阅读说明：本技术 一种蓝莓增殖培养基及其增殖培养方法 (Blueberry multiplication medium and multiplication culture method thereof ) 是由 杨姣 杨曙方 仪传冬 王�琦 赵荣芳 洪麒明 于 2020-08-04 设计创作，主要内容包括：本发明公开了一种蓝莓增殖培养基及其增殖培养方法,一种蓝莓增殖培养基包括初代培养基和继代培养基,所述初代培养基包括基本培养基、0.8～1.2mg/L的ZT、0.08～0.12mg/L的IBA、25～35mg/L的蔗糖以及5.5～7.5mg/L的琼脂,所述继代培养基包括基本培养基、0.4～0.6mg/L的ZT、0.04～0.06mg/L的IBA、25～35mg/L的蔗糖以及5.5～7.5mg/L的琼脂,一种蓝莓的增殖培养方法,包括a)培养基的制备；b)诱导侧芽萌发和c)增殖培养,所制备的培养基,制作成本低廉,在使用时,培苗成活率达到了100％,分化率为市售培养基的1.5倍,培养周期短,组培苗在培养50天后可达到8cm左右,茎段粗壮,不但增加了组培苗的利用率,也增加了后续移栽的成活率,利用组培培养的方法,可以在短期内大大提高增殖系数。(The invention discloses a blueberry multiplication culture medium and a multiplication culture method thereof, wherein the blueberry multiplication culture medium comprises a primary culture medium and a secondary culture medium, the primary culture medium comprises a basic culture medium, 0.8-1.2 mg/L ZT, 0.08-0.12 mg/L IBA, 25-35 mg/L sucrose and 5.5-7.5 mg/L agar, the secondary culture medium comprises a basic culture medium, 0.4-0.6 mg/L ZT, 0.04-0.06 mg/L IBA, 25-35 mg/L sucrose and 5.5-7.5 mg/L agar, and the multiplication culture method of blueberries comprises a) preparation of the culture medium; b) inducing lateral bud germination and c) enrichment culture, wherein the prepared culture medium is low in manufacturing cost, when the culture medium is used, the survival rate of the cultured seedlings reaches 100%, the differentiation rate is 1.5 times of that of a commercially available culture medium, the culture period is short, the tissue culture seedlings can reach about 8cm after being cultured for 50 days, the stem sections are thick and strong, the utilization rate of the tissue culture seedlings is increased, the survival rate of subsequent transplantation is increased, and the proliferation coefficient can be greatly increased in a short period by utilizing a tissue culture method.)

1. A blueberry multiplication culture medium is characterized in that: the culture medium comprises a primary culture medium and a secondary culture medium, wherein the primary culture medium comprises a basic culture medium, 0.8-1.2 mg/L ZT, 0.08-0.12 mg/L IBA, 25-35 mg/L sucrose and 5.5-7.5 mg/L agar, and the secondary culture medium comprises a basic culture medium, 0.4-0.6 mg/L ZT, 0.04-0.06 mg/L IBA, 25-35 mg/L sucrose and 5.5-7.5 mg/L agar.

2. The blueberry propagation medium as claimed in claim 1, wherein: the minimal medium comprises 675-825 mg/L NH₄NO₃445 to 545mg/L of K₂SO₄570-690 mg/L KNO₃280-340 mg/L MgSO (MgSO)₄·7H₂O, 125-155 mg/L KH₂PO₄250-310 mg/L Ca (NO)₃)₂·4H₂O, 175-215 mg/L CaCl₂·2H₂O, 5-7 mg/L H₃BO₃15-20 mg/L MnSO₄·H₂O, 7.5-9.5 mg/L ZnSO₄·7H₂O, 0.15-0.35 mg/L Na₂MoO₄·2H₂O, 0.15-0.35 mg/L CuSO₄·5H₂O, 35-40 mg/L of Na₂EDTA, 25-30 mg/L FeSO₄·7H₂O, inositol 80-120 mg/L, vitamin B0.4-0.6 mg/L₅0.4-0.6 mg/L pyridoxine hydrochloride, 0.8-1.2 mg/L thiamine hydrochloride, and 1.5-2.5 mg/L glycine.

