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Published on September 3, 2004; 10.1104/pp.104.046417

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Received May 13, 2004
Returned for revision July 6, 2004
Accepted July 13, 2004

The Elm1 (ZmHy2) Gene of Maize Encodes a Phytochromobilin Synthase

Ruairidh J.H. Sawers , Philip J. Linley , Jose F. Gutierrez-Marcos , Teegan Delli-Bovi , Phyllis R. Farmer , Takayuki Kohchi , Matthew J. Terry , and Thomas P. Brutnell *

Boyce Thompson Institute, Cornell University, Ithaca, New York 14853 (R.J.H.S., T.D.-B., T.P.B.); School of Biological Sciences, University of Southampton, Southampton SO16 7PX, United Kingdom (P.J.L., M.J.T.); Department of Plant Sciences, University of Oxford, Oxford OX1 3RB, United Kingdom (J.F.G.-M.); and Graduate School of Biostudies, Kyoto University, Kitashirakawa, Sakyo, Kyoto 606-8502, Japan (P.J.L., T.K.)

* Corresponding author; email: tpb8{at}cornell.edu.

The light insensitive maize (Zea mays) mutant elongated mesocotyl1 (elm1) has previously been shown to be deficient in the synthesis of the phytochrome chromophore 3E-phytochromobilin (P{Phi}B). To identify the Elm1 gene, a maize homolog of the Arabidopsis P{Phi}B synthase gene AtHY2 was isolated and designated ZmHy2. ZmHy2 encodes a 297-amino acid protein of 34 kD that is 50% identical to AtHY2. ZmHY2 was predicted to be plastid localized and was targeted to chloroplasts following transient expression in tobacco (Nicotiana plumbaginifolia) leaves. Molecular mapping indicated that ZmHy2 is a single copy gene in maize that is genetically linked to the Elm1 locus. Sequence analysis revealed that the ZmHy2 gene of elm1 mutants contains a single G to A transition at the 3' splice junction of intron III resulting in missplicing and premature translational termination. However, flexibility in the splicing machinery allowed a small pool of in-frame ZmHy2 transcripts to accumulate in elm1 plants. In addition, multiple ZmHy2 transcript forms accumulated in both wild-type and elm1 mutant plants. ZmHy2 splice variants were expressed in Escherichia coli and products examined for activity using a coupled apophytochrome assembly assay. Only full-length ZmHY2 (as defined by homology to AtHY2) was found to exhibit P{Phi}B synthase activity. Thus, the elm1 mutant of maize is deficient in phytochrome response due to a lesion in a gene encoding phytochromobilin synthase that severely compromises the P{Phi}B pool.




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