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Article

Knockout of SlMS10 Gene (Solyc02g079810) Encoding bHLH Transcription Factor Using CRISPR/Cas9 System Confers Male Sterility Phenotype in Tomato

1
Division of Horticultural Biotechnology, Hankyong National University, Anseong 17579, Korea
2
Institute of Genetic Engineering, Hankyong National University, Anseong 17579, Korea
3
Department of Chemistry, Hanyang University, Seoul 04763, Korea
4
Department of Horticulture, Sunchon National University, Suncheon 57922, Korea
5
Department of Crop Science, Chungbuk National University, Cheongju 28644, Korea
6
Tomato Research Center, Cheongju 28112, Korea
*
Author to whom correspondence should be addressed.
Plants 2020, 9(9), 1189; https://doi.org/10.3390/plants9091189
Received: 31 July 2020 / Revised: 26 August 2020 / Accepted: 9 September 2020 / Published: 11 September 2020
(This article belongs to the Special Issue Plant Systems and Application)
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. View Full-Text
Keywords: CRISPR/Cas9; bHLH transcription factor; male sterility; SlMS10 gene; tomatoes CRISPR/Cas9; bHLH transcription factor; male sterility; SlMS10 gene; tomatoes
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MDPI and ACS Style

Jung, Y.J.; Kim, D.H.; Lee, H.J.; Nam, K.H.; Bae, S.; Nou, I.S.; Cho, Y.-G.; Kim, M.K.; Kang, K.K. Knockout of SlMS10 Gene (Solyc02g079810) Encoding bHLH Transcription Factor Using CRISPR/Cas9 System Confers Male Sterility Phenotype in Tomato. Plants 2020, 9, 1189. https://doi.org/10.3390/plants9091189

AMA Style

Jung YJ, Kim DH, Lee HJ, Nam KH, Bae S, Nou IS, Cho Y-G, Kim MK, Kang KK. Knockout of SlMS10 Gene (Solyc02g079810) Encoding bHLH Transcription Factor Using CRISPR/Cas9 System Confers Male Sterility Phenotype in Tomato. Plants. 2020; 9(9):1189. https://doi.org/10.3390/plants9091189

Chicago/Turabian Style

Jung, Yu J., Dong H. Kim, Hyo J. Lee, Ki H. Nam, Sangsu Bae, Ill S. Nou, Yong-Gu Cho, Myong K. Kim, and Kwon K. Kang 2020. "Knockout of SlMS10 Gene (Solyc02g079810) Encoding bHLH Transcription Factor Using CRISPR/Cas9 System Confers Male Sterility Phenotype in Tomato" Plants 9, no. 9: 1189. https://doi.org/10.3390/plants9091189

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