Establishment of an Agrobacterium-mediated genetic transformation and CRISPR/Cas9-mediated mutagenesis of haploid inducer genes in Pak-choi plants (Brassica rapa ssp. chinensis).
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| Title: | Establishment of an Agrobacterium-mediated genetic transformation and CRISPR/Cas9-mediated mutagenesis of haploid inducer genes in Pak-choi plants (Brassica rapa ssp. chinensis). |
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| Authors: | Kim, Young-Cheon1,2 (AUTHOR), Thu, May Phyo1 (AUTHOR), Rahman, Falguni Maliha1 (AUTHOR), Yun, Young Jae1 (AUTHOR), Jang, Jin Hoon3,4 (AUTHOR), Lee, Ok Ran3,4 (AUTHOR), Lee, Jeong Hwan1,2 (AUTHOR) jhwanlee90@jbnu.ac.kr |
| Source: | Plant Biotechnology Reports. Apr2024, Vol. 18 Issue 2, p263-273. 11p. |
| Subjects: | Plant genetic transformation, Bok choy, Plant genes, Genetic transformation, CRISPRs, Genome editing |
| Geographic Terms: | Southeast Asia, East Asia |
| Abstract: | Pak-choi (Brassica rapa ssp. chinensis) is a popular vegetative crop in southern China, East Asia, and Southeast Asia. Owing to the threat of climate change, rapid breeding strategies for vegetable cultivars that are tolerant to abiotic and biotic stresses are required. Thus, the rapid fixation of useful agronomic traits using doubled haploid technology is urgent. The haploid-inducer gene is key to doubled haploidization. Two known CENH3 and pPLAIIγ genes, in which altered or partially deleted forms lead to haploid induction, were selected, and direct editing of Pak-choi CENH3 and pPLAIIγ genes (BcCENH3 and BcpPLAIIγ) was conducted using an Agrobacterium-mediated CRISPR/Cas9 system. First, BcCENH3 and BcpPLAIIγ genes were characterized by analyzing the spatial expression patterns and subcellular localization. The CENH3 expression levels in carpels and pPLAIIγ in various parts of Pak-choi flowers were higher than those of other parts. BcCENH3 and BcpPLAIIγ proteins targeted in the nucleus and plasma membrane, respectively. Whole plants were successfully regenerated from the shoot apical meristem (SAM) regions of Pak-choi seedlings using the optimized procedure and culture conditions. The regeneration results of SAM explants after Agrobacterium-mediated transformation of constructs expressing CRISPR/Cas9 and BcCENH3 or BcpPLAIIγ sgRNAs confirmed four independent BcCENH3-targeted transgenic lines with 2.1%, 1.8%, 1.8%, and 1.7% INDEL frequencies, and three independent BcpPLAIIγ-targeted transgenic lines with 24.5%, 33.7%, and 33.0% INDEL frequencies. Thus, our results suggested the possibility of developing transgenic Pak-choi lines by applying the CRISPR/Cas9 genome editing technology to BcCENH3 and BcpPLAIIγ as two haploid-inducer genes. [ABSTRACT FROM AUTHOR] |
| Copyright of Plant Biotechnology Reports is the property of Springer Nature and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.) | |
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| Header | DbId: egs DbLabel: Engineering Source An: 176471513 AccessLevel: 6 PubType: Academic Journal PubTypeId: academicJournal PreciseRelevancyScore: 0 |
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| Items | – Name: Title Label: Title Group: Ti Data: Establishment of an Agrobacterium-mediated genetic transformation and CRISPR/Cas9-mediated mutagenesis of haploid inducer genes in Pak-choi plants (Brassica rapa ssp. chinensis). – Name: Author Label: Authors Group: Au Data: <searchLink fieldCode="AR" term="%22Kim%2C+Young-Cheon%22">Kim, Young-Cheon</searchLink><relatesTo>1,2</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Thu%2C+May+Phyo%22">Thu, May Phyo</searchLink><relatesTo>1</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Rahman%2C+Falguni+Maliha%22">Rahman, Falguni Maliha</searchLink><relatesTo>1</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Yun%2C+Young+Jae%22">Yun, Young Jae</searchLink><relatesTo>1</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Jang%2C+Jin+Hoon%22">Jang, Jin Hoon</searchLink><relatesTo>3,4</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Lee%2C+Ok+Ran%22">Lee, Ok Ran</searchLink><relatesTo>3,4</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Lee%2C+Jeong+Hwan%22">Lee, Jeong Hwan</searchLink><relatesTo>1,2</relatesTo> (AUTHOR)<i> jhwanlee90@jbnu.ac.kr</i> – Name: TitleSource Label: Source Group: Src Data: <searchLink fieldCode="JN" term="%22Plant+Biotechnology+Reports%22">Plant Biotechnology Reports</searchLink>. Apr2024, Vol. 18 Issue 2, p263-273. 