Open access to science eprints
TheSparc provides both scientists and the general public free open access to scientific papers that are important for the survival of people and planet
The 'Golden Rice' - An Exercise in How Not to
ISIS-TWN Sustainable Science Audit #1
Evaluating science and technology for sustainability and social
ISIS believes science as much as scientists should be
socially and ecologically accountable, and has launched a sustainable
science audit project jointly with the Third World Network
See also The Golden Rice Scandal Unfolds
The golden rice - a GM rice engineered to
produce pro-Vitamin A - is being offered to the Third World as cure for
widespread vitamin A deficiency.
The audit uncovers fundamental deficiencies in all
aspects, from the scientific/social rationale to the science and
technology involved. It is being promoted in order to salvage a morally as
well as financially bankrupt agricultural biotech industry.
The scientific/social rationalization for the project
exposes a reductionist self-serving scientific paradigm that fails to see
the world beyond its own narrow confines. The golden rice is a
useless application. Some 70 patents have already been filed on the GM
genes and constructs used in making the golden rice. It is a
drain on public resources and a major obstruction to the implementation of
sustainable agriculture that can provide the real solutions to world
hunger and malnutrition.
Golden rice is not a second generation
GM crop as has been claimed. It involves standard first generation
technology, and carries some of the worst features in terms of hazards to
health and biodiversity. Rockefeller Foundation, the major funder of the
project by far has withdrawn support from it. The project should be
Key Words: Golden rice, vitamin A
deficiency, Green Revolution, sustainable science, GM technology, gene
patents, GM constructs
A gift-horse for the poor
A report in Financial Times states that the
creators of golden rice have struck a ground-breaking
deal with corporate giant AstraZeneca to give Third World farmers
free access to the grain while allowing it to be commercially exploited in
the developed world. The company will oversee the production of stable GM
line(s) and patenting, and take the lines through field trials and
commercial approval. While farmers in developed countries will have to pay
royalties, those in the Third World earning less than US$10 000 will not.
But will Third World farmers be allowed to save the seeds for replanting?
It did not say.
This golden rice, not yet available, is
already worth its weight in diamonds. The project was funded from four
sources of public finance totaling US$100 million: the philanthropic
Rockefeller Foundation, whose mission is to support scientific research
that specifically benefit the poor, the Swiss Federal Institute of
Technology, the European Community Biotech Program and the Swiss Federal
Office for Education and Science.
The announcement failed to mention that there are
already 70 patent claims on the genes, DNA sequences, and gene constructs
used to make the golden rice. Will the cost of paying royalties for the
previous 70 patent claims be added to the cost of the golden rice? Which
of the royalties on the seventy-odd patents would the Third World farmers
be absolved from paying? Rockefeller Foundation, the major funder by far,
has reportedly abandoned the project to " shift its agricultural
funding focus to support research that will have a more direct benefit to
The scientific/social rationale is fallacious
Many have commented on the absurdity of offering golden
rice as the cure for vitamin A deficiency when there are plenty of
alternative, infinitely cheaper sources of vitamin A or pro-vitamin A,
such as green vegetables and unpolished rice, which would be rich in other
essential vitamins and minerals besides. To offer the poor and
malnourished a high-tech golden rice tied up in multiple
patents, that has cost US$100 million to produce and may cost as much to
develop, is worse than telling them to eat cake.
Golden rice was engineered to produce
pro-vitamin A or b-carotene (the
substance that makes carrots orange) in the endosperm, i.e., the
part of the rice grain that remains after it has been polished. The
scientific paper started with a review of the literature to rationalize
why such GM rice is needed and of benefit for the Third World. The paper
was accompanied by an unusually long news feature entitled, The
Green Revolution Strikes Gold, which reinforced the rationalization
for the project, explaining the remarkable feat of technology involved and
stated that the scientists intend to make the golden rice "freely
available to the farmers who need it most." The last sentence in this
glowing report, however, gave the game away: "One can only hope that
this application of plant genetic engineering to ameliorate human misery
without regard to short-term profit will restore this technology to
What were the reasons for the scientists to embark on
the project? It is important to know, as these reasons may have been used
to persuade funders to support the project in the first place, and funders
ought to bear as much of the responsibility.
