Mosquito-repellent herbs are genus perennial plants of the genus Pelargonium. It not only naturally volatilizes citronellal, citronellol and other substances to effectively remove mosquitoes, but also emits lemon aroma to purify the air. Its main stem is soft and can be used to create a variety of bonsai for viewing. Mosquito repellent vanilla can normally bloom, but it cannot breed naturally. In practice, it has been proved that the survival rate of cuttings is low, and its function declines from generation to generation. The use of shoot-tip in vitro culture technology can effectively maintain the stability of insect-repellent herbs and increase the number of seedlings rapidly, which has the advantages of low cost, short time, and high propagation coefficient. (I) Materials and culture conditions 1. Materials: Mosquito repellent stem tips. 2. Culture conditions: MS is the basic medium. (1) Priming and proliferation medium: MS+ammonium nitrate 800 mg/L (the same unit below)+sodium dihydrogen phosphate, H2O150+gibberellin 1+KT8+IAA1; (2) Rooting medium: 1/2 MS+Fe salt 5ml+NAA0.2; medium was added sucrose 3%, agar 0.6%, pH 5.6, 121 °C, 0.11MPa sterilization 25 minutes. Cultivation temperature 25 (1) °C, light 12 hours / day, light intensity 2000Lx. (II) Growth and Differentiation 1. Acquisition of sterile materials: Select a robust insect repellent vanilla seedling, remove the shoot tip, and transplant it into a sterilized potting soil for indoor culture. After 15 to 20 days, shoot tips with a length of about 1.0 cm on the axillary buds were used as explants. Remove the unfolded young leaves, wash with water for 1 to 2 hours, place on a super-clean bench with 70% alcohol for 10 to 15 seconds, wash with sterile water 2 to 3 times, sterilize for 0.1 minutes with 0.1% high mercury salts, rinse again with sterile water 4 to 5 times. Under aseptic conditions, the surface of the material was blotted with sterile filter paper, and the tip of the stem was stripped under a double-dissecting microscope. The shoot tip with the two youngest leaflet primordia was inoculated on the initial culture medium. If browning occurs in time, it will be browned after 2 or 3 transfers. After 5 to 7 days of inoculation, shoots sprouted and grew to a length of 0.5 to 2 cm and clustered shoots with 1 to 2 leaves in 15 days. Subcultures were propagated after 25 days. 2. Axillary bud proliferation: The above-mentioned clustered buds were cut into buds with 1 or 2 buds, and the callus was removed from the roots and inoculated on the proliferation medium. After about 30 days of cultivation, the axillary buds of each bud germinate and grow about 3 cm, with clusters of 4 to 6 leaves. In this way, repeated subcultures are repeated. After each initial culture of cluster buds is propagated for about 15 generations, a large number of tissue culture seedlings can be obtained. 3. Rooting and hardening seedlings: under aseptic conditions, shoots with a height of about 3 cm in the shoots were cut and transferred to the rooting medium. After 20 days of cultivation, the adventitious roots began to form in large numbers. The average rooting rate per plant was more than 8 and the rooting rate was over 95%. . To make the test tube seedling gradually grow from heterotrophic growth to autotrophic growth, open the bottle and pour it into warm water of 20°C to a depth of about 1.5cm to moisturize and prevent dirt. After 4 to 6 days of hardening, remove carefully and wash the root agar for transplanting. 4. Transplantation: Before transplanting, peat soil and perlite were mixed in a 2:1 ratio and sterilized, and the climate was selected to be transplanted into the mixed matrix in a mild period of time. The rooting powder ABT 150 mg/L + fine soil 100 g/L was soaked for 20 minutes during transplanting. After easing the seedlings, the water solution of a large amount of 1/2MS elements is regularly chased. Attention should be paid to temperature, humidity, light and water and fertilizer management, and the survival rate of transplanting can reach 90% or more.
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