015b, Hosseini and
Ghorbani 2015), even against the most aggressive types (Ham etal. 2015). It is thought
that this protective effect of phytonutrients contained in fruits and vegetables is due to
the modulation of the expression of genes that are important in cancer‐related biologi-
cal and genetic pathways (de Kok etal. 2010).
Polyphenols, alkaloids, terpenes, saponins, amines and carbohydrates also possess
antidepressant activity and could be utilised as an alternative to conventional antide-
pressants (Bahramsoltani etal. 2015). Still on neurological diseases, compounds such as
flavonoids (Thapa and Chi 2015) have been shown to prevent and arrest neurodegen-
eration, impacting neurodegenerative diseases, like Alzheimer’s disease (Venkatesan
etal. 2015, Hügel 2015). Plant secondary metabolites, like ethanol and hexane extracts,
have also been identified as antimicrobial substances (for a recent review please see
Borges etal. 2015) and have been tested for their antimicrobial activity against multi‐
drug resistant bacteria, with positive results (Barreto etal. 2015).
Phytochemicals bioavailability is limited, since besides enduring food processing,
their release from food matrices to the organism is dependent on several factors (Bohn
etal. 2015). Their absorption occurs mainly in the microflora of the intestine and must
resist both liquid and solid phases of the digesta (Bohn et al . 2015). Moreover, it is
important to account that phytonutrients can eventually have toxic effects if consumed
in excess or combined with other incompatible supplements (de Kok etal. 2010).
5.3 Which Plant-Based Foods are the Best Known Sources
A diet composed of at least 400 g (corresponding to five portions) of fruits and vegeta-
bles a day (WHO 2015), which are rich in protein, micronutrients and dietary fibre, and
low in fat, is highly recommended and is part of the traditional diet in most Oriental
(Lee etal. 2013), Mediterranean and also some Nordic countries (Tennant etal. 2014).
However, most countries do not meet the fruit and vegetables intake recommendations
or even if the recommendations are met, the consumption of a limited number of foods
constraints a diverse phytonutrient intake, as happens in the United States (Murphy
etal. 2012). Some compounds, like vitamin A, vitamin C, vitamin E and ß‐carotene and
the food that provide them are widely known and consumed by the general public.
Carrots are the preferred source for vitamin A; oranges, tomatoes and sweet peppers
are the most common sources of vitamin C; for providing vitamin E, cereals, nuts and
seeds are the most consumed; and for ß‐carotene, green leafy, root and fruiting vegeta-
bles (García‐Closas etal. 2004, Sharma etal. 2014).
Phytonutritional Improvement of Crops
The Mediterranean diet has been considered has a health‐promoting diet for a long
time (Corella and Ordovás 2014). A recent review pointed certain fruits and vegetables
that can justify this potential effect in chronic disease control: broccoli, for their high
content in glucosinolate; dandelion, of which young leaves, roots and flower extracts
can be consumed, are rich in phenolics like hydroxycinnamic acid derivatives and in
flavonoids, chlorogenic and chicoric acids, luteolin and quercetin glycosides; garlic,
that not only contains sulphur compounds, but also enzymes, amino acids and miner-
als; and cocoa, which is rich in antioxidants, flavonoids and xanthines (Rodriguez‐
Other component of the Mediterranean diet that contributes to its high health
favourable effect is olive oil. This product has a highly diverse phenolic fraction, namely,
simple phenols, secoiridoids, lignans and flavones that possess the ability to decrease
LDL cholesterol and cell oxidation, microbial activity, markers of inflammation and of
platelet function, and increase HDL cholesterol, antioxidant capacity and bone health
(Cicerale etal. 2010), all of these contributing for an antiageing (Vazquez‐Martin etal.
2012) and anticarcinogenic (Coccina etal. 2014) effects.
Berries are another food rich in phytonutrients, particularly phenolics and flavonoids
(Wang and Lewers 2007) and carotenoids (Lashmanova et al. 2012), and have been
shown to have a powerful antioxidant effect (Liu etal. 2002, Wang and Lewers 2007)
and anticancer activity (Neto 2007). A recent study focused on the identification and
quantification of the phytochemicals present in Arbutus unedo berries and concluded
that these berries are a source of ω‐3 and ω‐6 fatty acids, phytosterols and tocopherols
(Fonseca etal. 2015). The main chemical components present in berries and their role
in regulating cellular processes have also been recently reviewed (Bishayee etal. 2015,
Mazzoni etal. 2016) and a putative chemopreventive role for the occurrence of cancers
and other chronic pathologies is suggested.
Certain beverages are widely associated with high phytonutrients content. For more
than two decades that an inverse relationship between red wine intake and the develop-
ment of coronary disease has been shown, and this is mostly due to the phenolic com-
pounds synthesised by red grapes, particularly resveratrol (Visioli etal. 2000).
Early reports showed that red wine has the ability to supress an oestrogen synthetase
(Eng et al. 2001) as well as the damage caused by 7,12‐dimethylbenz[a]anthracene
(Leung et al. 2009), both involved in breast cancer development; that the intake at
regular doses of red wine, expected in the Mediterranean diet, provides the sufficient
phytonutrients amounts to reduce cardiovascular disease incidence (Carluccio et al.
2003); and that grapes have not only high levels of phenylpropanoids but also of
melatonin, which is responsible for the regulation of physiological and pathological
conditions (Iriti and Faoro 2006). Despite all of these proven advantageous aspects of
wine light consumption, recent concern has been raised due to the correlation between
ethanol (independently from the type of beverage) and oral cancer (Varoni etal. 2015).
Other beverage widely associated with disease control is tea. Regarding its fermenta-
tion process, there are three tea types: green, black and oolong tea. Green tea isn’t fer-
mented and has high levels of a characteristic group of polyphenols, catechins. On the
contrary, since black and oolong tea are fermented, the natural simple polyphenols are
converted to more complex polyphenols, decreasing catechins content and increasing
caffeine content (Hayat etal. 2015). Despite the positive impact of black tea theaflavins
on health, for example, in the inhibition of tumour cell proliferation (Mujtaba and Dou
5 Strategies forEnhancing Phytonutrient Content inPlant-Based Foods 207
2012) and in the alleviation of cardiovascular disease (Cheang etal. 2015), the catechin
found in green tea—epigallocatechin gallate (EGCG)—is the most studied for its high
antioxidant capacity (Seeram etal. 2006) and cancer therapeutic properties (Bigelow
and Cardelli 2006; Zeng etal. 2014; Fang etal. 2015). Five to six cups per day of green
tea is the recommended dosage to consume sufficient amount of EGCG to act as a
therapeutic agent (Wolfram 2007). Green tea effectiveness was also shown in Parkinson’s
disease control (Dutta and Mohanakumar 2015), in bone mineral density increase in
elderly women (Devine etal. 2007), in inhibition of atherosclerosis associated inflam-
matory mediators (Cai etal. 2012), and in obesity control and low glycemic parameters
(Vernarelli and Lambert 2013, Lee etal. 2015).
A recent trend showcases edible flowers as essential components in gastronomy: as
ingredients in a recipe, for seasoning or garnish (Xiong etal. 2014). One notable exam-
ple is Moringa oleifera, which is fairly resistant to extreme weather and is native of India,
Pakistan, Asia Minor, Africa and Arabia (Anwar etal. 2007). The flowers from this tree
have high medicinal value and are commonly utilised as a main dish, as part of fruit
juices or as medicinal tea for cold treatment or weight loss (Anwar etal. 2007). The
antioxidant value of different plants was studied and Rosa spp. presented high levels of
polyphenolic content, as well as high antioxidant effect (Youwei etal. 2008). Further
studies with 10 different flowers show that Paeonia suffruticosa also have high antioxi-
dant capacity and phenolic content (Xiong etal. 2014).
On another context, food animals have been for decades treated with growth promot-
ing antibiotics, in order to increase production and as therapeutics or prophylactics.
However, a correlation between the administered antibiotics and foodborne human
pathogens antibiotic resistance has been found, and this is aggravated by the fact that,
as the manure utilised to amend soils is contaminated with antimicrobial resistance, the
environment is also a reservoir for this problem and is part of the transmission cycle
(Yang etal. 2015, Woolhouse etal. 2015). As such, the utilisation of phytonutrients as
animal feed additives is a viable alternative to antibiotics use, since they also have
growth promoting action and antimicrobial effect (Oh etal. 2013). Several plant‐derived
compounds have inclusively been patented as reviewed in Thormar (2012) and an
invitro study showed that specifically saponins, tannins and essential oils can reduce
methane potential by modulating the process of fermentation in the rumen (Cieslak
5.4 How Can We Enhance Phytonutrients?
5.4.1 Conventional Breeding
Plant breeding is one of the most used approaches to develop new varieties with specific
agronomic traits and improved nutritional qualities (Farnham etal. 1999, Unnevehr
etal. 2007). However, classical breeding approaches have many limitations because the
crossing can only be done between closely related specie or genus, and therefore it uses
available genetic diversity and existing traits to obtain new varieties. It also uses wild
relative species (WRS) with specific traits and transfers them to the new target varie-
ties although some WRS are not compatible for crossing (Farnham etal. 1999, Lemaux
2008). However, plant breeding and engineering researchers admit that WRS could be
Phytonutritional Improvement of Crops
used for potential breeding by introducing traits that have been lost or underutilised
during the domestication and subsequent breeding of these wild species (McCouch
etal. 2013). Despite these limitations, different successful attempts have been reported
to improve crops with phytonutrients. Lago etal. (2014) reported the development of
a new variety of maize rich in anthocyanins and high antioxidant properties, and Juhász
etal. (2014) obtained by preselecting potato individuals prior to breeding high vitamin
C, B5, and B6 contents bred tubers. The flesh of potatoes also contain carotenoids
mainly xanthophylls represented by lutein and zeaxanthin (Andre etal. 2007, Brown
2005, Brown etal. 1993). Nevertheless, screening of advanced breeding lines and varie-
ties of potatoes growing in different locations has shown that different cultivars contain
higher level of phenolics (Navarre etal. 2011), carotenoids (Brown etal. 2006), antho-
cyanins (Brown etal. 2003), as well as a good level of iron (Brown 2008). Other crops
have been improved by breeding to increase their nutritional values such as wheat with
higher proteins and fibres content (Baylan et al. 2013). Indeed, numerous attempts
have been made to enhance the phytochemical levels in crops using traditional breed-
ing strategies (Bouis et al. 2002, Mayer et al. 2006, Nestel etal. 2006). Nonetheless,
these strategies using multi‐generations crossing and back crossing is a time consum-
ing process, requires high trait genetic variations, and heritability as well (McGhie
5.4.2 Molecular Breeding
The technique of molecular breeding–called marker‐assisted breeding‐ is considered
one of the most powerful tools in modern biotechnology. Indeed, this technique is
based on the use of polymorphic single genes to facilitate the process of plant breeding,
and was first proposed by Sax in 1923. With the development of molecular biology, This
technique has found wide listeners amongst farmers due to the fact that it relies on
biological breeding processes rather than plant engineering by inserting external gene
in order to change the DNA of plants organism (Johnson 2004, Thompson etal. 2009).
Therefore, by using this technique the molecular marker can be detected in the new
seedlings, and consequently the presence or absence of the desired trait can be deter-
mined in the young plant and not delayed to the mature stage, and then reducing the
number of generation and saving years of time on crossing (Collard and Mackill 2008,
Lande and Thompson 1990).
Different crops have been or are being manipulated to have their phytonutrient con-
tents enhanced using molecular breeding as shown in Table 5.1.
5.4.3 Metabolic Engineering andGenetic Modification
Conventional agricultural approaches have show limitation in enhancing the nutritional
traits of food crops, however, with the development of molecular biology and related
techniques are making the exploitation to engineer crops with enhanced phytonutri-
ents progressing more rapidly (Hirschi 2009).
At the turn of the twenty‐first century, genetic engineering has known a rapid devel-
opment resulting from the progress made in molecular biology and the better under-
standing of the DNA and its functions in living organisms. Genetic engineering aims to
makeup the genome of a living organism in a laboratory using ‘recombinant DNA tech-
nology’ by inserting, altering, removing or switching off specific piece(s) of DNA
5 Strategies forEnhancing Phytonutrient Content inPlant-Based Foods 209
Table 5.1 Examples ofsome breeding programmes performed or being performed toenhance
thephytonutrient contents andmajor bioactive groups ofcompounds.
