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Analysis of indican synthesis pathway in Indigo plant (Polygonaceae).

Research Project

Project/Area Number 14540605
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field 植物生理
Research InstitutionOkayama university of science

Principal Investigator

MINAMI Yoshiko  Okayama university of science, Biochemistry, Assistant professor, 理学部, 助教授 (60192762)

Project Period (FY) 2002 – 2005
Project Status Completed (Fiscal Year 2005)
Budget Amount *help
¥3,700,000 (Direct Cost: ¥3,700,000)
Fiscal Year 2005: ¥400,000 (Direct Cost: ¥400,000)
Fiscal Year 2004: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 2003: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 2002: ¥1,300,000 (Direct Cost: ¥1,300,000)
KeywordsSecondary metabolism / Indican / Indigo / Tryptophan synthase / UDP-glucosyl transferase / cDNA cloning / Cytochrome P450 / 植物の2次代謝 / UDP-グルコシルトランスフェラーゼ / 植物の二次代謝 / トリプトファン合成経路
Research Abstract

Indican, one of the secondary metabolites, is stored in the leaves of Indigo plant (Polygonaceae). Probably, indican is synthesized from indole, an intermediate of tryptophan synthesis pathway. Indole is oxidized to indoxyl by cytochrome P450. Furthermore, indoxyl is converted to indican by UDP-glucosyltransferase. I predicted the presence of switching mechanism between tryptophan and indican synthesis pathways.
1.cDNA cloning of UDP-glucosyltransferase
A desired clone could not be gotten, although cDNA cloning was tried by some methods. By two-dimensional electrophoresis, the total proteins of callus were compared with those of leaves, the difference was analyzed. Still now, we continue the analysis.
2.cDNA cloning and analysis of cytochrome P450
ptP450 cDNA was isolated from cDNA library of indigo plant. The amino acid sequence of ptP450 is homologous to those of P450s, which react with some alkaloids including indole. Therefore, I tried the analysis of properties of recombinant ptP450 that was expressed in E.coll.
3.cDNA cloning and analysis of tryptophan synthase
Three cDNAs of tryptophan synthase β were amplified by RT-PCR. Now we examine the difference of expression by Northern blot.

Report

(5 results)
  • 2005 Annual Research Report   Final Research Report Summary
  • 2004 Annual Research Report
  • 2003 Annual Research Report
  • 2002 Annual Research Report

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Published: 2002-04-01   Modified: 2016-04-21  

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