Generation of early-flowering Chinese cabbage (Brassica rapa spp. pekinensis) through CRISPR/Cas9-mediated genome editing

Jeong, So Young; Ahn, Hyo-Min; Ryu, Jae‐Chun; Oh, Youngbin; Sivanandhan, Ganeshan; Won, Kanghee; Park, Young-Doo; Kim, Jin-Soo; Kim, Hye‐Ran; Lim, Yong Pyo; Kim, Sang-Gyu

doi:10.1007/s11816-019-00566-9

Cited by 43 publications

(24 citation statements)

References 28 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…A recent study reported a high mutation efficiency in genome editing of Chinese cabbage 39 and the ratio of the regenerated plantlets to the total number of cocultivated explants was relatively low (0.12–1.07%) compared with that in the present study (3%). Based on the T 1 generation plants, we knew that the mutations were stably inherited regardless of the presence of T-DNA.…”

Section: Discussioncontrasting

confidence: 84%

Brassica rapa orphan genes largely affect soluble sugar metabolism

Jiang

Zhan

et al. 2020

Hortic Res

View full text Add to dashboard Cite

Orphan genes (OGs), which are genes unique to a specific taxon, play a vital role in primary metabolism. However, little is known about the functional significance of Brassica rapa OGs (BrOGs) that were identified in our previous study. To study their biological functions, we developed a BrOG overexpression (BrOGOE) mutant library of 43 genes in Arabidopsis thaliana and assessed the phenotypic variation of the plants. We found that 19 of the 43 BrOGOE mutants displayed a mutant phenotype and 42 showed a variable soluble sugar content. One mutant, BrOG1OE, with significantly elevated fructose, glucose, and total sugar contents but a reduced sucrose content, was selected for in-depth analysis. BrOG1OE showed reduced expression and activity of the Arabidopsis sucrose synthase gene (AtSUS); however, the activity of invertase was unchanged. In contrast, silencing of two copies of BrOG1 in B. rapa, BraA08002322 (BrOG1A) and BraSca000221 (BrOG1B), by the use of an efficient CRISPR/Cas9 system of Chinese cabbage (B. rapa ssp. campestris) resulted in decreased fructose, glucose, and total soluble sugar contents because of the upregulation of BrSUS1b, BrSUS3, and, specifically, the BrSUS5 gene in the edited BrOG1 transgenic line. In addition, we observed increased sucrose content and SUS activity in the BrOG1 mutants, with the activity of invertase remaining unchanged. Thus, BrOG1 probably affected soluble sugar metabolism in a SUS-dependent manner. This is the first report investigating the function of BrOGs with respect to soluble sugar metabolism and reinforced the idea that OGs are a valuable resource for nutrient metabolism.

show abstract

Section: Discussioncontrasting

confidence: 84%

Brassica rapa orphan genes largely affect soluble sugar metabolism

Jiang

Zhan

et al. 2020

Hortic Res

View full text Add to dashboard Cite

show abstract

“…The designed LbCpf1-crRNA2 activity was two-fold higher than that of LbCpf1-crRNA1 based on the induced indel frequencies at the CaMLO2 gene loci. As previously reported with the distinct activities of designed guide RNAs in soybean, cabbage, and petunia [18,40,41], the two designed crRNAs of CaMLO2 exhibited differential editing efficacy based on unknown properties of the sequence context of a target gene. The indel patterns of targeted LbCpf1-crRNA1 complexes varied at the CaMLO2 gene locus in CM334 callus protoplasts, with several deletions of nucleotides (− 6, − 5, and − 10) in the first five ranked reads (Fig.…”

Section: Analysis Of Crispr/lbcpf1-rnp In Cm334 Callus Protoplastssupporting

confidence: 60%

A stable DNA-free screening system for CRISPR/RNPs-mediated gene editing in hot and sweet cultivars of Capsicum annuum

