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The Studies on Cryopreservation of Huaiqing Chrysanthemum (Endranthema Morifolium) Germplasm by Vitrification

Author: SongPingPing
Tutor: ZhaoXiTing
School: Henan Normal
Course: Botany
Keywords: Huai Chrysanthemum Germplasm Resources Cryopreservation By vitrification Relative survival Survival rate
CLC: S567.239
Type: Master's thesis
Year: 2010
Downloads: 50
Quote: 1
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Huai Chrysanthemum [Huaiqing chrysanthemum (Dendranthema morifolium) is one of the four pregnant drug is mainly produced in Henan Wenxian, Wu Zhi, fraternity (in ancient Huaiqing House under the jurisdiction of, hence the name), both ornamental and medicinal , Sibu three functions. But pregnant chrysanthemum germplasm resources still use the Daejeon planting saved a single way, together with its own characteristics, chrysanthemum is a cross-pollination of maize so that it can not guarantee purity, for a long time hybrid germplasm precious germplasm resources serious loss. Cryopreservation technique developed in the late 20th century, is the only viable long-term stability to the preservation of plant germplasm resources a good way, it can save space and avoid Following generation, pests and diseases and climate factors caused germplasm lost, and the device is simple, the operation simple. Based on this, we carry out the pregnant daisy vitrification method germplasm cryopreservation technology. Made the following findings: 1. Huai Chrysanthemum rapid propagation technology system. Huai Chrysanthemum Seedling occurrence of type two ways by the axillary branch type and buds for breeding, the axillary branches the best rapid propagation medium MS 6-BA 0.2 mg · L -1 of NAA 0.2 mg · L -1 ; multi-bud type (stems with buds and leaves) the best medium for the MS 6-BA 1 mg · L -1 NAA 2 mg · L -1 ; optimal medium for rooting MS Effect of PP333 0.5 mg · L -1 . 2. Establish a cryopreservation pregnant chrysanthemum germplasm resources by vitrification technology system. Following 40 d cultured plantlets exercise under 4 ℃ for 7 d under sterile conditions, cut the stems with buds about 0.5 cm, using MS medium supplemented with Glu 0.24 mol · L -1 Gly 0.4 mol · L -1 as a pre-culture medium at 4 ℃ constant 3 d, then the material with 60% PVS2 do the loading liquid loading at room temperature for 20 min, and then with modified The PVS2 (Gly 30% EG 15% DMSO 10%) 0 ° C dehydration 60 min, and quickly put into liquid nitrogen storage; immediately stored in liquid nitrogen for at least 1 d after the material is placed in a 40 ° C water bath to quickly thaw 1 to 3 min after the thaw in MS basic medium supplemented with 1.0 mol · L -1 and 1.2 mol · L -1 glucose gradient washing, every 10 min, then washed into the regeneration medium (with rapid propagation medium), incubated in the dark to start the formation of callus 15 d 30 d Go light culture, cultured callus began to turn green to 7 d to 25 d ~ 30 d when the buds begin to form and grow into normal plants 60 days after, the survival rate of up to 80% or more. 3. Saved regenerated seedlings stability analysis. Compared with the room temperature Miao, the vitrification cryopreservation pregnant chrysanthemum regenerated seedlings average internode length, plant height and number of leaves added value were no significant differences; blades soluble sugar, soluble protein and chlorophyll content were also no significant differences ; the relative conductivity of the regenerated plants leaves, superoxide anion and MDA content significantly reduce SOD and POD increased vitality isozymes analysis showed both POD, SOD zymography showed a high degree of consistency, but individual band width and brightness differences. Described above, Huai Chrysanthemum established vitrification the cryopreservation technology system, not only to maintain the stability of the morphological, physiological and zymography, but also enhance the ability of its anti-lipid peroxidation.

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CLC: > Agricultural Sciences > Crop > Economic crops > Medicinal crops > Herbal > Perennial > Other
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