Crop Compounds with bioactive properties References
Artichoke (Cynara cardunculus
Phenolics, in particular chlorogenic
Pandino etal. (2012)
Phenolics, in particular phenolic acids,
flavonoids, flavanols and ascorbic acid
Lee etal. (2014)
Cabbage and cauliflower
Glucosinolates, cartoenoids and
Padilla etal. (2007)
Carrot (Daucus carota L.) Carotenoids and phenolics, in
particular cholorogenic acid and
Celery (Apium graveolens L.) Phenolics Yao etal. (2010)
Cucumber (Cucumis sativus L.) Carotenoids, in particular β‐carotene Navazio and Simon
Eggplant (Solanum melongena L.) Phenolics, in particular chlorogenic
acid and antocyanins
Prohens etal. (2007)
Leek (Allium porrum L.) Phenolics, lutein, β‐carotene, ascorbic
acid and vitamin E
Lettuce (Lactuca sativa L.) Carotenoids, in particular β‐carotene
and lutein, and anthocyanins
Melon (Cucumis melo L.) Carotenoids Harel‐Beja etal.
Onion (Allium cepa L.) Phenolics, in particular flavonoids,
flavonols and anthocyanins, and
Yang etal. (2012)
Pepper (Capsicum annuum L.) Carotenoids, phenolics, and ascorbic
Pumpkin, squash and zucchini
Carotenoids, tocopherol, ascorbic acid de Carvalho etal.
Spinach (Spinacia oleracea L.) Lutein and phenolics Pandjaitan etal.
Table beet (Beta vulgaris subsp.
Betalains Gaertner etal.
Tomato (Solanum lycopersicum L.) Carotenoids, in particular lycopene,
phenolics, anthocyanins and ascorbic
Adalid etal. (2010)
Jones etal. (2003)
Watermelon (Citrullus lanatus
(Thunb.) Matsum. & Nakai)
Carotenoids, in particular lycopene,
and ascorbic acid
Yoo etal. (2012)
Mustard (Brassica juncea L.) Glucosinolates, total tocopherols and
Gupta etal. (2015)
Papaya (Carica papaya L.) Carotenoids and ascorbic acid Wall and Tripathi
Phytonutritional Improvement of Crops
containing the gene(s) of interest. As results, crops developed through genetic engi-
neering are commonly known as transgenic or genetically modified (GM) crops (Datta
2013, Desmond and Nicholl 1994). GE allows transferring specific and targeted genes
from close or distant related plant species to the targeted species, and therefore obtain-
ing a ‘new’ plant with desired agronomic traits. The two most interesting benefits of GE
are (i) the possibility to obtain a plant with specific agronomic traits difficult to obtain
in the case the trait is not present in the germplasm of the crop, and (ii) the long time
needed to introduce that trait in the targeted crop using conventional breeding
(Desmond and Nicholl 1994).
During the last decade, many commercial GE or GM crops have been made available
for farmers and delivered benefits in crop production and, moreover, a number of other
GM plants are still being developed to enhance their nutritional values including phy-
tonutrients. The many examples of phytonutrients‐enhanced crops are rice with higher
level of beta‐carotene, high‐lysine corn, maize with improved feed value and tomatoes
with high levels of flavonols amongst others. Table 5.2 below shows the different GM
and transgenic crops that have been developed or under development for higher phyto-
Undoubtedly, gene transfer systems have led to numerous developments and offered
clear insights into the regulation of gene expression and protein function in crops, and
living organisms as well. With the tremendous efforts made in the identification and
isolation of crop genes, a dramatic expansion in our understanding of gene structure
and function at the molecular level have been achieved, and new crops are released
5.5 Phenotyping for Phytonutrients at Different Levels
5.5.1 Low Throughput Techniques
Certain phytonutrients can be analysed using conventional techniques and the associ-
ated methods are well described (Table 5.3).
Table 5.1 (Continued)
Crop Compounds with bioactive properties References
Maize (Zea mays L.) Anthocyanins Lago etal. (2014)
Brassica spp. Glucoraphanin and other aliphatic
Stansell etal. (2015)
Sweet corn (Zea mays L.) and
broccoli (Brassica oleracea L. ssp.
Carotenoids and tocopherols. Ibrahim and Juvik
Strawberry (Fragaria × ananassa) Ellagic acid and ascorbic acid Atkinson etal.
Pepper (Capsicum annuum L.). Ascorbic acid Geleta and
Partly adapted from Plazas etal. (2014).
5 Strategies forEnhancing Phytonutrient Content inPlant-Based Foods 211
Table 5.2 Examples ofGM crops developed or inresearch withenhanced phytonutritional traits.
Crop Compounds with bioactive properties References
Tomato (Solanum lycopersicum) Higher levels of polyamines,
folate, phytoene, β‐carotene, lycopene,
Davuluri etal. (2005)
Díaz de la Garza
Enfissi etal. (2005)
Fraser etal. (2001)
Mehta etal. (2002)
Neelam etal. (2008)
Rosati etal. (2000)
Sestari etal. (2014)
Apple (Malus pumila) Stilbenes Szankowski etal.
Kiwi (Actinidia chinensis) Resveratrol Kobayashi etal.
Maize (Zea mays) Carotenoids, vitamin E, vitamin C,
Cahoon etal. (2003)
Chen etal. (2003)
Yu etal. (2000)
Soybean (Glycine max) Flavonoids Yu etal. (2003)
Canola (Brassica napus) Vitamin E, β‐carotene Shintani and
Potato (Solanum tuberosum)β‐carotene, lutein Ducreux etal. (2005)
Rice (Oryza sativa)β‐carotene, flavonoids and resveratrol Ye etal. (2000)
Shin etal. (2006)
(Fragaria × ananassa)
Vitamin C Agius etal. (2003)
Orange and other citrus
(Citrus × sinensis)
β‐carotene, anthocyanins Pons etal. (2014)
Sweetpotato (Ipomoea batatas) Anthocyanins, proanthocyanidin and
Park etal. (2015)
Dutt etal. (2016)
Spinach (Spinacia oleracea) Phytoecdysteroids, and
Cheng etal. (2010)
Lupin (Lupinus angustifolius) Methionine Molvig etal. (1997)
Phytonutritional Improvement of Crops
For example, the Folin‐Ciocalteu assay (Folin and Ciocalteau 1927) is widely employed
to determine total phenolic content in different types of samples, which is based in the
reduction of the phosphor‐molybdate heteropoly acids Mo(VI) centre in the heteropoly
complex to Mo(V). Through this method, root, leaf, stem or fruit fraction extracts can
be analysed using a standard curve to quantify the products and measuring samples at
765 nm with an UV‐visible spectrophotometer (Ramirez‐Sanchez etal. 2010, Jing etal.
2014, Chen etal. 2015).
Total flavonoids content can also be analysed using a methanolic extract of leaves or
roots of different plants. The aluminium chloride colourimetric method is often used
using a standard curve and reading the absorbance of the samples at 415 nm (Chang
et al. 2002). This protocol allows the quantification of flavonoids in diverse species,
such as Blumea eriantha (Gore and Desai 2014), Manihot esculenta (Omar etal. 2012)
or in medicinal plants of different genus, like Hibiscus, Premna, Mallotus, Trichosanthus,
Maharanga, Astilbe and Syzygium (Subedi etal. 2014).
For evaluating the antioxidant activity, the 1,1‐Diphenyl‐2‐picryl‐hydrazyl (DPPH)
method is conventionally used (Rivero‐Pérez etal. 2007). This method is also based in
the spectrophotometric measurement of samples (at 515 nm) and a change in the col-
our of the solution occurs when the DPPH free radical is reduced by hydrogen donation
(Omar etal. 2012). It has been widely applied and is an useful test to evaluate the anti-
oxidant capability of flowers, fruits and vegetables (Youwei etal. 2008, Omar etal. 2012,
Gore and Desai 2014, Subedi etal. 2014).
Anthocyanins and carotenoids are photosynthetic pigments, considered phytonutri-
ents, for their antioxidant action. Anthocyanins can be analysed with a spectrophoto-
metric differential pH method and applying a molar extinction coefficient of
pelargonidin‐3‐glucoside (Giusti and Wrolstad 2001, Brown etal. 2003) or of cyani-
din‐3‐glucoside chloride (Anisimoviené et al. 2013). Carotenoids can be quantified
accordingly to the method proposed by Sims and Gamon (2002), using spectrophoto-
metric readings and accounting the concentration of chlorophyll a, chlorophyll b and
anthocyanins (Pereira etal. 2014).
Determining foods’ mineral composition is essential in order to understand its nutri-
tional value. Atomic absorption spectroscopy was shown to be effective at detecting
several metals at parts per million level of concentration (Robinson 1960) and it is still
applied in mineral profiling of, for example, teas (Dambiec etal. 2013), medicinal plants
(Küçükbay and Kuyumcu 2014) or coffee (Oliveira etal. 2015).
Table 5.3 Conventional methods forphytonutrient phenotyping.
Analyte Method Reference
Phenolics Folin‐Ciocalteau Folin and Ciocalteau (1927)
Flavonoids Aluminium chloride Chang etal. (2002)
Antioxidant activity DPPH Rivero‐Pérez etal. (2007)
Anthocyanins pH differential Giusti and Wrolstad (2001)
Carotenoids Tris‐methanol extraction Sims and Gamon (2002)
Minerals Atomic absorption spectrometry Robinson (1960)
5 Strategies forEnhancing Phytonutrient Content inPlant-Based Foods 213
Although these techniques can be useful, their common feature is that the evaluation
of each phytonutrient can be performed at the tissue level, besides being time consum-
ing and having low representativeness. Hence, the increased demand for higher plant
yields and quality calls for the development of modern techniques that allow a more
efficient phenotyping and consequent use of food phytonutrients.
5.5.2 High‐Throughput Techniques
The development of plant phenotyping techniques has been associated with the meas-
uring of plant growth, architecture and composition (Fiorani and Schurr 2013), and
high‐throughput techniques have as main advantage the possibility to rapidly and
simultaneously determine a large profile of analytes, which can be the phytonutrients
themselves or the genes and proteins responsible for their production.
‘Omic’ technologies allow the characterisation of genetic diversity in plant systems
and have an important role in phytonutrient phenotyping. High‐throughput tools
accelerate the discovery of new genes and the functional characterisation of enzymes,
thus elucidating biochemical pathways or mineral trafficking players inside the plant
(Grusak 2002) (Figure 5.1).
Next‐generation sequencing (NGS) helps to establish the link between genotype and
phenotype regarding complex traits. Using NGS, the assembly of new data on genes and
their function/regulation, can reveal new molecular and useful phenotypic markers, for
example, in breeding programmes (Pawełkowicz etal. 2016). Genetic profiling, using
Whole Exome Sequencing
RNA Sequencing (RNA Seq)
Nuclear Magnetic Resonance (NMR)
Mass spectrometry (MS) coupled with:
Liquid Chromatography (LC)
Gas Chromatography (GC)
Matrix-assisted laser desorption
Figure 5.1 High‐throughput techniques relevant in phytonutrient phenotyping. (Seecolor plate
section for the color representation of thisfigure.)
Phytonutritional Improvement of Crops
single nucleotide polymorphisms (SNPs), started as a good way to understand absorp-
tion, circulation and the metabolism of phytochemicals, but its high sequencing cost
was a big disadvantage of this technology (de Kok etal. 2012).
Plant genomics gathers high‐throughput tools, like genotype‐by‐sequencing (GBS),
which is the next step in SNP discovery and genotyping, and uses restriction enzymes
to reduce genome complexity, together with multiplex NGS and enables the sequencing
of the whole genome (Elshire etal. 2011). Although GBS does not require a previous
genomic knowledge of the studied species, the obtained reads must be aligned to a ref-
erence genome using alignment tools and, since a large amount of data is produced,
adequate tools for data analysis must be applied. For example, association mapping like
genome‐wide association studies (GWAS) are statistical techniques for measuring the
strength between a marker locus and a natural variation in a target phenotype (Brachi
etal. 2011) and have been used to identify markers underlying natural variations in rice
nutritional quality traits, like phenolic and flavonoid content and antioxidant capacity
(Shao etal. 2011, Yang etal. 2014). Besides being utilised for SNPs discovery, GBS has
been also applied for linkage map construction and QTL identification for agronomi-
cally important traits (Bekele et al. 2013, Verma et al. 2015, Lee et al. 2016), where
GWAS can also be applied.
When compared to whole‐genome sequencing, the study of plant exomics through
exome sequencing allows the identification of protein‐coding genes in specific regions
of the genome, which reduces the time of analysis and the associated costs (Teer and
Mullikin 2010). Although these technologies have been mostly used in biomedical con-
text (Seaby et al. 2015), developing molecular markers to study the allelic variation
behind certain phenotypic traits through whole‐exome sequencing contributes to the
identification of thousands of exome SNPs and holds potential for practical crop
improvement strategies (Singh etal. 2012, Hashmi etal. 2015).