2020

Self Cite

View full text Add to dashboard Cite

Background DNA-free, clustered regularly interspaced short palindromic repeats (CRISPR)-associated protein (Cas) ribonucleoprotein (RNP)-based genome editing is a simple, convincing, and promising tool for precision crop breeding. The efficacy of designed CRISPR-based genome editing tools is a critical prerequisite for successful precision gene editing in crops. Results This study demonstrates that soil-grown leaf- or callus-derived pepper protoplasts are a useful system for screening of efficient guide RNAs for CRISPR/Cas9 or CRISPR/Cas12a (Cpf1). CRISPR/Cas9 or Cpf1 were delivered as CRISPR/RNP complexes of purified endonucleases mixed with the designed single guide RNA, which can edit the target gene, CaMLO2 in two pepper cultivars with whole genome sequenced, Capsicum annuum ‘CM334’ and C. annuum ‘Dempsey’. The designed guide RNAs (sgRNAs for Cas9 or crRNAs for Cpf1) are conserved for CaMLO2 in both CM334 and Dempsey and cleave CaMLO2 in vitro. CRISPR/Cas9- or /Cpf1-RNP complexes were transfected into purely isolated protoplasts of the hot pepper CM334 and sweet pepper Dempsey by PEG-mediated delivery. Targeted deep sequencing analysis indicated that the targeted CaMLO2 gene was differentially edited in both cultivars, depending on the applied CRISPR/RNPs. Conclusions Pepper protoplast-based CRISPR guide-RNA selection is a robust method to check the efficacy of designed CRISPR tools and is a prerequisite for regenerating edited plants, which is a critical time-limiting procedure. The rapid and convincing selection of guide RNA against a target genome reduces the laborious efforts for tissue culture and facilitates effective gene editing for pepper improvement.

show abstract

“…This multiplex gene knock-out study is very useful for gene functional studies in polyploid crops such as Brassica that contain many copies of the same genes as homeologs within the genome. The CRISPR-Cas9 system was also successfully applied in B. rapa for early-flowering trait genome editing [ 231 ]. In L. maculans , the chitin-binding gene LmCBP1 , which was highly expressed during infection in B. napus and functional analysis using CRISPR-Cas9, was shown to play a role in enhancing cell death for pathogen growth and also to be involved in tolerance towards H 2 O 2 during the plant immune response [ 232 ].…”

Section: Future Perspectivesmentioning

confidence: 99%

Understanding Host–Pathogen Interactions in Brassica napus in the Omics Era

Neik

Amas

Barbetti

et al. 2020

Plants

View full text Add to dashboard Cite

Brassica napus (canola/oilseed rape/rapeseed) is an economically important crop, mostly found in temperate and sub-tropical regions, that is cultivated widely for its edible oil. Major diseases of Brassica crops such as Blackleg, Clubroot, Sclerotinia Stem Rot, Downy Mildew, Alternaria Leaf Spot and White Rust have caused significant yield and economic losses in rapeseed-producing countries worldwide, exacerbated by global climate change, and, if not remedied effectively, will threaten global food security. To gain further insights into the host–pathogen interactions in relation to Brassica diseases, it is critical that we review current knowledge in this area and discuss how omics technologies can offer promising results and help to push boundaries in our understanding of the resistance mechanisms. Omics technologies, such as genomics, proteomics, transcriptomics and metabolomics approaches, allow us to understand the host and pathogen, as well as the interaction between the two species at a deeper level. With these integrated data in multi-omics and systems biology, we are able to breed high-quality disease-resistant Brassica crops in a more holistic, targeted and accurate way.

show abstract

Generation of early-flowering Chinese cabbage (Brassica rapa spp. pekinensis) through CRISPR/Cas9-mediated genome editing

Cited by 43 publications

References 28 publications

Brassica rapa orphan genes largely affect soluble sugar metabolism

Brassica rapa orphan genes largely affect soluble sugar metabolism

A stable DNA-free screening system for CRISPR/RNPs-mediated gene editing in hot and sweet cultivars of Capsicum annuum

Understanding Host–Pathogen Interactions in Brassica napus in the Omics Era

Contact Info

Product

Resources

About