42 Part I: Principles
used to transform tobacco explants. The results
showed that transgenic lines had a greater capacity
for overall biosynthesis of HGA and produced a
two-fold increase in vitamin E in the seeds. Vitamin
E content in leaves was not affected (Falk et al.
2003).
In another approach towards vitamin E enhance-
ment, Cahoon et al. (2003) reported the identifica-
tion and isolation of a novel monocot gene that
encodes HGGT, which is so far the only known
enzyme specific for the synthesis of tocotrienols.
These researchers found that the expression of the
barley HGGT enhanced the tocotrienol synthesis by
10- to 15-fold in the leaves of A. thalianaand by six-
fold in the seeds of corn. The barley HGGT cDNA
was placed under the control of the 35S CaMV pro-
moter and the nopaline synthase terminator. The
construct was inserted into the binary vector
pZS199 to generate plasmid pSH24. The plasmid
was then introduced into Agrobacteriumfor trans-
formation into tobacco and A. thaliana(Cahoon et
al. 2003).
A third way by which vitamin E content in plants
can be manipulated involves the last enzyme in the
final step of the tocotrienol and tocopherol biosyn-
thetic pathway, in which -tocotrienol and -toco-
pherol are converted to -tocotrienol and -toco-
pherol, respectively. This step is catalyzed by the
enzyme -tocopherol methyltransferase (-TMT)
(EC 2.1.1.95) (Fig. 3.5) (Shintani and DellaPenna
1998). -tocopherol has the highest oxidative prop-
erty among the members of the vitamin E family
(Kamal-Eldin and Appelqvist 1996). Unfortunately,
plant oils, which are the main dietary source of vita-
min E, contain only a fractional amount of -toco-
pherol but a high level of its precursor, -tocopherol.
Shintani and DellaPenna overexpressed endogenous
A. thaliana-TMT to enhance conversion of -
tocopherol into -tocopherol. They introduced the
-TMT cDNA construct under the control of a 35S
CaMV promoter in a binary vector into A. thaliana
plants by Agrobacterium-mediated transformation.
-tocopherol content of bioengineered seeds was
nine-fold greater than that of the wild-type seeds
(Shintani and DellaPenna 1998).
ESSENTIALMINERALS
To maintain a well functioning, healthy body, hu-
mans require 17 different essential minerals in their
diet. Minerals are inorganic ions found in nature and
cannot be made by living organisms. They can be
divided into two classes: macronutrients and micro-
nutrients. Macronutrients are the minerals that we
need in large quantity, including calcium, phospho-
rus, sodium, magnesium, chlorine, sulfur, and sili-
con. Micronutrients, or trace minerals, are the min-
erals that are required in small amounts, of which
iron is the most prevalent, followed by fluorine, zinc,
copper, cobalt, iodine, selenium, manganese, molyb-
denum, and chromium. Although a balanced con-
sumption of plant-based foods should naturally pro-
vide these nutrients, mineral deficiency, especially
of iron, is widespread among the world population.IronEven though iron is required in trace amounts, it is
the most widespread nutrient deficiency worldwide.
It is believed that about 30% of the world population
suffers from serious nutritional problems caused by
insufficient intake of iron (WHO 1992). Iron is an
important constituent of hemoglobin, the oxygen-
carrying component of the blood, and is also a part
of myoglobin, which helps muscle cells to store
oxygen. Low iron levels can cause the development
of iron deficiency anemia. In an anemic person the
blood contains a low level of oxygen, which result in
many health problems including infant retardation
(Walter et al. 1986), pregnancy complications (Mur-
phy et al. 1986), low immune function (Murakawa
et al. 1987), and tiredness (Basta et al. 1979). Iron is
present in food in both inorganic (ferric and ferrous)
and organic (heme and nonheme) forms. Heme iron,
which is highly bioavailable, is derived primarily
from the hemoglobin and myoglobin of flesh foods
such as meats, fish, and poultry (Taylor et al. 1986).
In humans, reduced iron (ferrous) is taken up more
readily than oxidized (ferric) iron. Several ap-
proaches have been used in the fight against iron
deficiency including nutraceutical supplementation,
food fortification, and various methods of food prep-
aration and processing (Maberly et al. 1994). So far,
none of these approaches has been successful in
eradicating iron deficiency, especially in developing
countries. A new tool in the fight against nutrient
deficiency is the use of biotechnology to improve
essential mineral nutrition in staple crops.
At this time, there are basically two ways in
which genetic engineering can be used for this pur-