Expression Characteristics of Rice OsGRAS39 Gene and Creation of Its Mutants Based on CRISPR/Cas9 Technique

doi:10.7668/hbnxb.20191602

Abstract

Abstract: In order to investigate the function of OsGRAS39 (LOC_Os10g22430.1) in rice, the expression patterns of OsGRAS39 in different tissues and under diverse abiotic stresses as well as phytohormone ABA and GA₃ treatments were analyzed by qRT-PCR; and the OsGRAS39 gene knockout mutants were constructed based on CRISPR/Cas9 system. The results of qRT-PCR analysis showed that OsGRAS39 was expressed in all tissues tested. The mRNA abundance of OsGRAS39 was highest in rice seedlings, followed by young roots and leaves, while lesser in leaf sheaths, panicles, roots and stems at the heading stage. The expression of OsGRAS39 was inhibited by high temperature, while induced by low temperature, NaCl and ABA. Under the treatment of 10% PEG6000, there was no significant difference in the expression level of OsGRAS39 at all treatment time points. Under the treatment of 20 μmol/L GA₃, the expression level of OsGRAS39 was down-regulated at 2-8 h and up-regulated at 24 h. In addition, the OsGRAS39 gene knockout vector pP1C.3-OsGRAS39-gRNA was successfully constructed by seamless cloning. Thirty transgenic rice lines were obtained by Agrobacterium-mediated genetic transformation. The sequencing results of target site showed that 8 out of 30 transgenic rice seedlings were found to have different mutation types, including 1 homozygous mutant, 5 biallelic mutants and 2 heterozygous mutants. In conclusion, OsGRAS39 may be involved in the regulation of rice resistance to high temperature, low temperature and NaCl stress. The OsGRAS39 knock-out mutants were successfully created.

Key words: Rice, OsGRAS39, Expression characteristics, CRISPR/Cas9, Gene editing

CLC Number:

Q78
S511.03

ZOU Jie. Expression Characteristics of Rice OsGRAS39 Gene and Creation of Its Mutants Based on CRISPR/Cas9 Technique[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2020, 35(6): 1-8. doi: 10.7668/hbnxb.20191602.

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