CRISPR/Cas9-mediated disruption of TaNP1 genes results in complete male sterility in bread wheat.

Journal of Genetics and Genomics = Yi Chuan Xue Bao
Jian LiXing Wang Deng

Abstract

Male sterile genes and mutants are valuable resources in hybrid seed production for monoclinous crops. High genetic redundancy due to allohexaploidy makes it difficult to obtain the nuclear recessive male sterile mutants through spontaneous mutation or chemical or physical mutagenesis methods in wheat. The emerging effective genome editing tool, CRISPR/Cas9 system, makes it possible to achieve simultaneous mutagenesis in multiple homoeoalleles. To improve the genome modification efficiency of the CRISPR/Cas9 system in wheat, we compared four different RNA polymerase (Pol) III promoters (TaU3p, TaU6p, OsU3p, and OsU6p) and three types of sgRNA scaffold in the protoplast system. We show that the TaU3 promoter-driven optimized sgRNA scaffold was most effective. The optimized CRISPR/Cas9 system was used to edit three TaNP1 homoeoalleles, whose orthologs, OsNP1 in rice and ZmIPE1 in maize, encode a putative glucose-methanol-choline oxidoreductase and are required for male sterility. Triple homozygous mutations in TaNP1 genes result in complete male sterility. We further demonstrated that any one wild-type copy of the three TaNP1 genes is sufficient for maintenance of male fertility. Taken together, this study provides an optimized C...Continue Reading

Citations

Apr 14, 2021·Transgenic Research·Jan G SchaartMarinus J M Smulders
Jun 15, 2021·Frontiers in Plant Science·Sunny AhmarKi-Hung Jung
Jul 22, 2021·Journal of Agricultural and Food Chemistry·Shakeel AhmadShaoqing Tang
Aug 8, 2021·International Journal of Molecular Sciences·Manjit SinghA Mark Cigan
Aug 25, 2021·Molecular Biology Reports·Mahpara KashtwariSajad Majeed Zargar

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