The first reason given is that the aleurone layer (in
unpolished rice) is usually removed by milling as it turns rancid on
storage, especially in tropical areas; and the remaining endosperm lacks
pro-vitamin A. The researchers are tacitly admitting that at least some
varieties of unpolished rice will have pro-vitamin A. The reason rice is
milled is to prolong storage for export, and to suit the tastes of the
developed world. So why not give the poor access to unpolished rice? A
proportion of every rice harvest could be kept unpolished and either given
freely to the poor, or sold at the cheapest prices. But the scientists
have not considered that possibility. Unpolished rice is fact part of the
traditional Asian diet until the Green Revolution when aggressive
marketing of white polished rice created a stigma of unpolished rice.
However, most rural communities still consume unpolished rice and now that
consumers have become aware of its nutritional value, unpolished rice is
becoming sought after.
"Predominant rice consumption", the
researchers claim, promotes vitamin A deficiency, a serious health problem
in at least 26 countries, including highly populated areas of Asia,
Africa, and Latin America. Some 124 million children worldwide are
estimated to be vitamin A deficient. (Actually, the latest figures quoted
in a press release from the International Rice Research Institute (IRRI)
is 250 million preschool children.) The scientists seem to be unaware that
people do not eat plain rice out of choice. The poor do not get enough to
eat and are undernourished as well as malnourished. The Food and
Agricultural Organization (FAO) started a project in 1985 to deal with
vitamin A deficiency using a combination of food fortification, food
supplements and general improvements in diets by encouraging people to
grow and eat a variety of green leafy vegetables. One main discovery is
that the absorption of pro-vitamin A depends on the overall nutritional
status, which in turn depends on the diversity of the food consumed.
"Predominant rice consumption" is most likely
to be accompanied by other dietary deficiencies. A recent study by the
Global Environmental Change Programme concludes that predominant
consumption of Green Revolution crops is responsible for iron deficiency
in an estimated 1.5 billion, or a quarter of the worlds population.
The worst affected areas are in rice-growing regions in Asia and
South-East Asia where the Green Revolution had been most successful in
increasing crop yield.
Research institutions such as IRRI have played the key
role in introducing Green Revolution crops to the Third World. IRRI was
founded in 1959 under an agreement forged by the Rockefeller and Ford
Foundations with the Philippine government, and its lease for operation
expires in 2003. At its recent 40th anniversary celebration, hundreds of
Filipino rice farmers protested against IRRI for introducing GM crops,
blaming IRRI, among other things, for promoting the Green Revolution and
causing massive loss of biological diversity in rice paddies throughout
It is clear that vitamin A deficiency is accompanied by
deficiencies in iron, iodine and a host of micronutrients, all of which
comes from the substitution of a traditionally varied diet with one based
on monoculture crops of the Green Revolution. The real cure is to
re-introduce agricultural biodiversity in the many forms of sustainable
agriculture already being practiced successfully by tens of millions of
farmers all over the world.
As the scientists know, clinical deficiency can be dealt
with by prescription of vitamin A pills, which are affordable and
immediately available. "Oral delivery of vitamin A is problematic",
they state. Judging from the reference cited they may be referring to the
well-known harmful effects of vitamin A overdose. But why would high
levels of pro-vitamin A rice in a staple food that people generally
consume in the largest amounts in a meal not also cause problems connected
with overdose? In particular, vitamin A poisoning has been known to
result from excessive b-carotene
intake in food.
