A weak allele of AtECB2, a member of the pentatricopeptide repeat motif superfamily, causes leaf virescence in Arabidopsis

Key message Through the study of a point mutation of AtECB2 , it is reconfirmed that AtECB2 plays an important role in the early development of chloroplast. AtECB2(EARLY CHLOROPLAST BIOGENESIS 2, At1g15510), a member of the pentatricopeptide repeat motif proteins (PPR) superfamily, and its loss of function mutation ecb2-1 causes seedling lethal, while a point mutation ecb2-2 causes delayed chloroplast development. Finding more AtECB2 weak alleles helps to understand the molecular mechanisms of AtECB2 . In this study, a leaf virescence mutant was identified from ethylmethane sulfonate (EMS) treated Arabidopsis Col-0 M2 mutants library. The mutation of this mutant was first confirmed as a recessive mutation of one gene through the phenotype of F1 and its F2 phenotype segregation of this mutant crossed with Col-0. The mutation of G1931A of AtECB2 is identified as the cause of this leaf virescence phenotype sequentially through positional cloning, whole genome resequencing, Sanger sequencing and complementation. Therefore, we named this weak allele of AtECB2 as ecb2-3. The chlorophyll content and photosystem II maximum photochemical efficiency of ecb2-3 are obviously lower than that of Col-0 and its complementation lines, respectively. The chloroplast development of ecb2-3 is also inferior to that of Col-0 and its complementation line at the observed time points using the transmission electron microscope. The RNA editing efficiency of three chloroplast gene sites ( accD C794 and C1568, ndhF C290) was observed much lower compared with that of Col-0 and its complementation line. In summary, AtECB2 plays an important role in early chloroplast biogenesis through related chloroplast gene editing regulation, and this weak mutant ecb2-3 may be useful material in dissecting the function of AtECB2 in the near future.