阅读说明：本技术 一种观赏水草南极杉的组织培养方法 (Tissue culture method of ornamental water plant Antarctic fir ) 是由 不公告发明人 于 2019-12-06 设计创作，主要内容包括：本发明涉及一种观赏水草南极杉的组织培养方法,包括以下步骤：(1)预处理；(2)将预处理的外植体切割成1-1.5cm长的外植体,切割后的每个外植体至少带一个腋芽；将切割后的外植体接入初代培养基进行初代培养；(3)分离初代培养基中的初代不定芽,使初代不定芽长度控制在2cm以内；将分离后的初代不定芽转入增殖培养基中培养；(4)将增殖培养基中长度大于3-4cm的植株转入生根培养基培养；(5)将带有完整根系的南极杉植株从生根培养基中取出,清水冲洗后移栽到生态缸中。该方法不受季节、温度、地域影响就能获得大量的新植株,快速建立南极杉无菌繁殖体系,为保持南极杉优良的种性提供技术支持,为广大的爱好者提供源源不断地无菌种苗。(The invention relates to a tissue culture method of ornamental float grass Antarctic fir, which comprises the following steps: (1) pre-treating; (2) cutting the pre-treated explants into 1-1.5cm long explants, each cut explant having at least one axillary bud; inoculating the cut explant into a primary culture medium for primary culture; (3) separating primary adventitious buds in the primary culture medium to control the length of the primary adventitious buds within 2 cm; transferring the separated primary adventitious bud into a multiplication culture medium for culture; (4) transferring the plant with the length of more than 3-4cm in the multiplication culture medium into a rooting culture medium for culture; (5) taking out the Antarctic fir plants with complete root systems from the rooting culture medium, washing with clear water, and transplanting into an ecological cylinder. The method can obtain a large number of new plants without being influenced by seasons, temperature and regions, quickly establish an Antarctic fir sterile propagation system, provide technical support for maintaining excellent seed properties of Antarctic fir, and provide continuous sterile seedlings for vast enthusiasts.)

1. A tissue culture method of ornamental float grass Antarctic fir is characterized by comprising the following steps:

(1) pretreatment: selecting healthy Antarctic fir plants without diseases and insect pests, and sterilizing and cleaning to obtain sterilized explants;

(2) primary culture: cutting the sterilized explants into 1-1.5cm long explants in a sterile culture dish, wherein each cut explant has at least one axillary bud; transferring the cut explant into a primary culture medium for primary culture for 10-20 days;

(3) and (3) proliferation culture: separating primary adventitious buds in the primary culture medium to control the length of the primary adventitious buds within 2 cm; transferring the separated primary adventitious bud into a proliferation culture medium for proliferation culture for 22-30 days;

(4) rooting culture: transferring the plant with the length of more than 3-4cm in the multiplication culture medium into a rooting culture medium for rooting culture;

(5) transplanting: taking out the Antarctic fir plants with complete root systems from the rooting culture medium, washing with clear water, and transplanting into an ecological cylinder.

2. The tissue culture method of ornamental float grass antarctic fir according to claim 1, characterized in that the specific methods of explant sterilization treatment and cleaning in step (1) are as follows: washing explant with running water for 30min, sterilizing with 75% alcohol for 15-30s, and washing for 3 times; sterilizing with 1% sodium hypochlorite for 10-20min, and washing with sterile water for 5 times.

3. The tissue culture method of ornamental float grass antarctic fir according to claim 1, wherein the primary culture medium is: MS culture medium, 6-10g/L agar, 10-30g/L cane sugar, 0.5-1.5mg/L kinetin and 0.05-0.5mg/L naphthylacetic acid.

4. The tissue culture method of ornamental float grass antarctic fir according to claim 1, wherein the multiplication medium is: MS culture medium, 6-10g/L agar, 10-30g/L cane sugar, 1-2.5mg/L6-BA and 0.5-1mg/L naphthylacetic acid.

5. The tissue culture method of ornamental float grass antarctic fir according to claim 1, wherein the rooting medium is: MS culture medium, 6-10g/L agar, 10-30g/L cane sugar and 0.1-0.5mg/L auxin.

6. The tissue culture method of ornamental float grass antarctic fir according to claim 5, wherein the rooting medium is: MS culture medium, 6-10g/L agar, 10-30g/L cane sugar and 0.1-0.5mg/L indoleacetic acid.

7. The tissue culture method of ornamental float grass antarctic fir according to claim 1, wherein the primary culture conditions are as follows: the illumination period is 14h/d, the temperature is 25 +/-2 ℃, and the illumination is 2500 Lux.

8. The tissue culture method of ornamental float grass antarctic fir according to claim 1, characterized in that the proliferation culture conditions are as follows: the illumination period is 14h/d, the temperature is 25 +/-2 ℃, and the illumination is 2500 Lux.

9. The tissue culture method of ornamental float grass antarctic fir according to claim 1, characterized in that the rooting culture conditions are as follows: the illumination period is 14h/d, the temperature is 25 +/-2 ℃, and the illumination is 2500 Lux.

Technical Field

The invention relates to the technical field of plant tissue culture, in particular to a tissue culture method of ornamental water plant Antarctic fir.

