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Transformation of alfalfa chloroplasts and expression of green fluorescent protein in a forage crop

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Abstract

The ability to transform chloroplasts in multiple species is important for improving agricultural traits. Chloroplast transformation of alfalfa (Medicago sativa L.), a useful forage plant with high market value, was achieved using a vector carrying aadA and gfp genes being introduced into the chloroplasts of alfalfa via particle bombardment using leaves and calli as explants. Resistant somatic embryos were generated and developed into plantlets from explants. The transformation efficiency was 1.3% for callus explants and 2.7% for leaf explants. PCR and Southern blotting analyses revealed that the foreign genes were integrated into the transformed chloroplast genome. The occurrence of GFP was further confirmed by fluorescence microscopy. Expression of foreign genes in alfalfa chloroplasts is therefore possible, and provides a novel means for genetic improvement of agronomically important traits and production of value-added proteins.

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Acknowledgment

This study was supported by Science and Technology Developmental Program (No. 20076021) and Postdoctoral Grant of Jilin Province.

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Authors and Affiliations

  1. Biotechnology Research Centre, Jilin Academy of Agricultural Sciences, 1363, Caiyu Street, Changchun, 130033, People’s Republic of China

    Zhengyi Wei, Yanzhi Liu, Chunjing Lin, Yunpeng Wang, Qin’an Cai, Yingshan Dong & Shaochen Xing

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  1. Zhengyi Wei
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  2. Yanzhi Liu
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  3. Chunjing Lin
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  4. Yunpeng Wang
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  5. Qin’an Cai
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  6. Yingshan Dong
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  7. Shaochen Xing
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Correspondence to Shaochen Xing.

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Zhengyi Wei and Yanzhi Liu contributed equally to this work.

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Wei, Z., Liu, Y., Lin, C. et al. Transformation of alfalfa chloroplasts and expression of green fluorescent protein in a forage crop. Biotechnol Lett 33, 2487–2494 (2011). https://doi.org/10.1007/s10529-011-0709-2

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  • DOI: https://doi.org/10.1007/s10529-011-0709-2

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