CN110250005B

CN110250005B - Cultivation method of new cymbidium goeringii variety

Info

Publication number: CN110250005B
Application number: CN201910662139.5A
Authority: CN
Inventors: 冷青云; 黄少华; 李贵雨; 王存
Original assignee: Tropical Crops Genetic Resources Institute CATAS
Current assignee: Tropical Crops Genetic Resources Institute CATAS
Priority date: 2019-07-22
Filing date: 2019-07-22
Publication date: 2023-04-07
Anticipated expiration: 2039-07-22
Also published as: CN110250005A

Abstract

The invention discloses a new cymbidium species cultivation method, which comprises the steps of (1) parent selection, (2) sterile sowing of seeds, (3) selection of excellent single plants and strain formation, (4) strain shape evaluation and the like, wherein the specificity, consistency and stability of strains are observed and evaluated according to the overall state, internode thickness, leaf shape and vein veins of the plants, and strains with specificity, consistent single plant performance and stable properties are screened to be new cymbidium species. The breeding method of the new variety has the advantages of high operability, strong variety stability and high breeding speed, can breed a large number of seedlings in a short time, and meets the commercial planting requirement.

Description

Cultivation method of new cymbidium goeringii variety

Technical Field

The invention relates to the technical field of new plant variety cultivation, relates to a cultivation method of a new plant variety of Orchidaceae cymbidium, and particularly relates to a cultivation method of a new cymbidium variety.

Background

All-grass of Broccoli (Ludisia discolor(ker-Gawl) A.Rich, also known as Nelumbo nucifera Gaertn, nereis, pinus martensii, etc., is a plant of the genus Haematococcus of the family Orchidaceae. Distributed in tropical subtropical regions such as Hainan, fujian, guangdong, guangxi, and Yunnan, and also distributed in southeast Asia such as Thailand, myanmar, vietnam, etc. The Chinese medicinal herb is compact in plant type, bright in vein veins of leaves, pure and white in flowers, and can be used for enjoying flowers and leaves, is often used as a small potted plant or planted on stones to be used as a garden landscape, and in addition, the angiopteris has the effects of nourishing yin and moisturizing lung, soothing nerves and strengthening spleen, clearing heat and cooling blood and the like, can treat symptoms such as neurasthenia, phthisis and hemoptysis, inappetence and the like, and is often used as a herbal medicine for treating lung diseases in folks. Due to the high ornamental and medicinal value, the market demand is increasing year by year in recent years.

At present, the market-sold cymbidium sinense is mainly obtained by field collection, because the variety has no large-scale commercial planting, no related variety report and unstable yield and quality. In addition, the wild collection of the cymbidium bicolor is the whole plant, the population of the collected place is sharply reduced, and the collected place cannot be recovered in a long period of time, the wild resources are seriously damaged by the plunder type collection, the danger-causing process of the species is accelerated, and the cymbidium bicolor is classified as a national secondary protection plant. Therefore, in order to develop the cymbidium goeringii industry and protect wild resources, the cultivation of new cymbidium goeringii varieties is particularly necessary.

Disclosure of Invention

The invention aims to provide a new variety cultivation method of cymbidium sinense, which has the advantages of high operability, strong variety stability and high propagation speed, can propagate a large number of seedlings in a short time and meets the commercial planting requirement.

In order to achieve the purpose, the technical scheme of the invention is as follows: the method for cultivating the new variety of the cymbidium sinense comprises the following steps:

(1) Selecting breeding parents: selecting strong plants, short and thick internodes, large leaves and obvious vein reticulate patterns as parents from wild collected cymbidium sources, transplanting the plants into a germplasm garden, carrying out alloplastic hybridization in the flowering season of 2-3 months, and picking for later use when the capsule seed coat is changed from green to brown but the seeds are not cracked;

(2) And (3) sterile sowing of seeds:

a. The picked cymbidium sinense seeds are subjected to surface disinfection for 30s on a sterile operation table by using 75% ethanol, are washed for 3-4 times by using sterile water, are transferred into 0.1% mercuric chloride solution for sterilization for 8-10 min, are shaken at intervals, are washed for 3-4 times by using sterile water and are inoculated into a germination culture medium;

b. culturing under dark conditions, wherein seeds swell and expand to break through seed coats within 5-8 days, milky round bulbs are formed within 13-15 days, fluff densely grows on the surfaces of the round bulbs along with the formation of the round bulbs, the round bulbs are cultured under the illumination condition at the moment, the round bulbs extend within 50-60 days, and leaves grow on the top ends of the round bulbs;

c. subculturing every 30 d for 2-3 times, and inoculating to rooting culture medium;

