| Summary: | 碩士 === 中興大學 === 生命科學系所 === 99 === The test material of this study is garlic (Allium sativum L.), and the purpose is to continue the previous studies on the protocols of the cryopreservation of garlic by vitrification. Other purpose is to explore the tissue culture protocol of garlic to increase the regrowth, proliferation, and the survival rate of garlic after cryopreservation and rewarming. Development suitable protocol for in vitro garlic culture helps for subsequent field planting to expand the use of plant genetic resource.
Cryprotectants at different pH treatments showed using PVS2 for cryopreservation, treated with LS for 60 minutes, PVS2 for 100 minutes, and unloading solution for 20 minutes, the results of treatment of PVS2 with pH 6.2 was the best. The survival rate and regeneration rate after cryopreservation were 70.8% and 40.0%, respectively; using PVS3 as cryoprotectant for cryopreservation, treated with LS for 50 minutes, PVS3 for 150 miuntes, and unloading solution for 40 minutes, pH 6.6 resulted in the best survival rate and regrowth rate of PVS3 cryopreservation. The survival rate and regeneration rate were 62.5% and 41.1%. The recovery medium test showed that MS + IAA 0.03 mg L-1 recovery medium had the best effect on the cryopreservation, treated with LS 60 for minutes, PVS2 for 100 minutes, unloading solution for 20 minutes. After cryopreservation, the survival rate and regrowth rate were 70.8% and 30.8%. For PVS3 cryopreservation, treated with LS for 50 minutes, PVS3 for150 minutes, unloading solution for 40 minutes, MS + kinetin 0.1 mg L-1 + IAA 0.03 mg L-1 recovery medium was the best, the survival rate after cryopreservation and the regrowth rate after cryopreservation were 85.0% and 26.9%. The cryopreservation of different varieties of garlic with PVS2 and PVS3 treatments had different results, some varieties suitable for PVS2, some for PVS3, and some varieties suitable for both. Most of the different cultivated garlic species’s survival rates were more than 50% and the regrowth rates were more than 30%. This result shows that the cryopreservation method has very high applicability.
Experimental results showed that for the purpose to use tissue culture of garlic to form bulbs, 0.4 M sucrose GMG media (MS + BA 0.5 mg L-1 + NAA 0.1 mg L-1) was the best. It had low hyperhydric rate (6.7%), high bulb formation rate (93.3%) and the highest proliferation rate and proliferation number per plant, with the value 53.3% and 2.07. It is the most suitable method. For normal tissue culture of garlic, the 0.1 M sucrose GMG media with 1 cm2 ventilation treatment had a higher rooting rate (66.7%) and no any hyperhydric rate (0%). It’s proliferation rate and the average proliferation number of per plant were 73.3% and 2.47. The result of the garlic rooting experiment showed that the medium, MS + IBA 1 mg L-1 has the best effect on rooting rate (86.7%).
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