Abstract
The chloroplast RNA splicing and ribosome maturation (CRM) domain proteins are involved in the splicing of chloroplast gene introns. Numerous CRM domain proteins have been reported to play key roles in chloroplast development in several plant species. However, the functions of CRM domain proteins in chloroplast development in rice remain poorly understood. In the study, we generated oscaf1 albino mutants, which eventually died at the seedling stage, through the editing of OsCAF1 with two CRM domains using CRISPR/Cas9 technology. The mesophyll cells in oscaf1 mutant had decreased chloroplast numbers and damaged chloroplast structures. OsCAF1 was located in the chloroplast, and transcripts revealed high levels in green tissues. In addition, the OsCAF1 promoted the splicing of group IIA and group IIB introns, unlike orthologous proteins of AtCAF1 and ZmCAF1, which only affected the splicing of subgroup IIB introns. We also observed that the C-terminal of OsCAF1 interacts with OsCRS2, and OsCAF1–OsCRS2 complex may participate in the splicing of group IIA and group IIB introns in rice chloroplasts. OsCAF1 regulates chloroplast development by influencing the splicing of group II introns.
Highlights
CAF1 and CAF2 could interact with CRS2, forming the CRS2–CAF1 and CRS2–CAF2 complexes, which participate in the splicing of chloroplast group II introns and the regulation of chloroplast development in maize [22]
We investigated the function of OsCAF1 in rice chloroplast development, and oscaf1 mutants were obtained using the CRISPR-Cas9 system
The results of the present study reveal the functions of OsCAF1 in the regulation of rice chloroplast development
Summary
Arabidopsis thaliana, maize, and rice chloroplast genomes all have only 1 group I intron, and 20, 17, and 17 group II introns, respectively [8] In plant chloroplasts, such introns have lost the capacity to self-splice in vivo and require nuclear gene-encoded proteins as co-factors to participate in splicing [8,12]. The CAF1 and CAF2 contain two CRM domains, which are required for the splicing of group IIB introns, including ndhA, ndhB, petB, petD, rpl, rps, trnG, and ycf in maize and A. thaliana [11,21]. CAF1 and CAF2 could interact with CRS2, forming the CRS2–CAF1 and CRS2–CAF2 complexes, which participate in the splicing of chloroplast group II introns and the regulation of chloroplast development in maize [22]. The results of the present study reveal the functions of OsCAF1 in the regulation of rice chloroplast development
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