Dataset Information

Comparison of efficiency and time to regeneration of Agrobacterium-mediated transformation methods in Medicago truncatula.

ABSTRACT:

Background

Tissue culture transformation of plants has an element of art to it, with protocols passed on between labs but often not directly compared. As Medicago truncatula has become popular as a model system for legumes, rapid transformation is critical, and many protocols exist, with varying results.

Results

The M. truncatula ecotypes, R108 and A17, were utilized to compare the effect of a modification to a previously used protocol based on shoot explants on the percentage of transformed plants produced from calli. This percentage was then compared to that of two additional transformation protocols based on root explants in the R108 ecotype. Variations in embryonic tissue sources, media components, time for transformation, and vectors were analyzed.

Conclusions

While no A17 transgenic plants were obtained, transgenic plantlets from the R108 ecotype were produced in as little as 4 months with a comparison of the two widely studied ecotypes under a single set of conditions. While the protocols tested gave similar results in percentage of transformed plants produced, considerations of labor and time to transgenics that vary between the root explant protocols tested were discovered. These considerations may influence which protocol to choose for introducing a single transgene versus creating lines with multiple mutations utilizing a CRISPR/Cas9 construct.

SUBMITTER: Wen L

PROVIDER: S-EPMC6394069 | biostudies-literature | 2019

REPOSITORIES: biostudies-literature

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Publications

Comparison of efficiency and time to regeneration of <i>Agrobacterium</i>-mediated transformation methods in <i>Medicago truncatula</i>.

Wen Li L Chen Yuanling Y Schnabel Elise E Crook Ashley A Frugoli Julia J

Plant methods 20190228

<h4>Background</h4>Tissue culture transformation of plants has an element of art to it, with protocols passed on between labs but often not directly compared. As <i>Medicago truncatula</i> has become popular as a model system for legumes, rapid transformation is critical, and many protocols exist, with varying results.<h4>Results</h4>The <i>M. truncatula</i> ecotypes, R108 and A17, were utilized to compare the effect of a modification to a previously used protocol based on shoot explants on the ...[more]

PMID: 30858871

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