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Production of Genetically Modified Grape (Vitis vinifera L.) Plants
| Content Provider | Semantic Scholar |
|---|---|
| Author | Soliman, Hemaid Ibrahim Ahemaidan |
| Copyright Year | 2018 |
| 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 vacuolartype 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 mgL1NAA 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 AgNHX1and 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. Keyword—Grape (Vitis vinifera L.), regeneration, transformation, Na+/H+ antiporter, PCR and northern blot hybridization. Abbreviations—2,4-D2,4-di-chloro-phenoxy-acetic-acid; nptII geneneomycin phosphotransferase(gene); BAPBenzylaminopurine; NAAα-Naphthalene Acetic Acid; LB Luria broth medium; KinKinetin;CTABCetyl-trimethylammonium-bromide; NN Nitsch and Nitsch medium; IBA Indole Butyric Acid; ODOptical density; PCRPolymerase chain reaction; RT-PCRReverse transcriptase polymerase chain reaction. |
| Starting Page | 111 |
| Ending Page | 120 |
| Page Count | 10 |
| File Format | PDF HTM / HTML |
| DOI | 10.22161/ijhaf.2.4.1 |
| Volume Number | 2 |
| Alternate Webpage(s) | http://aipublications.com/uploads/issue_files/1-IJHAF-JUL-2018-2-ProductionofGenetically.pdf |
| Alternate Webpage(s) | https://doi.org/10.22161/ijhaf.2.4.1 |
| Language | English |
| Access Restriction | Open |
| Content Type | Text |
| Resource Type | Article |