Background

Antarctic fir is a common ornamental aquatic weed. Antarctic fir is submerged herbaceous plant, Plantaginaceae, dicotyledonous plant, and emergent aquatic weed. In the wild condition, the water surface is beautiful like the flower sea every flowering phase, and the flowering condition can be seen even when the grass is cultivated in a jar. The aquatic weed can adapt to wide water quality change in water, can grow under weak light, and is preferably cultivated under strong light. Antarctic fir is morphologically comparable to green feather, with the difference that the former is acicular leaves and the latter is feathery leaves. The current breeding mode of Antarctic fir is cuttage and lateral bud breeding.

However, mature techniques for large-scale propagation of good-quality Antarctic fir plants are currently lacking.

Disclosure of Invention

Aiming at the technical problems in the prior art, the invention provides a tissue culture method of ornamental float grass Antarctic fir, which comprises the steps of inducing new seedlings of the Antarctic fir by adopting a proper culture environment and giving fixed illumination under the aseptic condition to gradually form an aseptic seedling system, and providing a large number of aseptic seedlings throughout the year in the later stage. The method is easy to implement, convenient to operate and high in yield, and solves the problems that Antarctic fir is difficult to propagate in an aquarium, the propagation speed is low and plant seedlings are difficult to obtain.

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

a tissue culture method of ornamental float grass Antarctic fir comprises the following steps:

(1) pretreatment: selecting healthy Antarctic fir plants without diseases and insect pests, and sterilizing and cleaning to obtain sterilized explants;

(2) primary culture: cutting the sterilized explants into 1-1.5cm long explants in a sterile culture dish, wherein each cut explant has at least one axillary bud; transferring the cut explant into a primary culture medium for primary culture for 10-20 days;

(3) and (3) proliferation culture: separating primary adventitious buds in the primary culture medium to control the length of the primary adventitious buds within 2 cm; transferring the separated primary adventitious bud into a proliferation culture medium for proliferation culture for 22-30 days;

(4) rooting culture: transferring the plant with the length of more than 3-4cm in the multiplication culture medium into a rooting culture medium for rooting culture;

(5) transplanting: taking out the Antarctic fir plants with complete root systems from the rooting culture medium, washing with clear water, and transplanting into an ecological cylinder.

Further, the specific method for sterilizing and cleaning the explant in the step (1) comprises the following steps: washing explant with running water for 30min, sterilizing with 75% alcohol for 15-30s, and washing for 3 times; sterilizing with 1% sodium hypochlorite for 10-20min, and washing with sterile water for 5 times.

Further, the primary medium is: MS culture medium, 6-10g/L agar, 10-30g/L cane sugar, 0.5-1.5mg/L kinetin and 0.05-0.5mg/L naphthylacetic acid.

Further, the proliferation medium is: MS culture medium, 6-10g/L agar, 10-30g/L cane sugar, 1-2.5mg/L6-BA and 0.5-1mg/L naphthylacetic acid.

Further, the rooting medium is: MS culture medium, 6-10g/L agar, 10-30g/L cane sugar and 0.1-0.5mg/L auxin.

Further, the rooting medium is: MS culture medium, 6-10g/L agar, 10-30g/L cane sugar and 0.1-0.5mg/L indoleacetic acid.

Further, the primary culture conditions are as follows: the illumination period is 14h/d, the temperature is 25 +/-2 ℃, and the illumination is 2500 Lux.

Further, the proliferation culture conditions are: the illumination period is 14h/d, the temperature is 25 +/-2 ℃, and the illumination is 2500 Lux.

Further, the rooting culture conditions are as follows: the illumination period is 14h/d, the temperature is 25 +/-2 ℃, and the illumination is 2500 Lux.

Therefore, compared with the prior art, the invention has the following beneficial technical effects:

(1) the tissue culture method of ornamental float grass Antarctic fir provided by the invention is characterized in that under the aseptic condition, a proper culture environment is adopted, fixed illumination is provided, the growth of the scion of the Antarctic fir is directly induced, a large number of new plants can be obtained without being influenced by seasons, temperature and regions, an Antarctic fir sterile propagation system is quickly established, technical support is provided for maintaining the excellent seed nature of the Antarctic fir, and continuous high-quality aseptic seedlings are provided for vast enthusiasts.

(2) The tissue culture formulas adopted in the tissue culture method are cooperated, the tissue propagation period is short, the propagation rate is high, the rooting rate of Antarctic fir seedlings aseptically cultured by the tissue culture method in natural water is 100%, the plant survival rate is more than 98%, the effective and rapid asexual propagation of Antarctic fir is realized, and the tissue culture method has important effects on promoting the production of ornamental aquatic weeds and the culture of new varieties and accelerating the rapid development of aquatic ornamental aquatic weed industry.

(3) The tissue culture method of the aseptic seedlings of the Antarctic fir is simple and easy to implement, convenient to operate and high in yield, and solves the problems that the propagation is difficult, the propagation speed is slow and plant seedlings are difficult to obtain in an aquarium.

Detailed Description

The following examples are presented to illustrate certain embodiments of the invention in particular and should not be construed as limiting the scope of the invention. The present disclosure may be modified from materials, methods, and reaction conditions at the same time, and all such modifications are intended to be within the spirit and scope of the present invention. Unless otherwise specified, the technical means used in the examples are conventional means well known to those skilled in the art. The reagents and biomaterials, if not specifically indicated, are commercially available.