(3) Selection and line formation of elite individuals:

a. after the aseptic seedling is cultured in a rooting culture medium for 90 days, 4 to 5 unfolded leaves exist;

b. according to the breeding targets of the overall plant state, internode thickness, leaf shape and vein, initially selecting and numbering individual plants with specificity by adopting a one-time individual plant selection method, and carrying out multiplication cultivation to form a plant line;

c. the proliferation culture method comprises the following steps: cutting a single plant into small sections of 2-3 cm, wherein each small section contains 2-3 internodes, and inoculating the small sections into a multiplication culture medium;

d. subculturing every 45 days for 4-6 times, inoculating to rooting culture medium when the plant is proliferated to more than 200 plants, culturing in the rooting culture medium for 60 days, and taking out of bottles for domestication and cultivation;

(4) Evaluating the shape of the strain:

a. cleaning the root culture medium of the rooted plants of the obtained strains, airing the rooted plants, wrapping and transplanting the roots with water moss to 105-hole trays for planting, and culturing in a greenhouse for 60 days;

b. the greenhouse environment is controlled at 24-26 ℃, the humidity is kept at 80-90 percent, and the shading degree is 80-85 percent;

c. watering the water-soluble fertilizer every 15 days after 30 days;

d. when 4-6 leaves grow out, observing and evaluating the specificity, consistency and stability of the strain according to the whole plant state, internode thickness, leaf shape and vein, and screening the strain with specificity, consistent single plant performance and stable character as a new variety of the cymbidium goeringii.

Further, the components of the germination culture medium comprise 1/2MS, 2g/L Huabao No. 1, 0.5-1 mg/L NAA, 30 g/L potato extract and 2g/L hydrolyzed casein.

Further, the subculture medium comprises 1/2MS, 1 g/L Huabao No. 1, 0.5-1 mg/L6-BA, 30 g/L potato extract and 0.5 g/L AC (activated carbon).

Further, the rooting medium comprises 1/2MS, 2g/L Huabao No. 1, 30 g/L potato extract and 0.5 g/L AC.

Further, the proliferation culture medium comprises 1/2MS, 1 g/L Huabao No. 1 and 1-2 mg/L6-BA.

Further, the water-soluble fertilizer is 20-20-20 parts of water-soluble fertilizer.

In the invention, 30 g/L of sucrose is added to all culture media without special specification, 7 g/L of agar powder is added to the solid culture medium, and the pH value is 5.8-6.2.

Drawings

FIG. 1 is a diagram of picking of the capsule seed coat of the present invention when the green color is changed into brown color but the seed is not cracked;

FIG. 2 is a view showing the seeds of the present invention swelling and breaking through the seed coat to form milky round bulb with dense fuzz on the surface;

FIG. 3 is a diagram showing the presence of 4 to 5 unfolded leaves behind 90d of Clerodendranthus spicatus of the present invention;

FIG. 4 is a diagram of the breeding of a new variety of the "Hongyu" of the cymbidium sinense of the invention;

FIG. 5 is a diagram of the breeding of a new variety of the black jade of the cymbidium sinense of the invention;

FIG. 6 is a diagram of the breeding of a new variety of Maolan "Maoyu" of the present invention.

Detailed Description

Example 1

Collecting a material with strong plants, short and thick internodes, large leaves and rich leaf vein reticulate patterns from the field as a parent, carrying out alloplastic selfing in the flowering season of 2 months, picking when the capsule seed coat changes from green to brown but the seed does not crack (figure 1), sterilizing the picked cymbidium sinense seed on a sterile operation table by using 75% ethanol for 30s, washing the seed with sterile water for 3 times, transferring the seed into 0.1% mercuric chloride solution for sterilization for 8 min, shaking the seed at intervals, and then, washing the seed with sterile water for 3 times and inoculating the seed into a germination culture medium; the components of the germination culture medium are 1/2MS +2.0 g/L Huabao No. 1 +0.5 mg/L NAA +30 g/L potato extract + 2g/L hydrolyzed casein. 7 d, the seeds are swollen and expanded to break through the seed coat, 15 d of the seeds form milky round bulbs, villi densely grow on the surfaces of the round bulbs along with the formation of the round bulbs (figure 2), 60 d of the round bulbs extend, leaves grow on the top of the round bulbs, the round bulbs are subjected to generation for 3 times in a culture medium and then inoculated into a rooting culture medium, and the generation culture medium is 1/2MS +1 g/L Huabao No. 1 +0.5 mg/L6-BA +30 g/L potato extract +0.5 g/L AC; the rooting culture medium is 1/2MS + 2g/L Huabao No. 1 +30 g/L potato extract +0.5 g/L AC, 4-5 unfolded leaves are arranged after 90d (figure 3), an excellent single plant is selected according to the aspects of the whole plant state, internode thickness, leaf shape, vein veins and the like and inoculated into a subculture multiplication culture medium, the components of the multiplication culture medium are 1/2MS +1 g/L Huabao No. 1 + 1.0 mg/L6-BA, the excellent single plant is multiplied for 4 times to a certain amount and then inoculated into a rooting culture medium, and the rooting culture medium is 1/2MS + 2g/L Huabao No. 1 +30 g/L potato extract +0.5 g/L AC (activated carbon). Taking out of bottles for domestication cultivation after rooting cultivation for 60 d. And (4) wrapping and transplanting the seedlings with the cleaned roots to 105-hole trays for planting by using the sphagna, and culturing in a greenhouse. The greenhouse environment is controlled at 24 ℃, the humidity is kept at about 90 percent, and the shading degree is 85 percent. 30 And watering the water-soluble fertilizer for 20-20-20 days after the step d every 15 days. When 4-6 leaves grow out, the specificity, consistency and stability of the strain are observed and evaluated from the aspects of the whole plant state, internode thickness, leaf shape, vein and the like, and the strain with specificity, consistent performance among single plants and stable character is selected as a new variety of the cymbidium bicolor.

