الفهرس 
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Abstract
The present study aimed best of all at developing reference biochemical and molecular fingerprints for the plant material to be used in regeneration and transformation experiment and at finding some biochemical or molecular marker at the biochemical and molecular level which might be used to identify faba bean cultivars resistant and susceptible to Botrytis fabae. In these experiments, nine cultivars, namely Giza 3, Giza 716, Sakha 1, Sakha 2, Nubariya 1 (resistance), Giza 843, Egypt 1 (moderate resistance) and Giza 40, Giza 429 (susceptible) SDS-PAGE and RAPD-PCR were analyzed.
Different regeneration systems (i.e. de novo regeneration system and direct shoot organogenesis) were tested with different Egyptian Vicia faba cultivars. The success of these experiments was the easy of an efficient regeneration protocol by means of direct shoot organogenesis.
Indirect regeneration system from etiolated epicotyl explants of some Egyptian faba bean cultivars forming calli ranged from 100% (for Egypt 1) and 81.9% (for Giza 716). The color of the proliferating callus ranged from creamy brown colour with high frequency (++), brown and green with high frequency (++) and brown colour with low frequency. It could be concluded that epicotyls explants of etiolated petioles proliferated callus but none of them succeeded in forming shoots.
The feasibility of the direct regeneration strategy developed in this work was initially evaluated by monitoring the number of regenerated shoots from embryo axis explants. It was observed that around 4-5 shoots regenerated from each explant. In this experiment the eight investigated faba bean cultivars can be ranked according to their response for direct organogenesis as follows (from high to low): Giza 843, Nubariya 1, Sakha 1, Sakha 2, Egypt 1, Egypt 2, Giza 716 and Giza 3.
The lethal concentration experiment was determined the selection effect of the herbicide PPT on the regeneration potential of embryo axes of faba bean (Vicia faba L.) cultivars. We found that, the lethal concentration of Giza 843 and Sakha 1is 1.5 mg/l; and for Giza 716, Giza 3 and Sakha 2 is 2 mg/l.
The present study was carried out with the objectives to optimize the transformation protocol for faba bean with gus (β-glucuronidase) as a reporter gene using Agrobacterium-mediated transformation. The bacterial density used for infection was critical. The highest densities tested (OD600 1-1.5) as well as prolonged infection (1/2 hr) gave the best results, i.e. number of GUS spots per explants. Although the Agrobacterium cell density OD600 1.0 showed the highest frequency of transient GUS expression.
The successful of these experiments is an efficient regeneration protocol by means of direct shoot regeneration then we use it for Agrobacterium transformation of meristematic cells (mature embryo axes).
The transformation experiments were performed using either EHA 105/pGIIvst carrying the Chit gene under a vst promoter and the selectable marker gene bar.
Nine transgenic clones were recovered from 3 different Faba bean cultivars (Sakha 2, Giza716 and Giza 843) carrying the Chit and bar genes. The proliferated shoot on selective medium were grafted under in vitro condition and finally transferred to the greenhouse.
The detection of the transgenes (Chitinase and bar genes) was studied by means of PCR analysis. In most of the recovered transgenic clones the foreign genes were active as predicted from the expression of the selectable marker bar gene (leaf paint test).