CN103004603A - Plant regeneration method for butterfly orchid - Google Patents

Abstract

The invention discloses a plant regeneration method of a Phalaenopsis variety, belonging to the field of Phalaenopsis cultivation. Select the axillary buds of the pedicels with a length of 2 to 5 cm on the Phalaenopsis plant as the explant material. After 2 to 3 weeks of induction culture at the shoot tip, the protocorm will gradually form in 25 to 30 days, and it will be continuously divided for subculture and accelerated propagation. , transplant 1 cm high seedlings into strong seedling medium for 3-6 months, induce rooting in 20-25 days and grow to 8-10 cm high before transplanting, the survival rate is as high as 95% or more; after the seedlings survive Strong growth, good growth. The invention can establish aseptic strains of high-quality Phalaenopsis varieties in a short period of time for large-scale industrial production, and can be produced every year, and the reproduction speed is fast, providing consumers with products with new, fresh, fragrant and colorful flowers in various colors and consistent shapes. ;Provide enterprises with high-tech production technology, make the plants consistent in size, facilitate flowering control and industrialized unified management, reduce production costs of enterprises, and bring huge economic benefits.

Description

Plant regeneration method of a kind of Phalaenopsis species

technical field

The invention belongs to the technical field of rapid propagation of Phalaenopsis plants, and in particular relates to a method for plant regeneration of Phalaenopsis varieties.

Background technique

Phalaenopsis, also known as Phalaenopsis, belongs to the genus Phalaenopsis, is a perennial single-stem epiphytic orchid, native to Asia, mainly distributed in the Philippines, the Malay Peninsula and Taiwan Province of my country. Phalaenopsis is a well-known cut flower species. Its plant shape is beautiful, with short stems, large leaves, one to several flower stems, arched, large flowers, and long-lasting flowering. The flower shape is like a butterfly flying, and the color is gorgeous. Its beautiful flowers and gorgeous colors are treasures of tropical orchids. It has the reputation of "Queen of Orchids". It is one of the most widely cultivated and popular species in the orchidaceae. There are more than 530 varieties through hybridization, but the ornamental value of native species is poor. Most of the varieties commercially used for large-scale production are hybrids, which have many varieties and long flowering periods, and are deeply loved by people. The scientific name of Phalaenopsis is "an orchid like a butterfly" according to the Greek original meaning. It can survive by absorbing nutrients in the air, and it belongs to the category of aerial orchids, which can be said to be a big family of tropical orchids. Its plants are very peculiar, neither stolons nor pseudobulbs. Each tree only grows a few thick broad leaves like spoons, which are stacked alternately on the base. The thick white aerial roots are exposed around the leaves, and some cling to the outer wall of the flower pot, which is very natural and wild. In the Spring Festival, a branch of flower stalks with a length of Yingchi will emerge from the leaf axils, and then bloom one by one. Each flower has 5 twilights, with lips inlaid in the middle.