Example 2

Collecting a material with strong plants, short and thick internodes, large leaves and rich leaf vein reticulate patterns from the field as a parent, carrying out alloplastic selfing in the flowering season of 3 months, picking when the capsule seed coat is changed from green to brown but the seed is not cracked (figure 1), disinfecting the picked cymbidium sanguineum seed on the surface of a sterile operation platform for 30s by using 75% ethanol, washing the seed with sterile water for 4 times, transferring the seed into 0.1% mercuric chloride solution for sterilization for 9 min, shaking the seed at intervals, and then inoculating the seed into a germination culture medium after washing the seed for 4 times with sterile water; the components of the germination culture medium are 1/2MS + 2g/L Huabao No. 1 +0.8mg/L NAA +30 g/L potato extract + 2g/L hydrolyzed casein. The seed swells and expands to break through the seed coat at 8 d, 14d forms a milky round bulb, the round bulb grows with dense villi on the surface (figure 2) along with the formation of the round bulb, 60 d round bulb extends, leaves grow on the top of the round bulb, the round bulb is inoculated into a rooting culture medium after being subcultured for 2 times in the culture medium, and the subculture medium is 1/2MS +1 g/L Huabao No. 1 +0.8 mg/L6-BA +30 g/L potato extract +0.5 g/L AC; the rooting culture medium is 1/2MS + 2g/L Huabao No. 1 +30 g/L potato extract +0.5 g/L AC, 4-5 unfolded leaves are arranged after 90d (figure 3), an excellent single plant is selected according to the aspects of the whole plant state, internode thickness, leaf shape, vein veins and the like and inoculated into a subculture multiplication culture medium, the components of the multiplication culture medium are 1/2MS +1 g/L Huabao No. 1 + 1.5 mg/L6-BA, and the excellent single plant is inoculated into the rooting culture medium after being multiplied for 5 times to a certain amount, and the rooting culture medium is 1/2MS + 2g/L Huabao No. 1 +30 g/L potato extract +0.5 g/L AC (activated carbon). Taking out of bottles for domestication cultivation after rooting cultivation for 60 d. And (4) wrapping and transplanting the seedlings with the cleaned roots to 105-hole trays for planting by using the sphagna, and culturing in a greenhouse. The greenhouse environment is controlled at 25 ℃, the humidity is kept at about 90 percent, and the shading degree is 85 percent. 30 And (4) watering the water soluble fertilizer for 20-20-20 days every 15 days after the d. When 4-6 leaves grow out, the specificity, consistency and stability of the strain are observed and evaluated from the aspects of the whole plant state, internode thickness, leaf shape, vein and the like, and the strain with specificity, consistent performance among single plants and stable character is selected as a new variety of the cymbidium bicolor.

