CN115251057B

CN115251057B - Method for inducing germination of phalaenopsis seedlings by using plant hormone composition

Info

Publication number: CN115251057B
Application number: CN202211030294.3A
Authority: CN
Inventors: 许申平; 张腾飞; 袁秀云; 张燕; 梁芳; 王默霏; 蒋素华; 牛苏燕; 崔波
Original assignee: Zhengzhou Normal University
Current assignee: Zhengzhou Normal University
Priority date: 2022-08-26
Filing date: 2022-08-26
Publication date: 2023-07-25
Anticipated expiration: 2042-08-26
Also published as: CN115251057A

Abstract

The invention belongs to the technical field of plant cultivation, and particularly relates to a method for inducing germination of phalaenopsis seedlings by using a plant hormone composition. The invention firstly provides a phytohormone composition which is formed by mixing 10.0-25.0 mg/L of DA-6, 1.0-5.0 mg/L of CPPU, 5.0-10.0 mg/L of 6-BA and 0.5-1.0 mg/L of NAA. According to the method provided by the invention, through adopting the plant hormone composition with a specific proportion and regulating and controlling the environmental conditions of the butterfly orchid seedlings, after the method is adopted, 12 seedlings (3 buds are arranged on each butterfly orchid pedicel and 4 seedlings are differentiated from each bud) can be induced per butterfly orchid, so that the method not only has a higher seedling yield, but also can induce and generate seedlings from the pedicel bases of different butterfly orchids. The method is simple to operate, can realize industrial cultivation of the butterfly orchid and improves ornamental quality of the butterfly orchid.

Description

Method for inducing germination of phalaenopsis seedlings by using plant hormone composition

Technical Field

The invention belongs to the technical field of plant cultivation, and particularly relates to a method for inducing germination of phalaenopsis seedlings by using a plant hormone composition.

Background

The butterfly orchid is a perennial tropical epiphytic CAM (Crassulaceae) plant of the genus Phalaenopsis (Phalaenopsis), is often used as a pot plant or cut flower, and is one of the most popular four-ocean orchids in the international flower market. The butterfly orchid is a single-stem aerial orchid, plants rarely develop on lateral branches, seeds of the butterfly orchid do not contain endosperm or other tissues, the germination rate is extremely low under natural conditions, and the factors directly restrict the large-scale production of the butterfly orchid.

At present, two main methods exist for propagation of butterfly orchid: one is sexual reproduction, namely, a seed sowing mode is adopted for hybridization and fructification, also called a sowing propagation method, and the method has high offspring mutation rate and is commonly used for breeding new varieties, but not for large-scale production; the other is to use advanced tissue culture technology to perform asexual propagation. In recent years, scholars at home and abroad have studied a great deal on the tissue culture technology of the butterfly orchid, and the explants of the butterfly orchid tissue culture induced protocorms comprise stem tips, leaves, peduncle axillary buds, peduncle internode segments and the like, so that the induced protocorms are further induced to realize rapid propagation. Wherein, the axillary buds of the butterfly orchid peduncles are used as explants, and a bud growing mode is adopted to become the most commonly used rapid propagation method of butterfly orchid seedlings.

Although tissue culture techniques have the advantage of rapid propagation of the seedling and the ability to maintain the superior properties of the female parent. However, not only is tissue culture under specific aseptic conditions, but also specific requirements are imposed on the technology, specific culture media are required to be prepared according to different varieties, and the cultured seedlings also need to be domesticated before transplanting.