Its growth habit likes high temperature, high humidity, and semi-shady environment, and the wintering temperature is not lower than 18 degrees. Because Phalaenopsis is native to tropical rainforest areas, it likes warmth and chills by nature. The optimum temperature for growth is 15-20°C, and it will stop growing below 10°C in winter, and it is easy to die below 5°C. If mass production is to be carried out in various parts of Lingnan, it is necessary to have cold-proof facilities and implement protective cultivation. If the family plants a small amount, it can survive the winter safely by moving it indoors immediately when it is cold. Most of its propagation adopts cell tissue culture, and the seedlings are transplanted through test tubes, and they can bloom after about two years. When the flowering period of some mother plants is over, sometimes the axillary buds on the pedicel will also grow and develop into daughter plants. When it grows roots, it can be cut from the pedicel for branch reproduction. The potted plant material should not use soil, but sphagnum moss, pumice, spinach chips, charcoal chips, etc., or directly fix the seedlings on the corrugated board (also known as snake wood), so that it can attach and grow by itself. This cultivation method is to imitate its original ecological environment. When it blooms, hang the whole board on the wall to watch, it really has a special charm. Phalaenopsis has a lot of aerial roots, and its root tips are emerald green and very sensitive. They must be carefully protected and must not be touched or damaged. Otherwise, it will be like breaking a person's mouth, and it will be difficult to eat normally. Phalaenopsis likes a humid and semi-shady environment, and requires the space to maintain a humidity of 50% to 70% frequently, and the pot should not be watered too much. If it is planted on the balcony of a home, it is best to sprinkle the ground three or four times a day on sunny days, so that the plants can absorb the evaporated water. Do not allow direct sunlight in summer and autumn. But the morning sun is the best for its growth and should be fully utilized. If there are too many cloudy and rainy days in spring, use a light tube to increase light for it at night to facilitate flowering in the future. Phalaenopsis has weak resistance to pests and diseases, and leaf spot and root rot often occur. It can be sprayed with a 1000-fold solution of the pesticide chlorothalonil or Daxianchong, once every seven or eight days, and sprayed three times in a row. These medicinal liquids leave white traces on the leaves, which do not need to be wiped off, so as to continue to play a bactericidal effect.

The flowers of Phalaenopsis are bright and eye-catching, including pure white and light yellow. Triple red, but also lavender, orange red and blue. There are many varieties with two or three colors, some seem to be embroidered with patterned stripes, and some are like sprayed with evenly colored dots. Each branch has seven or eight flowers, and there are more than twelve or three. sky. When all are in full bloom, it seems that a group of butterflies are flying gently. Its elegant and leisurely mood really makes people feel picturesque, dreamlike and dreamlike. Among them, the ones with yellow flowers are more expensive, and there is a yellow-flowered variety called "Tianhuang", which can be called a "superstar", and the bargain is very expensive. As for the orchid species, they are relatively rare. At the International Orchid Exposition in 1952 and 1953, the Phalaenopsis presented by Taiwan won gold medals for two consecutive years. In the 14th orchid exhibition held in Hong Kong in 1989, a Phalaenopsis with white petals and red lips presented by Mr. Hu Bingchi won the overall championship award. These outstanding awards have laid a solid foundation for the development of Phalaenopsis. Nowadays, the consumption of Phalaenopsis by people from various countries in Europe and the United States is constantly increasing, and Phalaenopsis is indispensable for all high-end banquets. Phalaenopsis of various colors represent different meanings. White Phalaenopsis represents pure love and precious friendship. Red Phalaenopsis represents good fortune and eternal unity. Red Phalaenopsis represents smooth career and happiness. Everything goes well and all wishes come true. The yellow Phalaenopsis represents a prosperous career and business, and the mini Phalaenopsis represents a happy angel and a prosperous life. the

In ancient my country, orchids, together with plum, bamboo, and chrysanthemum, were called the four famous flowers, and they were called "four gentlemen". The queen of orchids, Phalaenopsis, has fascinated countless poets and gentlemen with its outstanding demeanor. It adds infinite color and vitality to the home, and at the same time improves the air quality of the small environment, absorbs toxic waste gas, releases pleasant fresh air, and adds infinite dynamic vitality. Through the Orchid Exhibition, many people's minds have been cultivated, so that people unconsciously reject filth and evil, clean themselves, and experience the good atmosphere of consolidating roots, uniting and self-improvement, so as to achieve selfless dedication and spread fragrance in the world. . Gu has often become the favorite of literary masters, often seen in writing and ink, and has been handed down for generations. Phalaenopsis is also a "three-high product" of great commercial value. It has a wide range of medicinal and edible values. However, due to people's understanding of its research and cultivation, the mystery and value of orchid plants are far from being recognized. It is recognized that, in view of this, the present invention provides technical support for the development of my country's orchid industry through the research on plant regeneration and industrialization key technology development of various flower and color Phalaenopsis varieties.

