Editing Badh2 Gene to Improve the Fragrance of japonica Rice by CRISPR/Cas9 Technology

doi:10.7668/hbnxb.201751503

Abstract

Abstract: Rice is the main food crop planted in the world, and the fragrant rice breeding has always been a hotspot in the rice breeding. The aroma of rice is mainly controlled by betaine dehydrogenase gene(Badh2), located on chromosome 8. In this study, a non-fragrant japonica variety Dongnong 425 was used as the test material, and two CRISPR/Cas9 knockout vectors were constructed to edit Badh2. The two targets of the first vector(pYLCRISPR/Cas9-B1 -gRNA)were located on the second exon and the third exon, respectively, while the two targets of the second vector(pYLCRISPR/Cas9-B2 -gRNA)were both on the second exon. The Agrobacterium-mediated transformation was used to transform Dongnong 425, and the homozygous T₀ generation plants were successfully obtained. The T₁ plants derived from the homozygous T₀ generation plants were tested for T-DNA elements, and a total of eight homozygous mutant lines were obtained, which had different mutation types and non-transgenic components. At the same time, the seeds of the eight homozygous mutant lines were tested for fragrance by chewing method and potassium hydroxide immersion method. The results showed that the eight Badh2 lines had different degrees of fragrance, of which the fragrance of three lines edited by the vector pYLCRISPR/Cas9-B1 -gRNA were more intensive. The main agronomic traits of the eight homozygous mutants were investigated, indicating that the number of panicles per plant, the number of grains per panicle, the seed setting rate and the 1000-grain weight were not significantly different from those of the wild-type. In summary, this study successfully edited the Badh2 gene of Dongnong 425, and obtained the homozygous mutant lines with significantly improved fragrance and non-transgenic components, which provided the technical support for accelerating the breeding of fragrant japonica rice varieties.

Key words: japonica rice, CRISPR/Cas9, Badh2, Fragrance

CLC Number:

Q78
S511.03

SUN Huiyu, SONG Jia, WANG Jingguo, LIU Hualong, SUN Jian, MO Tianyu, XU Shanbin, ZHENG Hongliang, ZOU Detang. Editing Badh2 Gene to Improve the Fragrance of japonica Rice by CRISPR/Cas9 Technology[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2019, 34(4): 1-8. doi: 10.7668/hbnxb.201751503.

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