Cut–dip–budding delivery system enables genetic modifications in plants without tissue culture

Cao, Xuesong; Xie, Hua; Song, Meimei; Lu, Junfu; Ma, Pengtao; Huang, Boyu; Wang, Mugui; Tian, Yugang; Chen, Fan; Peng, Jun; Lang, Zhaobo; Li, Guofu; Zhu, Jian‐Kang

doi:10.1016/j.xinn.2022.100345

Cited by 74 publications

(67 citation statements)

References 29 publications

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“…For instance, A. rhizogenes has been used for achieving plant transformation under nonsterile conditions. Several recent transformation strategies have also employed A. rhizogenes for inducing the regeneration of plant organs 14,36 . However, A. rhizogenes potentially causes persistent abnormal plant growth owing to the presence of rol genes that are randomly inserted into the genome [37][38][39] .…”

Section: Discussionmentioning

confidence: 99%

Section: Mainmentioning

confidence: 99%

“…In another strategy, meristem identity regulators were used to promote the regeneration of transformed tissues without sterile operation procedures 13 . In a recent study, A. rhizogenes was used for directly inducing the regeneration of transgenic organs 14 . Additionally, numerous studies have employed various delivery intermediaries, including viruses and nanomaterials for plant transformation 6,15 .…”

Section: Mainmentioning

confidence: 99%

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A simple and efficientin plantatransformation method based on the active regeneration capacity of plants

Mei

Chen

Wang

et al. 2023

Preprint

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Plant genetic transformation strategies serve as essential tools for the genetic engineering and advanced molecular breeding of plants. However, the complicated operational protocol and low efficiency of the current transformation strategies restrict the genetic modification of most plant species. This paper describes the development of a Regenerative Activity-dependent in Planta Injection Delivery (RAPID) method based on the active regeneration capacity of plants. In this method, Agrobacterium tumefaciens was delivered to plant meristems via injection for inducing transfected renascent tissues. Stable transgenic plants were obtained by subsequent vegetative propagation of the positive renascent tissues. The method was successfully applied for the transformation of plants with strong regeneration capacity, including different genotypes of sweet potato (Ipomoea batatas), potato (Solanum tuberosum), and bayhops (I. pes-caprae). Compared to the traditional transformation methods, RAPID has a markedly high transformation efficiency (up to ~ 40%), shorter duration (less than 4 weeks), and does not require tissue culture procedures. The RAPID method therefore overcomes the limitations of traditional methods for achieving rapid in planta transformation, and can be potentially applied to a wide range of plant species that are capable of active regeneration.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Mainmentioning

confidence: 99%

Section: Mainmentioning

confidence: 99%

See 1 more Smart Citation

A simple and efficientin plantatransformation method based on the active regeneration capacity of plants

Mei

Chen

Wang

et al. 2023

Preprint

View full text Add to dashboard Cite

show abstract

“…For instance, A. rhizogenes has been used for achieving plant transformation under nonsterile conditions. Several recent transformation strategies have also employed A. rhizogenes for inducing the regeneration of plant organs 14,39 . However, A. rhizogenes potentially causes persistent abnormal plant growth owing to the presence of rol genes that are randomly inserted into the genome [40][41][42] .…”

Section: Discussionmentioning

confidence: 99%

A simple and efficient in planta transformation method based on the active regeneration capacity of plants

Mei

Chen

Wang³

et al. 2023

Preprint

View full text Add to dashboard Cite

Plant genetic transformation strategies serve as essential tools for the genetic engineering and advanced molecular breeding of plants. However, the complicated operational protocol and low efficiency of the current transformation strategies restrict the genetic modification of most plant species. This paper describes the development of a Regenerative Activity-dependent in Planta Injection Delivery (RAPID) method based on the active regeneration capacity of plants. In this method, Agrobacterium tumefaciens was delivered to plant meristems via injection for inducing transfected renascent tissues. Stable transgenic plants were obtained by subsequent vegetative propagation of the positive renascent tissues. The method was successfully applied for the transformation of plants with strong regeneration capacity, including different genotypes of sweet potato (Ipomoea batatas), potato (Solanum tuberosum), and bayhops (I. pes-caprae). Compared to the traditional transformation methods, RAPID has a markedly high transformation efficiency (up to 40%), shorter duration (less than 4 weeks), and does not require tissue culture procedures. The RAPID method therefore overcomes the limitations of traditional methods for achieving rapid in planta transformation, and can be potentially applied to a wide range of plant species that are capable of active regeneration.

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“…The embryogenic suspension line is currently the most widely used method. Recently, the genetic transformation system of sweetpotato was greatly optimized by the CDB method without tissue culture, by which the amylose content in tuberous root was changed by editing the GBSSI, SBEI, or SBEII gene 33 . Due to the complexity of genetic background, both research into and application of sweetpotato molecular breeding will face more difficulties.…”

Section: Introductionmentioning

confidence: 99%

Haploid-resolved and chromosome-scale genome assembly in hexa-autoploid sweetpotato (Ipomoea batatas(L.) Lam)

Yoon

Cao

Shirasawa

et al. 2022

Preprint

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Sweetpotato (Ipomoea batatas (L.) Lam) is the seventh most important food crop in the world by production quantity. Cultivated sweetpotato is a hexaploid (2n = 6x = 90), and its genome (B1B1B2B2B2B2) is quite complex due to polyploidy, self-incompatibility, and high heterozygosity. Here we established a haploid-resolved and chromosome-scale de novo assembly of autohexaploid sweetpotato genome sequences. Before constructing the genome, we created chromosome-scale genome sequences in I. trifida using a highly homozygous accession, Mx23Hm, with PacBio RSII and Hi-C reads. Haploid-resolved genome assembly was performed for a sweetpotato cultivar, Xushu18 by hybrid assembly with Illumina paired-end (PE) and mate-pair (MP) reads, 10X genomics reads, and PacBio RSII reads. Then, 90 chromosome-scale pseudomolecules were generated by aligning the scaffolds onto a sweetpotato linkage map. De novo assemblies were also performed for chloroplast and mitochondrial genomes in I. trifida and sweetpotato. In total, 34,386 and 175,633 genes were identified on the assembled nucleic genomes of I. trifida and sweetpotato, respectively. Functional gene annotation and RNA-Seq analysis revealed locations of starch, anthocyanin, and carotenoid pathway genes on the sweetpotato genome. This is the first report of chromosome-scale de novo assembly of the sweetpotato genome. The results are expected to contribute to genomic and genetic analyses of sweetpotato.

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Cut–dip–budding delivery system enables genetic modifications in plants without tissue culture

Cited by 74 publications

References 29 publications

A simple and efficientin plantatransformation method based on the active regeneration capacity of plants

A simple and efficientin plantatransformation method based on the active regeneration capacity of plants

A simple and efficient in planta transformation method based on the active regeneration capacity of plants

Haploid-resolved and chromosome-scale genome assembly in hexa-autoploid sweetpotato (Ipomoea batatas(L.) Lam)

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