Supersweet and waxy: meeting the diverse demands for specialty maize by genome editing

Dong, Le; Qi, Xiantao; Zhu, Jinjie; Liu, Changlin; Zhang, Xin; Cheng, Beijiu; Mao, Long; Xie, Chuanxiao

doi:10.1111/pbi.13144

Cited by 58 publications

(37 citation statements)

References 11 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…RNP delivered in plants [12] Arabidopsis thaliana, Lactuca sativa, Nicotiana attenuata, Oryza sativa Protoplasts PEG-mediated 5.7-40.0% [13] Malus domestica, Vitis vinifera Protoplasts PEG-mediated 0.1-6.9% [14] Petunia x hybrid Protoplasts PEG-mediated 2.4-21.0% [31] Triticum aestivum Protoplasts, immature embryos PEG-mediated, Biolistics 0.2-45.3% [15] Solanum tuberosum Protoplasts PEG-mediated 1.0-25.0% [32] Brassica oleracea, Brassica rapa Protoplasts PEG-mediated 0.1-24.5% [33] Oryza sativa Zygotes PEG-mediated 14.0-64.0% Zea mays Immature embryos Agrobacterium-mediated 12.0-74.0% [38] Zea mays Immature embryos Vector via Biolistics 60.0-98.0% [39] Zea mays Immature embryos Agrobacterium-mediated N.A [19] Zea mays Immature embryos Agrobacterium-mediated 5.0-100% [20] Zea mays Immature embryos Agrobacterium-mediated N.A [40] Zea mays Immature embryos Agrobacterium-mediated 90.0-100% [41] Zea mays Immature embryos Agrobacterium-mediated N.A [42] Zea mays Immature embryos Agrobacterium-mediated N.A [43] Zea mays Immature embryos Agrobacterium-mediated N.A [44] Zea mays Immature embryos Agrobacterium-mediated N.A [45] Zea mays Immature embryos Agrobacterium-mediated N.A [46] Zea mays Immature embryos Vector via biolistics N.A [47] Zea mays Immature embryos Agrobacterium-mediated 25.0-100% [48] Zea mays Immature embryos Agrobacterium-mediated N.A Present study Zea mays Protoplasts RNA via PEG-mediated 0.85-5.85% Note: N.A is 'not applicable' because transgenic plants were either selected using antibiotic marker genes or the analysis was not performed.…”

Section: Reference Plant Species Plant Materials Transfection Methods Gene-editing Efficiencymentioning

confidence: 99%

“…The few studies published on maize genome editing rely mostly on the stable transformation [ 17 , 18 , 19 , 20 ]. In the manuscript, we delivered Cas9-gRNA RNP into maize leaf protoplasts via polyethylene glycol (PEG)-calcium mediated transfection, and indicated that In/Del mutations occurred with relatively high efficiency of 0.85–5.85% among the PEG-calcium-treated protoplast.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

PEG-Delivered CRISPR-Cas9 Ribonucleoproteins System for Gene-Editing Screening of Maize Protoplasts

Sant’Ana

Caprestano

Nodari

et al. 2020

Genes

View full text Add to dashboard Cite

Clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 technology allows the modification of DNA sequences in vivo at the location of interest. Although CRISPR-Cas9 can produce genomic changes that do not require DNA vector carriers, the use of transgenesis for the stable integration of DNA coding for gene-editing tools into plant genomes is still the most used approach. However, it can generate unintended transgenic integrations, while Cas9 prolonged-expression can increase cleavage at off-target sites. In addition, the selection of genetically modified cells from millions of treated ones, especially plant cells, is still challenging. In a protoplast system, previous studies claimed that such pitfalls would be averted by delivering pre-assembled ribonucleoprotein complexes (RNPs) composed of purified recombinant Cas9 enzyme and in vitro transcribed guide RNA (gRNA) molecules. We, therefore, aimed to develop the first DNA-free protocol for gene-editing in maize and introduced RNPs into their protoplasts with polyethylene glycol (PEG) 4000. We performed an effective transformation of maize protoplasts using different gRNAs sequences targeting the inositol phosphate kinase gene, and by applying two different exposure times to RNPs. Using a low-cost Sanger sequencing protocol, we observed an efficiency rate of 0.85 up to 5.85%, which is equivalent to DNA-free protocols used in other plant species. A positive correlation was displayed between the exposure time and mutation frequency. The mutation frequency was gRNA sequence- and exposure time-dependent. In the present study, we demonstrated that the suitability of RNP transfection was proven as an effective screening platform for gene-editing in maize. This efficient and relatively easy assay method for the selection of gRNA suitable for the editing of the gene of interest will be highly useful for genome editing in maize, since the genome size and GC-content are large and high in the maize genome, respectively. Nevertheless, the large amplitude of mutations at the target site require scrutiny when checking mutations at off-target sites and potential safety concerns.

show abstract

Section: Reference Plant Species Plant Materials Transfection Methods Gene-editing Efficiencymentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