Among the numerous influencing factors of the butterfly orchid tissue culture, temperature is the primary environmental factor influencing the flowering of butterfly orchid. The butterfly orchid is in the growing and developing process, including a nutrition growing stage and a reproduction growing stage, and the butterfly orchid can be transferred into reproduction growth to carry out flower bud differentiation after a period of low-temperature stimulation after the nutrition growing stage, otherwise, the butterfly orchid can not bloom. The high temperature can inhibit the formation of flower buds of the butterfly orchid in the early growth stage, and the flower buds can be inhibited when the temperature is higher than 28 ℃. Therefore, the environmental temperature is often increased in the cultivation of butterfly orchid to prolong the vegetative growth stage of butterfly orchid, thereby obtaining high-quality flowering plants. When the environmental conditions are suitable, the flower buds break through leaf sheaths and sprout from axils, but the sprouted flower buds have different development directions according to the environmental conditions and the differentiation degree. Early researches show that when the flower bud length is less than 5cm, the butterfly orchid plant grows in an environment of 28 ℃, the flower bud can be reversed into a nutrition plant, or the flower bud is not opened but is infinitely elongated; when the flower bud length reaches 10cm, the flower petal primordia begin to differentiate, and when the butterfly orchid plant is placed in the environment of 28 ℃, the growth of the flower bud can be accelerated to bloom early.

The illumination also plays a very important role in the induction of flower buds of the butterfly orchid, wherein the illumination intensity is one of determining factors of the induction of flower buds of the butterfly orchid. The lower illumination intensity is beneficial to the nutrition growth of the butterfly orchid, and the higher illumination intensity is beneficial to the reproduction growth of the butterfly orchid. The flowering rate gradually increases along with the increase of illuminance within a certain range, and researches show that the butterfly orchid seedlings are respectively controlled under the weak illuminance (50 mu mol.m ^-2 ·s ^-1 ) Illuminance (100. Mu. Mol.m) ^-2 ·s ^-1 ) Intense light illuminance (200. Mu. Mol.m) ^-2 ·s ^-1 ) The flowering rates after 2 years are 12%, 77% and 98% respectively, and the stem extraction rate of the butterfly orchid is gradually increased, the growth speed of the pedicel is increased, the number of flowers is increased and the flowering period is advanced along with the illumination intensity.

Therefore, promotion of germination of butterfly orchid buds and seedlings by induction is an effective means. For example, chinese patent publication No. CN103988780a discloses a method for directly inducing adventitious buds of butterfly orchid, which can induce the production of adventitious buds and promote the rapid growth of plants by using hormone combination and anti-brown solution treatment. However, in the patent, the later induction period is required to be inoculated in a culture flask in a tissue culture chamber, the environment is severely required, the operation is complex, the growth period is long, simple, convenient, efficient and quick acquisition of the phalaenopsis seedlings cannot be realized, and the improvement of the seedling yield cannot be realized. Therefore, a simple and efficient butterfly orchid seedling cultivation technology is urgently needed.

In conclusion, the high temperature weak light is beneficial to the nutrition growth of the butterfly orchid. The inventor of the invention adopts a specific plant germination agent and a method for regulating and controlling conditions such as ambient temperature, illumination and the like so as to realize simple and efficient induction germination of specific phalaenopsis seedlings.

Disclosure of Invention

The invention aims to provide a method for inducing the germination of phalaenopsis seedlings by using a plant hormone composition, which can realize the rapid induction of the phalaenopsis seedlings by adopting the plant hormone composition with a specific proportion and regulating and controlling the environmental conditions of the phalaenopsis seedlings, has higher seedling yield and can induce the generation of the seedlings from the pedicel bases of the phalaenopsis of different varieties. The method is simple to operate, can realize industrial cultivation of the butterfly orchid and improves ornamental quality of the butterfly orchid.

In order to achieve the above purpose, the present invention adopts the following technical scheme:

a plant hormone composition for inducing phalaenopsis seedlings comprises DA-6 (diethyl aminoethyl hexanoate DA-6), CPPU (chlorpyribenzoxazole), NAA (naphthalene acetic acid) and 6-BA (benzylaminopurine), wherein the plant hormone composition is formed by mixing 10.0-25.0 mg/L of DA-6, 1.0-5.0 mg/L of CPPU, 5.0-10.0 mg/L of 6-BA and 0.5-1.0 mg/L of NAA.

