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Abstract
The utilization of male sterility into hybrid seed production reduces its cost and ensures high purity of tomato varieties because it does not produce pollen and has exserted stigmas. Here, we report on the generation of gene edited lines into male sterility phenotype by knockout of SlMS10 gene (Solyc02g079810) encoding the bHLH transcription factor that regulates meiosis and cell death of the tapetum during microsporogenesis in the tomato. Twenty-eight gene edited lines out of 60 transgenic plants were selected. Of these, eleven different mutation types at the target site of the SlMS10 gene were selected through deep sequencing analysis. These mutations were confirmed to be transmitted to subsequent generations. The null lines without the transferred DNA (T-DNA) were obtained by segregation in the T1 and T2 generations. In addition, we showed that the cr-ms10-1-4 mutant line exhibited dysfunctional meiosis and abnormal tapetum during flower development, resulting in no pollen production. RT-PCR analysis showed that the most genes associated with pollen and tapetum development in tomatoes had lower expression in the cr-ms10-1-4 mutant line compared to wild type. We demonstrate that modification of the SlMS10 gene via CRISPR/Cas9-mediated genome editing results in male sterility of tomato plants. Our results suggest an alternative approach to generating male sterility in crops.
Highlights
The tomato (Solanum lycopersicum L.) is a representative vegetable crop belonging to the Solanaceae family, and has high economic value in the market due to its high production and consumption worldwide
The results showed that SlMS10, OsUDT1, CabHLH and AtDYT1 were classified within the same clade (Figure 1 and Figure S1)
These results suggested that the transcription factors of tomato basic helix-loop-helix (bHLH) members showed very high homology regardless of origin
Summary
The tomato (Solanum lycopersicum L.) is a representative vegetable crop belonging to the Solanaceae family, and has high economic value in the market due to its high production and consumption worldwide. Plant male sterility is functionally unable to produce or release pollen grains because no anthers, microspores or male gametes are produced [3]. In Arabidopsis, several gene encoding transcription factors have been studied as regulators involved in pollen development such as AtDYT1, AtTDF1, AtAMS, AtbHLH10, AtbHLH89, AtbHLH91 and AtMYB103 [9,10,11,12,13,14]. The basic helix-loop-helix (bHLH) proteins play an important role in plant growth and development. A total of 152 bHLH transcription factors have been reported in the tomato genome [15,16,17,18,19,20]. The bHLH motif consists of two functionally distinct regions: the basic region for DNA binding and the HLH region for protein dimerization [21]
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