الفهرس 
INTRODUCTIONOnly 14 pages are availabe for public view
 |  | from | 90 |  |  |
| | | from | 90 | | |
Abstract
The experiments of the present study were carried out on three potato cultivars (Spunta, Diamont and Desiree) were performed during 2012-2016 at the agricultural genetic engineering research institute (AGERI).Two different types of explants (internode and leaf) were cultured for induction and plant regeneration. Then cloning the glucanase gene in pRI 201-AN plasmid which carrying kanamycin resistance (nptII) gene was perfomed and followed by transformation in agrobacterium tumefaciens strain LBA4404 which used for plant transfection. Nucleotide and amino acid sequences of transformed agrobacterium were analyzed. The putatively transgenic plants were confirmed using polymerase chain reaction (PCR) for genomic and cDNA.
The obtained results could be summarized as follow:
1- The explants were cultured onto MS media supplemented with compound of media 2.4-Dichlorophenoxy acetic acid(2.4-D).The highest value of Desiree leaf explants was 80% induced from develop callus in 3mg/l 2.4.D media.
2- When Data were analyzed after seven weeks of culture and analysis showed that the explants induction to develop shoot with culture media formulations and potato genotype. The best value of maximum shoot regeneration was 90.3 for Sponta in NAA and 4mg/L KIN media form internode and 88.4 for Desiree in BA, NAA and GA3 media from leaf proved to be more effective.
Potato transformation using Agrobacterium tumefaciens
After cloning the glucanase gene in pRI 201-AN plasmid followed by transformation in agrobacterium tumefaciens strain LBA4404 which used for plant transfection. Nucleotide and amino acid sequences of transformed agrobacterium were analyzed. The leaf potato of Desiree cultivars were transformed by agrobacterium tumefaciens. Harboring the pRI201-AN plasmid after exposure to 5 min the explant transferred into MS media.
During selection process successfully transformed calli continued to grow vigorously to produce shoot initiations, whereas the non-transformed ones failed to form shoot and eventually bleached and became necrotic within 3 weeks. Transformation efficiency was calculated as No. of shoots analysis by The SAS System, The GLM. Duncan’s Multiple Range Test for shoot proves the number of plants that show positive PCR and Rt –PCR result.