While core components of plastid protein import (Toc and Tic) and the principle of using N-terminal targeting sequences (NTS) are conserved, lineage-specific differences are known. Rhodophytes and glaucophytes carry a conserved NTS motif, which was lost in the green lineage that also added novel proteins to Toc and Tic. Here we compare the components of plastid protein import and generated RNA-Seq, pigment profile and trans-electron microscopy data based on high-light stress from representatives of the three archaeplastidal groups. In light of plastid protein targeting, we compare the response to high-light stress of archaeplastidal representatives based on RNA-Seq, pigment profile and trans-electron microscopy data. Like land plants, the chlorophyte Chlamydomonas reinhardtii displays a broad respond to high-light stress, not observed to the same degree in the glaucophyte Cyanophora paradoxa or the rhodophyte Porphyridium purpureum. We find that only the green lineage encodes a conserved duplicate of the outer plastid membrane protein channel Oep80, namely Toc75 and suggest that the ability to respond to high-light stress entailed evolutionary changes in protein import, including the departure from phenylalanine-based targeting and the introduction of a green-specific Toc75 next to other import components unique to Chloroplastida. One consequence of relaxed NTS specificity was the origin of dual-targeting of plastid derived proteins to mitochondria and vice versa, using a single ambiguous NTS. Changes in the plastid protein import enabled the green lineage to import proteins at a more efficient rate, including those required for high-light stress response, a prerequisite for the colonization of land.

Major changes in plastid protein import and the origin of the Chloroplastida