Specifically, the plant hormone composition is prepared by the following method:

1) Dissolving DA-6 in water to obtain a solution A; dissolving CPPU and NAA in an organic solvent I to obtain a solution B; dissolving 6-BA in NaOH solution (or KOH solution) to obtain solution C;

2) Mixing the solution A, B, C to obtain a mixed solution, wherein the concentration of DA-6 is 10.0-25.0 mg/L, the concentration of CPPU is 1.0-5.0 mg/L, the concentration of 6-BA is 5.0-10.0 mg/L, and the concentration of NAA is 0.5-1.0 mg/L;

3) Heating and melting the semi-solid plant hormone carrier, placing the semi-solid plant hormone carrier into the mixed solution in the step 2), uniformly mixing, and cooling to room temperature to obtain a solid product, namely the plant hormone composition.

Specifically, the organic solvent I is absolute ethanol, acetone or methanol.

Specifically, the ratio of the semi-solid phytohormone carrier to the mixed solution in the step 3) is (7-10) g:1ml.

Further preferably, the semi-solid phytohormone carrier is lanolin or vaseline, the vaseline has good waterproof effect, and the vaseline can be used as semi-solid lipid phytohormone carrier instead of lanolin when being used in spraying equipment.

The application of the plant hormone composition in the induction of the germination of the phalaenopsis seedlings, specifically, the application of the plant hormone composition in the reproductive growth stage of the phalaenopsis can induce the generation of the seedlings from the root parts of the pedicel.

Specifically, the butterfly orchid is a total inflorescence variety with more peduncle axillary buds, and specifically includes: "big capsicum", "Fule sunset" or "Neishan girl".

The method for inducing the germination of the phalaenopsis seedlings by using the plant hormone composition comprises the following steps:

(1) Pretreatment: applying fertilizer 1-2 times per week in the flowering period of butterfly orchid to ensure that the plants have sufficient nutrition for the induction of the stem bud seedlings in the later period;

(2) Induction treatment: after the butterfly orchid flowers wither, cutting off the withered flowers from the basal parts of the pedicel, stripping off bracts covered by axillary buds of the pedicel, exposing bud points in internodes, applying the plant hormone composition once every 4 days, and applying for 4-6 times;

(3) Post-induction growth management: light-shielding treatment in the first month of induced growth, the illumination intensity is less than 50mu mol.m ^-2 ·s ^-1 Adjusting the ambient temperature to 28-32deg.C, applying fertilizer during the period, and 1-2 times per week;

expanding and differentiating axillary buds of the flower stalks in the second month, removing a covered sunshade net on the 30 th day of cutting inflorescences, and carrying out light treatment, wherein the maximum light intensity is not more than 15000Lux; adjusting the ambient temperature to 24-30 ℃; and fertilizer is applied 1-2 times per week.

Specifically, the phalaenopsis in the step (1) is a total inflorescence variety with more peduncles and axillary buds, for example, the following: "big capsicum", "Fule sunset" or "Neishan girl".

Specifically, in the step (1), the butterfly orchid is a mature seedling flowering plant of 2.5 inches, 2.8 inches or 3.5 inches.

Specifically, the number of applications of the phytohormone composition in step (2) is proportional to the number of differentiated germinated plants: if the application is carried out for 1-2 times, only one seedling grows up between each node; if applied more than 4 times, 4-6 seedlings are differentiated per internode.

Preferably, the phytohormone composition of step (2) is applied by the following specific procedures: the plant hormone composition is uniformly smeared on exposed internode nodes, each bud node is smeared with 4-6mg, 3 pedicel axillary buds are smeared once every 4 days, namely, the 1 st day, the 5 th day, the 9 th day, the 13 th day and the 17 th day of the inflorescence pruning are smeared for 5 times.

Preferably, the phytohormone composition of step (2) is applied by the following specific procedures: the plant hormone composition is uniformly smeared on exposed internode nodes, each bud node is smeared with 4-6mg, each plant is smeared with 1 pedicel axillary bud, and each plant is smeared for 2 times once every 4 days, namely, the 1 st day and the 5 th day of cutting off inflorescences respectively.

Specifically, the light-shading treatment in the step (3) is to cover plants with a material with a sun-shading rate of 100%.

