关键词: 水稻, 单碱基基因编辑, 抗除草剂, 乙酰乳酸合成酶

Abstract:

Objective This study edited the acetyl lactate synthase (ALS) gene at designated locus of Ningjing 56, Ningxia salt-tolerant variety, and created rice germplasm resources with herbicide resistance characteristics. Method Using the cytosine base editor BE3, single base editing vector OsALS171-SpG-eBE3 and OsALS190-SpG-eBE3 were constructed with ALS gene 171 and 190 as target sites. Gene edited plants were obtained through Agrobacterium mediated transformation method and target sites were genotyped with Sanger sequencing. A rapid herbicide tolerance system was established and tolerance level was identified in edited herbicide materials. Result Through T₁ target sites sequencing, Two ALS^P171Fmutant homozygous lines and one homozygous strain of ALS^{E187K R190H}, ALS^E187K, ALS^V188I, ALS^R190H, and ALS^D201Nmutation types were obtained. Compared to wild-type material, Two ALS^P171F mutant homozygous strains have strong tolerance to bispyribac-sodium(BS), while ALS^{E187K R190H}, ALS^E187K, ALS^V188I, ALS^R190H, and ALS^D201Nmutant homozygous plants have some tolerances to BS, but the tolerance is not significant. Conclusion The gene edited materials can maintain stable inheritance, using the cytosine base editor BE3 to obtain materials with herbicide resistance characteristics, and the ALS^P171Fmutation demonstrates strong resistance to bispyribac-sodium, which is a priority site for cultivating herbicide resistant rice.

Key words: rice, single base gene editing, herbicide-resistant, acetolactate synthase