PEG-Delivered CRISPR-Cas9 Ribonucleoproteins System for Gene-Editing Screening of Maize Protoplasts

Sant’Ana

Caprestano

Nodari

et al. 2020

Genes

View full text Add to dashboard Cite

show abstract

“…In a consumer-driven market, the demand for sweet, waxy, and baby corn variety is increasing day by day. Keeping this fact in mind, Dong et al (2019) have developed super sweet and waxy corn variety by mutating SH2 and WX genes involved in starch and sugar metabolism (Dong et al, 2019). An exciting study opens a new horizon of trait modifications in crops such as maize by showing one step genome editing can be possible during haploid induction, suggesting that gene-editing attempts will rise in maize in the near future (Kelliher et al, 2019).…”

Section: Maizementioning

confidence: 99%

Emerging tools and paradigm shift of gene editing in cereals, fruits, and horticultural crops for enhancing nutritional value and food security

Tiwari

Trivedi

Pandey

2020

Food and Energy Security

View full text Add to dashboard Cite

Gene editing using sequence‐specific nucleases, particularly CRISPR/Cas ribonucleoprotein, has drawn enormous attention in plant research in recent years. Nearly a decade ago, Cas9 protein was initially discovered for a role in adaptive immunity in bacteria. Owing to vast potential, a large number of reports came out in a short span of time, comprising the identification of Cas protein from different bacterial sources, new Cas9 variants with reduced off‐targets, multiplexing, base editing, prime editing, and RNA manipulation in plants. Studies revealed that CRISPR/Cas‐based gene editing can play a major role in ensuring food security via developing resilient commercial crops with improved yield and nutritional value. Use of the CRISPR/Cas9 system for creating mutation in genes and regulatory regions of promoter generated a number of alleles with variable phenotypes, which can serve as an excellent genetic resource in the breeding program. In this review, we provide a recent overview of state‐of‐art discoveries in the CRISPR/Cas system comprised of new Cas proteins, modifications of existing Cas9, refinements in CRISPR/Cas‐induced gene editing, applications, and outcome emphasizing on major cereals and horticultural crops. We also highlight the current global policy framework for the regulation of gene‐edited crops.

show abstract

“…Waxy maize, also known as sticky maize, has high economic, nutritional, and processing value 1 , 2 . The starch in the endosperm of waxy maize is nearly 100% amylopectin, which confers the sticky quality to maize grains.…”

Section: Introductionmentioning

confidence: 99%

“…It is mainly consumed as food in Asia, and is also an important ingredient in the textile and paper industries. Waxy maize was first discovered in China in 1908, and was later reported in other locations of Asia 2 . The results of several studies suggest that the southwest region of China, particularly Yunnan Province and its surrounding areas, is the central origin of Chines waxy maize 3 , 4 .…”

Section: Introductionmentioning

confidence: 99%

Sequence polymorphism of the waxy gene in waxy maize accessions and characterization of a new waxy allele

Luo

Shi

Yang

et al. 2020

Sci Rep

View full text Add to dashboard Cite

Waxy maize has many excellent characteristics in terms of its nutritional and economic value. In recent decades, the waxy maize germplasm has increased dramatically as a result of different selection methods. We collected 200 waxy maize inbred accessions from different origins to study their genetic diversity and phylogenetic relationships, and to identify new waxy mutations. A simple sequence repeat (SSR) analysis revealed wide genetic diversity among the 200 waxy maize accessions. The maize accessions were clustered into three groups. We sequenced the waxy gene from the first to the 14th exon. Nucleotide variation analysis of 167 waxy maize and 14 flint maize lines revealed some nucleotide differences in the waxy gene among different waxy maize groups, and much narrower nucleotide diversity in waxy maize than in flint maize. In a phylogenetic analysis, waxy maize carrying the same mutation allele clustered together, and waxy maize carrying different mutation alleles distributed in different groups; waxy maize was intermixed with flint maize in each branch, and wx-D7 waxy maize separated significantly from waxy maize lines carrying wx-D10, wx-124 and wx-hAT mutant alleles. The wx-hAT was a new waxy mutation identified in this study. It consisted of a 2286-bp transposon inserted into the middle of exon three of the waxy gene. A PCR marker specific for the wx-hAT allele was developed. These results will be useful for the utilization and preservation of the waxy maize germplasm, and the PCR marker has potential uses in waxy maize breeding programs.

show abstract

Supersweet and waxy: meeting the diverse demands for specialty maize by genome editing

Cited by 58 publications

References 11 publications

PEG-Delivered CRISPR-Cas9 Ribonucleoproteins System for Gene-Editing Screening of Maize Protoplasts

PEG-Delivered CRISPR-Cas9 Ribonucleoproteins System for Gene-Editing Screening of Maize Protoplasts

Emerging tools and paradigm shift of gene editing in cereals, fruits, and horticultural crops for enhancing nutritional value and food security

Sequence polymorphism of the waxy gene in waxy maize accessions and characterization of a new waxy allele

Contact Info

Product

Resources

About