Contents of the invention

The object of the present invention is to provide a plant regeneration method of a Phalaenopsis variety, which solves the problem of slow plant propagation speed, low seed germination rate, serious viral disease, inconsistent plant growth cycle, uneven flowering time in the prior art, and cannot be controlled in large quantities. The flowering period directly affects the quality, sales quantity and price of orchids, as well as storage and transportation. The measures taken by the present invention can make high-quality Phalaenopsis varieties establish aseptic strains in a short period of time to carry out mass industrial production, and can be produced every year, with fast reproduction speed, and provide consumers with new, fresh, fragrant and bright colors. Products with consistent flower shapes; provide enterprises with high-tech production technology to make their plants consistent in size, which is convenient for flowering period control and unified management of industrialization, and brings huge benefits for reducing production costs of enterprises and providing high-quality products to stabilize market sales. economic benefits.

To achieve the above object, the present invention adopts the following technical solutions:

The plant regeneration method of a kind of Phalaenopsis variety, selects the plant of Phalaenopsis variety, cuts the pedicel axillary bud stem segment with a length of 2 to 5 cm from a strong mother plant as explant material, and induces and cultivates the stem tip for 2 to 3 weeks , the growth point starts to turn green, the stems and leaves start to grow first, and then the base of the growth point begins to hypertrophy, and the protocorm gradually forms in 25 to 30 days. Newly formed seedlings with a height of 1 cm are moved into a larger culture container on a strong seedling medium and cultivated for 3 to 6 months. After 20 to 25 days of induced rooting, they grow to a height of 8 to 10 cm before transplanting. Specifically include the following steps:

(1) Material collection method: Select several Phalaenopsis varieties with no diseases and insect pests introduced from Taiwan, and cut the pedicel axillary bud stem section with a length of 2 to 5 cm from the strong mother plant as the explant material;

(2) Preparation of medium:

Bud induction medium: MS +1.0mg/L BA + 1.0mg/L NAA +6.5 g/L Ag +30 g/L Su +2-2.5 g/L Ac +50 g/L coconut water +1-3 g /L Peptone, the pH value is 5.6;

Cluster bud induction medium: 1/2MS +3.0mg/L BA + 0.5mg/L NAA +6.5 g/L Ag +30 g/L Su +2-2.5 g/L Ac +150 g/L coconut milk +1 -3 g/L Peptone, pH 5.6;

Bud Proliferation Medium: 1/2MS+2.0mg/L BA + 0.3g/L NAA + 6.5 g/L Ag +30 g/L Su +2-2.5 g/L Ac +100 g/L Banana Juice+1- 3 g/L Hyponex1+1-3 g/L Peptone, pH 5.6;

Rooting medium: 1/2 MS + 0.5mg/L IBA +0.3mg/L NAA +6.5 g/L Ag +30 g/L Su +2-2.5 g/L Ac, pH 5.6;

(3) Material treatment: Cut the flower stalks of several Phalaenopsis orchids with side buds and no pests and diseases into 2~3cm segments, rinse them with tap water, and then soak them in saturated bleach supernatant for 15 minutes, stirring constantly during soaking; After soaking, the side buds of the pedicels are dripped under tap water for 1 hour, then drained, sterilized and shaken with 75% alcohol in an ultra-clean workbench for 3-5 seconds, poured out the alcohol, and added 0.1% chlorine Treat the mercury solution for 8-12 minutes, pour the mercury solution into the waste mercury bottle, rinse it with sterile water for 3-4 times, and dry the surface moisture with sterile filter paper;

(4) Induction culture: cut the peduncle after strict disinfection with a scalpel into 1 bud and 1 segment and inoculate it in the bud induction medium;

(5) Culture conditions: the temperature of the culture room is 25±2°C, the light time is 12h/d, and the light intensity is 1500-2000lx;

(6) Cluster bud induction culture: Cut the induced buds that were induced and grown robustly into small pieces of 0.5 cm, and inoculate them in the bud induction medium respectively; after 13 to 15 days, the shoot tips begin to expand and appear light green hemispherical , induce the protocorm after 3 months;