Although SNP discovery is simple and effective in diploid species, the genomic com-
plexity of polyploid plants, that have multiple homologous gene copies, hampers SNP
identification (Chopra et al. 2015). In this scenario, plant transcriptomics can give
information on genetic diversity and analysis. High‐throughput RNA sequencing tech-
nology (RNA‐seq) developed by Illumina has enabled the understanding of biological
pathways, through their transcriptional information given by millions of short sequence
reads. It gathers numerous advantages since RNA‐seq does not depend on genomic
sequence description of the target species, enables gene expression quantification (even
of low‐abundance transcripts), permits the simultaneous identification of different
types of transcripts (isoforms, promoters, transcription start sites and alternative splic-
ing sites), and its output is in the form of base pair resolution (Mata‐Pérez etal. 2015).
RNA‐seq can be used in phytonutrient phenotyping, at the tissue level, since expression
results can provide information on genes relevant in phytonutrient and pathway regula-
tion (Santos etal. 2013, Li and Lan 2015); on transcription factors related to secondary
metabolism pathways, namely the regulation of the biosynthesis of carotenoids in
potato (Li etal. 2015a), and ascorbic acid, carotenoids and flavonoids in tomato (Ye
etal. 2015); and on the identification of transcripts related to pathways involved in the
synthesis of anticancer compounds (Annadurai etal. 2013).
However, understanding the regulation of certain compounds in plants cannot rely
solely on one type of analysis and, nowadays, a combined approach is being adopted.
For example, combining gene expression analysis with metabolite analysis facilitates the
5 Strategies forEnhancing Phytonutrient Content inPlant-Based Foods 215
interpretation between key regulators and compound accumulation profile results
(Wen etal. 2015). As previously mentioned, plant metabolites comprise amino acids,
lipids and carbohydrates (primary metabolites) and polyphenols, alkaloids, terpenes,
polyketides and hormones (secondary metabolites). Hence, plant metabolomics can not
only help to establish the function of transcription factors suspected to control plant
metabolic pathways, but also to identify and quantify plant phytonutrients, contribut-
ing to the characterisation of the potential health benefits of a plant. This has been
achieved using two main techniques, namely, nuclear magnetic resonance (NMR) spec-
troscopy and mass spectrometry (MS) coupled with gas chromatography (GS/MS) or
liquid chromatography (LC/MS) (Guo etal. 2011).
When utilising NMR, the analysis is performed at the plant tissue level and the sam-
ple doesn’t need to be destroyed and can be recovered, resulting in low effort for sample
preparation; also, it is possible to quantify without using internal standard (Wishard
2008). This technique is utilised to analyse the pharmacological properties of medicinal
plants due to the fact that it enables the identification of both primary and secondary
plant metabolites, as well as new metabolites (Holmes etal. 2006). This technique was
applied to study of the primary and secondary metabolites of leaves and seeds of the
papaya plant (Gogna etal. 2015); to identify markers for nutrient deficiency (Lima etal.
2014) and to assess seed metabolomic diversity (Harrigan etal. 2015) in soybean; or to
analyse the unknown phytochemical composition of different plants, such as a highly
consumed Egypt‐native desert palm fruit, Hyphaene thebaica (Farag and Paré 2013)
and the guggul plant, which has proven anti‐inflammatory, antirheumatic, hyppocho-
lesterolemic properties (Bhatia etal. 2015).
The technique most utilised to detect volatile organics is GC/MS. With this type of
technology, in a single sample extract is possible to identify hundreds of metabolites.
Also, by adding time‐of‐flight‐MS to GC (GC‐TOF‐MS) is possible to reach faster scan
times (resolving co‐eluting peaks) and higher sample throughput (Fernie and Schauer
2008). In a recent work, GC‐MS analyses were performed to understand how the differ-
ences in metabolite composition in response to climate factors might influence tea
quality and flavour and detected several chemical families, such as hydrocarbons; oxy-
genated monoterpenes, diterpenes, sesquiterpenes and heterocycles; monoterpene and
sesquiterpene hydrocarbons; aliphatic alcohols, aldehydes, ketones and esters; acids;
and nitrogen‐ and sulphur‐containing compounds (Kowaksick et al. 2014). On the
other hand, when analysing secondary metabolites in which plants are rich, LC/MS
seems to be the most suitable method (Guo etal. 2011), although it does not resolve the
problem for co‐eluting entities (Fernie and Schauer 2008). High‐performance liquid
chromatography (HPLC) is frequently utilised in phytonutrient phenotyping, for exam-
ple, the potential of sea buckthorn as a source of nutrients was evaluated and amongst
the most prevalent phenolic compounds and flavonoids were gallic, caffeic, p‐coumaric,
and ferulic acids and myricetin, kaempferol, naringin, quercetin and isorhamnetin
(Fatima etal. 2015); and also, the variation in secondary metabolites between different
developing stages of pear fruits was studied using this technique, and these data can aid
in the development of phytonutrient enhancement strategies in this crop (Oikawa etal.
2015). Furthermore, when compared to HPLC, ultra‐performance liquid chromatogra-
phy (UPLC) provides better peak separation and higher reproducibility of retention
time (Guo etal. 2011) and, when coupled with Q‐TOF, its sensitivity increases (Farag
and Paré 2013). This technique has been applied in metabolite profiling of Hyphaene
Phytonutritional Improvement of Crops
thebaica (Farag and Paré 2013) and Ligustrum lucidum Ait (Guo etal. 2011) fruits, and
it is also being associated with commercial metabolite software packages to automati-
cally profile flavonoids, allowing an automatic determination of the identity of the com-
pounds (Gu etal. 2015).
Mass spectrometry methods also allow analysing a specific part of the metabolome
—the proteins. Plant proteomics can be defined as the high‐throughput study of these
molecules. Proteomic profiling is an advantageous tool in the improvement of crop
nutritional value as, with it, one can investigate the genotypic variability of two closely
related cultivars with regard to: (i) the biochemical composition (Shekhar etal. 2015,
Gupta etal. 2015); (ii) quantifying protein abundances and correlating this factor with
the expression of certain phenotypes (Morton etal. 2016); (iii) or even performing a
targeted analysis using the mass spectrometry technique Selected Reaction Monitoring
(SRM), that enables detection and quantification of preselected peptides (Chawade
Lipids are a subset of the metabolome and fewer studies on plant lipidomics are avail-
able (Han and Gross 2003; German etal. 2007). Nowadays, lipidomic studies can be
used in the production of food with optimised composition, through a detailed charac-
terisation of lipids and the gathering of quantitative information on lipid class, head-
group and acyl group combination (González‐Thuillier etal. 2015). For example, this
technique has been applied in the establishment of the lipid profile of functional foods,
like edible macroalgae (Van Ginneken etal. 2011) and olive oils (Alves etal. 2016).
Despite of the advantages that each one of these techniques comprises, research stud-
ies are currently combining them, in order to surpass their individual constraints. For
example, NMR and HPLC‐MS methods were combined in order to identify a larger
number of metabolites in Allium species (Soininen etal. 2014). Both gas‐ and liquid‐
chromatography can be simultaneously applied, as recently performed in rice seeds
where their antioxidant properties were compared to the metabolites content in differ-
ent rice cultivars (Kim etal. 2014) and in red grape berries, to study the influence of
climate changes in metabolite profile (Ayenew etal. 2015). Furthermore, integrating
metabolomics results with transcriptomic and proteomic data gives information about
the function of a certain gene in a given metabolic pathway and also about the tran-
scriptional control of the various enzymatic steps of that pathway (Tohge etal. 2015).
This is being frequently applied, for example, in the study of the carotenoid pathway in
maize kernels engineered for higher carotenoid biosynthesis (Decourcelle etal. 2015);
to understand the transcriptional regulation in phenylpropanoid pathway or in antho-
cyanin metabolism in red grape berries (Ayenew etal. 2015); to correlate the expression
of genes involved in flavonoid biosynthesis with the accumulation of phenolics in sea
buckthorn developing berries (Fatima et al. 2015); and also to identify differentially
expressed proteins and metabolites between two soybean cultivars with contrasting
seed coat colour (Gupta etal. 2015).
5.6 The Future Ahead/Concluding Remarks
Plant phytonutrients are receiving increasing attention by scientists and the general
public alike due to their perception as important anti‐disease agents. As phenotyping
is an emerging field and is being applied mostly in the evaluation of growth, yield and
5 Strategies forEnhancing Phytonutrient Content inPlant-Based Foods 217
plant disease resistance, it is important to explore the tools presented in this chapter
in the study of other less explored features, namely, nutrient use efficiency and phyto-
nutrient characterisation, in order to produce high‐quality food with increased phy-
tonutrient value. The technological development that is being leveraged by the
increased worldwide nutritional need could not be enough facing the predicted chal-
lenges ahead: population increase, drastic climate changes and lack of available arable
land (Carvalho and Vasconcelos 2013).
Supplementation through diet is well accepted, and the use of nutraceuticals (or food‐
derived products with positive effect on health) is increasing. However, it is also largely
associated with lack of information, since these health benefits are not yet fully under-
stood from a scientific point of view and their regulation, regarding legislation and
distribution, is also ambiguous (Ahmad etal. 2011). Combining efforts in order to bet-
ter understand the mechanisms of action of such products and use breeding and new
‘omic techniques to increase food phytonutrient content might hold the answer for the
future in agriculture.
This work was supported by National Funds from FCT through projects UID/
Multi/50016/2013 and PTDC/AGRPRO/3972/2014, and PhD scholarship SFRH/
Adalid, AM, Roselló, S, & Nuez, F 2010, ‘Evaluation and selection of tomato accessions
(Solanum section Lycopersicon) for content of lycopene, β‐carotene and ascorbic acid’,
Journal of Food Composition and Analysis, vol. 23, pp. 613–618.
Agius, F, Gonzalez‐Lamothe R, Caballero JL, Munoz‐Blanco J, Botella MA, & Valpuesta, V
2003, ‘Engineering increased vitamin C levels in plants by overexpression of a
D‐galacturonic acid reductase’, Nature Biotechnology, vol. 21, pp. 177–181.
Agrawal, AA 2007, ‘Macroevolution of plant defense strategies’, Trends in Ecology and
Evolution, vol. 22, pp. 103–109.
Ahmad, MF, Ashraf, SA, Ali Ahmad, F, Ansari, JA, & Siddiquee, MRA 2011,
‘Nutraceuticalmarket and its regulation’, American Journal of Food Technology, vol. 6,
Alves, E, Melo, T, Rey, F, Moreira, ASP, Domingues, P, & Domingues, MR 2016, ‘Polar lipid
profiling of olive oils as a useful tool in helping decipher their unique fingerprint’,
LWT–Food Science and Technology, vol. 74, pp. 371–377.
Andre, CM, Ghislain, M, Bertin, P, Oufir, M, Herrera, Mdel R, Hoffmann, L, Hausman, JF,
Larondelle, Y, & Evers, D 2007, ‘Andean potato cultivars (Solanum tuberosum L.) as a
source of antioxidant and mineral micronutrients’, Journal of Agricultural and Food
Chemistry, vol. 55, pp. 366–378.
Anisimoviené, N, Jankauskiené, J, Jodinskiené, M, Bendokas, V, Stanys, V, & Siksnianas, T
2013, ‘Phenolics, antioxidative activity and characterization of anthocyanins in berries of
blackcurrant interspecific hybrids’, Acta Biochimica Polonica, vol. 60, pp. 767–772.
Phytonutritional Improvement of Crops
Annadurai, RS, Neethiraj, R, Jayakumar, V, Damodaran, AC, Rao, SN, Katta, MAVSK,
Gopinathan, S, Sarma, SP, Senthilkumar, V, Niranjan, V, Gopinath, A, &
Mugasimangalam, RC 2013, ‘De novo transcriptome assembly (NGS) of Curcuma longa
L. rhizome reveals novel transcripts related to anticancer and antimalarial terpenoids’,
PLoS ONE, vol. 8, e56217.
Anwar, F, Latif, S, Ashraf, M, & Filani, AH 2007, ‘Moringa oleifera: A food plant with
multiple medicinal uses’, Phytotherapy Research, vol. 21, pp. 17–25.
Appel, HM 1993, ‘Phenolics in ecological interactions: The importance of oxidation’,
Journal of Chemical Ecology, vol. 19, pp. 1521–1552.
Atkinson, CJ, Dodds, PAA, Ford, YY, Le Mière, J, Taylor, JM, Blake, PS, & Paul, N 2006,
‘Effects of Cultivar, Fruit Number and Reflected Photosynthetically Active Radiation on
Fragaria x ananassa Productivity and Fruit Ellagic Acid and Ascorbic Acid
Concentrations’, Annals of Botany, vol. 97, pp. 429–441.
