Plant Molecular Biology

, Volume 62, Issue 3, pp 325–337 | Cite as

Rice Chlorina-1 and Chlorina-9 encode ChlD and ChlI subunits of Mg-chelatase, a key enzyme for chlorophyll synthesis and chloroplast development

  • Haitao Zhang
  • Jinjie Li
  • Jeong-Hoon Yoo
  • Soo-Cheul Yoo
  • Sung-Hwan Cho
  • Hee-Jong Koh
  • Hak Soo Seo
  • Nam-Chon Paek
Article

Abstract

Photosynthetic organisms exhibit a green color due to the accumulation of chlorophyll pigments in chloroplasts. Mg-protoporphyrin IX chelatase (Mg-chelatase) comprises three subunits (ChlH, ChlD and ChlI) and catalyzes the insertion of Mg2+ into protoporphyrin IX, the last common intermediate precursor in both chlorophyll and heme biosyntheses, to produce Mg-protoporphyrin IX (MgProto). Chlorophyll deficiency in higher plants results in chlorina (yellowish-green) phenotype. To date, 10 chlorina (chl) mutants have been isolated in rice, but the corresponding genes have not yet been identified. Rice Chl1 and Chl9 genes were mapped to chromosome 3 and isolated by map-based cloning. A missense mutation occurred in a highly conserved amino acid of ChlD in the chl1 mutant and ChlI in the chl9 mutant. Ultrastructural analyses have revealed that the grana are poorly stacked, resulting in the underdevelopment of chloroplasts. In the seedlings fed with aminolevulinate-dipyridyl in darkness, MgProto levels in the chl1 and chl9 mutants decreased up to 25% and 31% of that in wild-type, respectively, indicating that the Mg-chelatase activity is significantly reduced, causing the eventual decrease in chlorophyll synthesis. Furthermore, Northern blot analysis indicated that the nuclear genes encoding the three subunits of Mg-chelatase and LhcpII in chl1 mutant are expressed about 2-fold higher than those in WT, but are not altered in the chl9 mutant. This result indicates that the ChlD subunit participates in negative feedback regulation of plastid-to-nucleus in the expression of nuclear genes encoding chloroplast proteins, but not the ChlI subunit.

Keywords

Rice chlorina-1 chlorina-9 Map-based cloning Mg-chelatase OsChlD subunit OsChlI subunit 

Abbreviations

chl1

chlorina-1

chl9

chlorina-9

aa

amino acid

ChlH

Mg-chelatase H subunit

ChlI

Mg-chelatase I subunit

ChlD

Mg-chelatase D subunit

ALA

5-aminolevulinate

Proto

protoporphyrin IX

MgProto

Mg-Protoporphyrin IX

MgProtoME

Mg-Protoporphyrin IX monomethyl ester

MgProto(ME)

MgProto + MgProtoME

DP

2,2′-dipyridyl

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Notes

Acknowledgements

We thank the Rice Genome Research Program (Japan) for providing cDNA stocks and the AIMS (USA) for T-DNA insertional mutant stocks. This research was supported by a grant (CG3131) from Crop Functional Genomics Center of the 21st century Frontier Research Program funded by the Ministry of Science and Technology (MOST) and Rural Development Administration (RDA) of Republic of Korea. Mr. H. Zhang was supported by a fellowship from the Korea Research Foundation Grant Funded by the Korea Government (MOEHRD) (KRF-2004-211-410039). Mr. J.-H. Yoo and S.-H.␣Cho were supported by Brain Korea 21 Project of Korean Research Foundation.

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Copyright information

© Springer Science+Business Media B.V. 2006

Authors and Affiliations

  • Haitao Zhang
    • 1
  • Jinjie Li
    • 1
  • Jeong-Hoon Yoo
    • 1
  • Soo-Cheul Yoo
    • 1
  • Sung-Hwan Cho
    • 1
  • Hee-Jong Koh
    • 1
  • Hak Soo Seo
    • 1
  • Nam-Chon Paek
    • 1
  1. 1.Department of Plant ScienceSeoul National UniversitySeoulRepublic of Korea

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