Two-step seedling quick propagation method for hemerocallis tissue culture by using tender pedicel as explant
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A tissue culture and explant technology, applied in the fields of botany equipment and methods, horticultural methods, plant regeneration, etc., can solve the problems of cumbersome steps and high production costs, and achieve the goal of reducing steps, reducing production costs, and satisfying large-scale production. Effect
Inactive Publication Date: 2010-12-01
浙江清华长三角研究院生物技术与医药研究所
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Problems solved by technology
At present, the group cultivation of Hemerocallis seedlings is mainly completed through three stages: "callus induction, adventitious bud production → bud proliferation → tissue cul
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Embodiment 1
[0026] (1) preparation of culture medium,
[0027] (1) Preparation:
[0028] ①Induction and differentiation medium: use MS as the basic medium, add 4mg of 6-BA, 1mg of 2,4-D, 0.5mg of NAA, 4.5g of agar and 30g of sucrose per liter, and adjust the pH value to 5.8;
[0029] ② Rooting medium: use 1 / 2MS as the basic medium, add 1.0mg of NAA, 4.5g of agar and 15g of sucrose per liter, and adjust the pH to 5.8;
[0030] (2) Packing: Heat the prepared culture medium until transparent, and pack it into clean culture bottles while hot, about 70ml per bottle;
[0031] (3) Sterilization: Seal the mouth of the culture bottle as soon as possible after the culture medium is subpackaged, and put it in an autoclave in time for sterilization. Sterilization conditions are: 121° C., 0.1 MPa for 20 minutes. After the sterilized culture medium is taken out from the pressure cooker, it is placed on a flat table and allowed to cool and solidify naturally.
[0032] (2) Sterilization of explants: ...
Embodiment 2
[0038] When the explants are sterilized, the pedicels are soaked in 75% alcohol for 10 seconds, and then soaked in 0.1% mercuric chloride solution for 10 minutes. Other steps are the same as in Example 1. Using this sterilization method, the contamination rate of the explants was 33% after 15 days, which was higher than that in Example 1.
Embodiment 3
[0040] When the explants are sterilized, the pedicels are soaked in 75% alcohol for 10 seconds, and then soaked in 0.1% mercuric chloride solution for 20 minutes. Other steps are the same as in Example 1. Using this sterilization method, the contamination rate of explants was 2.4% after 15 days, which was lower than that in Example 1. However, the induction rate was 25%, and the late proliferation rate was 1.3, both lower than that of Example 1, and the tissue culture seedlings grew thin and difficult to take root.
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Abstract
The invention discloses a two-step seedling quick propagation method for hemerocallis tissue culture by using tender pedicel as explant. The method comprises the following steps of: sterilizing the tender hemerocallis pedicel serving as the explant, then cutting the pedicel into sections of 1 to 1.5 centimeters, and inoculating the cut pedicel to an induction and differentiation culture medium; transferring sterile seedlings with the height of 4 to 6 centimeters and over 4 leaves to a rooting culture medium after 2.5 to 3 months to perform rooting culture medium; and when each tissue culture seedling grows over 3 roots and the adventitious root reaches a length of 2 to 4 centimeters, opening a bottle cap, hardening the seedlings for 3 to 4 days indoors, taking the tissue culture seedlings out of a culture bottle, washing the culture medium, and transplanting the seedlings to the sterilized culture medium, wherein the induction and differentiation culture medium: MS is used as a basic culture medium; 2 to 10 mg of 6-benzyladenine (6-BA), 0 to 2 mg of 2,4-dichlorphenoxyacetic acid (2,4-D), 0.2 to 1 mg of alpha-naphthylacetic acid (NAA), 4.5 g of agar and 30 g of cane sugar are added into each liter of basic culture medium; and the pH value of the culture medium is adjusted to 5.8. The rooting culture medium: 1/2MS is used as a basic culture medium; 0.1 to 2.0 mg of NAA, 4.5 g of agar and 15 to 30 g of cane sugar are added into each liter of basic culture medium; and the pH value of the culture medium is adjusted to 5.8. The method can massively propagate good-quality hemerocallis tissue culture seedlings in short time so as to meet the requirement of large-scale production, and reduces the steps of preparing the culture medium and transferring the bottle, so the method is simple and feasible and reduces the production cost.
Description
technical field [0001] The invention relates to a simple and rapid propagation method for two-step seedling formation of daylily by tissue culture, in particular to tissue culture of daylily with young flower stalks as explants. Background technique [0002] Hemerocallis (Hemerocallis.sp) is a perennial herbaceous plant of Liliaceae. This plant is "viewed as a flower, eaten as a vegetable, and used as a medicine". It has wide application, strong adaptability, easy cultivation, simple management, quick income, high benefit, Features such as long economic life. All of these are of great significance for beautifying the environment, improving people's health, developing mountain economy, increasing exports, etc., and the market demand for them continues to increase. In nature, plants of this genus can be propagated by sowing, branching, etc., due to the strong heterosis of this plant, it is difficult to maintain good traits in seed propagation, and some varieties have low seed...
Claims
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