Selectable marker independent transformation of recalcitrant maize inbred B73 and sorghum P898012 mediated by morphogenic regulators BABY BOOM and WUSCHEL2
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Discriminatory co-expression of maize BBM and WUS transcriptional factor genes promoted somatic embryogenesis and efficient Agrobacterium -mediated transformation of recalcitrant maize inbred B73 and sorghum P898012 genotypes without use of a selectable marker gene. The use of morphogenic regulators to overcome barriers in plant transformation is a revolutionary breakthrough for basic plant science and crop applications. Current standard plant transformation systems are bottlenecks for genetic, genomic, and crop improvement studies. We investigated the differential use of co-expression of maize transcription factors BABY BOOM and WUSCHEL2 coupled with a desiccation inducible CRE/lox excision system to enable regeneration of stable transgenic recalcitrant maize inbred B73 and sorghum P898012 without a chemical selectable marker. The PHP78891 expression cassette contains CRE driven by the drought inducible maize RAB17M promoter with lox P sites which bracket the CRE, WUS, and BBM genes. A constitutive maize UBI M promoter directs a ZsGreen GFP expression cassette as a reporter outside of the excision sites and provides transient, transgenic, and developmental analysis. This was coupled with evidence for molecular integration and analysis of stable integration and desiccation inducible CRE-mediated excision. Agrobacterium-mediated transgenic introduction of this vector showed transient expression of GFP and induced somatic embryogenesis in maize B73 and sorghum P898012 explants. Subjection to desiccation stress in tissue culture enabled the excision of CRE, WUS, and BBM, leaving the UBI M::GFP cassette and allowing subsequent plant regeneration and GFP expression analysis. Stable GFP expression was observed in the early and late somatic embryos, young shoots, vegetative plant organs, and pollen. Transgene integration and expression of GFP positive T0 plants were also analyzed using PCR and Southern blots. Progeny segregation analysis of primary events confirmed correlation between functional GFP expression and presence of the GFP transgene in T1 plants generated from self pollinations, indicating good transgene inheritance. This study confirms and extends the use of morphogenic regulators to overcome transformation barriers.
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Overexpression of developmental regulators such as the maize BABY BOOM (BBM) and WUSCHEL 2 (WUS2) genes has increased the transformation efficiency in sorghum, rice, and sugarcane.
“Overexpression of developmental regulators such as the maize BABY BOOM (BBM) and WUSCHEL 2 (WUS2) genes has increased the transformation efficiency in sorghum, rice, and sugarcane (Lowe et al., 2016;Mookkan et al., 2017).”
From Genetic resources and precise gene editing for targeted improvement of barley abiotic stress tolerance (2023) · cites this paper as Mookkan et al., 2017
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“Overexpression of developmental regu‐ lators such as the maize BABY BOOM ( BBM ) and WUSCHEL 2 ( WUS2 ) genes has increased the trans‐ formation efficiency in sorghum, rice, and sugarcane (Lowe et al., 2016; Mookkan et al., 2017).”
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- Authors
- Mookkan, M., Nelson-Vasilchik, K., Hague, J., Zhang, Z. J., & Kausch, A. P.
- Affiliations
- University of Missouri (US)
- University of Rhode Island (US)
- Corresponding author
- Zhang, Zhanyuan J.
- Kausch, Albert P.
- Venue
- Plant Cell Reports
- Published
- 2017-07-05
- Volume
- 36
- Issue
- 9
- Pages
- 1477-1491
- ISSN
- 0721-7714 (ISSN-L)
- 1432-203X
- PMC
- PMC5565672
- OpenAlex
- W2732418005
- Cited by
- 162
- References
- 57
- Citations by year
- 2026: 15 · 2025: 11 · 2024: 21 · 2023: 16 · 2022: 23 · 2021: 29 · 2020: 22 · 2019: 13 · 2018: 9 · 2017: 2
- FWCI
- 5.14
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Locations
- Plant Cell Reports (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Journal of Media Literacy Education (submittedVersion, cc-by)
- Funders
- National Science Foundation Plant Genome (1444478)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Plant tissue culture and regeneration
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biotechnology › Transgenic Plants and Applications
- Life Sciences › Agricultural and Biological Sciences › Plant Science › Genetically Modified Organisms Research
- Keywords
- marker-free transformation, somatic embryogenesis, baby boom, desiccation stress, Agrobacterium-mediated transformation, transgene integration, transgene inheritance, GFP reporter, plant regeneration
- MeSH terms
- Zea mays, Genetic Markers, Genotype, Models, Genetic, Plant Proteins, Promoter Regions, Genetic, Transcription Factors, Transformation, Genetic, Agrobacterium tumefaciens, Gene Expression Regulation, Plant, Plants, Genetically Modified, Sorghum, Green Fluorescent Proteins, Droughts
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