Generation of herbicide tolerance traits and a new selectable marker in wheat using base editing

Zhang, Rui; Liu, Jin-Xing; Chai, Zhuangzhuang; Chen, Sha; Bai, Yang; Zong, Yuan; Chen, Kunling; Li, Jiayang; Jiang, Linjian; Gao, Caixia

doi:10.1038/s41477-019-0405-0

Cited by 218 publications

(125 citation statements)

References 23 publications

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“…2 and Table 3). Antibiotic screening is frequently used in most transformation studies, such as kanamycin, hygromycin or herbicide [26,51,52]. However, the dosage of these substances might influence the regeneration rate and further break the balance of transgenic/non-transgenic seeds.…”

Section: Discussionmentioning

confidence: 99%

A convenient, rapid and efficient method for establishing transgenic lines of Brassica napus

Zhang

Liu

et al. 2020

Plant Methods

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Background: Brassica napus is an important oilseed crop that offers a considerable amount of biomass for global vegetable oil production. The establishment of an efficient genetic transformation system with a convenient transgenic-positive screening method is of great importance for gene functional analysis and molecular breeding. However, to our knowledge, there are few of the aforementioned systems available for efficient application in B. napus. Results: Based on the well-established genetic transformation system in B. napus, five vectors carrying the red fluorescence protein encoding gene from Discosoma sp. (DsRed) were constructed and integrated into rapeseed via Agrobacterium-mediated hypocotyl transformation. An average of 59.1% tissues were marked with red fluorescence by the visual screening method in tissue culture medium, 96.1% of which, on average, were amplified with the objective genes from eight different rapeseed varieties. In addition, the final transgenic-positive efficiency of the rooted plantlets reached up to 90.7% from red fluorescence marked tissues, which was much higher than that in previous reports. Additionally, visual screening could be applicable to seedlings via integration of DsRed, including seed coats, roots, hypocotyls and cotyledons during seed germination. These results indicate that the highly efficient genetic transformation system combined with the transgenic-positive visual screening method helps to conveniently and efficiently obtain transgenic-positive rapeseed plantlets. Conclusion: A rapid, convenient and highly efficient method was developed to obtain transgenic plants, which can help to obtain the largest proportion of transgene-positive regenerated plantlets, thereby avoiding a long period of plant regeneration. The results of this study will benefit gene functional studies especially in high-throughput molecular biology research.

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Section: Discussionmentioning

confidence: 99%

A convenient, rapid and efficient method for establishing transgenic lines of Brassica napus

Zhang

Liu

et al. 2020

Plant Methods

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“…A popular application of BE has been to introduce base pair changes in target genes that confer herbicide resistance in crops such as cotton, maize, rice, watermelon and wheat. Wheat strains with tolerance to multiple herbicides such as sulfonylurea-, imidazolinoneand aryloxyphenoxy propionate-type herbicides have recently been generated by BE [24] .…”

Section: Trait Improvements Developed By Bementioning

confidence: 99%

Genome-edited crops: how to move them from laboratory to market

Kunling¹,

Gao²

2020

Front. Agr. Sci. Eng.

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“…Precise C-to-T or A-to-G changes could be achieved on a desired DNA sequence by base editors, without the formation of DSBs andinduced NHEJ or MMEJ events that eliminates further re-arrangements and without the need for HDR that is restricted to S-and G2-phases of cells. Thus, base-editing represents a powerful and promising tool for pointmutagenesis that can be exploited in crop development, as seen in the manipulation of genes associated with agronomical traits (Hua et al 2018;Yan et al 2018;Kang et al 2018;Endo et al 2019;Zhang et al 2019).…”

Section: Base Editorsmentioning

confidence: 99%

“…Cytidine deaminases have been used to edit genomes of rice, wheat, maize, tomato, potato and watermelon (Zong et al 2017;Lu and Zhu 2017;Ren et al 2017Ren et al , 2018Ren et al , 2019Shimatani et al 2017;Zong et al 2018Tian et al 2018;Endo et al 2019;Hua et al 2019b;Veillet et al 2019;Zhang et al 2019). Optimizations for increasing targeting flexibility, improving editing purity and expanding PAM recognition are provided in Table 1.…”

Section: Base Editorsmentioning

confidence: 99%

“…Loss-of-function phenotypes can also be achieved by introduced missense mutations or mutations that alter RNA splicing sites as demonstrated in Arabidopsis (Kang et al 2018;Li et al 2019b). Gainof-function phenotypes were reported such as dominant semi-dwarf phenotype by editing SLR1 in rice and herbicide resistance by editing ALS in rice and wheat (Shimatani et al 2017;Zong et al 2018;Zhang et al 2019) or editing ACC in rice (Li et al 2018b). In principle, base editors could also be applied to target upstream open reading frames (uORFs) for improved protein translation, as well as to target promoter regions to introduce quantitative traits, as recently demonstrated with the use of Cas9 nuclease (Rodríguez-Leal et al 2017;Zhang et al 2018).…”

Section: Base Editorsmentioning

confidence: 99%

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CRISPR-Cas nucleases and base editors for plant genome editing

et al. 2019

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Clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR-associated protein (Cas) and base editors are fundamental tools in plant genome editing. Cas9 from Streptococcus pyogenes (SpCas9), recognizing an NGG protospacer adjacent motif (PAM), is a widely used nuclease for genome editing in living cells. Cas12a nucleases, targeting T-rich PAMs, have also been recently demonstrated in several plant species. Furthermore, multiple Cas9 and Cas12a engineered variants and orthologs, with different PAM recognition sites, editing efficiencies and fidelity, have been explored in plants. These RNA-guided sequence-specific nucleases (SSN) generate double-stranded breaks (DSBs) in DNA, which trigger non-homologous end-joining (NHEJ) repair or homology-directed repair (HDR), resulting in insertion and deletion (indel) mutations or precise gene replacement, respectively. Alternatively, genome editing can be achieved by base editors without introducing DSBs. So far, several base editors have been applied in plants to introduce C-to-T or A-to-G transitions, but they are still undergoing improvement in editing window size, targeting scope, off-target effects in DNA and RNA, product purity and overall activity. Here, we summarize recent progress on the application of Cas nucleases, engineered Cas variants and base editors in plants.

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Generation of herbicide tolerance traits and a new selectable marker in wheat using base editing

Cited by 218 publications

References 23 publications

A convenient, rapid and efficient method for establishing transgenic lines of Brassica napus

A convenient, rapid and efficient method for establishing transgenic lines of Brassica napus

Genome-edited crops: how to move them from laboratory to market

CRISPR-Cas nucleases and base editors for plant genome editing

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