(7) Proliferation culture: Cut the well-grown protocorm bodies of the above induction culture into small pieces, and inoculate them in the bud proliferation medium for subculture; subculture once every 50-60 days, and the multiplication factor is 10-13. The corm is placed on the medium containing low concentration of cytokinin and continues to be cultured for 60 days, and after 60 days, small buds grow out successively, becoming a complete protocorm; after 95 days, most of them grow into small seedlings with 2 to 3 leaves; a side bud of a pedicel germinates to form a small seedling , and then use the leaves of the seedlings to induce the protocorm to proliferate, the test tube seedlings are planted out of the bottle, and the growth characteristics are stable, and the method of protocorm multiplication greatly improves the propagation speed, and realizes the industrial production of Phalaenopsis;

(8) Rooting induction: when the clustered shoots grow to a height of 2 cm, the shoots are divided into individual plants and transferred to different rooting media;

(9) Transplanting of seedlings: When the test-tube seedlings grow to 3-4cm high, have 3-5 roots, 5-7 leaves, and 2-3cm in length, the seedlings can be hardened, and the test-tube seedlings can be hardened under natural light. The seedlings are 5 to 7 days old, and the bottle cap is opened to harden the seedlings for 2 to 3 days to enhance the adaptability of the test tube seedlings to the outdoor environment; then take out the seedlings from the culture bottle, wash the root medium, and wrap the roots with clean water moss Plant in plug trays, keep the temperature at 25~30°C, and the air humidity at 85%, for 25~30 days, when new leaves and roots grow, spray KH ₂ PO ₄ solution with a mass fraction of 0.3%~0.5% , once every 7 days, the survival rate reached 95%.

The beneficial effects of the present invention are: the present invention can establish aseptic strains of high-quality Phalaenopsis varieties in a short period of time for large-scale industrial production, and can be produced every year, and the reproduction speed is fast, providing consumers with fresh, fresh and fragrant varieties of various colors. , products with the same shape of bright flowers; provide enterprises with high-tech production technology to make the plants consistent in size, facilitate flowering control and unified management of industrialization, and reduce production costs for enterprises and provide high-quality products to stabilize market sales. huge economic benefits.

Detailed ways

The present invention will be further described below in conjunction with embodiment.

Example 1:

(1) Material collection method: Select several Phalaenopsis varieties with no diseases and insect pests introduced from Taiwan, and cut the pedicel axillary bud stem section with a length of 2 to 5 cm from the strong mother plant as the explant material;

(2) Preparation of medium:

Bud induction medium: MS +1.0mg/L BA + 1.0mg/L NAA +6.5 g/L Ag +30 g/L Su +2-2.5 g/L Ac +50 g/L coconut water +1-3 g /L Peptone, the pH value is 5.6;

Cluster bud induction medium: 1/2MS +3.0mg/L BA + 0.5mg/L NAA +6.5 g/L Ag +30 g/L Su +2-2.5 g/L Ac +150 g/L coconut milk +1 -3 g/L Peptone, pH 5.6;

Bud Proliferation Medium: 1/2MS+2.0mg/L BA + 0.3g/L NAA + 6.5 g/L Ag +30 g/L Su +2-2.5 g/L Ac +100 g/L Banana Juice+1- 3 g/L Hyponex1+1-3 g/L Peptone, pH 5.6;

Rooting medium: 1/2 MS + 0.5mg/L IBA +0.3mg/L NAA +6.5 g/L Ag +30 g/L Su +2-2.5 g/L Ac, pH 5.6;

