Transgenic melon (cucumis melo L.) and potential for expression of novel proteins important to food industry

Abstract

Melon (Cucumis melo L.) is an important fruit crop cultivated widely in every region of the world. Our laboratory is targeting this species for production of novel proteins important to food industry. Prior to expression of protein of interest in transgenic melon an efficient genetic transformation system has to be developed. In this context we are testing a wide variety of promoters fused to reporter gene for β‐glucuronidase (GUS) for expression specifically in melon fruits. In this study in melon, salicylic acid‐inducible promoter region of pathogenesis‐related protein gene (PR1a) of tobacco fused to β‐glucuronidase (GUS) gene was introduced into melon via Agrobacterium‐mediated gene transfer using a binary vector system. Gene transfer was effective when Agrobacterium virulence factors like acetosyringone (100 μM) and low pH (5.2) were provided during the co‐culture step. Transformed shoots were recovered from benzyladenine‐induced cut cotyledons using kanamycin gene as a selective marker. Regeneration of shoots from cotyledons was stimulated by providing 10 mM proline in the shoot organogenesis medium. Southern and Northern blot analysis of transformants confirmed the presence of β‐glucuronidase gene in two selected clones J‐3 and PR‐G. The transformants also showed high β‐glucuronidase activity after salicylic acid treatment. Thiamine, a previously known inducer of pathogenesis‐related protein, stimulated β‐glucuronidase in J‐3 but not PR‐G melon transformants tested in this study. These studies showed that tobacco PR1a promoter region can be expressed in melon and it was stimulated by salicylic acid. This indicates the potential to use the promoter region of tobacco PR1a for genetic improvement of melon for specific food processing‐related characteristics or for expression of novel food‐related proteins. The promoter region could be used to drive specific target genes under stress or salicylic acid induced conditions.