Medicinal Resources, tissue Culture and Rapid Propagation of Bletilla striata

Medicinal Resources, tissue Culture and Rapid Propagation of Bletilla striata

After tuber propagation, Baiji is easy to be infected by diseases, insect pests and viruses, resulting in weak growth, serious diseases and insect pests, reduced yield and other degradation phenomena. Baiji has a large number of seeds and does not spread the virus, but it is not easy to germinate. After aseptic germination through seed culture, a large number of disease-and insect-free plants can be obtained by tissue culture and rapid propagation, and then the survival rate of transplanted seedlings can be improved by seedling training. This method can not only obtain a large number of healthy seedlings, but also avoid species degradation caused by repeated tuber propagation.

1. Aseptic sowing

White and mature uncracked capsules were collected as materials for tissue culture aseptic explants. Carry out aseptic sowing according to the following steps: ① capsule aseptic treatment: wash the surface dust off the capsule with tap water, soak in 75% alcohol and wipe and disinfect for 1 minute, rinse with sterile water twice, soak in 0.1% mercuric chloride for 8 minutes for 10 minutes, rinse with aseptic water for 5-6 times, remove and use aseptic filter paper to absorb excess water. Aseptic treatment of ② medium: the medium of 1/2MS+NAA 0.5mg/L was sterilized at high temperature and high pressure, then cooled and set aside; ③ inoculation and culture: on the ultra-clean worktable, the sterilized capsule was cut open with a sterile scalpel, and the seeds were evenly inoculated on the surface of the medium with aseptic tweezers; ④ put the inoculated culture flask in the culture room at (25 ±2) ℃, light intensity of 1500 to 2000 Ix, light for 12 hours a day.

2. Aseptic germination

After 1 week of seed culture, the seeds began to expand and germinate, green dots could be seen in 2 weeks, and then the protocorms gradually germinated into yellow. the protocorms gradually turned green and differentiated into leaf primordia, and leaves grew after 1 month.

three. Proliferation of tissue culture plantlets

The rootless tissue culture seedlings with 2-3 leaves obtained by aseptic germination were used as the proliferation material of tissue culture seedlings. Take MS+ white sugar 3% + Agar powder 0.7% + activated carbon 0.1%+6-BA 1.0mg/L+NAA 0.2mg/L as culture medium, sterilize at high temperature and high pressure, then cool and set aside. The rootless tissue culture seedlings were transferred to the medium for culture. The culture conditions were as follows: temperature (25 ±3) ℃, light intensity 1500 ~ 2000 lx, light 12 hours per day.

4. Rooting induction of tissue culture plantlets

The single plant of multiplying seedlings was used as rooting induction material, and the culture medium was 2 MS+ sugar 2% + Agar powder 0.7% + activated carbon 0.l%+NAA 0.2 ~ 0.5mg/L. The rooting number was 3-5 and the rooting rate was 100% [Jishan Huayao] under the conditions of temperature (25 ±3) ℃, light intensity 1500-2000 lx and light 12 hours per day.

5. Seedling refining and transplanting

The tissue culture seedlings with induced rooting were used as materials for seedling refinement and transplanting. The specific operation steps are as follows: ① refining seedling treatment: in order to obtain robust tissue culture tubers and improve the survival rate of domestication and transplanting, the tissue culture seedlings induced by rooting need to be placed in the open room, and then transferred to the greenhouse for 4 days. ② acclimation and transplanting: one hundred bulbs of white and tissue culture seedlings with weight greater than 43g after seedling refining treatment were selected as acclimation and transplanting materials, and domesticated seedlings were domesticated on a loose, breathable and water-retaining medium, such as sawdust or vermiculite + bark (1:1) substrate; ③ spraying nutrient solution: organic water-soluble fertilizer or foliar fertilizer was sprayed on the transplanted tissue culture seedlings to increase recombination seedlings and promote the growth of white and tissue culture seedlings (figure).