(3) Material treatment: Cut the flower stalks of several Phalaenopsis orchids with side buds and no pests and diseases into 2~3cm segments, rinse them with tap water, and then soak them in saturated bleach supernatant for 15 minutes, stirring constantly during soaking; After soaking, the side buds of the pedicels are dripped under tap water for 1 hour, then drained, sterilized and shaken with 75% alcohol in an ultra-clean workbench for 3-5 seconds, poured out the alcohol, and added 0.1% chlorine Treat the mercury solution for 8-12 minutes, pour the mercury solution into the waste mercury bottle, rinse it with sterile water for 3-4 times, and dry the surface moisture with sterile filter paper;

(4) Induction culture: cut the peduncle after strict disinfection with a scalpel into 1 bud and 1 segment and inoculate it in the bud induction medium;

(5) Culture conditions: the temperature of the culture room is 25±2°C, the light time is 12h/d, and the light intensity is 1500-2000lx;

(6) Cluster bud induction culture: Cut the induced buds that were induced and grown robustly into small pieces of 0.5 cm, and inoculate them in the bud induction medium respectively; after 13 to 15 days, the shoot tips begin to expand and appear light green hemispherical , induce the protocorm after 3 months;

(7) Proliferation culture: Cut the well-grown protocorm bodies of the above induction culture into small pieces, and inoculate them in the bud proliferation medium for subculture; subculture once every 50-60 days, and the multiplication factor is 10-13. The corm is placed on the medium containing low concentration of cytokinin and continues to be cultured for 60 days, and after 60 days, small buds grow out successively, becoming a complete protocorm; after 95 days, most of them grow into small seedlings with 2 to 3 leaves; a side bud of a pedicel germinates to form a small seedling , and then use the leaves of the seedlings to induce the protocorm to proliferate, the test tube seedlings are planted out of the bottle, and the growth characteristics are stable, and the method of protocorm multiplication greatly improves the propagation speed, and realizes the industrial production of Phalaenopsis;

(8) Rooting induction: when the clustered shoots grow to a height of 2 cm, the shoots are divided into individual plants and transferred to different rooting media;

(9) Transplanting of seedlings: When the test-tube seedlings grow to 3-4cm high, have 3-5 roots, 5-7 leaves, and 2-3cm in length, the seedlings can be hardened, and the test-tube seedlings can be hardened under natural light. The seedlings are 5 to 7 days old, and the bottle cap is opened to harden the seedlings for 2 to 3 days to enhance the adaptability of the test tube seedlings to the outdoor environment; then take out the seedlings from the culture bottle, wash the root medium, and wrap the roots with clean water moss Plant in plug trays, keep the temperature at 25~30°C, and the air humidity at 85%, for 25~30 days, when new leaves and roots grow, spray KH ₂ PO ₄ solution with a mass fraction of 0.3%~0.5% , once every 7 days, the survival rate reached 95%.

The above descriptions are only preferred embodiments of the present invention, and all equivalent changes and modifications made according to the scope of the patent application of the present invention shall fall within the scope of the present invention.

Claims (2)

1. the plant regeneration method of a butterfly orchid variety, it is characterized in that: the plant of choosing butterfly orchid variety, cutting length from healthy and strong maternal plant is that 2 ~ 5 centimetres bennet axillary bud stem section is explant material, the cultivation of inducing through 2 ~ 3 weeks of stem apex, it is green that growing point begins to be, cauline leaf begins first growth, then the base portion of growing point begins hypertrophy, formed gradually protocorm in 25 ~ 30 days, the protocorm that forms is by constantly cutting apart, carry out the subculture accelerates multiplication until obtain requirement, cultivated 3 ~ 6 months on the culture vessel strong seedling culture base that the seedling immigration of 1 centimetre of high new formation is larger, length can be transplanted to 8 ~ 10 centimetres high after the root induction in 20～25 days.

