title: Genetic stability developed for β-carotene synthesis in BR29 rice line using dihaploid homozygosity.
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.
authors.affiliation: Department of Botany, Plant Molecular Biology and Biotechnology Laboratory, University of Calcutta, Kolkata, India.
authors.forename: Karabi
authors.initials: K
authors.lastname: Datta
authors.affiliation: Central Rice Research Institute, Cuttack, Odisha, India.
authors.forename: Gayetri
authors.initials: G
authors.lastname: Sahoo
authors.affiliation: Department of Botany, Goa University, Goa, India.
authors.forename: Sellappan
authors.initials: S
authors.lastname: Krishnan
authors.affiliation: Department of Botany, Plant Molecular Biology and Biotechnology Laboratory, University of Calcutta, Kolkata, India.
authors.forename: Moumita
authors.initials: M
authors.lastname: Ganguly
authors.affiliation: Department of Botany, Plant Molecular Biology and Biotechnology Laboratory, University of Calcutta, Kolkata, India; Divisions of Crop Science, Indian Council of Agricultural Research (ICAR), Krishi Bhawan, New Delhi, India.
authors.forename: Swapan K
authors.initials: SK
authors.lastname: Datta
mesh_terms: D015139:Blotting, Southern; D002851:Chromatography, High Pressure Liquid; D005818:Genetic Engineering; D006720:Homozygote; D008359:Mannose-6-Phosphate Isomerase / Q000235:genetics / Q000378:metabolism*; D012275:Oryza / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism*; D030821:Plants, Genetically Modified / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D011003:Ploidies; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012639:Seeds / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D019207:beta Carotene / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D010940:Plant Proteins; D012333:RNA, Messenger; D019207:beta Carotene; D008359:Mannose-6-Phosphate Isomerase