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Plant and Cell Physiology, 2003, Vol. 44, No. 5 463-472
© 2003 Oxford University Press

Characterization of a Rice Chlorophyll-Deficient Mutant Using the T-DNA Gene-Trap System

Ki-Hong Jung1, Junghe Hur1, Choong-Hwan Ryu1, Youngju Choi2, Yong-Yoon Chung2, Akio Miyao3, Hirohiko Hirochika3 and Gynheung An1,4

1 National Research Laboratory of Plant Functional Genomics, Division of Molecular and Life Sciences, Pohang University of Science and Technology (POSTECH), Pohang, 790-784 Republic of Korea
2 School of Life Sciences and Biotechonology, Korea University, Seoul, 136-701 Republic of Korea
3 Department of Molecular Genetics, National Institute of Agrobiological Resources, Tsukuba, Ibaraki, 305-8602 Japan

We have previously generated a large pool of T-DNA insertional lines in rice. In this study, we screened those T-DNA pools for rice mutants that had defective chlorophylls. Among the 1,995 lines examined in the T2 generation, 189 showed a chlorophyll-deficient phenotype that segregated as a single recessive locus. Among the mutants, 10 lines were ß-glucuronidase (GUS)-positive in the leaves. Line 9-07117 has a T-DNA insertion into the gene that is highly homologous to XANTHA-F in barley and CHLH in Arabidopsis. This OsCHLH gene encodes the largest subunit of the rice Mg-chelatase, a key enzyme in the chlorophyll branch of the tetrapyrrole biosynthetic pathway. In the T2 and T3 generations, the chlorina mutant phenotypes are co-segregated with the T-DNA. We have identified two additional chlorina mutants that have a Tos17 insertion in the OsCHLH gene. Those phenotypes were co-segregated with Tos17 in the progeny. GUS assays and RNA blot analysis showed that expression of the OsCHLH gene is light inducible, while TEM analysis revealed that the thylakoid membrane of the mutant chloroplasts is underdeveloped. The chlorophyll content was very low in the OschlH mutants. This is the first report that T-DNA insertional mutagenesis can be used for functional analysis of rice genes.

4 Corresponding author: E-mail, genean@postech.ac.kr; Fax, +82-54-279-2199.


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