AnAgrobacterium-mediated transformation via organogenesis regeneration of a facultative CAM plant, the common ice plantMesembryanthemum crystallinumL

Abstract

The common ice plant, Mesembryanthemum crystallinum L. provides a useful model for the study of environmentally induced photosynthetic conversion and abiotic stresses tolerance. However, a procedure for the production of transgenic ice plant, which is essential for functional genomics, has not been fully established. Here we tested the factors on the transformation of cotyledonary nodes excised from the ice plant seedlings such as thidiazuron (TDZ), NaCl and phytosulfokine (PSK), a peptidyl plant growth factor using Agrobacterium tumefaciens strains EHA101 and EHA105 harboring binary vector plasmids pBI7EGFP and pCAMBIA1302, respectively. The established procedure is as follows: the explants (cotyledonary nodes) were co-cultivated with Agrobacterium for 3 days, and the explants were cultured in the medium with 0.5 mg l −1 kinetin and 100 mg l −1 carbenicillin for 72 h, and they were cultured in the medium with 0.5 mg l −1 kinetin and 100 nM PSKfor 4 weeks. Thidiazuron and NaCl enhanced the production of multiple adventitious shoot formation during regeneration but reduced the transformation efficiency due to the vitrification of adventitious shoots. PSK was effective in the production of healthy adventitious shoots. The transformation frequency at the stage of whole plants was 0.6% and 4.6% per inoculated cotyledonary nodes using the Agrobacterium strain EHA101 (pBI7EGFP) and EHA105 (pCAMBIA1302), respectively. ARTICLE HISTORY

Modeling Agrobacterium-Mediated Gene Transformation of Tobacco (Nicotiana tabacum)—A Model Plant for Gene Transformation Studies

“…The LB4404 Agrobacterium strain was more efficient than the AGL1 strain in the gene transformation of foxtail millet ( Setaria italic L.) (Sood et al, 2020 ). Host–pathogen interaction is an important factor in Agrobacterium -mediated T-DNA delivery, as it can significantly affect the survival rate and regeneration activity of the transformed explants (Agarie et al, 2020 ). The transcription levels of the virulence ( vir ) genes in the induction medium can affect the transformation efficiency of Agrobacterium tumefaciens strains, as high transcriptional levels of vir genes were important for successful transformation (Wang et al, 2020 ).…”

Bringing CAM photosynthesis to the table: Paving the way for resilient and productive agricultural systems in a changing climate

“…The recent publication of M . crystallinum transformation protocols ( Hwang et al., 2019 ; Agarie et al., 2020 ), coupled with new genome-editing and synthetic biology techniques, can help accelerate the functional characterization of genes essential for enhancing stress resilience and WUE in major crops.…”

Transcription Factor McHB7 Improves Ice Plant Drought Tolerance through ABA Signaling Pathway

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