2. the plant regeneration method of butterfly orchid variety according to claim 1 is characterized in that: may further comprise the steps:

(1) method of drawing material: choose the plant without some pattern butterfly orchid varieties of damage by disease and insect of introducing from Taiwan, cutting length from healthy and strong maternal plant is that 2 ~ 5 centimetres bennet axillary bud stem section is explant material;

(2) preparation of medium:

Bud inducing culture: MS+1.0mg/L BA +1.0mg/L NAA+6.5 g/L Ag+30 g/L Su+2-2.5 g/L Ac+50 g/L coconut milk+1-3 g/L Peptone, the pH value is 5.6;

Clump bud inducing culture: 1/2MS+3.0mg/L BA +0.5mg/L NAA+6.5 g/L Ag+30 g/L Su+2-2.5 g/L Ac+150 g/L coconut milk+1-3 g/L Peptone, the pH value is 5.6;

Bud proliferated culture medium: 1/2MS+2.0mg/L BA +0.3g/L NAA+6.5 g/L Ag+30 g/L Su+2-2.5 g/L Ac+100 g/L bananas juice+1-3 g/L Hyponex1+1-3 g/L Peptone, the pH value is 5.6;

Root media: 1/2 MS+0.5mg/L IBA+0.3mg/L NAA+6.5 g/L Ag+30 g/L Su+2-2.5 g/L Ac, the pH value is 5.6;

(3) material processed: will with lateral bud be cut into the sections of 2 ~ 3cm without some pattern Moth orchid bennets of damage by disease and insect, with the running water flushing, in saturated bleaching powder supernatant, soak 15min again, to constantly stir during immersion; Bennet lateral bud after the immersion drains after 1 hour in lower punching of running water, in superclean bench, after swaying for 3 ~ 5 seconds, the alcohol disinfecting of mass concentration as 75% pour out alcohol, adding mass fraction and be 0.1% mercuric chloride solution processed 8 ~ 12 minutes, pour mercury solution into useless mercury bottle, after sterile water punching 3 ~ 4 times, blot surface moisture with sterilized filter paper;

(4) induce cultivation: the bennet after strict sterilization is cut into 1 section material of 1 bud with scalpel is seeded in the bud inducing culture;

(5) condition of culture: culturing room's temperature is 25 ± 2 ℃, light application time 12h/d, intensity of illumination 1500～2000lx;

(6) the clump bud is induced cultivation: above-mentioned induced bud of inducing cultivation, robust growth is cut into the fritter of 0.5cm, is seeded in respectively from the bud inducing culture; Stem apex begins to expand after 13 ~ 15 days, is light green hemispherical, induces the protocorm caulome after 3 months;

(7) propagation is cultivated: the above-mentioned good protocorm caulome of incubation growth of inducing is cut into fritter, be seeded in respectively bud proliferated culture medium relaying culture; 50 ~ 60 days subcultures once, the propagation multiple is 10 ~ 13, protocorm is placed on the medium that contains the low concentration basic element of cell division to continue to cultivate after 60 days to grow successively budlet, becomes complete protocorm; After 95 days, major part grows up to the seedling of 2 ~ 3 leaves; Bennet sprouting of lateral bud forms seedling, and the blade of recycling seedling is induced the protocorm shoot proliferation, and test-tube plantlet Planting out of test-tube growth traits is stable, and the method by Protocorm Multiplication greatly improves reproduction speed, realizes the industrialization production of Moth orchid;

(8) root induction: when Multiple Buds grows to 2cm when high, bud is divided into individual plant, transfers in the different root medias;

(9) little transplantation of seedlings: to grow to 3～4cm high when test-tube plantlet, 3～5 roots arranged, 5～7 blades, during length 2～3cm, just can carry out hardening, test-tube plantlet was placed on the natural daylight lower refining seedling 5～7 days, open bottle cap and carried out first hardening 2～3 days, to strengthen test-tube plantlet to the adaptive capacity of outdoor environment; Then take out seedling from blake bottle, clean the root medium, plant in the dish of cave with clean water tongue parcel root, keep 25 ~ 30 ℃ of temperature, air humidity is 85%, 25 ~ 30 days, when young leaves, when newly root grows, sprays mass fraction and be 0.3% ~ 0.5% KH ₂PO ₄Solution, once survival rate reached 95% in per 7 days.