Example 3

Collecting a material with strong plants, short and thick internodes, large leaves and rich leaf vein reticulate patterns from the field as a parent, carrying out alloplastic selfing in the flowering season of 2 months, picking when the capsule seed coat is changed from green to brown but the seed is not cracked (figure 1), disinfecting the picked cymbidium sanguineum seed on the surface of a sterile operation platform for 30s by using 75% ethanol, washing the seed with sterile water for 4 times, transferring the seed into 0.1% mercuric chloride solution for sterilization for 10 min, shaking the seed at intervals, and then inoculating the seed into a germination culture medium after washing the seed for 4 times with sterile water; the germination medium comprises 1/2MS + 2g/L Huabao No. 1 + 1mg/L NAA +30 g/L potato extract and 2g/L hydrolyzed casein. The seeds are swollen and expanded to break through the seed coat after 7 d, a milky round bulb is formed after 15 d, villi densely grow on the surface of the round bulb along with the formation of the round bulb (figure 2), the round bulb is elongated after 60 d, leaves grow on the top of the round bulb, the round bulb is inoculated into a rooting culture medium after 3 generations in the culture medium, and the subculture medium is 1/2MS +1 g/L Huabao No. 1 +1 mg/L6-BA +30 g/L potato extract +0.5 g/L AC; the rooting culture medium is 1/2MS + 2g/L Huabao No. 1 +30 g/L potato extract +0.5 g/L AC, 4-5 unfolded leaves are arranged after 90d (figure 3), an excellent single plant is selected according to the aspects of the whole plant state, internode thickness, leaf shape, vein veins and the like and inoculated into a subculture multiplication culture medium, the components of the multiplication culture medium are 1/2MS +1 g/L Huabao No. 1 +2.0 mg/L6-BA, the excellent single plant is multiplied for 4 times to a certain amount and then inoculated into a rooting culture medium, and the rooting culture medium is 1/2MS + 2g/L Huabao No. 1 +30 g/L potato extract +0.5 g/L AC (activated carbon). Taking out of bottles for domestication cultivation after rooting cultivation for 60 d. And (4) wrapping and transplanting the seedlings with the cleaned roots to 105-hole trays for planting by using the sphagna, and culturing in a greenhouse. The greenhouse environment is controlled at 26 ℃, the humidity is kept at about 90 percent, and the shading degree is 85 percent. 30 And (4) watering the water soluble fertilizer for 20-20-20 days every 15 days after the d. When 4-6 leaves grow out, observing and evaluating the specificity, consistency and stability of the strain from the aspects of plant overall state, internode thickness, leaf shape, vein and the like, and screening the strain with specificity, consistent single plant expression and stable character as a new variety of the cymbidium sinense.

FIG. 4 is a diagram of the breeding of a new variety of the "Hongyu" of the cymbidium sinense of the invention; FIG. 5 is a diagram of the breeding of a new variety of the Heiyu orchid of the invention; FIG. 6 is a diagram of the breeding of a new variety of Maolan "Maoyu" of the present invention.

The above disclosure is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the scope of the present invention, therefore, the present invention is not limited by the appended claims.

Claims

1. A cultivation method of a new species of cymbidium sinense is characterized by comprising the following steps:

(1) Collecting strong plant, short and thick internode, large leaf and rich leaf vein reticulate pattern materials from the field as parents, carrying out alloplastic selfing in the flowering season of 2 months, and picking when the capsule seed coat is changed from green to brown but the seeds are not cracked;

(2) Sterilizing the surface of picked cymbidium sinense seeds with 75% ethanol for 30s on an aseptic operation platform, washing with sterile water for 3 times, transferring to 0.1% mercuric chloride solution for sterilization for 8 min, shaking occasionally, washing with sterile water for 3 times, and inoculating to a germination culture medium; the components of the germination culture medium are 1/2MS +2.0 g/L Huabao No. 1 +0.5 mg/L NAA +30 g/L potato extract + 2g/L hydrolyzed casein;

(3) The seeds are swollen and expanded to break through the seed coat after 7 d, a milky round bulb is formed after 15 d, fluff is densely grown on the surface of the round bulb along with the formation of the round bulb, the round bulb is elongated after 60 d, leaves grow on the top of the round bulb, the round bulb is inoculated into a rooting culture medium after being subcultured for 3 times in the culture medium, and the subculture medium is 1/2MS +1 g/L Huabao No. 1 +0.5 mg/L6-BA +30 g/L potato extract +0.5 g/L AC; the rooting culture medium is 1/2MS + 2g/L Huabao No. 1 +30 g/L potato extract +0.5 g/L AC, and 4-5 unfolded leaves exist after 90 days;

(4) Selecting excellent single plants according to the aspects of plant integral state, internode thickness, leaf shape and leaf vein, inoculating the excellent single plants into a subculture multiplication culture medium, wherein the components of the multiplication culture medium are 1/2MS +1 g/L Huabao No. 1 + 1.0 mg/L6-BA, and inoculating the excellent single plants into a rooting culture medium after the excellent single plants are multiplied to a certain amount for 4 times, wherein the rooting culture medium is 1/2MS + 2g/L Huabao No. 1 +30 g/L potato extract +0.5 g/L AC;

(5) Rooting culture for 60 d, taking out of bottles for domestication and cultivation, wrapping and transplanting seedlings with cleaned roots to 105-hole trays for planting by using water moss, placing in a greenhouse for cultivation, controlling the greenhouse environment at 24 ℃, keeping the humidity at about 90%, keeping the shading degree at 85%, pouring 20-20-20 water soluble fertilizer every 15 d after 30 d, observing and evaluating the specificity, consistency and stability of strains in terms of the whole state, internode thickness, leaf shape and vein pattern of the plants when the plants grow 4-6 leaves, and screening the strains with specificity, consistent performance among single strains and stable properties to be new species of the cymbidium bicolor.