The chloroplast ycf7 (petL) open reading frame of Chlamydomonas reinhardtii encodes a small functionally important subunit of the cytochrome b₆f complex

The small chloroplast open reading frame ORF43 (ycf7) of the green unicellular alga Chlamydomonas reinhardtii is cotranscribed with the psaC gene and ORF58. While ORF58 has been found only in the chloroplast genome of C. reinhardtii, ycf7 has been conserved in land plants and its sequence suggests that its product is a hydrophobic protein with a single transmembrane α helix. We have disrupted ORF58 and ycf 7 with the aadA expression cassette by particle-gun mediated chloroplast transformation. While the ORF58::aadA transformants are indistinguishable from wild type, photoautotrophic growth of the ycf 7::aadA transformants is considerably impaired. In these mutant cells, the amount of cytochrome b₆f complex is reduced to 25-50% of wild-type level in mid-exponential phase, and the rate of transmembrane electron transfer per b₆f complex measured in vivo under saturating light is three to four times slower than in wild type. Under subsaturating light conditions, the rate of the electron transfer reactions within the b₆f complex is reduced more strongly in the mutant than in the wild type by the proton electrochemical gradient. The ycf 7 product (Ycf 7) is absent in mutants deficient in cytochrome b₆f complex and present in highly purified b₆f complex from the wild-type strain. Ycf7-less complexes appear more fragile than wild-type complexes and selectively lose the Rieske iron-sulfur protein during purification. These observations indicate that Ycf7 is an authentic subunit of the cytochrome b₆f complex, which is required for its stability, accumulation and optimal efficiency, We therefore propose to rename the ycf 7 gene petL.