Assembly of LHCA5 into PSI blue shifts the far-red fluorescence emission in higher plants.

In higher plants, the PSI core complex is associated with light-harvesting complex I (LHCI), forming the PSI-LHCI super-complex. In vascular plants, four major antenna proteins (LHCA1-4) are assembled in the order of LHCA1, LHCA4, LHCA2, and LHCA3 into a crescent-shaped LHCI, while LHCA5 and LHCA6 are minor antenna proteins. By contrast, in moss and green algae, LHCA5 or LHCA5-like protein functions as one of the major antenna proteins by residing at the second site of LHCI. In order to learn the effect of binding different LHCA proteins, i.e. LHCA4 or LHCA5, within the PSI-LHCI super-complex on photosynthetic properties of plants, we constructed LHCA5 overexpression plants with a wild type (WT) background and an lhca4 mutant background in Arabidopsis thaliana. The results showed that: (i) there are little difference in phenotype, pigment composition and chlorophyll fluorescence parameters between the transgenic Arabidopsis and their corresponding background materials; (ii) in spite of a small amount of LHCA5, the LHCA5-included PSI-LHCI super-complex can be obtained by extracting samples incubated with anti-FLAG M2 Affinity Gel, in which LHCA5 is found to substitute for LHCA4 as analyzed by immunoblotting analysis; (iii) the replacement of LHCA4 with LHCA5 within PSI-LHCI super-complex leads to a blue shift in low temperature fluorescence emission, suggesting a decrease in far-red absorbance. These results provide new clues for understanding the position and function of LHCA4 and LHCA5 during the evolution of green plants from aquatic to terrestrial lifestyles.