<i>Agrobacterium tumefaciens</i>
            transformation of
            <i>Medicago truncatula</i>
            cell suspensions

Iantcheva, Anelia; Revalska, Miglena

doi:10.1002/9781119409144.ch150

Cited by 2 publications

(1 citation statement)

References 36 publications

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“…There is a protocol for the generation of transgenic plants of M. truncatula cv. Jemalong 2HA which employs liquid cultures and the procedure is similar to our protocol (Iantcheva et al 2014 ; Iantcheva and Revalska 2019 ). In this work, Medicago cells were co-cultured with Agrobacterium and positive transformants were screened based on GFP or GUS activity.…”

Section: Discussionmentioning

confidence: 99%

A simplified protocol for Agrobacterium-mediated transformation of cell suspension cultures of the model species Medicago truncatula A17

Ferreira

Rebelo

Abranches

2023

Plant Cell Tiss Organ Cult

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This manuscript describes a unique protocol for the rapid transformation of Medicago truncatula A17 cell suspension cultures mediated by Agrobacterium tumefaciens. Medicago cells were collected on day 7 of the growth curve, which corresponded to the beginning of the exponential phase. They were then co-cultured with Agrobacterium for 3 days before being spread onto a petri dish with appropriate antibiotic selection. The Receptor Binding Domain of the Spike protein of SARS-CoV-2 was used as a model to develop this protocol. The presence of the transgene was assessed using PCR, and the integrity of the product was evaluated by SDS-PAGE and Western-blotting.

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Section: Discussionmentioning

confidence: 99%

A simplified protocol for Agrobacterium-mediated transformation of cell suspension cultures of the model species Medicago truncatula A17

Ferreira

Rebelo

Abranches

2023

Plant Cell Tiss Organ Cult

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Cellular and Molecular Tools for the Investigation of Somatic Embryogenesis in Medicago Species

Iantcheva

2023

Genome Size and Genetic Homogeneity of Regenerated Plants: Methods and Applications

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The chapter presents the knowledge accumulated on the recent investigation of somatic embryogenesis (SE) in genera Medicago. The role of 2,4-D in the process of induction of embryogenic potential in diploid Medicago and its transport by the combined action of auxin transporters or diffusion of dissociated molecules is discussed. Among the many methods for studying the process, this chapter is focused on cellular and molecular tools – flow cytometry, assessment of expression level of SE related transcripts of key genes of auxin inducible process and different PCR techniques. Our recent studies on the process of SE in M. truncatula are focused on the role of the two genes MtLAX3 (an auxin transmembrane transporter) and a transcriptional factor MtARF-B3 (an auxin response factor, containing a B3-binding domain). The transcription profiles of these genes are evaluated and their expression patterns are assessed during indirect somatic embryogenesis – steps of callus formation, embryogenic zone formation and the stages of globular, torpedo and cotyledonary embryos. The localization of expression during the process of SE is traced by the β-glucuronidase reporter gene (GUS) under the control of the promoters of these genes. Inverse PCR (IPCR) and Transposon display (TD) are techniques which evaluate transposition and new retrotransposon copies in the investigated mutant lines, and we used these methods as markers for the efficiency of the induction phase of the process of SE. The use of all these methods turns light on a better understanding of the process of somatic embryogenesis in the model species Medicago truncatula and other annual medics.

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Agrobacterium tumefaciens transformation of Medicago truncatula cell suspensions

Cited by 2 publications

References 36 publications

A simplified protocol for Agrobacterium-mediated transformation of cell suspension cultures of the model species Medicago truncatula A17

A simplified protocol for Agrobacterium-mediated transformation of cell suspension cultures of the model species Medicago truncatula A17

Cellular and Molecular Tools for the Investigation of Somatic Embryogenesis in Medicago Species

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