11p. – Name: Subject Label: Subjects Group: Su Data: <searchLink fieldCode="DE" term="%22Plant+genetic+transformation%22">Plant genetic transformation</searchLink><br /><searchLink fieldCode="DE" term="%22Bok+choy%22">Bok choy</searchLink><br /><searchLink fieldCode="DE" term="%22Plant+genes%22">Plant genes</searchLink><br /><searchLink fieldCode="DE" term="%22Genetic+transformation%22">Genetic transformation</searchLink><br /><searchLink fieldCode="DE" term="%22CRISPRs%22">CRISPRs</searchLink><br /><searchLink fieldCode="DE" term="%22Genome+editing%22">Genome editing</searchLink> – Name: SubjectGeographic Label: Geographic Terms Group: Su Data: <searchLink fieldCode="DE" term="%22Southeast+Asia%22">Southeast Asia</searchLink><br /><searchLink fieldCode="DE" term="%22East+Asia%22">East Asia</searchLink> – Name: Abstract Label: Abstract Group: Ab Data: Pak-choi (Brassica rapa ssp. chinensis) is a popular vegetative crop in southern China, East Asia, and Southeast Asia. Owing to the threat of climate change, rapid breeding strategies for vegetable cultivars that are tolerant to abiotic and biotic stresses are required. Thus, the rapid fixation of useful agronomic traits using doubled haploid technology is urgent. The haploid-inducer gene is key to doubled haploidization. Two known CENH3 and pPLAIIγ genes, in which altered or partially deleted forms lead to haploid induction, were selected, and direct editing of Pak-choi CENH3 and pPLAIIγ genes (BcCENH3 and BcpPLAIIγ) was conducted using an Agrobacterium-mediated CRISPR/Cas9 system. First, BcCENH3 and BcpPLAIIγ genes were characterized by analyzing the spatial expression patterns and subcellular localization. The CENH3 expression levels in carpels and pPLAIIγ in various parts of Pak-choi flowers were higher than those of other parts. BcCENH3 and BcpPLAIIγ proteins targeted in the nucleus and plasma membrane, respectively. Whole plants were successfully regenerated from the shoot apical meristem (SAM) regions of Pak-choi seedlings using the optimized procedure and culture conditions. The regeneration results of SAM explants after Agrobacterium-mediated transformation of constructs expressing CRISPR/Cas9 and BcCENH3 or BcpPLAIIγ sgRNAs confirmed four independent BcCENH3-targeted transgenic lines with 2.1%, 1.8%, 1.8%, and 1.7% INDEL frequencies, and three independent BcpPLAIIγ-targeted transgenic lines with 24.5%, 33.7%, and 33.0% INDEL frequencies. Thus, our results suggested the possibility of developing transgenic Pak-choi lines by applying the CRISPR/Cas9 genome editing technology to BcCENH3 and BcpPLAIIγ as two haploid-inducer genes. [ABSTRACT FROM AUTHOR] – Name: AbstractSuppliedCopyright Label: Group: Ab Data: <i>Copyright of Plant Biotechnology Reports is the property of Springer Nature and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract.</i> (Copyright applies to all Abstracts.) |
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| RecordInfo | BibRecord: BibEntity: Identifiers: – Type: doi Value: 10.1007/s11816-024-00898-1 Languages: – Code: eng Text: English PhysicalDescription: Pagination: PageCount: 11 StartPage: 263 Subjects: – SubjectFull: Plant genetic transformation Type: general – SubjectFull: Bok choy Type: general – SubjectFull: Plant genes Type: general – SubjectFull: Genetic transformation Type: general – SubjectFull: CRISPRs Type: general – SubjectFull: Genome editing Type: general – SubjectFull: Southeast Asia Type: general – SubjectFull: East Asia Type: general Titles: – TitleFull: Establishment of an Agrobacterium-mediated genetic transformation and CRISPR/Cas9-mediated mutagenesis of haploid inducer genes in Pak-choi plants (Brassica rapa ssp. chinensis). Type: main BibRelationships: HasContributorRelationships: – PersonEntity: Name: NameFull: Kim, Young-Cheon – PersonEntity: Name: NameFull: Thu, May Phyo – PersonEntity: Name: NameFull: Rahman, Falguni Maliha – PersonEntity: Name: NameFull: Yun, Young Jae – PersonEntity: Name: NameFull: Jang, Jin Hoon – PersonEntity: Name: NameFull: Lee, Ok Ran – PersonEntity: Name: NameFull: Lee, Jeong Hwan IsPartOfRelationships: – BibEntity: Dates: – D: 01 M: 04 Text: Apr2024 Type: published Y: 2024 Identifiers: – Type: issn-print Value: 18635466 Numbering: – Type: volume Value: 18 – Type: issue Value: 2 Titles: – TitleFull: Plant Biotechnology Reports Type: main |
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