A kind of stem segment induction plant regeneration medium and its tissue culture rapid propagation method
A technique of regeneration medium and rooting medium, which is applied in the field of regeneration medium for stem segment induction of Trichosanthes and its tissue culture rapid propagation, can solve the problems of low rooting rate and poor rooting quality of Trichosanthes tissue culture, and achieve low cost , high rooting rate and simple formula
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example 1
[0037] Example 1: Axillary bud induction test
[0038] 1. Test materials: The test materials are collected from the four-year-old Phytophthora seedlings, and the young stems that grow vigorously and are free from diseases and insect pests or the stems after hydroponic cultivation are selected as explant materials.
[0039] After washing the harvested stems of the hairy stems, they were sterilized with 75% alcohol for 30 s on the ultra-clean workbench, rinsed with sterile water for 3 times, and sterilized with 0.1% HgCl sterilant. 2 The solution is sterilized for 4 to 5 minutes. After sterilization, rinse with sterile water for 5 to 6 times, cut into 1.5-1.8 cm stem segments, and each stem segment has at least one axillary bud, ready for inoculation.
[0040] Experimental design: A single factor completely randomized design was adopted, with 20 bottles per treatment, 1 explant was inoculated in each bottle, and repeated 3 times.
[0041] 2. The period of explant collection: In...
example 2
[0044] Example 2: Test tube plantlet proliferation and culture test
[0045] 1. Combination of BA and IBA: The induced axillary buds were transferred to MS and improved MS basic medium, and different concentrations of BA and IBA were added. After 4 weeks, the proliferation rate and proliferation coefficient of the test-tube plantlets were counted, and the effect of the combination of BA and IBA was studied. Effects on proliferation of capillary in vitro plantlets.
[0046] 2. BA and IBA, GA 3 Combination: Transfer the induced axillary buds to MS and modified MS basic medium, and add different concentrations of BA, IBA, GA 3 After 4 weeks, the proliferation rate and proliferation coefficient of the test tube seedlings were counted, and different concentrations of BA, IBA, and GA were studied. 3 Effects of combinations on the proliferation of capillary in vitro plantlets.
[0047] 3. Culture conditions: the culture temperature is 25 ± 2°C, the light intensity is 2500 lx ~ 300...
example 3
[0048] Example 3: Test tube seedling rooting culture test
[0049] 1. Test material: use the test-tube seedlings with a height of > 2 cm as the test material, and transfer them to the rooting medium for rooting culture.
[0050] 2. Experimental design: 1 / 2MS and 1 / 4MS were used as the basic medium, and different concentrations of IBA or NAA (0.05, 0.1, 0.2, 0.3, 0.5, 1.0 mg·L -1 ), the basic medium without any auxin was used as the control. After four weeks, observe and record the rooting situation, count the rooting rate and the number of roots, and study the influence of NAA or IBA on the rooting of test-tube seedlings.
[0051] A single-factor completely randomized design was adopted, with 20 bottles per treatment and repeated 3 times.
[0052] 3. Culture conditions: the culture temperature is 25 ± 2°C, the light intensity is 2500 lx ~ 3000 lx, and the light time is 14 ~ 16 h / d; all media are added with 15 g L of sucrose -1 , agar 6.5 g·L -1 .
[0053] 4. Data statisti...
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