Gene transfer utilizing pollen-tubes of Albuca nelsonii and Tulbaghia violacea

Aloka Kumari,P Baskaran ,J Van Staden

doi:10.1590/1984-70332017v17n3a35

Aloka Kumari, P Baskaran + Show 1 more

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https://doi.org/10.1590/1984-70332017v17n3a35

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Abstract

Abstract Developing a tissue culture-independent genetic transformation system would be an interesting technique for gene transfer in valuable medicinal and horticultural plants. Efficient gene delivery (Agrobacterium tumefaciens strain LBA 4404: harbouring PBI121 plasmid) was achieved with Km-resistant pollen grains (pollen tube technique) and were found to be GUS-positive for Albuca nelsonii (31.3%) and Tulbaghia violacea (32.6%). The Km-resistance (95.6% for A. nelsonii and 86.7% for T. violacea) and GUS-positive (100% for A. nelsonii and 97.5% for T. violacea) putative transgenic seedlings in vitro were obtained with 200 mg L-1 Km. The in vitro plants were obtained from leaf explants of putative transgenic seedlings and were confirmed to be Km-resistant and GUS-positive (T. violacea, 73.7% and A. nelsonii, 80.5%). The plants were successfully acclimatized in the greenhouse. We describe a tissue culture-independent gene transfer technique with high efficiency clonal transgenic plant production for A. nelsonii and T. violacea. This can also be applied to biotechnological crop improvement of the same species and potentially to other plants.

Highlights

Tulbaghia violacea (Alliaceae), commonly known as wild garlic, is cultivated for both ornamental and medicinal use (Reinten et al 2011)
Gene delivery by pollen transformation Pollen grains were inoculated on BWK medium with different concentrations of Km to investigate the optimal concentration of Km for transformation studies
Pollen tube growth was completely inhibited at 200 mg L-1 Km for T. violacea and 600 mg L-1 Km for A. nelsonii except the controls (Figure 1A, B, C, D). This would allow for efficient gene delivery via the pollen tube pathway

Summary

Introduction

Tulbaghia violacea (Alliaceae), commonly known as wild garlic, is cultivated for both ornamental and medicinal use (Reinten et al 2011). It is a clumpforming perennial with narrow leaves, and produces a large cluster of fragrant, violet flowers from mid-summer to autumn (Harvey 1837). Albuca nelsonii (Hyacinthaceae) is a perennial ornamental and traditional medicinal plant, which is commonly known as Nelson’s slime lily (Ascough and Van Staden 2010). The medicinal and ornamental value of T. violacea and A. nelsonii suggests their use in studies of pollen transformation as bulbous model plants to obtain a higher efficacy of gene transfer

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