Levels of phytoene and β-carotene in transgenic honeydew melon (Cucumis melo L. inodorus)

Abstract

Using a previously reported optimized Agrobacterium tumefaciens-mediated transformation protocol, a watermelon phytoene synthase-C (PSY-C) gene was introduced into the elite honeydew breeding line ‘150’. Putative transformants were selected on kanamycin-containing medium (from 150 to 50 mg·l−1) and presence of the transgene was confirmed using polymerase chain reaction (PCR). Moreover, Southern blot analysis confirmed integration of the transgene and revealed presence of one to two copies of the PSY-C transgene in the different transgenic lines. In addition, reverse transcription (RT)-PCR analysis revealed transcript levels of the transgene in different tissues of these transgenic lines. Using flow cytometric analysis, it was found that all T0 transgenic plants were tetraploid. Moreover, altered phenotypes (color change) were observed for rind tissues of transgenic lines. Based on high performance liquid chromatography (HPLC), β-carotene content and phytoene accumulation in fruit of transgenic lines were 32-fold and 11 μg·g−1 FW higher than the levels found in the control plants.

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Abbreviations

PCR:

Polymerase chain reaction

HPLC:

High performance liquid chromatography

PG:

Polygalacturonase

PSY:

Phytoene synthase

T0 :

Regenerated transgenic plant from the original inoculated explant

T1 :

The progeny of the original T0 transgenic plants

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Acknowledgments

This research was supported by the USDA-CSREES (2008-34402-19195, 2009-34402-19831) and USDA-NIFA (2010-34402-20875), “Designing Foods for Health” grants, through the Vegetable and Fruit Improvement Center, Texas AgriLife Research. We wish to thank Drs. Jim Giovannoni and Li Li at Cornell University for providing binary vectors and technical advice. We also appreciate Dr. Yong Hun Chi’s help in providing E. coli for gene construction and technical advice.

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Correspondence to Kevin M. Crosby.

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Ren, Y., Bang, H., Lee, E.J. et al. Levels of phytoene and β-carotene in transgenic honeydew melon (Cucumis melo L. inodorus). Plant Cell Tiss Organ Cult 113, 291–301 (2013). https://doi.org/10.1007/s11240-012-0269-8

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Keywords

  • Honeydew (Cucumis melo L. inodorus)
  • Phytoene synthase
  • Polygalacturonase
  • Phytoene
  • β-Carotene
  • Tetraploid
  • Flesh color