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ARS Home » Pacific West Area » Pullman, Washington » WHGQ » Research » Publications at this Location » Publication #386069

Research Project: Genetic Improvement of Wheat and Barley for Environmental Resilience, Disease Resistance, and End-use Quality

Location: Wheat Health, Genetics, and Quality Research

Title: High-efficiency plastome base editing in rice with TAL cytosine deaminase

item LI, RIQING - University Of Missouri
item CHAR, SI NIAN - University Of Missouri
item LIU, BO - University Of Missouri
item LIU, HUA - University Of Missouri
item Li, Xianran
item YANG, BING - University Of Missouri

Submitted to: Molecular Plant
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 7/11/2021
Publication Date: 9/6/2021
Citation: Li, R., Char, S., Liu, B., Liu, H., Li, X., Yang, B. 2021. High-efficiency plastome base editing in rice with TAL cytosine deaminase. Molecular Plant. 14(9):1412-1414.

Interpretive Summary: Base editing is a no-destructive strategy to alter DNA molecular without inducing double strand break. It has been successfully applied in editing nuclear genome in plants, however, base editing chloroplasts, an essential organelle genome encoded important genes involving photosynthesis, has not been established. This study developed the base editing system for chloroplasts using rice as a model. Results indicated a high efficiency of based editing, which established a new strategy to engineer crops for better performances.

Technical Abstract: Chloroplasts, the organelles responsible for photosynthesis in plants, contain their own DNA, so-called plastome. However, plastome editing has not yet been established, although nuclear genome editing has been widely used for basic and applied research in plant biology and agriculture. Here we report the establishment and validation of an efficient plastome base editing system in rice. The plastome cytosine base editor (cpDdCBE) consists of a pair of designer TAL (transcription activator-like) effector DNA binding domains each tagged N-terminally with a chloroplast transition peptide and fused C-terminally with a split-half of the cytidine deaminase domain DddA along with a UGI (uracil glycosylase inhibitor). Transgenic rice callus and stable plants showed efficient C•G to T•A conversions in the chlorophyll biosynthesis gene psaA induced by the designer cpDdCBEs. The C to T changes led to the formation of stop codons and inactivation of psaA as confirmed by Sanger sequencing the relevant PCR-amplicons, producing albino plants at about sixty-four percent among 33 independent T0 rice lines. Our results demonstrate the feasibility of genome editing in chloroplasts and open doors to plastome engineering for crop improvement.