A CRISPR-Cas9-Derived Male Sterility System for Tomato Breeding
Ming Zhou,
Lei Deng,
Guoliang Yuan,
Wei Zhao,
Mingyang Ma,
Chuanlong Sun,
Minmin Du,
Chuanyou Li,
Changbao Li
Affiliations
Ming Zhou
State Key Laboratory of Vegetable Biobreeding, Beijing Institute of Vegetable Science, Beijing Academy of Agriculture and Forestry Sciences, Beijing 100097, China
Lei Deng
State Key Laboratory of Plant Genomics, National Centre for Plant Gene Research, Institute of Genetics and Developmental Biology, Innovative Academy for Seed Design, Chinese Academy of Sciences, Beijing 100101, China
Guoliang Yuan
State Key Laboratory of Vegetable Biobreeding, Beijing Institute of Vegetable Science, Beijing Academy of Agriculture and Forestry Sciences, Beijing 100097, China
Wei Zhao
State Key Laboratory of Plant Genomics, National Centre for Plant Gene Research, Institute of Genetics and Developmental Biology, Innovative Academy for Seed Design, Chinese Academy of Sciences, Beijing 100101, China
Mingyang Ma
State Key Laboratory of Vegetable Biobreeding, Beijing Institute of Vegetable Science, Beijing Academy of Agriculture and Forestry Sciences, Beijing 100097, China
Chuanlong Sun
State Key Laboratory of Plant Genomics, National Centre for Plant Gene Research, Institute of Genetics and Developmental Biology, Innovative Academy for Seed Design, Chinese Academy of Sciences, Beijing 100101, China
Minmin Du
Beijing Key Laboratory of Growth and Developmental Regulation for Protected Vegetable Crops, College of Horticulture, China Agricultural University, Beijing 100193, China
Chuanyou Li
State Key Laboratory of Plant Genomics, National Centre for Plant Gene Research, Institute of Genetics and Developmental Biology, Innovative Academy for Seed Design, Chinese Academy of Sciences, Beijing 100101, China
Changbao Li
State Key Laboratory of Vegetable Biobreeding, Beijing Institute of Vegetable Science, Beijing Academy of Agriculture and Forestry Sciences, Beijing 100097, China
Male sterility can reduce cost and enable high seed purity during hybrid seed production. However, the commercial application of male sterility in hybrid seed production has not been widely used in tomatoes. CRISPR/Cas9-mediated gene editing can facilitate acceleration for the practical application of male sterility in hybrid seed production. Here, by using the CRISPR-Cas9 system, two genes DYSFUNCTIONAL TAPETUM1 (SlDYT1) and Glutathione S-transferase (SlGSTAA), which underly the two closely linked loci Male sterile 10 (Ms10) and Anthocyanin absent (AA), were knocked out simultaneously in two tomato parental lines. The generated dyt1gstaa double mutants developed green hypocotyl owing to anthocyanin deficiency and exhibited stable male sterility. Up to 92% effectiveness in selecting male sterility was achieved using green hypocotyl as a morphological marker, and thereafter an efficient and stable propagation strategy of male sterility with the aid of the morphological marker selection was developed. Furthermore, dyt1gstaa-derived hybrid seeds were produced and found to have comparable yield, weight, and germination rate with the corresponding WT-derived F1 seeds. The dyt1gstaa system not only increased hybrid seed purity to 100% but also facilitated its rapid and cost-effective determination. Moreover, this system was discovered to have no evident side effects on important agronomic traits. This study suggested that our CRISPR/Cas9-created dyt1gstaa system can be deployed in tomato hybrid seed production.
