Resistance of the CRISPR-Cas13a Gene-Editing System to Potato Spindle Tuber Viroid Infection in Tomato and <i>Nicotiana benthamiana</i>
Ying Wei Khoo,
Qingsong Wang,
Shangwu Liu,
Binhui Zhan,
Tengfei Xu,
Wenxia Lv,
Guangjing Liu,
Shifang Li,
Zhixiang Zhang
Affiliations
Ying Wei Khoo
State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China
Qingsong Wang
State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China
Shangwu Liu
Institute of Industrial Crops, Heilongjiang Academy of Agricultural Sciences, Harbin 150086, China
Binhui Zhan
State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China
Tengfei Xu
Department of Fruit Science, College of Horticulture, China Agricultural University, Beijing 100193, China
Wenxia Lv
Inner Mongolia Zhongjia Agricultural Biotechnology Co., Ltd., Ulanqab 011800, China
Guangjing Liu
Inner Mongolia Zhongjia Agricultural Biotechnology Co., Ltd., Ulanqab 011800, China
Shifang Li
State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China
Zhixiang Zhang
State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China
Gene-editing technology, specifically the CRISPR-Cas13a system, has shown promise in breeding plants resistant to RNA viruses. This system targets RNA and, theoretically, can also combat RNA-based viroids. To test this, the CRISPR-Cas13a system was introduced into tomato plants via transient expression and into Nicotiana benthamiana through transgenic methods, using CRISPR RNAs (crRNAs) targeting the conserved regions of both sense and antisense genomes of potato spindle tuber viroid (PSTVd). In tomato plants, the expression of CRISPR-Cas13a and crRNAs substantially reduced PSTVd accumulation and alleviated disease symptoms. In transgenic N. benthamiana plants, the PSTVd levels were lower as compared to wild-type plants. Several effective crRNAs targeting the PSTVd genomic RNA were also identified. These results demonstrate that the CRISPR-Cas13a system can effectively target and combat viroid RNAs, despite their compact structures.
