| Rehmannia glutinosa Libosch. belongs to Scrophulariaceae, was one kind of famous medicinal materials, and has high medical value. It used the root, stem, petiole, and leaf of good Rehmannia glutinosa Libosch.variety "Bei jing 5"planted in shaanxi, studied the tissue culture technology of Rehmannia glutinosa Libosch., established the rapid propagation and plant regeneration systems, investigated the structure of leaf of its normal and hyperhydric shoot, and studied the occurrence and their control methods of hyperhydric shoots. The main results were as follows:1.Using the root of Rehmannia glutinosa Libosch. as materials to study the rapid propagation system, the main results showed:(1) It can lower the pollution ratio, improve the ratio of germination after the root pasteurized with 75% alcohol from25 to 30s and 0.1%Hgcl2 from 17 to 20min. The shoots were cut down from the bottom when they reached 0.5-1cm, and transferred to the MS medium supplemented with BA 0.5mg/L, NAA 0.1mg/L, the shoots had better growth ability. The test-tube seedlings were well after culture for 5-6 weeks.(2) The test-tube seedlings were cut into several stems with axillary buds and cultured on MS medium supplemented with BA 0.5 mg/L, NAA 0.1mg/L, on which a few buds may multiplied; and the MS medium supplemented with BA 0.5 mg/L, NAA 0.1mg/L was most suitable for buds multiplication, on which the buds multiplication ratio was about 6.5 than 4-5 weeks later. According to the result, the rapid propagation system of Rehmannia glutinosa Libosch. was set up, and a lot of test-tube seedlings can be obtained. 2. With different explants including stem, petiole, leaf of the test-tube seedlings, the research established the plant regeneration system of Rehmannia glutinosa Libosch., the main results showed:(1)When the stem and the petiole were cultured on MS medium supplemented with BA 1.0 mg/L, NAA 0.5 mg/L, the induction ratio of callus was 100% and 95.6% respectly. The differentiation ratio of callus was about 41.7% and the callus formed adventitious buds when they were cultured on MS medium supplemented with BA 3.0 mg/L for 1-2 subculture(2)A few emerald green or green callus were formed when the leaves were cultured on MS medium supplemented with BA 0.5mg/L, NAA 0.3mg/L, and the induction ratio of callus was 96%. Adventitious buds were induced from callus of leaf on MS medium supplemented with BA1.0 mg/L, NAA0.1 mg/L, and after 25 days of culture the differentiation ratio of adventitious buds from leaf callus was about 44.4%; while the leaves were cultured on MS medium with different combinations and concentration of 2,4-D/BA, 2,4-D/KT and NAA/2,4-D, the induction ratio of callus can reach 100%, and most of the callus was gray and yellow, the structure was loose and difficult to form adventitious buds, but it can easy form the root from the surface of callus.(3) Adventitious buds were regenerated directly from the leaf of Rehmannia glutinosa Libosch., the most suitable medium was MS medium supplemented with BA3.0 mg/L, NAA0.1 mg/L, and the differentiation ratio of adventitious buds was about 77.5%. The differentiation of adventitious buds from leaf was influenced by the orientation and the explant location, the differentiation ratio of adventitious buds can increase obviously when the abaxial of upper infant leaf was on the medium under illumination culture(4)The shoots regenerated from different explants that reached 1-2cm were detached and transferred to the modified MS medium supplemented with NAA0.05 mg/L for rooting, after 15 to 20 days of culture the differentiation ratio of root was about 100%. When the plantlets were transferred to the culture matrix compounded equally by the humus and the sand for 2-3 weeks of cultivation, the survival ratio of plantlets can reach 96.7%. 3. The structure of normal and hyperhydric plantlets was scanned by electron scanning microscopy, optics microscopy and transmission electron microscopy. The structure of the leaf for the normal plantlets was similar to that of plants found in th...
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