Specifically, in the step (3), after 4 months of induced growth, rooting liquid is applied to promote rooting of plant seedlings, and when the root system grows to 2-4 cm, the seedlings can be cut off from the mother plant and planted independently.

Specifically, in the method for inducing the germination of the butterfly orchid seedlings, the fertilizer applied is 'Huaduo No. 1', and the fertilizer is applied by using 3000 times of the liquid of 'Huaduo No. 1' as the fertilizer.

Compared with the prior art, the invention has the advantages that:

the method for applying the plant hormone composition and combining environmental condition control is adopted to quickly induce the butterfly orchid seedlings, the selected material is the butterfly orchid after flowering (complete inflorescence), waste is avoided in material utilization, and after the method provided by the invention is adopted, 12 seedlings (3 buds are arranged on each butterfly orchid pedicel and 4 seedlings are differentiated from each bud) can be induced by each butterfly orchid, so that the method has higher single-plant seedling yield.

The method has the advantages of simple operation method, high seedling yield and good application value.

Drawings

FIG. 1 is a photograph of an upper bloom flower cut from the base of a pedicel at the 4 th to 5 th knots up during induction in example 1;

FIG. 2 is a photograph showing bud points in internodes of example 1, which were obtained by peeling off bracts covered by axillary buds of butterfly orchid stems during induction;

FIG. 3 is a photograph showing the expansion of the axillary buds of butterfly orchid flower stems smeared for 2 times in example 1 and an anatomic diagram, wherein the left figure is a photograph showing the expansion of the axillary buds of butterfly orchid flower stems, and the right figure is an anatomic diagram;

FIG. 4 is a photograph showing the axillary buds of the inflorescence of the spread 2 times butterfly orchid in example 1 grown into seedlings;

FIG. 5 is a photograph showing differentiation of seedlings after 4 applications in example 1;

FIG. 6 is a photograph of a seedling root induced by the method described in example 1 and a stem extracted after 6 months of inflorescence pruning;

FIG. 7 is a photograph showing the result of induction of individual seedlings in example 2;

FIG. 8 is a photograph showing the result of induction of butterfly orchid in comparative example 3.

Detailed Description

The present invention will be described in further detail with reference to examples, but embodiments of the present invention are not limited thereto. Unless specifically stated otherwise, the reagents, methods and apparatus employed in the present invention are those conventional in the art. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art.

The following experiments were conducted in 2021 at the institute of orchid engineering, zhengzhou institute of education.

Example 1

The embodiment provides a method for rapidly inducing butterfly orchid seedlings by combining phytohormone smearing and environmental condition control after the butterfly orchid flowers, which comprises the following specific steps:

1. formulation of phytohormone compositions

The plant hormone composition comprises DA-6, CPPU, NAA and 6-BA.

The specific preparation method comprises the following steps: 30mg of DA-6 was dissolved in 50mL of pure water to obtain solution A;

5mg of CPPU and 1mg of NAA are dissolved in 50mL of absolute ethyl alcohol to obtain solution B;

10mg of 6-BA was dissolved in 100mL of a 0.1N NaOH solution to give solution C;

heating and melting 1.8kg of lanolin in a water bath mode, adding the three solutions after melting, fully and uniformly stirring, and cooling to obtain a brown yellow pasty product, namely the plant hormone composition.

2. Rapid induction of butterfly orchid seedlings

The common butterfly orchid variety is adopted, and the total inflorescence variety with more peduncles and axillary buds is preferable; in this example, 3.5 inch mature seedlings were selected as the best flowering plants, and specific induction was carried out as follows:

(1) Pretreatment: selecting butterfly orchid 'big capsicum' as a test object, and supplying 3000 times of liquid 'Huaduo No. 1' as fertilizer in the flowering period for 1 week for 1 time so as to ensure that plants have sufficient nutrition for the induction of later-stage pedicel bud seedlings;