Ayenew, B, Degu, A, Manela, N, Perl, A, Shamir, MO, & Fait, A 2015, ‘Metabolite profiling
and transcript analysis reveal specificities in the response of a berry derived cell culture
to abiotic stresses’, Frontiers in Plant Science, vol. 6, pp. 728.
Bahramsoltani, R, Farzaei, MH, Farahani, MS, & Rahimi, R 2015, ‘Phytochemical
constituents as future antidepressants: a comprehensive review’, Reviews in the
Neurosciences, vol. 26, pp. 699–719.
Balyan, HS, Gupta, PK, Kumar, S, Dhariwal, R, Jaiswal, V, Tyagi, S, Agarwal, P, Gahlaut, V,
& Kumari, S 2013, ‘Genetic improvement of grain protein content and other health‐
related constituents of wheat grain’, Plant Breeding, vol. 132, pp. 446–457.
Baranski, R, Allender, C, & Klimek‐Chodacka, M 2012, ‘Towards better tasting an more
nutritious carrots: Carotenoid and sugar content variation in carrot genetic resources.
Food Research International, vol. 47, pp. 182–187
Barreto, HM, Coelho, KM, Ferreira, JH, Dos Santos, BH, de Abreu, AP, Coutinho, HD, da
Silva, RA, de Sousa, TO, Citó, AM, & Lopes, JA 2015, ‘Enhancement of the antibiotic
activity of aminoglycosides by extracts from Anadenanthera colubrine (Vell.) Brenan var.
cebil against multi‐drug resistant bacteria’, Natural Product Research, vol. 30,
Beecher, GR 1999, ‘Phytonutrients’ role in metabolism: Effects on resistance to
degenerative processes’, Nutrition Reviews, vol. 57, pp. 3–6.
Bekele, WA, Wieckorst, S, Friedt, W, & Snowdon, RJ 2013, ‘High‐throughput genomics in
sorghum: from whole‐genome resequencing to a SNP screening array’, Plant
Biotechnology Journal, vol. 11, pp. 1112–1125.
Benkeblia, N (ed) 2012, OMICs Technologies: Tools for food science. CRC Press, Boca Raton.
Bernaert, N, de Paepe, D, Bouten, C, de Clerq, H, Stewart, D, van Bockstaele, E, de Loose,
M, & van Droogenbroeck, B 2012, ‘Antioxidant capacity, total phenolic and ascorbate
content as a function of the genetic diversity of leek (Allium ampeloprasum var.
porrum)’, Food Chemistry, vol. 134, pp. 669–677.
Bhatia, A, Bharti, SK, Tripathi, T, Mishra, A, Sidhu, OP, Roy, R, & Nautiyal, CS 2015,
‘Metabolic profiling of Commiphora wightii (guggul) reveals a potential source for
pharmaceuticals and nutraceuticals’, Phytochemistry, vol. 110, pp. 29–36.
Bigelow, RLH, & Cardelli, JA 2006, ‘The green tea catechins, (‐)‐Epigallocatechin‐3‐gallate
(EGCG) and (‐)‐Epicatechin‐3‐gallate (ECG), inhibit HGF/Met signalling in
immortalized and tumorigenic breast epithelial cells’, Oncogene, vol. 25, pp. 1922–1930.
5 Strategies forEnhancing Phytonutrient Content inPlant-Based Foods 219
Birt, DF 2006, ‘Phytochemicals and cancer prevention: from epidemiology to mechanism
of action’, Journal of the American Dietetic Association, vol. 106, pp. 20–21.
Birt, DF, Hendrich, S, & Wang, W 2001, ‘Dietary agents in cancer prevention: flavonoids
and isoflavonoids’, Pharmacology & Therapeutics, vol. 90, pp. 157–177.
Bishayee, A, Haskell, Y, Do, C, Siveen, KS, Mohandas, N, Sethi, G, & Stoner, GD 2015,
‘Potential benefits of edible berries in the management of aerodigestive and
gastrointestinal tract cancers: preclinical and clinical evidence’, Critical Reviews in Food
Science and Nutrition, vol. 56, pp. 1753–1775.
Bohn, T, McDougall, GJ, Alegría, A, Alminger, M, Arrigoni, E, Aura, AM, Brito, C, Cilla, A,
El, SN, Karakaya, S, Martínez‐Cuesta, MC, & Santos, CN 2015, ‘Mind the gap–deficits
in our knowledge of aspects impacting the bioavailability of phytochemicals and their
metabolites–a position paper focusing on carotenoids and polyphenols’, Molecular
Nutrition & Food Research, vol. 59, pp. 1307–1323.
Borges, A, Saavedra, MJ, & Simões, M 2015, ‘Insights on antimicrobial resistance, biofilms
and the use of phytochemicals as new antimicrobial agents’, Current Medicinal
Chemistry, vol. 22, pp. 2590–2614.
Bouis HE 2002, ‘Plant breeding: a new tool for fighting micronutrient malnutrition’, Journal
of Nutrition, vol. 132, pp. 491S–494S.
Brachi, B, Morris, GP, & Borevitz, JO 2011, ‘Genome‐wide association studies in plants: the
missing heritability is in the field’, Genome Biology, vol. 12, pp. 232.
Brown, CR, Edwards, CG, Yang, CP, & Dean, BB 1993, ‘Orange flesh trait in potato:
Inheritance and carotenoid content’, Journal of the American Society for Horticultural
Science, vol. 118, pp. 145–150.
Brown, CR, Wrolstad, R, Durst, R, Yang, CP, & Clevidence, B 2003 ‘Breeding studies in
potatoes containing high concentrations of anthocyanins’, American Journal of Potato
Research, vol. 80, pp. 241–250.
Brown, CR, Culley, D, Yang, CP, Durst, R, & Wrolstad, R 2005, ‘Variation of anthocyanin
and carotenoid contents and associated antioxidant values in potato breeding lines’,
Journal of the American Society for Horticultural Science, vol. 130, pp. 174–180.
Brown CR, Kim, TS, Ganga, Z, Haynes, K, De Jong, D, Jahn, M, Paran, I, & De Jong, W
2006, ‘Segregation of total carotenoid in high level potato germplasm and its relationship
to beta‐carotene hydroxylase polymorphism’, American Journal of Potato Research,
vol.83, pp. 365–372.
Brown, CR 2005, ‘Antioxidants in potato’, American Journal of Potato Research, vol. 62,
Brown, CR 2008, ‘Breeding for Phytonutrient Enhancement of Potato’, American Journal of
Potato Research, vol. 85, pp. 298–307.
Cahoon EB, Hall SE, Ripp KG, Ganzke TS, Hitz WD, & Coughlan SJ 2003, ‘Metabolic
redesign of vitamin E biosynthesis in plants for tocotrienol production and increased
antioxidant content’, Nature Biotechnology, vol. 21, pp. 1082–1087.
Cai, Y, Kurita‐Ochiai, T, Hashizume, T, & Yamamoto, M 2012, ‘Green tea
epigallocatechin‐3‐gallate attenuates Porphyromonas gingivalis‐induced atherosclerosis’,
Pathogens and Disease, vol. 67, pp. 76–83.
Cárdeno, A, Sánchez‐Hidalgo, M, Rosillo, MA, Alarcón de la Lastra, C 2013, ‘Oleuropein, a
secoiridoid derived from olive tree, inhibits the proliferation of human colorectal cancer
cell through downregulation of hif‐1alpha’, Nutrition and Cancer, vol. 65, pp. 147–156.
Phytonutritional Improvement of Crops
Carluccio, MA, Siculella, L, Ancora, MA, Massaro, M, Scoditti, E, Storelli, C, Visioli, F,
Distante, A, & Caterina, RD 2003, ‘Olive oil and red wine antioxidant polyphenols
inhibit endothelial activation. Antiatherogenic properties of Mediterranean diet
phytochemicals’, Arteriosclerosis, Thrombosis, and Vascular Biology, vol. 23,
Carvalho, SMP, & Vasconcelos, MW 2013, ‘Producing more with less: strategies and novel
technologies for plant based food biofortification’, Food Research International, vol. 54,
Cassidy, A & Kay, CD 2010, Phytochemicals in Nutrition and Metabolism, 2nd edn,
Wiley‐Blackwell, Oxford, UK, pp. 339–352.
Chaithongyot, S, Asgar, A, Senawong, G, Yowapuy, A, Lattmann, E, Sattayasai, N, &
Senawong, T 2015, ‘Anticancer effects of Curcuma C20‐Dialdehyde against colon and
cervical cancer cell lines’, Asian Pacific Journal of Cancer Prevention, vol. 16,
Chang, CC, Yang, MH, Wen, HM, & Chern, JC 2002, ‘Estimation of total flavonoid content
in propolis by two complementary colorimetric methods’, Journal of Food and Drug
Analysis, vol. 10, pp. 178–182.
Chawade, A, Alexandersson, E, Bengtsson, T, Andreasson, E, & Levander, F 2016, ‘Targeted
proteomics approach for precision plant breeding’, Journal of Proteome Research, vol. 15,
Cheang, WS, Ngai, CY, Tam, YY, Tian, XY, Wong, WT, Zhang, Y, Lau, CW, Chen, ZY, Bian,
ZX, Huang, Y, & Leung, FP 2015, ‘Black tea protects against hypertension‐assiciated
endothelial dysfunction through alleviation of endoplasmic reticulum stress’, Scientific
Reports, vol. 15, pp. 10340.
Chen, PX, Tang, Y, Marcone, MF, Pauls, PK, Zhang, B, Liu, R, & Tsao, R 2015,
‘Characterization of free, conjugated and bound phenolics and lipophilic antioxidants in
regular‐ and non‐darkening cranberry beans (Phaseolus vulgaris L.)’, Food Chemistry,
vol. 185, pp. 298–308.
Chen Z, Young TE, Ling J, Chang SC, & Gallie DR 2003, ‘Increasing vitamin C content of
plants through enhanced ascorbate recycling’, Proceedings of the National Academy of
Sciences of the USA, vol. 100, pp. 3525–3530.
Cheng DM, Yousef GG, & Lila MA 2010, ‘Variation in Phytoecdysteroid Accumulation in
Seeds and Shoots of Spinacia oleracea L. Accessions’, HortScience, vol. 45, pp. 1634–1638
Chopra, R, Burow, G, Farmer, A, Mudge, J, Simpson, CE, Wilkins, TA, & Baring, MR 2015,
Next‐generation transcriptome sequencing, SNP discovery and validation in four market
classes of peanut, Arachis hypogaea L .’, Molecular Genetics and Genomics, vol. 290,
Cicerale, S, Lucas, L, & Keast, R 2010, ‘Biological activities of phenolic compounds present
in virgin olive oil’, International Journal of Molecular Sciences, vol. 11, pp. 458–479.
Cieslak, A, Szumacher‐Strabel, M, Stochmal, A, & Oleszek, W 2013, ‘Plant components
with specific activities against rumen methanogens’, Animal, vol. 7, pp. 253–265.
Close, CD, & McArthur, C 2002, ‘Rethinking the role of many plant phenolics–protection
from photodamage not herbivores?’, Oikos, vol. 99, pp. 166–172.
Coccina, A, Bastianelli, D, Mosca, L, Monticolo, R, Panuccio, I, Carbone, A, Calogero, A,
Lendaro, E 2014, ‘Extra virgin olive oil phenols suppress migration and invasion of T24
human bladder cancer cells through modulation of matrix metalloproteinase‐2’,
Nutrition and Cancer, vol. 66, pp. 946–954.
5 Strategies forEnhancing Phytonutrient Content inPlant-Based Foods 221
Collard BCY, & Mackill DJ 2008, ‘Marker‐assisted selection: an approach for precision
plant breeding in the twenty‐first century’, Philosophical Transactions of the Royal
Society B, vol 363, pp. 557–572.
Constans, J, Bennetau‐Pelissero, C, Martin, JF, Rock, E, Mazur, A, Bedel, A, Morand, C, &
Bérard, AM 2015 ‘Marked antioxidant effect of orange juice intake and its
phytomicronutrients in a preliminary randomized cross‐over trial on mild
hypercholesterolemic men’, Clinical Nutrition, vol. 34, pp. 1093–1100.
Corella, D, & Ordovás, JM 2014, ‘How does the Mediterranean diet promote
cardiovascular health? Current progress toward molecular mechanisms’, Bioessays,
vol.36, pp. 526–537.
Dambiec, M, Polechonska, L, & Klink, A 2013, ‘Levels of essential and non‐essential
elements in black teas commercialized in Poland and their tranfers to tea infusion’,
Journal of Food Composition and Analysis, vol. 31, pp. 62–66.
