Application of cryopreservation techniques for preservation of potato (Solanum tuberosum) gerplasms
2007
Zarghami, Reza | Najafi , Pouya | Dabir Ashrafi, Omid
Cryopreservation technology is highly importance for long term conservation of plant germoplasms. is. This technology is useful to conserve plant materials on condition that no genetical, morphological and physiological variations occurs in plant. Potato tubers were tipped and treated by thermotherapy for 4 weeks. Meristems were removed and placed in a liquid medium on a paper bridge. ELIZA test indicated that the resulted plantlets are healthy. The purpose of this project was to determine the best culture medium for high plantlet and rooting regeneration. Hormone-free MS was identified to be the best medium. To multiply healthy plantlets, plantlets were first cut into different segments, each having one bud and a total of 200 single nodes were placed on 4 culture media with different concentrations of plant growth regulators. The results indicated that MS medium with 0.25 mg/l of gibberlice acid, 0.1 mg/l NAA and 2 mg/l of calcium pantothenate was the best medium for shooting and hormone-free medium was the best for rooting. The results also showed that the best density in 250 ml flasks was 6 to 9 explants of single nodes. Two technique of preculture –dehydration and vitrification were studied for freezing. The results showed that the response of various varieties to the applied methods were different and adding abscisic acid and proline increased the regeneration rate after freezing. Aluminum foil was more suitable for speeding up the stages of freezing and refreezing compared to plastic container. The best concentrations for glycerol and sorbitol for vitrification was 7 molar. AFLP molecular markers, Flow cytometry and cytological studies were used to investigate genetic stability. Twenty- three descriptors were used to study morphological stability and free proline, soluble carbohydrates, soluble proteins and a and b chlorophyll were measured for physiological stability which all of them confirmed the lack of genetical, morphological or physiological variation.
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