3. The blueberry propagation medium as claimed in claim 1, wherein: the pH values of the primary culture medium and the secondary culture medium are both adjusted to 5.0-5.3.

4. A proliferation culture method of blueberries is characterized by comprising the following steps:

a) preparation of a culture medium: adding 0.8-1.2 mg/L ZT, 0.08-0.12 mg/L IBA, 25-35 mg/L sucrose and 5.5-7.5 mg/L agar into a basic culture medium to obtain a primary culture medium, and adding 0.4-0.6 mg/L ZT, 0.04-0.06 mg/L IBA, 25-35 mg/L sucrose and 5.5-7.5 mg/L agar into the basic culture medium to obtain a secondary culture medium;

b) inducing lateral bud germination: cleaning and disinfecting a blueberry explant, and then inoculating the blueberry explant into the primary culture medium prepared in the step a) for induced differentiation to obtain a blueberry primary tissue culture seedling;

c) and (3) proliferation culture: trimming the blueberry primary tissue culture seedlings prepared in the step b), and then inoculating the blueberry primary tissue culture seedlings into the subculture medium prepared in the step a) for subculture to obtain blueberry subculture tissue culture seedlings.

5. The propagation culture method of blueberries as claimed in claim 4, wherein: in the step b), the induced differentiation conditions of the blueberry explants are dark culture for the first 6-8 days, the temperature is 20-24 ℃, then the light culture is changed, the illumination intensity is 2500-3000 lx, the illumination time per day is 10-14 h, the temperature is 23-27 ℃, and the total culture time is 35-45 days.

6. The propagation culture method of blueberries as claimed in claim 4, wherein: in the step c), trimming the primary tissue culture seedlings of the blueberries to 1.3-1.7 cm.

7. The propagation culture method of blueberries as claimed in claim 4, wherein: in the step c), the subculture condition of the primary tissue culture seedlings of the blueberries is light culture, the illumination intensity is 2800-3200 lx, the illumination time is 14-18 h every day, the temperature is 23-27 ℃, and the culture time is 40-60 days.

[ technical field ] A method for producing a semiconductor device

The invention relates to the technical field of blueberry multiplication, in particular to the technical field of a blueberry multiplication culture medium and a multiplication culture method thereof.

[ background of the invention ]

Blueberries, belonging to the family of ericaceae, are plants of the genus Vaccinium, originating in North America, and are perennial shrub berry trees. Blueberries have extremely high economic and nutritional values, so that the blueberries are concerned by people. The blueberry contains nutrient elements such as fructose, fiber and vitamins, is rich in compounds such as antioxidants, folic acid and anthocyanin which are beneficial to removing free radicals in a human body and inhibiting and reversing aging of the human body, and is listed as one of five health foods of human beings by the international food and agriculture organization.

Along with the continuous improvement of the international demand of blueberries, the cultivation area of the blueberries in the world continuously rises, and the demand of people on the blueberry seedlings also increases year by year. The artificial cultivation of the blueberries in China starts late, the overall scale is small, the increasing requirements of consumers cannot be met, and the problems of weak tissue culture seedlings, low differentiation rate, long culture period, low utilization rate and low subsequent transplanting survival rate also exist. Therefore, how to obtain enough blueberry seedlings quickly and enlarge the blueberry cultivation area becomes an important problem to be solved urgently.

[ summary of the invention ]

The invention aims to solve the problems in the prior art, provides a blueberry multiplication culture medium and a multiplication culture method thereof, and solves the problems of weakness, low differentiation rate, long culture period, low utilization rate and low subsequent transplanting survival rate of blueberry tissue culture seedlings.