(2) Induction treatment: after the butterfly orchid flowers wither and have no ornamental value, cutting off flowers with upper part withered from the 4 th to 5 th sections upwards from the basal part of the flower stalks (shown in figure 1), stripping off bracts covered by axillary buds of the butterfly orchid flower stalks, exposing bud points in internodes (shown in figure 2), uniformly smearing the plant hormone composition on the exposed internode nodes by using cotton swabs, smearing about 5mg on each bud node, smearing 3 axillary buds of the flower stalks every 4 days, and 10 a.m: 00, namely, cutting off the inflorescence on the 1 st day, the 5 th day, the 9 th day, the 13 th day and the 17 th day, and smearing for 5 times;

fig. 3 is a photograph showing the expansion of axillary buds of butterfly orchid flower stem smeared for 2 times in example 1 and an anatomic diagram, and it can be seen from the figure that the axillary buds are expanded at this time, and the microscopic structure of paraffin sections is observed to show the shape of leaf buds. FIG. 4 is a photograph showing that 2 times of the spread of the inflorescence peduncles of the butterfly orchid in example 1 grow into seedlings, and it can be seen from the figure that two leaves are also grown on the axillary buds at this time to form seedlings. FIG. 5 is a photograph showing the differentiation of seedlings by applying 4 times in example 1, and it can be seen from the photograph that axillary buds are differentiated, and 6 seedlings are differentiated;

(3) First month growth management: light-shielding treatment is required in the first month of induced growth, and the illumination intensity is 30 mu mol.m ^-2 ·s ^-1 The daytime temperature is higher than or equal to 28 ℃ and lower than 32 ℃, 3000 times of liquid of Huaduo No. 1 is used as fertilizer to be supplied during the period, 1 time is carried out in 1 week, plant materials are covered by a sunshade net with 100% of sunshade rate, and the ambient temperature is adjusted to be 30 ℃ in the daytime and 28 ℃ at night;

(4) Second month growth management: in the second month, the axillary bud point of the pedicel can be seen to expand and differentiate, the covered sunshade net is removed on the 30 th day of cutting the inflorescence, the normal fertilizer and water management is completed, the illumination intensity is maintained to be natural illumination, and the maximum light intensity is not more than 15000Lux; humidity is 70-80%; the temperature of day and night is 28/24 ℃; 3000 times of liquid of Huaduo No. 1 is used as fertilizer to be supplied for 1 week and 1 time;

(5) From the second month, 50% carbendazim wettable powder is applied once a month, the wettable powder is diluted 500-800 times for spraying, and a 2000-time liquid of the cyromazine or the fast flutter is adopted for preventing and controlling red spiders or scale insects, a 3000-time liquid of the delphagostimulant is adopted for preventing and controlling white flies, after normal maintenance for 4 months, 100-150 times of trypan rooting liquid can be sprayed (or the common rooting liquid can be purchased for spraying according to the requirement), the wettable powder is sprayed once a week, the wettable powder is sprayed for 3 times altogether, the plant seedlings are promoted to root (the butterfly orchid is a plant with the same root, the root can be differentiated without spraying the rooting liquid, but the time for differentiating the root can be relatively long), and when the root system is 2-4 cm long, the seedlings can be cut off a parent plant and planted independently, and normal maintenance is carried out.

TABLE 1 Phalaenopsis Capsicum annuum flower stalk axillary bud direct induced seedling condition

Number of plants/plant	Number of axillary buds of pedicel/number of axillary buds of pedicel	Total number of differentiated shoots/number of differentiated shoots	Differentiation coefficient	Yield of single plant
					50	150	600	4	1200％

The induction results are shown in Table 1 and FIG. 6, and it can be seen from Table 1 and FIG. 6 that, after 6 months of pruning the inflorescence, the method described in example 1 can successfully induce seedlings after flowering of the butterfly orchid "big capsicum", each axillary bud is differentiated at a minimum into 2 shoots after induction, at a maximum into 6 shoots, and an average mature 3.5 inch plant gives seedlings 12 plants, and each seedling and mother plant have new inflorescences extracted. At this time, the induced seedlings are cut off and planted separately, so that the mother plant is not damaged, a large number of high-quality seedlings can be obtained, and the method is simple and easy to implement and has excellent application value.

