Biochemical & Molecular Basis of Omega-3 Biosynthesis in Plants

Bhalani H N and D N Vakharia

DEPARTMENT OF BIOCHEMISTRY
COLLEGE OF AGRICULTURE
JUNAGADH AGRICULTURAL UNIVERSITY,
JUNAGADH-362001
Gujarat-India
What is Omega-3?
Omega-3 fatty acids are a family of essential unsaturated fatty acids that have in common a
final carbon–carbon double bond in the ω-3(Omega) position; i.e. the third bond from the methyl
end of the fatty acid chain. Some essential ω-3 fatty acids are α-Linolenic acid, Steridonic acid,
Eicosapentanoic acid, and Docosahexenoic acid.
Why Omega-3?
It plays an important role to prevent or treat many diseases like blood pressure, skin, bi-polar
disorder, infant development, diabetes, cancer, systemic lupus erytheematosus (SLE), rheumatoid
arthritis, osteoporosis, and macular degeneration. The major source of Omega-3 is fish,
microorganisms, and genetically modified foods.
Actually fish are not able to synthesis Omega-3 in to their body but it can obtained from
their diet which they consume specially microalgae. Microalgae are the richest source of ω-3 fatty
acid in the world but cultivation and use of these microalgae is very costly.
Fatty acid biosynthesis:
Acetyl-CoA produced from photosynthate or stored carbohydrates material react with
enzyme acetyl-CoA carboxylase, or fatty acid synthatase and produce long chain of fatty acid and by
the action of enzyme desaturases and elongases it can be converted in to the very long chain
polyunsaturated fatty acid. Key enzymes regulating PUFA in plants is mainly series of desaturase
andgseriesdofbelongases.
Need for genetic modifications:
Both γ-Linolenic acid (GLA) and Steridonic acid (SDA) are produced naturally in certain
plants. GLA is found in borage and primrose seed, while hemp seed and black current contain both
SDA and GLA. These plants are difficult to cultivate and are relatively poor yielder. Thus there is
considerable interest in producing GLA and SDA in oilseed crops.
Case studies:
Reddy and Thomas, (1996) isolated Δ6-desaturase gene from the Cyanobacterium
Synechocystis, and expressed in tobacco plants which represented only about 1% GLA and 1-3%
SDA. While Sayanova et al., (1999) isolated a borage Δ6-desaturase cDNA, and expression of this
cDNA in tobacco plants led to the accumulation of approximately 13% GLA and 10% SDA in leaf
tissue, and it also accumulated low level of GLA in mature seed tissue.
Biosynthesis of very long chain polyunsaturated fatty acid (VLCPUFAs):
Abbadi et al., (2004) constructed cDNAs from the protein sequences of VLCPUFA-
producing organisms and used for transformation into Nicotiana tabacum and Linum usitatissimum
by constructing four various combination A, B, C, D. in transformed progeny C & D gives
successful expression but transformant one Show high proportion of C18 PUFAs and and
disproportionately low levels of C20-PUFA. Homozygous transformant three progeny accumulated
the remarkable level VLCPUFAs which were absent in the wild type.

Expression of VLCPUFAs:
 Qi. (2004) achieved synthesis of VLCPUFAs in Arabidopsis thaliana by sequential transfer
and expression of three genes encoding Δ9- elongase activity from Isochrysis galbana, Δ8-desaturase
from Euglena gracilis, and a Δ5-desaturase from Mortierella alpina, respectively. The coding
regions of these three genes were placed in CaMV 35S promoter of the plant binary vectors
pCB302-1, pBECKS and pCAMBIA 1300 and A. thaliana were subjected to agrobacterium
mediated transformation resulted in sucessesful accumulation of Arachidonic acid and
Ecosapentanoic acid.

Rapid expression of seed-specific constructs for DHA production:

Petrie et.al., (2010) constructed three binary vectors for the assay of green fluorescence
protein are having transcription factor LEC2, green fluoresce protein (GFP), P19 viral suppressor
respectively and another construct pjP3057 was made for insertion of desired gene. In contrast,
infiltration of FP1: GFP and 35S:P19 alone resulted in minimal GFP expression through Western
blot confirmed that its expression was markedly increased in the presence of LEC2. The construct
pJP3057 was then stably-transformed in A. thaliana (ecotype Columbia). The transformant 2
progeny plants showed synthesis of DHA ranging from 0.2% to2.4% along with higher expression
of Δ5 Elongase activity.

Synthesis and accumulation of stearidonic acid:

Lopez et al., (2009) Δ-6 Desaturase From Primula vialii cloned using USP seed specific
promoter from Vica faba and introduce into A. thaliana and linseed via Agrobacterium.
Another same T- DNA binary vector was also constructed (PUFA38) containing primmula
desaturase and used to transformed in both species using Kanamycin resistance gene as a
selectable markers. Their results showed that arabidopsis line containing SDA in T1 range from
1.0%-3.3% of TFA and linseed line containing SDA range starting 7.1%-15.4% of TFA.

Is there any side effect from taking Omega-3?

Only a very small percentage of people actually experienced side effects from consuming
ω-3s. These side effects include gastrointestinal upset, nausea, decline of blood sugar levels, fishy
after-taste and increase in LDL cholesterol for people with high triglycerides.

Conclusions:
Linoleic acid & α-Linolenic acid are the precursor for the VLCPUFAs which are present in
fish, microalgae and fungi. Regulatory gene of producing polyunsaturated fatty acid (ω-3) can be
transferred in to the crops using appropriate molecular techniques to produce transgenic plants to
increase the production of Omega-3 and Omega-6. This fatty acid prevents many chronic diseases
like cancer, autoimmune disease and cardiovascular diseases.
References:
Abbadi A., Domergue F., Bauer J., Napier J., Welti R., Zhringer U., Cirpus P. and Heinz E. (2004).
The Plant Cell, 16: 2734–2748.
Lopez N. R., Haslam R. P., Caleron M. V., Larson T. R., Graham I. A., Napier J. A. and Sayanova
O. (2009). Plant Biotechnology Journal, 7:704–716.
Petrie J.R., Shrestha P., Liu Q., Mansour M., Wood C., Zhou X. R., Nichols P.D., Green A.G.
and Surinder Singh. (2010). Plant Methods, 6:1-6.
Qi, B. (2004). ISB News Report at http://www.isb.vt.edu.
Reddy, A. S. and Thomas, T. L. (1996). Nature Biotechnology, 14:639-642.
Sayanova O., Davies G.M., Smith M.A., Stobart A.K., Christie W., Shewry P.R., and Napier J.A.
(1999). Journal of Experimental Botany, 50:1647-1652
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