بررسي روش مناسب توليد و كشت پروتوپلاست براي بازايي گياه كامل چغندرقند

Investigation on suitable sugar beet protoplast source and culture conditions to obtain regeneration

مهمترين جنبه هاي كاربردي كشت پروتوپلاست در چغندرقند شامل انتقال ژنهاي جديد و توليد هيبريدهاي غيرجنسي مي باشد. بررسي قابليت باززايي از كشت پروتوپلاست هدف تحقيق حاضر را در بر مي گيرد.گياهچه هاي كشت شده در شرايط استريل از گياهان مادري رقم 9597 تهيه شدند. دو روش توليد پروتوپلاست بكار گرفته شد : 1) جداسازي پروتوپلاست از بافت مزوفيل برگ 2)جداسازي از كشت سوسپانسيون سلولي. پس از خالص سازي پروتوپلاست ها، كشت آنها در اولين محيط غذايي (K8P) انجام پذيرفت. ميكروكالوس هاي حاصل به محيط هاي غذايي مختلف شامل MS, PGoB و محيط N6 حاوي هورمون هاي مختلف انتقال داده شدند. انتقال كالوس ها جهت جوانه زايي به محيط PGoB و N6 حاوي يك ميلي گرم در ليتر ژيبرليك اسيد (GA3) بنزيل آمينوپورين (BAP) منتقل شده بودند، توانايي توليد كالوس هاي باززا را از خود نشان دادند.

The most important applications of sugar beet protoplast culture are production of transgenic plants and somatic hybridization. Fortunately, to date many obstacles which were involved with sugar beet protoplast culture, have successfully been removed in Iran. The production of whole plant via protoplast culture as the most final step towards using this technique is the base of this investigation. The maternal plants were in vitro cultured seedlings of a monogerm sugar beet line (9597).Protoplast isolation was carried out by two methods L1) from leaf mesophyll tissue, (2) through cell suspension culture. Purified protoplosts were cultured in K8p medium as the first nutrient medium. The obtained microcalli were transferred onto several nutrient media : PGo, MS and N6 containing different phytohormones. The subsequent calli were transferred onto PGoB and N6 media for further development. Finally, the protoplasts which had been obtained from cell suspension culture and their subsequent microcalli and calli were transferred onto N6 medium containing I mg/l BAP and 1 mg/1 GA3, revealed to be regenerative.