Finally, why is it necessary to genetic engineer rice? "Because
no rice cultivars produce [pro-vitamin A] in the endosperm, recombinant
technologies rather than conventional breeding are required." This is
the conclusion to the whole fallacious reasoning process. It amounts to
this: rice is polished, which removes pro-vitamin. A, therefore a hundred
million dollars (much of it tax-payers money) are needed to put
pro-vitamin A into polished rice. A more likely explanation is that the
geneticists are looking for funding to do their research, and have
constructed, as best they could, a series of rationalizations for why they
should be supported. Neither the scientists nor the funders have looked
further beyond the technology to peoples needs and aspirations, or
to what the real solutions are.
The science and technology is standard first generation
It took ten years to engineer b-carotene
into polished rice because rice naturally does not have the metabolic
pathway to make it in the endosperm, perhaps for good biological reasons.
Immature rice endosperm makes the early precursor,
geranylgeranyl-diphosphate (GGPP). In order to turn GGPP into
b-carotene, four metabolic reactions
are needed, each catalyzed by a different enzyme. Enzyme 1, phytoene
synthase converts GGPP to phytoene, which is colorless. Enzymes 2 and 3,
phytoene desaturase and z-carotene
desaturase, each catalyzes the introduction of two double-bonds into the
phytoene molecule to make lycopene, which is red in colour. Finally,
Enzyme 4, lycopene b-cyclase turns
lycopene into b-carotene. Hereafter,
the enzymes will be referred to by numbers only. Thus, a total of four
enzymes have to be engineered into the rice in such a way that the enzymes
are expressed in the endosperm. Some very complicated artificial gene
constructs have to be made. The gene constructs are made in units called
expression cassettes (see Box 1)
The gene expression cassette a
unit of transgenic construct
The gene for each enzyme never goes in alone. It has
to be accompanied by a special piece of genetic material (DNA), the
promoter, which signals the cell to turn the gene on, ie, to
transcribe the DNA gene sequence into RNA. At the end of the gene,
there has to be another signal, a terminator, to mark the RNA
so it can be translated into protein. To target the protein to the
endosperm, an extra bit of DNA, a transit sequence, is
required. The resulting expression cassette for each gene is as
Typically, each bit of the construct: promoter,
transit sequence, gene and terminator is from a different source.
Several expression cassettes are usually linked in series, or stacked
in the final construct.
In order to select for the plant cells that have taken
up the foreign genes and gene-constructs, golden rice makes
use of a standard antibiotic resistance gene coding for hygromycin
resistance, also equipped with its own promoter and terminator. All these
expression cassettes have to be introduced into the rice plant cells. One
simplification available is that the reactions catalyzed by two of the
enzymes, 2 and 3, could be done by a single bacterial enzyme, lets
call it enzyme 2-3, so a total of four expression cassettes have to be
introduced, one for each of three enzymes and the fourth for the
antibiotic resistance marker.
Unlike natural genetic material which consists of stable
combinations of genes that have co-existed for billions of years,
artificial constructs consists of combinations that have never existed,
not in billions of years of evolution. Artificial gene-constructs are
well-known to be structurally unstable, which means they tend to break and
join up incorrectly, and with other bits of genetic material, resulting in
new unpredictable combinations. This process of breaking and joining of
genetic material is referred to as recombination. The more
complicated the construct, the more it tends to break and rearrange or
form new combinations. The instability of the construct means that it is
seldom inserted into the plant genome in its intended form. The inserts
are generally rearranged, with parts deleted, or repeated.
In order to make many copies of the construct and to
facilitate entry into plant cells, the construct is spliced into an
artificial vector, which is generally made from genetic parasites that
live inside cells. The artificial vector also enables the construct to be
efficiently smuggled into the plant cell and to jump into the genome of
the plant cell. The vector used in the case of the golden rice
is the one most widely used since the beginning of plant genetic
engineering. It is derived from the T-DNA, part of the
tumor-inducing (Ti) plasmid (a genetic parasite) of the soil
bacterium, Agrobacterium. The Ti plasmid naturally invades
plant cells, inserting the T-DNA into the plant cell genome, and
causing the cell to develop into a plant tumor or gall. The artificial
gene construct is spliced in between the left and right borders of the
T-DNA vector. The borders of the T-DNA are hotspots
for recombination, ie, they have a pronounced tendency to break and join
up, which is ultimately why the vector can invade the plants genome
and carry its hitch-hiker gene construct along with it.
