الفهرس 
REVIEW OF LITERATUREOnly 14 pages are availabe for public view
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Abstract
SUMMARY
This investigation was carried out at the Agriculture Genetic Engineering Research Institute (AGERI), ARC, Giza-Egypt and the Department of Genetics, Faculty of Agriculture, Ain Shams University, during the period 2012 - 2017. Two lines and one cultivar of potato (Solanum tuberosum L) were used in this study.
Potato is the plant belonging to Solanaceae species, a tuberous crop with economic value and nutritional, infected with many diseases, including fungal and bacterial diseases and viral.
The most important fungal disease is early blight, which is caused by Alternaria solani. This fungus spends a large proportion of potatoes for injury to plant the crop at all stages of growth and tubers during storage.
Through the Agricultural Genetic Engineering Applications can be obtained on the plants transgenic resistance to this fungus by genetic transfer of chitinase gene inside the potato plants to the ability of this enzyme to analyze chitin of cell wall .Through this analytical influence fungus cannot complete its life cycle and thus It can get rid of the pathogen effect of this fungus.
During this study, genetic modification potato plants PVY 5 and PVY 15 lines and cultivar Desriee through genetic transfer the chitinase gene using Agrobacterium LBA4404 line containing plasmid pRI 201-AN which carries chitinase gene to be transferred to potato leaves which used in the study, the genetic transformation protocol was applied on Desriee, PVY 5 and PVY 15 leaves. After successive growth stages of transformed leaves and access to developed a complete shoot is conducting confirmatory testing by polymerase chain reaction to make sure shoots are transgenic and resistance to early blight disease.
The performed steps can be summarized which used to produce genetically modified plants as follows:
1. Design primers to add sequence of NdeI and SalI restriction enzymes to chitinase gene by polymerase chain reaction technique.
2. Gene transfer inside pRI 201-AN plasmid by cutting and pasting steps, then transformed into Agrobacterium LBA 4404.
3. Leaves of PVY 5 & PVY 15 lines and Desriee cultivar were transformed by Agrobacterium.
4. After completing bacterial infection steps, the leaves were incubated on callus induction media containing 5.0 mg 2-4, D / L for 5-10 days in the dark, then transferred to regeneration media containing 1.0 mg Indole Acidic Acid /L and 1.0 mg Benzyl Adenine/ L and 10 mg Giberillic Acid /L for 72 days in the lighting, after regeneration and elongation steps; shoots were calculated and analysis to compare the strains.
5. Shoots were tested by extracting DNA for each shoot and tested by polymerase chain reaction technique to identify genetically modified shoots.
The obtained results could be summarized as follow:
1- After 3 weeks of transformation steps to leaves of PVY5 and PVY15 lines and Desiree cultivar by LBA4404 Agrobacterium which contained CHI gene cloned in pRI 201-AN plasmid, The callus induction results were a 53.3%, 77.1% and 100% respectively.
2- After 9-10 weeks, regeneration results of PVY5 and PVY15 lines and Desiree cultivar were 71.4%, 73.5% and 96.6% respectively.
3- After 72 days, The screening result of the transgenic plants of Desriee cultivar by using CHI primer was 5 transgenic plants at expected size was 798 bp.
4- The result of detection 5 transgenic plants of Desriee cultivar by using kanamycine primer was at expected size 650 bp.
5- The screening result of the transgenic shoots of PVY5 line by using CHI primer was one transgenic shoot at expected size was 798 bp.
6- The result of detection one transgenic shoot of PVY5 line by using kanamycine primer was at expected size 650 bp.
7- The screening result of the transgenic shoots of PVY15 line by using CHI primer was one transgenic shoot at expected size was 798 bp.
8- The result of detection one transgenic shoot of PVY15 line by using kanamycine primer was at expected size 650 bp.