Generation of CRISPR/Cas9-mediated mutants in Monochasma savatieri using a hairy root system

NWIPB OpenIR

	Generation of CRISPR/Cas9-mediated mutants in Monochasma savatieri using a hairy root system
	Bai, Chen; Cao, Yingping; Zhao, Siyi; Wu, Zhenying; Dai, Shaojun; Wang, Honglun; Fu, Chunxiang
	2023
发表期刊	INDUSTRIAL CROPS AND PRODUCTS
卷号	191
摘要	Monochasma savatieri Franch. ex Maxim is a perennial medicinal plant with antibiotic properties. However, the current M. savatieri resources cannot meet the increasing market demand. Thus, an advanced bioengineering technology platform is required for breeding novel and high-yielding M. savatieri germplasm/cultivars with high value. In the present study, a hairy root system induced by Agrobacterium rhizogenes, which is often used as a model to study root function and bioactive compound production, was established for M. savatieri for the first time. The hairy root induction frequencies of the different explants ranged from 4.6% to 68.3% after infection with A. rhizogenes strain LBA9402. Furthermore, many co-transformed hairy root lines using the binary vector pEG100-35S::GFP were generated. Fluorescence microscopy revealed that GFP was expressed in 72.9% of the transgenic hairy root lines. After six months, the co-transformed hairy root lines were able to grow into trans-genic plantlets on medium containing cytokinin. In addition, to construct a gene editing system for M. savatieri, the tRNA-target-gRNA cassette was cloned into the pRGEB31 vector with AtU6-sgRNA and hSpCas9 backbones. The cellulose synthase-like D (CSLD2/3) genes were used as markers to assess the gene editing efficiency of the CRISPR/Cas9 vector in M. savatieri. The results showed that the CSLD2/3 sequences were altered in 11 out of 59 co-transformed hairy root lines. This study successfully established a genome editing platform for M. savatieri, for the first time, using the A. rhizogenes-mediated co-transformation system. This will enable the development of novel M. savatieri germplasm/cultivars for facility cultivation and exploitation of its medicinal value in the future.
收录类别	SCIE
文献类型	期刊论文
条目标识符	http://210.75.249.4/handle/363003/61719
专题	中国科学院西北高原生物研究所
推荐引用方式 GB/T 7714	Bai, Chen,Cao, Yingping,Zhao, Siyi,et al. Generation of CRISPR/Cas9-mediated mutants in Monochasma savatieri using a hairy root system[J]. INDUSTRIAL CROPS AND PRODUCTS,2023,191.
APA	Bai, Chen.,Cao, Yingping.,Zhao, Siyi.,Wu, Zhenying.,Dai, Shaojun.,...&Fu, Chunxiang.(2023).Generation of CRISPR/Cas9-mediated mutants in Monochasma savatieri using a hairy root system.INDUSTRIAL CROPS AND PRODUCTS,191.
MLA	Bai, Chen,et al."Generation of CRISPR/Cas9-mediated mutants in Monochasma savatieri using a hairy root system".INDUSTRIAL CROPS AND PRODUCTS 191(2023).