Three different constructs were made. The first consists
of the expression cassettes of enzyme 1 from daffodils and enzyme 2-3 from
the plant bacterial pathogen, Erwinia uredovora, together with the
expression cassette of an antibiotic resistance marker gene that codes for
hygromycin resistance. Another antibiotic resistance gene (coding for
kanamycin resistance) is also present, albeit lacking a promoter.
Hygromycin and kanamycin are both aminoglycoside antibiotics that inhibit
protein synthesis. The resistance genes originate from bacteria and
generally have specificities for more than one aminoglycoside antibiotic.
This first construct is the most complicated, but it still does not have
all the required enzymes. Enzyme 1 and the hygromycin resistance gene are
both equipped with a promoter from the cauliflower mosaic virus (CaMV),
which is especially hazardous (see below).
The second construct consists of the expression
cassettes of enzyme 1 and enzyme 2-3 as in the first, but without any
antibiotic resistance marker genes. The third construct consists of the
expression cassette of enzyme 4, again from daffodil, stacked with the
hygromycin-resistance marker-gene cassette. The strategy of separating the
genes for the enzymes and antibiotic resistance marker into two different
constructs is that it overcomes some of the problems of structural
instability: the more cassettes stacked, the more unstable is the
Each construct was spliced into a T-DNA vector,
and two transformation experiments were carried out. In the first
experiment, 800 immature rice embryos were inoculated with the vector
containing the first construct, and hygromycin was used to select for
resistant plants that have taken up the vector, resulting in 50 GM plants.
In the second experiment, 500 immature embryos were inoculated with a
mixture of the vectors containing the second and third construct
respectively. Selection with hygromycin gave rise to 60 GM plants that
have taken up the third construct, but only twelve of these had taken up
the second construct as well. The transformation process is well-known to
be random, as there is no way to target the foreign genes to precise
locations in the genome. There could be more than one site of insertion in
a single cell. Furthermore, as mentioned earlier, the actual inserts are
likely to be rearranged, or subject to deletions or repetitions. Hence
each transformed cell will have its own distinctive pattern of insert(s),
and each GM plant, which comes from the single transformed cell, will
differ from all the rest.
Note that the GM plants from the first experiment will
not have the full complement of enzymes required to make
b-carotene, and should give red
endosperm from the lycopene present. Only the GM plants from the second
experiment which have taken up both vectors would possess all the
enzymes needed, and give orange-colored endosperm.
Uncontrollable technology and unpredictable outcomes raise questions on
Unexpectedly, transgenic plants from both transformation
experiments gave orange polished grains. Chemical analyses confirmed that
only b-carotene, in varying amounts,
was found in all lines, but no lycopene. This suggests that enzyme 4 may
be present in rice endosperm normally, or it could be induced by lycopene,
to turn all of the lycopene into b-carotene.
Lutein and zeaxanthin, two other products derived from lycopene, were also
identified in varying amounts besides b-carotene.
All of these were absent from non-GM rice.
In addition, many other uncharacterized, unidentified
products were found, which differ from one line to another. What is the
nutritional value of the other products? Are any of the known and unknown
products harmful? Without thorough chemical analyses and toxicity tests,
it is impossible to tell. This highlights the unpredictable,
uncontrollable nature of the technology.