Datta, A 2013, ‘Genetic engineering for improving quality and productivity of crops’,
Agriculture & Food Security, vol. 2, pp. 15.
Davuluri, GR, van Tuinen, A, Fraser, PD, Manfredonia, A, Newman, R, Burgess, D,
Brummell, DA, King, SR, Palys, J, Uhlig, J, Bramley, PM, Pennings, HMJ, & Bowle, C
2005, ‘Fruit‐specific RNAi‐mediated suppression of DET1 enhances carotenoid and
flavonoid content in tomatoes’, Nature Biotechnology, vol. 23, pp. 890–895.
de Carvalho, LMJ, Gomes, PB, Godoy, RLD, Pacheco, S, do Monte, PHF, de Carvalho, JLH,
Nutti, MR, Neves, ACL, Vieira, ACRA, & Ramos, SRR 2012, ‘Total carotenoid content,
α‐carotene and β‐carotene, of landrace pumpkins (Cucurbita moschata Duch):
Apreliminary study’, Food Research International, vol. 47, pp. 337–340.
Decourcelle, M, Perez‐Fons, L, Baulande, S, Steiger, S, Couvelard, L, Hem, S, Zhu, C,
Capell, T, Christou, P, Fraser, P, & Sandmann, G 2015, ‘Combined transcript, proteome,
and metabolite analysis of transgenic maize seeds engineered for enhanced carotenoid
synthesis reveals pleotropic effects in core metabolism’, Journal of Experimental Botany,
vol. 66, pp. 3141–3150.
de Kok, MCM, de Waard, P, Wilms, LC, & van Breda, SGJ 2010, ‘Antioxidative and
antigenotoxic properties of vegetables and dietary phytochemicals: The value of
genomics biomarkers in molecular epidemiology’, Molecular Nutrition Food Reasearch,
vol. 54, pp. 208–217.
de Kok, MCM, van Breda, SGJ, Briedé, JJ 2012, ‘Genomics‐based identification of
molecular mechanisms behind the cance preventive action of phytochemicals: potential
and challenges’, Current Pharmaceutical Biotechnology, vol. 13, pp. 255–264.
DellaPenna, D 2007, ‘Biofortification of plant‐based food: enhancing folate levels by
metabolic engineering’, Proceedings of the National Academy of Sciences of the USA,
vol.104, pp. 3675–3676.
Devine, A, Hodgson, JM, Dick, IM, & Prince, RL 2007, ‘Tea drinking is associated with
benefits on bone density in older women’, American Journal of Clinical Nutrition, vol.86,
Desmond, S & Nicholl, T 1994, An Introduction to Genetic Engineering, Cambridge
Díaz de la Garza, R, Quinlivan, EP, Klaus, SM, Basset, GJ, Gregory, JF, & Hanson, AD 2004,
‘Folate biofortification in tomatoes by engineering the pteridine branch of folate
synthesis’, Proceedings of the National Academy of Sciences of the USA, vol. 101,
Phytonutritional Improvement of Crops
Dillard, CJ, & German, BJ 2000, ‘Phytochemicals: nutraceuticals and human health’, Journal
of the Science of Food and Agriculture, vol. 80, pp. 1744–1756.
Drewnowski, A, & Gomez‐Carneros, C 2000, ‘Bitter taste, phytonutrients, and the
consumer: a review’, American Journal of Clinical Nutrition, vol. 72, pp. 1424–1435.
Ducreux, LJM, Morris, WL, Hedley, PE, Shepherd, T, Davies, HV, Millam, S, & Taylor,
MA 2005, ‘Metabolic engineering of high carotenoid potato tubers containing
enhanced levels of b‐carotene and lutein’, Journal of Experimental Botany, vol. 56,
Dutt, M, Stanton, D, & Grosser, JW 2016, ‘Ornacitrus: Development of Genetically
Modified Anthocyanin‐expressing Citrus with Both Ornamental and Fresh Fruit
Potential’, Journal of the American Society for Horticultural Science, vol. 141, pp. 54–61
Dutta, D, Mohanakumar, KP 2015, ‘Tea and Parkinson’s disease: Constituents of tea
synergize with antiparkinsonian drugs to provide better therapeutic benefits’,
Neurochemistry International, vol. 89, 181–190.
Elshire, RJ, Glaubitz, JC, Sun, Q, Poland, JA, Kawamoto, K, Buckler, ES, & Mitchell, SE
2011, ‘A robust, simple genotyping‐by‐sequencing (GBS) approach for high diversity
species’, PLoS ONE, vol. 6, e19379.
Enfissi, EMA, Fraser, PD, Lois, LM, Boronat, A, Schuch, W, & Bramley, PM 2005,
‘Metabolic engineering of the mevalonate and non‐mevalonate isopentenyl diphosphate‐
forming pathways for the production of health‐promoting isoprenoids in tomato’, Plant
Biotechnology Journal, vol. 3, pp. 17–27.
Eng, ET, Williams, D, Mandava, U, Kirma, N, Tekmal, RR, & Chen, S 2001, ‘Suppression of
aromatase (estrogen synthetase) by red wine phytochemicals’, Breast Cancer Research
and Treatment, vol. 67, pp. 133–146.
Fang, CY, Wu, CC, Hsu, HY, Chuang, HY, Huang, SY, Tsai, CH, Chang, Y, Tsao, GSW,
Chen, CL, & Chen, JY 2015, ‘EGCG inhibits proliferation, invasiveness and tumor
growth by up‐regulation of adhesion molecules, suppression of gelatinases activity, and
induction of apoptosis in nasopharyngeal carcinoma cells’, International Journal of
Molecular Sciences, vol. 16, pp. 2530–2558.
Farag, MA, & Paré, PW 2013, ‘Phytochemical analysis and anti‐inflammatory potential of
Hyphaene thebaica L. fruit’, Journal of Food Science, vol. 78, pp. C1503‐C1508.
Farnham, MW, Simon, PW, Stommel, JR 1999, ‘Improved Phytonutrient Content through
Plant Genetic Improvement’, Nutrition Reviews, vol. 657, pp. 19–26.
Fatima, T, Kesari, V, Watt, I, Wishart, D, Todd, JF, Schroeder, WR, Paliyath, G, & Krishna, P
2015, ‘Metabolite profiling and expression analysis of flavonoid, vitamin C and
tocopherol biosynthesis genes in the antioxidant‐rich sea buckthorn (Hippophae
rhamnoids L.)’, Phytochemistry, vol. 118, pp. 181–191.
Fernie, AR, & Schauer, N 2008, ‘Metabolomics‐assisted breeding: a viable option for crop
improvement?’, Trends in Genetics, vol. 25, pp. 39–48.
Fiorani, F, Schurr, U 2013, ‘Future scenarios for plant phenotyping’, Annual Review of Plant
Biology, vol. 64, pp. 267–291.
Folin, O, & Ciocalteau, V 1927, ‘On tyrosine and tryptophane determinations in proteins’,
Journal of Biological Chemistry, vol. 73, pp. 627–650.
Fonseca, DFS, Salvador, AC, Santos, SAO, Vilela, C, Freire, CSR, Silvestre, AJD, & Rocha,
SM 2015, ‘Bioactive phytochemicals from wild Arbutus unedo L. berries from different
locations in Portugal: quantification of lipophilic components’, International Journal of
Molecular Sciences, vol. 16, pp. 14194–14209.
5 Strategies forEnhancing Phytonutrient Content inPlant-Based Foods 223
Fraser, PD, Romer, S, Kiano, JW, Shipton, CA, Mills, PB, Drake, R, Schuch, W, & Bramley,
PM 2001, ‘Elevation of carotenoids in tomato by genetic manipulation’, Journal of the
Science of Food and Agriculture, vol. 81, pp. 822–827.
García‐Closas, R, Berenguer, A, José Tormo, M, José Sánchez, M, Quirós, JR, Navarro, C,
Arnaud, R, Dorronsoro, M, Dolores Chirlaque, M, Barricarte, A, Ardanaz, E, Amiano, P,
Martinez, C, Agudo, A, & González, CA 2004, ‘Dietary sources of vitamin C, vitamin E
and specific carotenoids in Spain’, British Journal of Nutrition, vol. 91, pp. 1005–1011.
Geleta, LF, & Labuschagne, MT 2006, ‘Combining ability and heritability for vitamin C and
total soluble solids in pepper (Capsicum annuum L.)’, Journal of the Science of Food and
Agriculture, vol. 86, pp. 1317–1320.
German, JB, Gillies, LA, Smilowitz, JT, Zivkovic, AM, & Watkins, SM 2007, ‘Lipidomics
and lipid profiling in metabolomics’, Current Opinion in Lipidology, vol. 18, pp. 66–71.
Giusti, MM, & Wrolstad, RE 2001, ‘Anthocyanins: Characterization and measurement with
UV‐visible spectroscopy’, in RE Wrolstad (ed.), Current Protocols in Food Analytical
Chemistry, John Wiley & Sons, New York.
Gogna, N, Hamid, N, & Dorai, K 2015, ‘Metabolomic profiling of the phytomedicinal
constituents of Carica papaya L. leaves and seeds by 1H NMR spectroscopy and multivariate
statistical analysis’, Journal of Pharmaceutical and Biomedical Analysis, vol. 115, pp. 74–85.
González‐Thuillier, I, Salt, L, Chope, G, Penson, S, Skeggs, P, Tosi, P, Powers, SJ, Ward, JL,
Wilde, P, Shewry, PR, & Haslam, RP 2015, ‘Distribution of lipids in the grain of wheat
(cv. Hereward) determined by lipidomic analysis of milling and pearling fractions’,
Journal of Agricultural and Food Chemistry, vol. 63, pp. 10705–10716.
Gore, M, & Desai, NS 2014, ‘Characterization of phytochemicals and evaluation of
anti‐cancer potential of Blumea eriantha D C ’, Physiology and Molecular Biology of
Plants, vol. 20, pp. 475–486.
Grusak, MA 2002, ‘Phytochemicals in plants: genomics‐assisted plant improvement for
nutritional and health benefits’, Current Opinion in Biotechnology, vol. 13, pp. 508–511.
Grusak, MA & DellaPenna, D 1999, ‘Improving the nutrient composition of plants to
enhance human nutrition and health’, Annual Review of Plant Physiology and Plant
Molecular Biology, vol. 50, pp. 133–161.
Gu, WY, Li, N, Leung, ELH, Zhou, H, Luo, GA, Liu, L, & Wu, JL 2015, ‘Metabolites software‐
assisted flavonoid hunting in plants using ultra‐high performance liquid chromatography‐
quadrupole‐time of flight mass spectrometry’, Molecules, vol. 20, pp.3955–3971.
Guo, N, Yu, Y, Ablajan, K, Li, L, Fan, B, Peng, J, Yan, H, Ma, F, & Nie, Y 2011, ‘Seasonal
variations in metabolite profiling of the fruits of Ligustrum lucidum Ait.’, Rapid
Communications in Mass Spectrometry, vol. 25, pp. 1701–1714.
Gupta, C, & Prakash, D 2014, ‘Phytonutrients as therapeutic agents’, Journal of
Complementary and Integrative Medicine, vol. 11, pp. 151–169.
Gupta, S, Sangha, MK, Kaur, G, Banga, S, Gupta, M, Kumar, H, & Banga, SS 2015, ‘QTL
analysis for phytonutrient compounds and the antioxidant molecule in mustard
(Brassica juncea L.)’, Euphytica, vol. 201, pp 345–356.
Ham, SL, Nasrollahi, S, Shah, KN, Soltisz, A, Paruchuri, S, Yun, YH, Luker, GD, Bishayee,
A, & Tavana, H 2015, ‘Phytochemicals potently inhibit migration of metastatic breast
cancer cells’, Integrative Biology, vol. 7, pp. 792.
Han, X, & Gross, RW 2003, ‘Global analyses of cellular lipidomes directly from crude
extracts of biological samples by ESI mass spectrometry: a bridge to lipidomics’, Journal
of Lipid Research, vol. 44, pp. 1071–1079.
Phytonutritional Improvement of Crops
Harborne, JB 1991, ‘The chemical basis of plant defense’, in: RT Palo & CT Robbins (eds.),
Plant Defenses Against Mammalian Herbivory, CRC Press, Boca Raton (FL).
