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Production of fertile transgenic lentil (Lens culinaris Medik) plants using particle bombardment

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Summary

A reproducible system for gene transfer in lentil through particle bombardment is presented. Lentil cotyledonary nodes excised from germinated seedlings were bombarded with a plasmid containing a mutant acetolactate synthase gene (ALS) from tobacco conferring resistance to sulfonylurea herbicides. Putative transgenic shoots regenerated on Murashige and Skoog medium supplemented with 6-benzylaminopurine (BA) and chlorsulfuron (5 nM for first 4 wk followed by 2.5 nM for the remainder of the culture period) were micrografted and successfully transferred to soil. T0 and selfed progeny plants were screened using metsulfuron herbicide leaflet painting. The non-transformed escapes died and transformed plants survived the test. The surviving plants were phenotypically normal and produced viable seeds. The presence and stable transmission of the transgene into genomic DNA of screened T1 transformants was confirmed by PCR and Southern hybridization. This method for producing transformed plants will allow new opportunities for lentil breeding to produce improved cultivars.

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Authors and Affiliations

  1. Department of Biological Sciences, University of Calgary, 2500 University Dr. N.W., T2N1N4, Calgary, AB, Canada

    Anju Gulati

  2. Crop Development Centre, University of Saskatchewan, 51 Campus Drive, S7N5A8, Saskatoon, SK, Canada

    Pat Schryer

  3. Department of Botany and Plant Sciences, College of Natural and Agricultural Sciences, University of California, Batchelor Hall 3110, 92521, Riverside, CA

    Alan McHughen

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  1. Anju Gulati
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  2. Pat Schryer
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  3. Alan McHughen
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Correspondence to Pat Schryer.

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Gulati, A., Schryer, P. & McHughen, A. Production of fertile transgenic lentil (Lens culinaris Medik) plants using particle bombardment. In Vitro Cell.Dev.Biol.-Plant 38, 316–324 (2002). https://doi.org/10.1079/IVP2002303

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  • DOI: https://doi.org/10.1079/IVP2002303

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