In order to achieve the purpose, the invention is realized by the following technical scheme:

the blueberry multiplication culture medium comprises a primary culture medium and a secondary culture medium, wherein the primary culture medium comprises a basic culture medium, 0.8-1.2 mg/L of ZT, 0.08-0.12 mg/L of IBA, 25-35 mg/L of cane sugar and 5.5-7.5 mg/L of agar, and the secondary culture medium comprises a basic culture medium, 0.4-0.6 mg/L of ZT, 0.04-0.06 mg/L of IBA, 25-35 mg/L of cane sugar and 5.5-7.5 mg/L of agar.

Preferably, the minimal medium comprises 675-825 mg/L NH₄NO₃445 to 545mg/L of K₂SO₄570-690 mg/L KNO₃280-340 mg/L MgSO (MgSO)₄·7H₂O, 125-155 mg/L KH₂PO₄250-310 mg/L Ca (NO)₃)₂·4H₂O, 175-215 mg/L CaCl₂·2H₂O, 5-7 mg/L H₃BO₃15-20 mg/L MnSO₄·H₂O, 7.5-9.5 mg/L ZnSO₄·7H₂O, 0.15-0.35 mg/L Na₂MoO₄·2H₂O, 0.15-0.35 mg/L CuSO₄·5H₂O, 35-40 mg/L of Na₂EDTA, 25-30 mg/L FeSO₄·7H₂O, inositol 80-120 mg/L, vitamin B0.4-0.6 mg/L₅0.4-0.6 mg/L pyridoxine hydrochloride, 0.8-1.2 mg/L thiamine hydrochloride, and 1.5-2.5 mg/L glycine.

Preferably, the primary culture medium and the secondary culture medium are both adjusted to pH 5.0-5.3.

A proliferation culture method of blueberries comprises the following steps:

a) preparation of a culture medium: adding 0.8-1.2 mg/L ZT, 0.08-0.12 mg/L IBA, 25-35 mg/L sucrose and 5.5-7.5 mg/L agar into a basic culture medium to obtain a primary culture medium, and adding 0.4-0.6 mg/L ZT, 0.04-0.06 mg/L IBA, 25-35 mg/L sucrose and 5.5-7.5 mg/L agar into the basic culture medium to obtain a secondary culture medium;

b) inducing lateral bud germination: cleaning and disinfecting a blueberry explant, and then inoculating the blueberry explant into the primary culture medium prepared in the step a) for induced differentiation to obtain a blueberry primary tissue culture seedling;

c) and (3) proliferation culture: trimming the blueberry primary tissue culture seedlings prepared in the step b), and then inoculating the blueberry primary tissue culture seedlings into the subculture medium prepared in the step a) for subculture to obtain blueberry subculture tissue culture seedlings.

Preferably, in the step b), the induced differentiation conditions of the blueberry explants are dark culture for 6-8 days, the temperature is 20-24 ℃, then, light culture is changed, the illumination intensity is 2500-3000 lx, the illumination time per day is 10-14 h, the temperature is 23-27 ℃, and the total culture time is 35-45 days.

Preferably, in the step c), the blueberry primary tissue culture seedlings are trimmed to 1.3-1.7 cm.

Preferably, in the step c), the subculture condition of the primary tissue culture seedlings of the blueberries is light culture, the illumination intensity is 2800-3200 lx, the illumination time is 14-18 h per day, the temperature is 23-27 ℃, and the culture time is 40-60 days.

The invention has the beneficial effects that: the culture medium prepared by the invention has low manufacturing cost, when in use, the survival rate of the cultured seedlings reaches 100 percent, the differentiation rate is 1.5 times of that of the commercial culture medium, the culture period is short, the tissue culture seedlings can reach about 8cm after being cultured for 50 days, the stem sections are thick and strong, the utilization rate of the tissue culture seedlings is increased, the survival rate of subsequent transplantation is also increased, the propagation coefficient can be greatly improved in a short period by utilizing the tissue culture method, the propagation time is shortened, and the excellent characteristics of the variety are kept.

[ detailed description ] embodiments