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Sadat City University (2015)

Molecular Studies on Salinity and Drought Stress Gene (s) in Some Natural Plant

Ahmed, Mohamed Zoelfakar Sayed

Titre : Molecular Studies on Salinity and Drought Stress Gene (s) in Some Natural Plant

Auteur : Ahmed, Mohamed Zoelfakar Sayed.

Etablissement de soutenance : Sadat City University

Grade : Doctor of Philosophy in Gentic Engineering and biotechnology 2015

Résumé partiel
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) usingNanoDROP 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.

Présentation étendue (EULC)

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