Molecular analyses of the GM inserts were not done in
any detail. Nevertheless, judging from the evidence presented, there are
the usual signs of deletions, rearrangements and multiple repeats of the
constructs inserted due to structural instability of the constructs and
the tendency for recombination. There is no guarantee that any of the
plants will give stable progeny in successive generations. The instability
of GM lines is well-known, and is a continuing problem for the industry.
Inserted genes can lose their activities or become lost altogether in
subsequent generations. There is nothing in golden rice to
distinguish it from standard first generation GM plants with all the
well-known defects and hazards.
Golden rice is no technical improvement and more unsafe
Golden rice exhibits all the undesirable,
hazardous characteristics of existing GM plants, and in added measure on
account of the increased complexity of the constructs and the sources of
genetic material used. The hazards are highlighted below.
- It is made with a combination of genes and genetic material from
viruses and bacteria, associated with diseases in plants, and from other
- The gene constructs are new, and have never existed in billions of
years of evolution
- Unpredictable by-products have been generated due to random gene
insertion and functional interaction with host genes, which will differ
from one plant to another.
- Over-expression of transgenes linked to viral promoters, such as that
from CaMV, exacerbates unintended metabolic effects as well as
instability (see below). There are at least two CaMV promoters in each
transgenic plant of the golden rice, one of which is linked
to the antibiotic resistance marker gene.
- The transgenic DNA is structurally unstable, leading to instability
of the GM plants in subsequent generations, multiplying unintended,
- Structural instability of transgenic DNA increases the likelihood of
horizontal gene transfer and recombination.
- Instability of transgenic DNA is enhanced by the CaMV promoter, which
has a recombination hotspot, thereby further increasing the potential
for horizontal gene transfer.
- The CaMV promoter is promicuous in function and works efficiently in
all plants, in green algae, yeast and E. coli. The spread of
genes linked to this promoter by ordinary cross-pollination or by
horizontal gene transfer will have enormous impacts on health and
biodiversity. In particular, the hygromycin resistance gene linked to it
may be able to function in bacteria associated with infectious diseases.
- Horizontal transfer of transgenic DNA from GM plants into soil fungi
and bacteria has been demonstrated in laboratory experiments. Recent
evidence suggests that it has also taken place in a field-trial site for
GM sugar-beets, in which transgenic DNA persisted in the soil for at
least two years afterwards.
- Prof. Hans-Hinrich Kaatz from the University of Jena, has just
presented new evidence of horizontal gene transfer within the gut of bee
larvae. Pollen from GM rapeseed tolerant to the herbicide glufosinate
were fed to immature bee larvae. When the microorganisms were isolated
from the gut of the larvae and examined for the presence of the gene
conferring glufosinate resistance, it was found in some of the bacteria
as well yeast cells.
- All cells including those of human beings are now known to take up
genetic material. While natural (unmanipulated) genetic material is
simply broken down to supply energy, invasive pieces of genetic material
may jump into the genome to mutate genes. Some insertions of foreign
genetic material may also be associated with cancer.
- Horizontal transfer of genes and constructs from the golden
rice will spread transgenes, including antibiotic resistance genes
to bacterial pathogens, and also has the potential to create new viruses
and bacteria associated with diseases.
In conclusion, the golden rice project was a
useless application, a drain on public finance and a threat to health and
biodiversity. It is being promoted in order to salvage a morally as well
as financially bankrupt agricultural biotech industry, and is obstructing
the essential shift to sustainable agriculture that can truly improve the
health and nutrition especially of the poor in the Third World. This
project should be terminated immediately before further damage is done.
The golden rice possesses all the usual
defects of first generation transgenic plants plus multiple copies of the
CaMV promoter which we have strongly recommended withdrawing from use on
the basis of scientific evidence indicating this promoter to be especially
unsafe. A growing number of scientists (318 scientists from 39 countries
to-date) are calling for a global moratorium on the environmental releases
of GMOs until and unless they can be shown to be safe.
I am grateful to Joe Cummins for helpful comments and
for supplying key references in preparing this audit.