Efficient production of transgenic melon via Agrobacterium-mediated transformation.

Oriental melon (Cucumis melo L. var. makuwa) is an important fruit for human consumption. However, this plant species is one of the most recalcitrant to genetic transformation. The lack of an efficient in vitro system limits the development of a reproducible genetic transformation protocol for Oriental melon. In this study, an efficient transgenic production method for Agrobacterium-mediated transformation using cotyledon explants of Oriental melon was developed. Cotyledon explants were pre-cultivated for two days in the dark, and the optimal conditions for transformation of melon were determined to be a bacteria concentration of OD600 0.6, inoculation for 30 min, and two days of co-cultivation. Transgenic melon plants were produced from kanamycin-resistant shoots. A total of 11 independent transgenic plants were regenerated with a transformation efficiency of 0.8% of the inoculated explants. The transgenic plants were phenotypically normal and fully fertile, which might be a consequence of the co-cultivation time.

[1]  T. J. Fang,et al.  Transgenic lines of melon (Cucumis melo L. var. makuwa cv. ‘Silver Light’) expressing antifungal protein and chitinase genes exhibit enhanced resistance to fungal pathogens , 2013, Plant Cell, Tissue and Organ Culture (PCTOC).

[2]  Y. Chung,et al.  An efficient selection and regeneration protocol for Agrobacterium-mediated transformation of oriental melon (Cucumis melo L. var. makuwa) , 2012, Plant Cell, Tissue and Organ Culture (PCTOC).

[3]  B. Patil,et al.  Agrobacterium-mediated transformation and shoot regeneration in elite breeding lines of western shipper cantaloupe and honeydew melons (Cucumis melo L.) , 2011, Plant Cell Tissue and Organ Culture.

[4]  J. Aarrouf,et al.  Histological study of organogenesis in Cucumis melo L. after genetic transformation: why is it difficult to obtain transgenic plants? , 2011, Plant Cell Reports.

[5]  C. Chiang,et al.  Transgenic watermelon lines expressing the nucleocapsid gene of Watermelon silver mottle virus and the role of thiamine in reducing hyperhydricity in regenerated shoots , 2011, Plant Cell, Tissue and Organ Culture (PCTOC).

[6]  R. Khan,et al.  Stable integration and expression of wasabi defensin gene in “Egusi” melon (Colocynthis citrullus L.) confers resistance to Fusarium wilt and Alternaria leaf spot , 2010, Plant Cell Reports.

[7]  S. Yeh,et al.  Generation of transgenic oriental melon resistant to Zucchini yellow mosaic virus by an improved cotyledon-cutting method , 2009, Plant Cell Reports.

[8]  D. Cantliffe,et al.  Melon Fruits : Genetic Diversity , Physiology , and Biotechnology Features , 2008 .

[9]  K. Hirschi,et al.  Agrobacterium-mediated transformation of bottle gourd (Lagenaria siceraria Standl.) , 2005, Plant Cell Reports.

[10]  H. Ezura,et al.  Efficient plant regeneration and Agrobacterium-mediated transformation via somatic embryogenesis in melon (Cucumis melo L.) , 2004 .

[11]  R. Grumet,et al.  Agrobacterium tumefaciens mediated transformation and regeneration of muskmelon plants , 1990, Plant Cell Reports.

[12]  M. Fuchs,et al.  Transgenic Melon and Squash Expressing Coat Protein Genes of Aphid-borne Viruses do not Assist the Spread of an Aphid Non- transmissible Strain of Cucumber Mosaic Virus in the Field , 2004, Transgenic Research.

[13]  R. Serrano,et al.  Transfer of the yeast salt tolerance gene HAL1 to Cucumis melo L. cultivars and in vitro evaluation of salt tolerance , 2004, Transgenic Research.

[14]  J. M. Lasa,et al.  Agrobacterium-mediated transformation of commercial melon (Cucumis melo L., cv. Amarillo Oro) , 2004, Plant Cell Reports.

[15]  D. Kenigsbuch,et al.  A melon genotype with superior competence for regeneration and transformation , 2003 .

[16]  Jorge Adolfo Silva,et al.  MELON REGENERATION AND TRANSFORMATION USING AN APPLE ACC OXIDASE ANTISENSE GENE , 2001 .

[17]  J. Pech,et al.  A reliable system for the transformation of cantaloupe charentais melon (Cucumis melo L. var. cantalupensis) leading to a majority of diploid regenerants , 2000 .

[18]  M. Szwacka,et al.  Transformation of cucumber Cucumis sativus L. , 1997 .

[19]  J. Pech,et al.  Expression of ACC oxidase antisense gene inhibits ripening of cantaloupe melon fruits , 1996, Nature Biotechnology.

[20]  N. Chua,et al.  Transformation of Melon (Cucumis melo L.) and Expression from the Cauliflower Mosaic Virus 35S Promoter in Transgenic Melon Plants , 1991, Bio/Technology.

[21]  M. Ziv Vitrification: morphological and physiological disorders of in vitro plants , 1991 .

[22]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .