title: Accumulation of phytoene, a colorless carotenoid by inhibition of phytoene desaturase (PDS) gene in Dunaliella salina V-101.
abstract: The aim of this work was to study the accumulation of phytoene in Dunaliella salina V-101 by down-regulating its phytoene desaturase (PDS) gene expression using RNA interference and Antisense technology. RNAi and antisense constructs were introduced into the Dunaliella cells by Agrobacterium-mediated transformation. Among thirty-two transformants, six showed positive down-regulation of PDS expression with RNAi construct and five positive transformants were obtained using antisense construct. Characterization of PDS suppression was carried out using semi-quantitative RT-PCR and quantitative determination of phytoene as well as other carotenoids by HPLC. Both the RNAi and antisense lines showed a significant decrease in the expression levels of phytoene desaturase and carotenoid content compared to wild type cells. The RNAi line #5 showed maximum Phytoene content (108.34±22.34µg/100mg DCW) compared to other transgenic lines. These phytoene-accumulating phenotypes exhibited slower growth rates and were found to be sensitive to high light conditions.
authors.affiliation: School of Bio-Sciences and Technology, VIT University, Vellore 632 014, Tamil Nadu, India.
authors.forename: Ramachandran
authors.initials: R
authors.lastname: Srinivasan
authors.affiliation: School of Bio-Sciences and Technology, VIT University, Vellore 632 014, Tamil Nadu, India.
authors.forename: S
authors.initials: S
authors.lastname: Babu
authors.affiliation: School of Bio-Sciences and Technology, VIT University, Vellore 632 014, Tamil Nadu, India. Electronic address: gothandam@gmail.com.
authors.forename: K M
authors.initials: KM
authors.lastname: Gothandam
mesh_terms: D002338:Carotenoids; D000460:Chlorophyta / Q000235:genetics; D010088:Oxidoreductases
chemical_list: D002338:Carotenoids; C100185:(all-E) phytoene; D010088:Oxidoreductases; C022398:phytoene dehydrogenase
keywords: Carotenoids; Dunaliella salina; Phytoene; Phytoene desaturase; Transgenic lines