Harel‐Beja, R, Tzuri, G, Portnoy, V, Lotan‐Pompan, M, Lev, S, Cohen, S, Dai, N, Yeselson,
L, Meir, A, Libhaber, SE, Avisar, E, Melame, T, van Koert, P, Verbakel, H, Hofstede, R,
Volpin, H, Oliver, M, Fougedoire, A, Stahl, C, Fauve, J, Copes, B, Fei, Z, Giovannoni, J,
Ori, N, Lewinsohn, E, Sherman, A, Burger, J, Tadmor, Y, Schaffer, AA, & Katzir, N 2010,
‘A genetic map of melon highly enriched with fruit quality QTLs and EST markers,
including sugar and carotenoid metabolism markers’, Theoretical and Applied Genetics,
vol. 121, pp. 511–533.
Harrigan, GG, Skogerson, K, MacIsaac, S, Bickel, A, Perez, T, & Li, X 2015, ‘Application of
1H NMR profiling to assess seed metabolomics diversity. A case study on soybean era
population’, Journal of Agricultural and Food Chemistry, vol. 63, pp. 4690–4697.
Hashmi, U, Shafqat, S, Khan, F, Majid, M, Hussain, H, Kazi, AG, John, R, & Ahmad, P 2015,
‘Plant exomics: concepts, applications and methodologies in crop improvement’, Plant
Signaling & Behavior, vol. 10, e976152-2.
Hayat, K, Iqbal, H, Malik, U, Bilal, U, & Mushtaq, S 2015, ‘Tea and its consumption:
benefits and risks’, Critical Reviews in Food Science and Nutrition, vol. 55, pp. 939–954.
Hirschi, KD 2009, ‘Nutrient Biofortification of Food Crops’, Annual Review of Nutrition,
vol. 29, pp. 401–421.
Holmes, E, Tang, H, Wang, Y, Seger, C 2006, ‘The assessment of plant metabolite profiles
by NMR‐based methodologies’, Planta Medica, vol. 72, pp. 771–785.
Hosseini, A, & Ghorbani, A 2015, ‘Cancer therapy with phytochemicals: evidence from
clinical studies’, Avicenna Journal of Phytomedicine, vol. 5, pp. 84–97.
Hügel, HM 2015, ‘Brain food for Alzheimer‐free ageing: focus on herbal medicines’,
Advances in Experimental Medicine and Biology, vol. 863, pp. 95–116.
Hung, CH, Chan, SH, Chu, PM, & Tsai, KL 2015, ‘Quercetin is a potent anti‐atherosclerotic
compound by activation of DIRT1 signalling under oxLDL stimulation’, Molecular
Nutrition and Food Research, vol. 59, pp. 1905–1917.
Ibrahim, KE, & Juvik, JA 2009, ‘Feasibility for improving phytonutrient content in vegetable
crops using conventional breeding strategies: Case study with carotenoids and
tocopherols in sweet corn and broccoli’, Journal of Agricultural and Food Chemistry,
vol.57, pp. 4636–4644.
Iriti, M, & Faoro, F 2006, ‘Grape phytochemicals: A bouquet of old and new nutraceuticals
for human health’, Medical Hypotheses, vol. 67, pp. 833–838.
Jing, P, Song, LH, Shen, SQ, Zhao, SJ, Pang, J, & Qian, BJ 2014, ‘Characterization of
phytochemicals and antioxidant activities of red radish brines during lactic acid
fermentation’, Molecules, vol. 19, pp. 9675–9688.
Jones, CM, Mes, P, & Myers, JR 2003, ‘Characterization and Inheritance of the
Anthocyanin fruit (Aft) Tomato’, Journal of Heredity, vol. 94, pp. 449–456.
Johnson, R 2004, ‘Marker‐assisted selection’, in Janick J (ed.), Plant Breeding Review, vol. 24,
Part 1, John Wiley & Sons, pp. 293–309,
Juhász, Z, Dancs, G, Marincs, F, Vossen, M, Allefs, S, & Bánfalvi, Z 2014, ‘Vitamin C, B5, and
B6 contents of segregating potato populations detected by GC‐MS: a method facilitating
breeding potatoes with improved vitamin content’, Plant Breeding, vol. 133, pp. 515–520.
Kang, NJ, Lee, KW, Kim, BH, Bode, AM, Lee, HJ, Heo, YS, Boardman, L, Limburg, P, Lee,
HJ, & Dong, Z 2011, ‘Coffee phenolic phytochemicals suppress colon cancer metastasis
by targeting MEK and TOPK’, Carcinogenesis, vol. 32, pp. 921–928.
5 Strategies forEnhancing Phytonutrient Content inPlant-Based Foods 225
Kim, GR, Jung, ES, Lee, S, Lim, SH, Ha, SH, & Lee, CH 2014, ‘Combined mass
spectrometry‐based metabolite profiling of different pigmented rice (Oryza sativa L.)
seeds and correlation with antioxidant activities’, Molecules, vol. 19, pp. 15673–15686.
Kobayashi, S, Ding, CK, Nakamura, Y, Nakajima, I, & Matsumoto, R 2000, ‘Kiwifruits
(Actinidia deliciosa) transformed with a Vitis stilbene syn‐ thase gene produce piceid
(resveratrol‐glucoside)’, Plant Cell Reports, vol. 19, pp. 904–910
Kochian, LV, & Garvin, DF 1999, ‘Agricultural approaches to improving phytonutrient
content in plants: an overview’, Nutrition Reviews, vol. 57, pp. S13‐S18.
Kowalsick, A, Kfoury, N, Robbat Jr, A, Ahmed, S, Orians, C, Griffin, T, Cash, SB, & Stepp,
JR 2014, ‘Metabolite profiling of Camellia sinensis by automated sequential,
multidimensional gas chromatography/mass spectrometry reveals strong monsoon
effects on tea constituents’, Journal of Chromatography A, vol. 1370, pp. 230–239.
Küçükbay, FZ, & Kuyumcu, E 2014, ‘Determination of elements by atomic absorption
spectrometry in medicinal plants employed to alleviate common cold symptoms’, Guang
Pu Xue Yu Guang Pu Fen Xi, vol 34, pp. 2548–2556.
Lago, C, Cassani, E, Zanzi, C, Landoni, M, Trovato, R, & Pilu, R 2014, ‘Development and
study of a maize cultivar rich in anthocyanins: coloured polenta, a new functional food’,
Plant Breeding, vol. 133, pp. 210–217.
Lamb, JJ, Konda, VR, Quig, DW, Desai, A, Minich, DM, Bouillon, L, Chang, J, His, A, Lerman,
RH, Kornberg, J, Bland, JS, & Tripp, ML 2011, ‘A program consisting of a phytonutrient‐rich
medical food and an elimination diet ameliorated fibromyalgia symptoms and promoted
toxic‐element detoxification in a pilot trial’, Alternative Therapies, vol. 17, pp. 36–44.
Lande, R, & Thompson, R 1990, ‘Efficiency of marker‐assisted selection in the
improvement of quantitative traits’, Genetics, vol. 124, pp. 743–756.
Lashmanova, KA, Kuzivanova, OA, & Dymova, OV 2012, ‘Northern berries as a source of
carotenoids’, Acta Biochimica Polonica, vol. 59, pp. 133–134.
Lee, HS, Cho, YH, Park, J, Shin, HR, & Sung, MK 2013, ‘Dietary intake of phytonutrients in
relation to fruit and vegetable consumption in Korea’, Journal of the Academy of
Nutrition and Dietetics, vol. 113, pp. 1194–1199.
Lee, J, Izzah, NK, Choi, BS, Joh, HJ, Lee, SC, Perumal, S, Seo, J, Ahn, K, Jo, EJ, Choi, GJ,
Nou, IS, Yu, Y, & Yang, TJ 2016, ‘Genotyping‐by‐sequencing map permits identification
of clubroot resistance QTLs and revision of the reference genome assembly in cabbage
(Brassica oleracea L.)’, DNA Research, vol. 23, pp. 29–41.
Lee, LS, Choi, JH, Sung, MJ, Hur, JY, Hur, HJ, Park, JD, Kim, YC, Gu, EJ, Min, B, & Kim, HJ
2015, ‘Green tea changes serum and liver metabolomics profiles in mice with high‐fat
diet‐induced obesity’, Molecular Nutrition and Food Research, vol. 59, pp. 784–794.
Lemaux, PG 2008, ‘Genetically engineered plants and foods: A scientist’s analysis of the
issues (Part I)’, Annual Review of Plant Biology, vol. 59, pp. 771–812.
Leung, HY, Yung, LH, Shi, G, Lu, A, & Leung, LK 2009, ‘The red wine polyphenol
resveratrol reduces polycyclic aromatic hydrocarbon‐induced DNA damage in
MCF‐10A cells’, British Journal of Nutrition, vol. 102, pp. 1462–1468.
Li, W, & Lan, P 2015, ‘Genome‐wide analysis of overlapping genes regulated by iron
deficiency and phosphate starvation reveals new interactions in Arabidopsis roots’, BMC
Research Notes, vol. 8, pp. 555.
Li, R, Zhai, H, Kang, C, Liu, D, He, S, & Liu, Q 2015a, ‘De Novo transcriptome sequencing
of the orange‐fleshed sweet potato and analysis of differentially expressed genes related
to carotenoid biosynthesis’, International Journal of Genomics, vol. 2015, 843802.
Phytonutritional Improvement of Crops
Li, YH, Niu, YB, Sun, Y, Zhang, F, Liu, CX, Fan, L, & Mei, QB 2015b, ‘Role of
phytochemicals in colorectal cancer prevention’, World Journal of Gastroenterology,
vol.21, pp. 9262–9272.
Liang, Z, Cheng, L, Zhong, GY, & Liu, RH 2014, ‘Antioxidant and antiproliferative activities
of twenty‐four Vitis vinifera grapes’, PLoS One, vol. 9, e105146.
Lima, MRM, Diaz, SO, Lamego, I, Grusak, MA, Vasconcelos, MW, & Gil, AM 2014,
‘Nuclear magnetic resonance metabolomics of iron deficiency in soybean leaves’, Journal
of Proteome Research, vol. 13, pp. 3075–3087.
Liu, M, Li, XQ, Weber, C, Lee, CY, Brown, J, & Liu, RH 2002, ‘Antioxidant and
antiproliferative activities of raspeberries’, Journal of Agricultural and Food Chemistry,
vol. 50, pp. 2926–2930.
Martin, C 2013, ‘The interface between plants metabolic engineering and human health’,
Current Opinion in Biotechnology, vol. 24, pp. 344–353.
Martin, C, Butelli, E, Petroni, K, & Tonelli, C 2011, ‘How can Research on plants contribute
to promoting human health?’, The Plant Cell, vol. 23, pp. 1685–1699.
Mata‐Pérez, C, Sánchez‐Calvo, B, Begara‐Morales, JC, Luque, F, Jiménez‐Ruiz, J, Padilla,
MN, Fierro‐Risco, J, Valderrama, R, Fernández‐Ocaña, A, Corpas, FJ, & Barroso, JB
2015, ‘Transcriptomic profiling of linolenic acid‐responsive genes in ROS signalling
from RNA‐seq data in Arabidopsis’, Frontiers in Plant Science, vol. 6, pp. 122.
Mathew, S, Abraham, TE, & Zakaria, ZA 2015, ‘Reactivity of phenolic compounds towards
free radicals under in vitro conditions’, Journal of Food Science and Technology, vol. 52,
Matthews, PD, Luo, RB, & Wurtzel, ET 2003, ‘Maize phytoene desaturase and ζ‐carotene
desaturase catalyse a poly‐Z desaturation pathway: implications for genetic engineering
of carotenoid content among cereal crops’, Journal of Experimental Botany, vol. 54,
Mayer, JE, Pfeiffer, WH, & Beyer, P. 2008, ‘Biofortified crops to alleviate micronutrient
malnutrition’, Current Opinion in Plant Biology, vol. 11, pp. 166–70
Mazzoni, L, Perez‐Lopez, P, Giampieri, F, Alvarez‐Suarez, JM, Gasparrini, M, Forbes‐
Hernandez, TY, Quiles, JL, Mezzetti, B, & Battino, M 2016, ‘The genetic aspects of berries:
from field to health’, Journal of the Science of Food and Agriculture, vol. 96, pp.365–371.
McCouch, S, Baute, GJ, Bradeen, J, Bramel, P, Bretting, PK, Buckler, E, Burke, JM, Charest,
D, Cloutier, S, Cole, G, Dempewolf, H, Dingkuhn, M, Feuillet, C, Gepts, P, Grattapaglia,
D, Guarino, L, Jackson, S, Knapp, S, Langridge, P, Lawton‐Rauh, A, Lijua, Q, Lusty, C,
Michael, T, Myles, S, Naito, K, Nelson, RL, Pontarollo, R, Richards, CM, Rieseberg, L,
Ross‐Ibarra, J, Rounsley, S, Hamilton, RS, Schurr, U, Stein, N, Tomooka, N, van der
Knaap, E, van Tassel, D, Tol, J, Valls, J, Varshney, RK, Ward, J, Waugh, R, Wenzl, P, &
Zamir, D 2013, ‘Agriculture: Feeding the future’, Nature, vol. 499, pp. 23–24.
