Chinese Journal OF Rice Science >
Construction of osarf7 Mutants in Rice Based on CRISPR/Cas9 Technology and Investigation on Their Agronomic Traits
Received date: 2021-01-17
Revised date: 2021-04-22
Online published: 2022-05-11
【Objective】The purpose is to offer a deep insight into the biological function of OsARF7 and its effect on agronomic traits during the growth and development stage of rice. 【Method】CRISPR/Cas9 technology was employed to edit the gene sequence of OsARF7 and to obtain the mutants of japonica variety Zhonghua 11. The agronomic traits of the mutants were investigated in a field experiment. 【Result】Twenty-two T0 transgenic lines were obtained through Agrobacterium-mediated infection of japonica rice Zhonghua 11 callus, and twenty positive transgenic lines were distinguished by screening the gene encoding hygromycin. For T2 transgenic lines, the regions of genomic DNA including two target sites were detected and identified by PCR amplifying and sequencing, and fifteen homozygous mutations were confirmed. Gene expression results displayed that the expression levels of OsARF7, OsPIN1b, OsPIN2, and OsLAX2 in the osarf7 mutants were significantly lower than those in the wild type. In comparison with the wild-type plants, osarf7 mutants showed a significant decrease in the productive panicle percentage. Among them, the osarf7 mutants with more tillers than the wild type showed a similar effective panicle number to the wild type, and the osarf7 mutants with the same tiller number as the wild type presented a decrease in the effective panicle number per plant in paddy field conditions, resulting in increased non-productive tillers and decreased effective panicle percentage. 【Conclusion】OsARF7 was involved in the emergence and development of tiller bud dominated by auxin, and the successful construction of osarf7 mutants plays a positive role in revealing the biological function of OsARF7 and the mechanism of tiller formation and development dominated by auxin.
Key words： rice; gene editing; OsARF7; CRISPR/Cas9; auxin
LI Zhaowei, SUN Congying, LING Donglan, ZENG Huiling, ZHANG Xiaomei, FAN Kai, LIN Wenxiong . Construction of osarf7 Mutants in Rice Based on CRISPR/Cas9 Technology and Investigation on Their Agronomic Traits[J]. Chinese Journal OF Rice Science, 2022 , 36(3) : 237 -247 . DOI: 10.16819/j.1001-7216.2022.210117
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