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        Molecular cloning and expression analysis of the first two key genes through 2-C-methyl-D-erythritol 4-phosphate (MEP) pathway from Pyropia haitanensis(Bangiales, Rhodophyta)

        Yu Du,Jian Guan,Ruijun Xu,Xin Liu,Weijie Shen,Yafeng Ma,Yuan He,Songdong Shen 한국조류학회I 2017 ALGAE Vol.32 No.4

        Pyropia haitanensis (T. J. Chang et B. F. Zheng) N. Kikuchi et M. Miyata is one of the most commercially usefulmacroalgae cultivated in southeastern China. In red algae, the biosynthesis of terpenoids through 2-C-methyl-Derythritol4-phosphate (MEP) pathway can produce a direct influence on the synthesis of many biologically importantmetabolites. In this study, two genes of cDNAs, 1-deoxy-D-xylulose-5-phosphate synthase (DXS) and 1-deoxy-Dxylulose-5-phosphatereductase (DXR), which encoding the first two rate-limiting enzymes among MEP pathway werecloned from P. haitanensis. The cDNAs of P. haitanensis DXS (PhDXS) and DXR (PhDXR) both contained complete openreading frames encoding polypeptides of 764 and 426 amino acids residues, separately. The expression analysis showedthat PhDXS was significant differently expressed between leafy thallus and conchocelis as PhDXR been non-significant. Additionally, expression of PhDXR and its downstream gene geranylgeranyl diphosphate synthase were both inhibitedby fosmidomycin significantly. Meanwhile, we constructed types of phylogenetic trees through different algae and higherplants DXS and DXR encoding amino acid sequences, as a result we found tree clustering consequences basically in linewith the "Cavalier-Smith endosymbiotic theory." Whereupon, we speculated that in red algae, there existed only completeMEP pathway to meet needs of terpenoids synthesis for themselves; Terpenoids synthesis of red algae derivativesthrough mevalonate pathway came from two or more times endosymbiosis of heterotrophic eukaryotic parasitifer. Thisstudy demonstrated that PhDXS and PhDXR could play significant roles in terpenoids biosynthesis at molecular levels. Meanwhile, as nuclear genes among MEP pathway, PhDXS and PhDXR could provide a new way of thinking to researchthe problem of chromalveolata biological evolution.

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        Molecular cloning and expression analysis of the first two key genes through 2-C-methyl-D-erythritol 4-phosphate (MEP) pathway from Pyropia haitanensis (Bangiales, Rhodophyta)

        Du, Yu,Guan, Jian,Xu, Ruijun,Liu, Xin,Shen, Weijie,Ma, Yafeng,He, Yuan,Shen, Songdong The Korean Society of Phycology 2017 ALGAE Vol.32 No.4

        Pyropia haitanensis (T. J. Chang et B. F. Zheng) N. Kikuchi et M. Miyata is one of the most commercially useful macroalgae cultivated in southeastern China. In red algae, the biosynthesis of terpenoids through 2-C-methyl-D-erythritol 4-phosphate (MEP) pathway can produce a direct influence on the synthesis of many biologically important metabolites. In this study, two genes of cDNAs, 1-deoxy-D-xylulose-5-phosphate synthase (DXS) and 1-deoxy-D-xylulose-5-phosphate reductase (DXR), which encoding the first two rate-limiting enzymes among MEP pathway were cloned from P. haitanensis. The cDNAs of P. haitanensis DXS (PhDXS) and DXR (PhDXR) both contained complete open reading frames encoding polypeptides of 764 and 426 amino acids residues, separately. The expression analysis showed that PhDXS was significant differently expressed between leafy thallus and conchocelis as PhDXR been non-significant. Additionally, expression of PhDXR and its downstream gene geranylgeranyl diphosphate synthase were both inhibited by fosmidomycin significantly. Meanwhile, we constructed types of phylogenetic trees through different algae and higher plants DXS and DXR encoding amino acid sequences, as a result we found tree clustering consequences basically in line with the "Cavalier-Smith endosymbiotic theory." Whereupon, we speculated that in red algae, there existed only complete MEP pathway to meet needs of terpenoids synthesis for themselves; Terpenoids synthesis of red algae derivatives through mevalonate pathway came from two or more times endosymbiosis of heterotrophic eukaryotic parasitifer. This study demonstrated that PhDXS and PhDXR could play significant roles in terpenoids biosynthesis at molecular levels. Meanwhile, as nuclear genes among MEP pathway, PhDXS and PhDXR could provide a new way of thinking to research the problem of chromalveolata biological evolution.

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