McGhie, TK, & Currie, AJ 2008, ‘Effect of pre and post‐harvest technologies on the health
promoting properties of fruit and vegetables’, in: FA Tomas‐Barberan & MI Gil (eds.),
Improving the Health‐promoting Properties of Fruit and Vegetable Products, 1st ed, CRC
Press LLC, Boca Raton, FL, pp. 301–325.
Mehta, RA, Cassol, T, Li, N, Ali, N, Handa, AK, & Mattoo, AK 2002, ‘Engineered
polyamine accumulation in tomato enhances phytonutrient content, juice quality, and
vine life’, Nature Biotechnology, vol. 20, pp. 613–618.
Molvig, L, Tabe, LM, Eggum, BO, Moore, AE, Craig, S, Spencer, D, & Higgins, TJV 1997,
‘Enhanced methionine levels and increased nutritive value of seeds of transgenic lupins
5 Strategies forEnhancing Phytonutrient Content inPlant-Based Foods 227
(Lupinus angustifolius L.) expressing a sunflower seed albumin gene’, Proceedings of the
National Academy of Science of the USA, vol. 94, pp. 8393–8398.
Morton, KJ, Jia, S, Zhang, C, & Holding, DR 2016, ‘Proteomic profiling of maize opaque
endosperm mutants reveals selective accumulation of lysine‐enriched proteins’, Journal
of Experimental Botany, vol. 67, pp. 1381–1396.
Mou, B 2005, ‘Genetic variation of beta‐carotene and lutein contents in lettuce’, Journal of
the American Society for Horticultural Science, vol. 130, pp. 870–876.
Mujtaba, T, Dou, QP 2012, ‘Black tea polyphenols inhibit tumor proteasome activity’,
Invivo, vol. 26, pp. 197–202.
Murphy, MM, Barraj, LM, Herman, D, Bi, X, Cheatham, R, & Randolph, RK 2012,
‘Phytonutrient intake by adults in the United Stated in relation to fruit and vegetable
consumption’, Journal of the Academy of Nutrition and Dietetics, vol. 112, pp. 222–229.
Muzhingi, T, Palacios‐Rojas, N, Miranda, A, Cabrera, ML, Yeum, KJ, & Tang, G 2016,
‘Genetic variation of carotenoids, vitamin E and phenolic compounds in Provitamin
Abiofortified maize’, Journal of the Science of Food and Agriculture, doi: 10.1002/
Nakamura, Y, Yogosawa, S, Izutani, Y, Watanabe, H, Otsuji, E, & Sakai, T 2009, ‘A
combination of indol‐3‐carbinol and genistein synergistically induces apoptosis in
human colon cancer HT‐29 cells by inhibiting Akt phosphorylation and progression of
autophagy’, Molecular Cancer, vol. 8, pp. 100.
Navarre, DA, Pillai, SS, Shakya, R, & Holden, MJ 2011, ‘HPLC profiling of phenolics in
diverse potato genotypes’, Food Chemistry, vol. 127, pp. 34–41.
Navazio, JP, & Simon, PW 2001, ‘Diallel analysis of high carotenoid content in cucumbers’,
Journal of the American Society for Horticultural Science, vol. 126, pp. 100–104.
Neelam, A, Cassol, T, Mehta, RA, Abdul‐Baki, AA, Sobolev, AP, Goyal, RK, Abbott, J,
Segre, AL, Handa, AK, & Mattoo, AK 2008, ‘A field‐grown transgenic tomato line
expressing higher levels of polyamines reveals legume cover crop mulch‐specific
perturbations in fruit phenotype at the levels of metabolite profiles, gene expression, and
agronomic characteristics’, Journal of Experimental Botany, vol. 59, pp. 2337–2346.
Nestel, P, Bouis, HE, Meenakshi, JV, Pfeiffer, W 2006, ‘Biofortification of staple food crops’,
Journal of Nutrition, vol. 136, pp. 1064–1067.
Neto, CC 2007, ‘Cranberry and blueberry: evidence for protective effects against cancer
and vascular diseases’, Molecular Nutrition and Food Research, vol. 51, pp. 652–664.
Nunes, AR, Alves, MG, Moreira, PI, Oliveira, PF, & Silva, BM 2014, ‘Can tea consumption
be a safe and effective therapy against diabetes mellitus‐induced neurodegeneration?’,
Current Neuropharmacology, vol. 12, pp. 475–489.
Oh, J, Hristov, AN, Lee, C, Cassidy, T, Heyler, K, Varga, GA, Pate, J, Walusimbi, S,
Brzezicka, E, Toyokawa, K, Werner, J, Donkin, SS, Elias, R, Dowd, S, & Bravo, D 2013,
‘Immune and production responses of dairy cows to postruminal supplementation with
phytonutrients’, Journal of Dairy Science, vol. 96, pp. 7830–7843.
Oikawa, A, Otsuka, T, Nakabayashi, R, Jikumaru, Y, Isuzugawa, K, Murayama, H, Saito, K,
& Shiratake, K 2015, ‘Metabolic profiling of developing pear fruits reveals dynamic
variation in primary and secondary metabolites, including plant hormones’, PLoS ONE,
vol. 10, e0131408.
Oliveira, M, Ramos, S, Delerue‐Matos, C, & Morais, S 2015, ‘Espresso beverages of pure
origin coffee: Mineral characterization, contribution for mineral intake and geographical
discrimination’, Food Chemistry, vol. 177, pp. 330–338.
Phytonutritional Improvement of Crops
Omar, NF, Hassan, SA, Yusoff, UK, Abdullah, NAP, Wahab, PEM, & Sinniah, R 2012,
‘Phenolics, flavonoids, antioxidant activity and cyanogenic glycosides of organic and
mineral‐base fertilized cassava tubers’, Molecules, vol. 17, pp. 2378–2387.
Padilla, G, Cartea, ME, Velasco, P, de Haro, A & Ordás, A 2007, ‘Variation of glucosinolates
in vegetable crops of Brassica rapa’, Phytochemistry, vol. 68, pp. 536–545.
Pandino, G, Lombardo, S, & Mauromicale, G 2011, ‘Chemical and morphological
characteristics of new clones and commercial varieties of globe artichoke (Cynara
cardunculus var. scolymus)’, Plant Foods for Human Nutrition, vol. 66, pp. 291–297.
Pandjaitan, N, Howard, LR, Morelock, T, & Gil, MI 2005, ‘Antioxidant capacity and
phenolic content of spinach as affected by genetics and maturation’, Journal of
Agriculture and Food Chemistry, vol. 53, pp. 8618–8623.
Park, SH, Kim, YH, Kim, SH, Jeong, YJ, Kim, CY, Lee, JS, Bae, JY, Ahn, MJ, Jeong, JC, Lee,
HS, & Kwak, SS 2015, ‘Overexpression of the IbMYB1 gene in an orange‐fleshed sweet
potato cultivar produces a dual‐pigmented transgenic sweet potato with improved
antioxidant activity’, Physiologia Plantarum, vol. 153, pp. 525–537.
Pawełkowicz, M, Zieliński, K, Zielińska, D, Pląder, W, Yagu, K, Wojcieszek, M, Siedlecka, E,
Bartoszewski, G, Skarzyńska, A, & Przybecki, Z 2016, ‘Next generation sequencing and
omics in cucumber (Cucumis sativus L.) breeding directed research’, Plant Science,
vol.242, pp. 77–88.
Percival, M 1997, ‘Phytonutrients and detoxification’, Clinical Nutrition Insights, vol. 5,
Pereira, MP, Santos, C, Gomes, A, & Vasconcelos, MW 2014, ‘Cultivar variability of iron
uptake mechanisms in rice (Oryza sativa L.)’, Plant Physiology and Biochemistry, vol. 85,
Plazas, M, Vilanova, S, Andújar, I, Gramazio, P, Herraiz, FJ, Raigón, MD, Soler, S, Figàs,
MR, Rodríguez‐Burruezo, R, Fita, A, Borràs, D, & Prohens J 2014, ‘Breeding vegetables
with improved bioactive properties’, Bulletin UASVM Horticulture, vol. 71, pp. 165–172.
Pons, E, Alquiezar, B, Rodriguez, A, Martorell, P, Genoves, S, Ramon, D, Rodrigo, MJ,
Zacarias, L, & Pena, L 2014, ‘Metabolic engineering of b‐carotene in orange fruit increases
its in vivo antioxidant properties’, Plant Biotechnology Journal, vol. 12, pp. 17–27.
Prohens, J, Rodríguez‐Burruezo, A, Raigón, MD, & Nuez, F 2007, ‘Total phenolics
concentration and browning susceptibility in a collection of different varietal types and
hybrids of eggplant: implications for breeding for higher nutritional quality and reduced
browning’, Journal of the American Society for Horticultural Science, vol. 132,
Ramirez‐Sanchez, I, Maya, L, Ceballos, G, & Villarreal, F 2010, ‘Fluorescent detection of
(‐)‐epicatechin in microsamples from cacao seeds and cocoa products: comparison with
Folin‐Ciocalteu method’, Journal of Food Composition and Analysis, vol. 23, pp. 790–793.
Rechkemmer, G 2001, ‘Funktionelle Lebensmittel‐Zukunft de Ernahrung oder Marketing‐
Strategie’, Forschungereport Sonderheft, vol. 1, pp. 12–15.
Riccioni, G, Speranza, L, Pesce, M, Cusenza, S, D’Orazio, N, & Glade, MJ 2012, ‘Novel
phytonutrient contributors to antioxidant protection against cardiovascular disease’,
Nutrition, vol. 28, pp. 605–610.
Rivero‐Pérez, MD, Muñiz, P, & González‐Sanjosé, ML 2007, ‘Antioxidant profile of red
wines evaluated by total antioxidant capacity, scavenger activity, and biomarkers of
oxidative stress methodologies’, Journal of Agricultural and Food Chemistry, vol. 55,
5 Strategies forEnhancing Phytonutrient Content inPlant-Based Foods 229
Robinson, JW 1960, ‘Atomic Absorption Spectroscopy’, Analytical Chemistry, vol. 32,
Rocheford, TR, Wong, JC, Egesel, CO, & Lambert, RJ 2002, ‘Enhancement of vitamin E
levels in corn’, The Journal of the American College of Nutrition, vol. 21, pp. 191S–198S.
Rodríguez‐Burruezo, A, Raigón, MD, Prohens, J, & Nuez, F 2012, ‘Characterization for
bioactive compounds of Spanish pepper landraces’, Acta Horticulturae, vol. 918,
Rodriguez‐Casado, A 2016, ‘The health potential of fruits and vegetables phytochemicals:
notable examples’, Critical Reviews in Food Science and Nutrition, vol. 56, pp.
Ronco, A, Stefani, ED, Boffetta, P, Deneo‐Pellegrini, H, Mendilaharsu, M, & Leborgne, F
1999, ‘Vegetables, fruits, and related nutrients and risk of breast cancer: a case‐control
study in Uruguay’, Nutrition and Cancer, vol. 35, pp. 111–119.
Rosati, C, Aquilani, R, Dharmapuri, S, Pallara, P, Marusic, C, Tavazza, R, Bouvier, F,
Camara, B, & Giuliano, G 2000, ‘Metabolic engineering of beta‐ carotene and lycopene
content in tomato fruit’, Plant Journal, vol. 24, pp. 413–419.
Saito, K 2013, ‘Phytochemical genomics–a new trend’, Current Opinion in Plant Biology,
vol. 16, pp. 373–380.
Santos, CS, Silva, AI, Serrão, I, Carvalho, AL, & Vasconcelos, MW 2013, ‘Transcriptomic
analysis of iron deficiency related genes in the legumes’, Food Research International,
vol.54, pp. 1162–1171.
Sax, K 1923, ‘The association of size differences with seed coat pattern and pigmentation in
Phaseolus vulgaris’, Genetics, vol. 8, pp. 552–560.
Seaby, EG, Pengelly, RJ, & Ennis, S 2015, ‘Exome sequencing explained: a practical guide to
its clinical application’, Briefings in Functional Genomics, pii: elv054.
Seeram, NP, Henning, SM, Niu, Y, Lee, R, Scheuller, HS, & Heber, D 2006, ‘Catechin and
caffeine content of gree tea dietary supplements and correlation with antioxidant
capacity’, Journal of Agricultural and Food Chemistry, vol. 54, pp. 1599–1603.
