IJHAF | Production of Genetically Modified Grape (Vitis vinifera L.) Plants

Indexing and Abstracting

Production of Genetically Modified Grape (Vitis vinifera L.) Plants

Hemaid Ibrahim Ahemaidan Soliman

International Journal of Horticulture, Agriculture and Food science(IJHAF), Vol-2,Issue-4, July - August 2018, Pages 111-120, 10.22161/ijhaf.2.4.1

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Abstract:

Grape (Vitis vinifera L.) is one of the most economically important fruits in the world. High salinity stress adversely affects plant growth and limits agricultural production worldwide. This study describes a successful method of somatic embryogenesis using in vitro-derived leaf explants and introduction of a vacuolar-type Na+/H+ antiporter gene from a halophytic plant, Atriplex gmelini (AgNHX1) confers salt tolerance to grape cv. Superior Seedless using the Agrobacterium-mediated transformation. Callus embryogenic was induced on NN medium 2.0 mgL-1 2,4-D, 0.5 mgL-1 BAP and 0.5 mgL-1 NAA. Subsequent subculture of callus on NN medium containing 1.5 mgL-1 BAP, 0.5 mgL-1kinetin and 0.5 mgL-1NAA induced shoot organogenesis after eight weeks of culture. The leaf explants were co-cultivated with Agrobacterium strain LBA4404 harbouring the binary vector pBI121 which contained the AgNHX1 and nptII genes and putative transgenic plants were produced. The presence and stable integration of AgNHX1 gene in transgenic plants was conﬁrmed by PCR and northern blot hybridization. The transgenic grape plants overexpressing the AgNHX1 gene showed a strong tolerance to salt stress under 250 mM NaCl, whereas non-transgenic plants died under the same conditions. Salt tolerance assays followed by salt treatments showed that the transgenic plants overexpressing AgNHX1 could survive under conditions of 250 mM NaCl for 4 weeks while the non-transgenic plants died under the same conditions. These results indicate that overexpression of the Na+/H+ antiporter gene in grape plants significantly improves their salt tolerance.

Keywords:

Grape (Vitis vinifera L.), regeneration, transformation, Na+/H+ antiporter, PCR and northern blot hybridization.

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