الفهرس 
Plant material speciesOnly 14 pages are availabe for public view
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Abstract
Molecular Studies on Salinity and Drought Stress Gene (s) in Some Natural Plants” This work was carried out to isolate abiotic stress related genes (salinity and drought stress) from some desert plants. Six plants were selected to characterize and study the vacuolar Na+/H+ antiporter gene in them. To achieve this work it was essential to establish a suitable and more efficient technique to isolate total RNA with high quality yield from these plants with high level of phenolic compound, polysaccharides and secondary metabolites. The results are summarized in the following points: 1. Plants materials were collected from North West Coast of Marsa Matruh, Egypt and stored at - C such as: Group I. (plant species from family of Amaranthaceae): Atriplex halimus (salt bush), Suaeda pruinosa (Suwaid) and Salsola vermiculata (El-rawsa). Group II. (Plant species from different families): Capparis orientalis (Lasaf/Kabbar), Lycium shawii (Awsaj), and Zygophyllum album L. (El-retrat). 2. TissueLyser II machine with liquid nitrogen (LN2) was used as a fast suitable method for homogenization to get fine powder, avoid browning and RNA degradation during extraction. 3. Two modified methods were investigated: the first one based on non Kit using a commercial reagent (TRIzol® Reagent) with RNeasy Plant Mini Kit for RNA cleanup and the second one is the RNeasy Plant Mini Kit plus polyethylene glycol (PEG-6000) for RNA isolation to identify the most suitable for cDNA application. 4. High quality, quantity and yield of total RNA were evaluated by absorbance at 260 nm (A260/A230 and A260/A280 ratios) using
Summary
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NanoDROP spectrophotometer (NanoDrop, Technologies Inc.)., electrophoreses on 1.2% agarose gel, and reverse transcription RT-PCR (one step RT-PCR) to amplify 1 Kb of 18S ribosomal RNA were found. The two evaluated methods proved to be successful for obtaining total RNA from all plants under this study with high quality and quantity. 5. Oligo (dT)20 and M-MLV RT (SuperScript III Reverse Transcriptase) was used for cDNA synthesis reaction. 6. One pair of oligonucleotide primer specific with touchdown PCR program (TD-PCR) was used to amplify a novel full length of ORF vacuolar Na+/H+ antiporter gene of plant species (Atriplex halimus (AhNHX1), and two parts (P1 & P2) from Suaeda pruinosa (SpNHX1) and Salsola vermiculata (SvNHX1) genes. 7. One pair of oligonucleotide primer degenerate with touchdown PCR program (TD-PCR) was used to amplification fragment about 600 bp in the middle regions of ORF vacuolar Na+/H+ antiporter gene of species (Atriplex halimus, Suaeda pruinosa, Salsola vermiculata, Capparis orientalis, Lycium shawii and Zygophyllum album. 8. Only touchdown PCR program (TD-PCR) of cDNAs succeed to gave high yield of amplify a Na+/H+ antiporter gene and increase specificity, sensitivity. 9. In group I: Sequence analysis showed that 1668 bp (AhNHX1), (KJ452341.1) full length of ORF represent fragment for Atriplex halimus, 1662 bp (SpNHX1), (KJ452342.1) two partial fragment of ORF was represent for Suaeda pruinosa, while two partial fragment were obtained, about 797 bp (SvNHX1), (KJ452343.1), and 603 bp (SvNHX1), (KJ452344.1) was from Salsola verimiculata.