Sestari, I, Zsögön, A, Rehder, GG, de Lira Teixeira, L, Hassimotto, NMA, Purgatto, E,
Benedito, VA, & Pereira Peres, LE 2014, ‘Near‐isogenic lines enhancing ascorbic acid,
anthocyanin and carotenoid content in tomato (Solanum lycopersicum L. cv Micro‐Tom)
as a tool to produce nutrient‐rich fruits’, Scientia Horticulturae, vol 175, pp.111–120.
Sharma, S, Sheehy, T, & Kolonel, L 2014, ‘Sources of vegetables, fruits and vitamins A, C
and E among five ethnic groups: results form a multiethnic cohort study’, European
Journal of Clinical Nutrition, vol. 68, pp. 384–391.
Shao, Y, Jin, L, Zhang, G, Lu, Y, Shen, Y, & Bao, J 2011, ‘Association mapping of grain color,
phenolic content, flavonoid content and antioxidant capacity in dehulled rice’,
Theoretical and Applied Genetics, vol. 122, pp. 1005–1016.
Shekhar, S, Mishra, D, Buragohain, AK, Chakraborty, S, & Chakraborty, N 2015,
‘Comparative analysis of phytochemicals and nutrient availability in two contrasting
cultivars of sweet potato (Ipomoea batatas L.)’, Food Chemistry, vol. 173, pp. 957–965.
Shewmaker, CK, Sheehy, JA, Daley, M, Colburn, S, & Ke, DY 1999, ‘Seed‐specific
overexpression of phytoene synthase: increase in carotenoids and other metabolic
effects’, The Plant Journal, vol. 20, pp. 401–412.
Shin, Y, Park, H, Yim, S, Baek, N, Lee, C, An, G, & Woo Y 2006, ‘Transgenic rice lines
expressing maize C1 and R‐S regulatory genes produce various flavonoids in the
endosperm’, Plant Biotechnology Journal, vol. 4, pp. 303–315.
Phytonutritional Improvement of Crops
Shintani, D, & DellaPenna, D 1998, ‘Elevating the vitamin E content of plants through
metabolic engineering’, Science, vol. 282, pp. 2098–2100.
Sims, DA, & Gamon, JA 2002, ‘Relationships between leaf pigment content and spectral
reflectance across a wide range of species, leaf structures and developmental stages’,
Remote Sensing of Environment, vol. 81, pp. 337–354.
Singh, D, Singh, PK, Chaudhary, S, Mehla, K, & Kumar, S 2012, ‘Exome sequencing and
advances in crop improvement’, Advances in Genetics, vol. 79, pp. 87–121.
Soininen, TH, Jukarainen, N, Auriola, SOK, Julkunen‐Tiitto, R, Karjalainen, R, &
Vepsäläinen, JJ 2014, ‘Quantitative metabolite profiling of edible onion species by NMR
and HPLC‐MS’, Food Chemistry, vol. 165, pp. 499–505.
Stansell, S, Cory, W, Couillard, D, & Farnham, M 2015, ‘Collard landraces are novel sources
of glucoraphanin and other aliphatic glucosinolates’, Plant Breeding, vol. 134,
Stark‐Lorenzen, P, Nelke, B, Hanssler, G, Muhlbach, HP, & Thomzik, JE 1997, ‘Transfer of a
grapevine stilbene synthase gene to rice (Oryza sativa L.)’, Plant Cell Reports, vol. 16,
Subedi, L, Timalsena, S, Duwadi, P, Thapa, R, Paudel, A, Parajuli, K 2014, ‘Antioxidant
activity and phenol and flavonoid contents of eight medicinal plants from Western
Nepal’, Journal of Traditional Chinese Medicine, vol. 34, pp. 584–590.
Szankowski, I, Briviba, K, Fleschhut, J, Schonherr, J, Jacobsen, HJ, & Kiesecker, H 2003,
‘Transformation of apple (Malus domestica Borkh.) with the stilbene synthase gene from
grapevine (Vitis vinifera L.) and a PGIP gene from kiwi (Actinidia deliciosa)’, Plant Cell
Reports, vol. 22, pp. 141–149.
Teer, JK, & Mullikin, JC 2010, ‘Exome sequencing: the sweet spot before whole genomes’,
Human Molecular Genetics, vol. 19, pp. R145–R151.
Tennant, DR, Davidson, J, & Day, AJ 2014, ‘Phytonutrient intakes in relation to European
fruit and vegetable consumption patterns observed in different food surveys’, British
Journal of Nutrition, vol. 112, pp. 1214–1225.
Thapa, A, & Chi, EY 2015, ‘Biflavonoids as potential small molecule therapeutics for
Alzheimer’s Disease’, Advances in Experimental Medicine and Biology, vol. 863,
Thomson, MJ, Ismail, AM, McCouch, SR, & Mackill, DJ 2009, ‘Marker assisted breeding’,
in: Abiotic Stress Adaptation in Plants, A Pareek, SK Sopory, & HJ Bohnert (eds.),
Springer, Amsterdam, pp 451–469.
Thormar, H 2012, ‘Patented non‐antibiotic agents as animal feed additives’, Recent Patents
on Food, Nutrition & Agriculture, vol. 4, pp. 155–168.
Tohge, T, Scossa, F, & Fernie, AR 2015, ‘Integrative approaches to enhance understanding
of plant metabolic pathway structure and regulation’, Plant Physiology, vol. 169,
Unnevehr, L, Pray, C, & Paarlberg, R 2007, ‘Addressing micronutrient deficiencies:
Alternative interventions and technologies’, AgBioForum, vol. 10, pp. 124–134.
Van Ginneken, VJT, Helsper, JPFG, de Visser, W, van Keulen, H, & Brandenburg, WA 2011,
‘Polyunsaturated fatty acids in various macroalgal species from north Atlantic and
tropical seas’, Lipids in Health and Disease, vol. 10, pp. 104.
Varoni, EM, Lodi, G, & Iriti, M 2015, ‘Ethanol versus phytochemicals in wine: oral cancer
risk in a light drinking perspective’, International Journal of Molecular Sciences, vol. 16,
5 Strategies forEnhancing Phytonutrient Content inPlant-Based Foods 231
Vazquez‐Martin, A, Fernández‐Arroyo, S, Cufí, S, Oliveras‐Ferraros, C, Lozano‐Sánchez, J,
Vellón, L, Micol, V, Joven, J, Segura‐Carretero, A, & Menendez, JA 2012, ‘Phenolic
secoiridoids in extra virgin olive oil impede fibrogenic and ocogenic epithelial‐to‐
mesenchymal transition: extra virgin olive oil as a source of novel antiaging
phytochemicals’, Rejuvenation Research, vol. 15, pp. 3–21.
Venkatesan, R, Ji, E, & Kim, SY 2015, ‘Phytochemicals that regulate neurodegenerative
disease by targeting neurotrophins: a comprehensive review’, BioMed Research
International, vol. 2015, 814068.
Verma, S, Gupta, S, Bandhiwal, N, Kumar, T, Bharadwaj, C, & Bhatia, S 2015, ‘High‐density
linkage map construction and mapping of seed trait QTLs in chickpea (Cicer arietinum
L.) using Genotyping‐by‐Sequencing (GBS)’, Scientific Reports, vol. 5, 17512.
Vernarelli, JA, & Lambert, JD 2013, ‘Tea consumption is inversely associated with weight
status and other markers for metabolic syndrome in US adults’, European Journal of
Nutrition, vol. 52, pp. 1039–1048.
Visioli, F, Borsani, L, & Galli, C 2000, ‘Diet and prevention of coronary heart disease: the
potential role of phytochemicals’, Cardiovascular Research, vol. 47, pp. 419–425.
Wall, MM, & Tripathi, S 2013, ‘Papaya Nutritional Analysis’, in: R Ming & PH Moore (eds.),
Plant Genetics and Genomics of Papaya, Springer, New York, pp. 377–390.
Wang, SY, & Lewers, KS 2007, ‘Antioxidant capacity and flavonoid content in wild
strawberries’, Journal of the American Society for Horticultural Science, vol. 132, pp.629–637.
Wen, Y, Zhong, G, Gao, Y, Lan, Y, Duan, C, & Pan, Q 2015, ‘Using the combined analysis of
transcripts and metabolites to propose key genes for differential terpene accumulation
across two regions’, BMC Plant Biology, vol. 15, 240.
WHO (2015) Helathy diet. Fact sheet N° 394. http://www.who.int/mediacentre/factsheets/
fs394/en/ (last accessed in: October 15, 2015)
Wishard, DS 2008, ‘Quantitative metabolomics using NMR’, Trends in Analytical
Chemistry, vol. 27, pp. 228–237.
Wolfram, S 2007, ‘Effects of green tea and EGCG on cardiovascular and metabolic health’,
Journal of the American College of Nutrition, vol. 26, pp. 373S‐388S.
Woolhouse, M, Ward, M, van Bunnik, B, & Farrar, J 2015, ‘Antimicrobial resistance in
humans, livestock and the wider environment’, Philosophical Transactions of the Royal
Society B, vol. 370, 20140083.
Yang, J, Meyers, KJ, van der Heide, J, & Liu, RH 2012, ‘Varietal differences in phenolic
content and antioxidant and anti proliferative activities of onions’, Journal of Agriculture
and Food Chemistry, vol. 52, pp. 6787–6793.
Yang, Q, Wang, R, Ren, S, Szoboszlay, M, & Moe, LA 2015, ‘Practical survey on antibiotic‐
resistant bacterial communities in livestock manure and manure‐amended soil’, Journal
of Environmental Science and Health, Part B 0, 1–10.
Yang, W, Guo, Z, Huang, C, Duan, L, Chen, G, Jiang, N, Fang, W, Feng, H, Xie, W, Lian, X,
Wang, G, Luo, Q, Zhang, Q, Liu, Q, & Xiong, L 2014, ‘Combining high‐throughput
phenotyping and genome‐wide association studies to reveal natural genetic variation in
rice’, Nature Communications, vol. 5, 5087.
Yao, Y, Sang, W, Zhou, M, & Ren, G 2010, ‘Phenolic composition and antioxidant activities
of 11 celery cultivars’, Journal of Food Science, vol. 75, pp. C9‐C13.
Ye, J, Hu, T, Yang, C, Li, H, Yang, M, Ijaz, R, Ye, Z, & Zhang, Y 2015, ‘Transcriptome
profiling of tomato fruit development reveals transcription factors associated with
ascorbic acid, carotenoid and flavonoid biosynthesis’, PLoS ONE, vol. 10, e0130885.
Phytonutritional Improvement of Crops
Ye, X, Al‐Babili, S, Kloti, A, Zhang, J, Lucca, P, Beyer, P, & Potrykus, I 2000, ‘Engineering
the provitamin A (beta‐carotene) biosynthetic pathway into (carotenoid‐free) rice
endosperm’, Science, vol. 287, pp. 303–305.
Yoo, KS, Bang, H, Lee, EJ, Crosby, K, & Patil, BS 2012, ‘Variation of carotenoid, sugar, and
ascorbic acid concentrations in watermelon genotypes and genetic analysis’, Horticulture,
Environment, and Biotechnology, vol. 53, pp. 552–560.
Youwei, Z, Jinlian, Z, & Yonghong, P 2008, ‘A comparative study on the free radical
scavenging activities of some fresh flowers in southern China’, LWT–Food Science and
Technology, vol. 41, pp. 1586–1591.
Yu, O, Jung, W, Shi, J, Croes, RA, Fader, GM, McGonigle, B, & Odell, JT 2000, ‘Production
of the isoflavones genistein and daidzein in non‐legume dicot and monocot tissues’,
Plant Physiology, vol. 124, pp. 781–794.
Yu, O, Shi, J, Hession, AO, Maxwell, CA, McGonigle, B, & Odell, JT 2003, ‘Metabolic
engineering to increase isoflavone biosynthesis in soybean seed’, Phytochemistry, vol. 63,
Xiong, L, Yang, J, Jiang, Y, Li, B, Hu, Y, Zhou, F, Mao, S, & Shen, C 2014, ‘Phenolic
compounds and antioxidant capacities of 10 common edible flowers from China’, Journal
of Food Science, vol. 79, pp. C517‐C525.
Zeng, L, Holly, JM, & Perks, CM 2014, ‘Effects of physiological levels of the green tea
extract epigallocatechin‐3‐gallate on breast cancer cells’, Frontiers in Endocrinology,
vol. 5, 61.
Zhao, J 2007, ‘Nutraceuticals, nutritional therapy, phytonutrients, and phytotherapy for
improvement of human health: A perspective on plant biotechnology application’, Recent
Patents on Biotechnology, vol. 1, pp. 75–97.