Biofortification of rice by targeted genetic engineering.

Globally, malnutrition, including both overt nutrient deficiencies as well as diet-related chronic diseases (e.g., heart disease, cancer stroke and diabetes) is responsible for more deaths than any other cause accounting for over 20 million mortalities annually(Kennedy et al., 2003; World Health Organization and Food and Agriculture Organization, 2003). Malnutrition also contributes to increased morbidity, disability, stunted mental and physical growth and reduced national socio-economic development (World Health Organization and Food and Agriculture Organization, 2003).  Micronutrient malnutrition alone afflicts over two billion people mostly among resource-poor families in developing countries with iron, iodine, Folic acid, zinc and vitamin A deficiencies most prevalent (Kennedy et al., 2003). 

Biofortification of Rice is a method of breeding rice variety to increase their nutritional value. This can be done either through conventional selective breeding, or through genetic engineering.

Conventional breeding has some problem as such either the desire gene should be or not introduce into the targeted rice plant. So , Genetic engineering is the way to confirm that the transgenes would be inserted into the targeted rice plant, because of the use of the marker gene with the transgenes.

Biofortification of Rice differs from ordinary fortification because it focuses on making rice foods more nutritious as the rice plants are growing, rather than having nutrients added to the rice foods when they are being processed. This is an improvement on ordinary fortification when it comes to providing nutrients for the rural poor, who rarely have access to commercially fortified foods.  As such, biofortification of rice is seen as an upcoming strategy for dealing with deficiencies of micronutrients in the developing world.

How can economically, nutritional rice plant food can be produced through genetic engineering?

The aim of the study is to explore the valuable nutrients into the rice plants through genetic engineering.

It has chosen the quantitative research method and prospective type of descriptive survey to carry out the answer of the research question. This study had been aimed to explore the state of biofortification of rice throw genetic engineering. 

Oryza sativa ssp. japonica is being transformed with the plasmid vector containing four transgenes. The transformation vector is constructed by cloning into pCAMBIA 1300 (Cambia, Canberra, Australia) the AtNAS, Pvferritin, fphytase and pmi genes encoding Arabidopsis thaliana NAS (At5g04950), Phaseolus vulgaris- ferritin (X58274), Aspergillus fumigates strain Af293 phytase (AFUA_4G08630) and Escherichia coli phosphomannose-isomerase, pmi- gene, respectively. The AtNAS and pmi genes are placed under the control of 35S promoters; the Pvferritin and Afphytase genes are placed under the transcriptional control of endosperm-specific globulin promoters. The pmi gene is used as a selectable marker. The transformation, selection and plant regeneration are conducted according to Lucca et al. (2001b) following a mannose selection regime. In brief, immature embryos are used as starting material and gene transfer is mediated by Agrobacterium tumefaciens strain LBA4404 (Hoekema et al., 1984). A total of 150 putative primary transformants is obtained after transformation and selection on mannose, of which 20 plants are regenerated and analysed by Southern blot as described by Poletti (2006). In order to exclude the possibility that null segregants dilute the results, all plants are checked by PCR for the presence of the transgenes prior to hydroponic culture using the following primers: Afphy-f, 5-AGCTGTCCGTGTCGAGTAAG-3; Afphy-r, 5-TGGAGACTAGAGTCGAGTTAG-3;Actin-f,5-TATGGTTGGGATGGGACA-3,Actin-r,5-AGCACGGCTTGAATAGCG-3.

In addition, RNA is extracted from the leaves of transgenic (NFP) and WT plants during the vegetative growth phase according to Poletti (2006). The transcription of the transgenes in the NFP line was checked by real time PCR.