FsHemF is Involved in the Formation of Yellow Forsythia Leaves by Regulating Chlorophyll Synthesis in Response to Light Intensity.

The leaves of Forsythia koreana 'Suwon Gold' are yellow under natural light condition and can revert to green when the light intensity is reduced. To understand the molecular mechanism of leaf color changes in response to light intensity, we compared the chlorophyll content and precursor content between yellow- and green-leaf Forsythia under shade and light-recovery conditions. We identified the conversion of coproporphyrin III (Coprogen III) to protoporphyrin IX (Proto IX) as the primary rate-limiting step of chlorophyll biosynthesis in yellow-leaf Forsythia. Further analysis of the activity of the enzymes that catalyze this step and the expression pattern of the chlorophyll biosynthesis-related genes under different light intensities revealed that the negatively regulated expression of FsHemF by light intensity was the major cause affecting the leaf color change in response to light intensity in yellow-leaf Forsythia. To further understand the cause of differential expression pattern of FsHemF in yellow- and green-leaf lines, we compared the coding sequence and promoter sequence of FsHemF between yellow- and green-leaf Forsythia. We found that one G-box light-responsive cis-element was absent in the promoter region of green-leaf lines. To investigate the functional role of FsHemF, we performed virus-induced gene silencing (VIGS) of FsHemF in green-leaf Forsythia, which leads to yellowing leaf veins, decreased chlorophyll b content, and inhibition of chlorophyll biosynthesis. The results will assist in elucidating the mechanism of yellow-leaf Forsythia in response to light intensity.