Abstract
Obtaining transgenic crop lines with stable levels of carotenoids is highly desirable. We addressed this issue by employing the anther culture technique to develop dihaploid lines containing genes involved in β-carotene metabolism. First, we used Agrobacterium- mediated transformation to develop primary transgenic plants containing the β-carotene biosynthetic genes, phytoene synthase (psy) and phytoene desaturase (crtI), which were engineered for expression and accumulation in the endosperm. Transgenic plants were recovered by selecting for the expression of the phosphomannose isomerase (pmi) gene. Dihaploid plants in addition to haploid and tetraploid plant were generated from anther cultures of these primary transgenic plants. In addition to anatomical features of stomata, pollen of different ploidy-plants, molecular analyses confirmed the stable integration of the genes in the anther culture-derived dihaploid plants, and the yellow color of the polished seeds indicated the accumulation of carotenoids in the endosperm. High performance liquid chromatography (HPLC) analysis of the carotenoid extract further confirmed the levels of β–carotene accumulation in the endosperms of the transgenic dihaploid rice seeds.
Highlights
As a dominant cereal crop, rice serves as the staple food for more than half the world’s population
A total of 278 plants were found to be positive based on Polymerase Chain Reaction (PCR) and Southern analysis
The regeneration of fertile b-carotene producing dihaploid plants which occurs via direct embryogenesis involving the spontaneous doubling of the chromosomes
Summary
As a dominant cereal crop, rice serves as the staple food for more than half the world’s population. In Asian countries, rice accounts for 30–72% of the energy intake of the people. Micronutrient concentrations in rice are not sufficient to meet the recommended daily dietary allowances (RDA) to sustain good health and minimize the occurrence of diet-related chronic diseases. Because of high per capita consumption of rice, increasing its nutritive value may have a significant positive impact on the health of the rice consuming population. Transgenic methods are one option for improving the micronutrient content of rice [1,2]. Vitamin A deficiency, which affects the rod cells of the retina, proper immune system function and many other physiological functions in the human body, is a major nutritional problem facing
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