Example 2

1. formulation of phytohormone compositions

The plant hormone composition comprises DA-6, CPPU, NAA and 6-BA.

The specific preparation method comprises the following steps: 10mg of DA-6 was dissolved in 50mL of pure water to obtain solution A;

1mg of CPPU and 0.5mg of NAA are dissolved in 50mL of absolute ethyl alcohol to obtain solution B;

5mg of 6-BA was dissolved in 100mL of a 0.1N NaOH solution to give solution C;

2. Rapid induction of butterfly orchid seedlings

In the embodiment 2 of the invention, 3.5 inch mature seedling flowering plants are selected as the best, and the specific induction implementation process is as follows:

(1) Pretreatment: selecting butterfly orchid 'Fule sunset' as a test object, and feeding 3000 times of liquid 'Huaduo No. 1' as fertilizer in the flowering period for 1 week for 1 time to ensure that plants have sufficient nutrition for the induction of later-stage pedicel buds;

(2) Induction treatment: after the butterfly orchid flowers wither and have no ornamental value, cutting off the flowers with upper portion withered from the 4 th to 5 th knots upwards from the basal part of the flower stalks, stripping off the bracts covered by the axillary buds of the butterfly orchid flower stalks, exposing the bud points in the knots, uniformly smearing the plant hormone composition on the exposed knots of the knots by using a cotton swab, wherein each plant is smeared with 1 axillary bud of the flower stalks once every 4 days, and the method is characterized in that the method comprises the following steps: 00, namely, cutting off the inflorescence on the 1 st day and the 5 th day, and smearing for 2 times;

(3) First month growth management: the first month of induced growth requires light-shielding treatment, and the illumination intensity is lower than 30 mu mol.m ^-2 ·s ^-1 The daytime temperature is higher than or equal to 28 ℃ and lower than 32 ℃, 3000 times of liquid of Huaduo No. 1 is used as fertilizer to be supplied during the period, 1 time is carried out in 1 week, plant materials are covered by a sunshade net with 100% of sunshade rate, and the ambient temperature is adjusted to be 30 ℃ in the daytime and 28 ℃ at night;

(5) From the second month, 50% carbendazim wettable powder is applied once a month, the wettable powder is diluted 500 times for spraying, and a 2000-time liquid of the cyromazine or the fast flutter is adopted to prevent and control red spiders or scale insects, a 3000-time liquid of the delphagostimulant is adopted to prevent and control white flies, after normal maintenance for 4 months, 100 times of trypan rooting liquid can be sprayed once a week, the wettable powder is sprayed for 3 times in total, plant seedling rooting is promoted, the phalaenopsis is a root-bearing plant, when the root system grows to 2-4 cm, the seedlings can be separated from a mother plant, and the white flies are planted independently and normally maintained.

TABLE 2 Phalaenopsis Fule sunset flower peduncle axillary bud direct induced seedling condition

Number of plants/plant	Number of axillary buds of pedicel/number of axillary buds of pedicel	Total number of differentiated shoots/number of differentiated shoots	Differentiation coefficient	Yield of single plant
					50	50	50	1	100％

The induction results are shown in Table 2 and FIG. 7, and as shown in Table 2, after cutting the inflorescence for 4 months, the method described in example 2 can successfully induce seedlings after the Phalaenopsis flower "Fule sunset", each axillary bud is differentiated into 1 tender bud after induction (as shown in FIG. 7), 3 seedlings are obtained by an average mature 3.5 inch plant, and as can be seen from FIG. 7, axillary buds have germinated and grown into complete plants with leaves and roots.

From the results of examples 1 and 2, it can be seen that the number of induced seedlings is positively correlated with the maturity of the mother plant, the number of peduncle axillary buds possessed by the mother plant, and the number of times of applying hormone.

In general, after the methods described in examples 1 and 2 are used, the single plant yield induced by the phalaenopsis seedlings is over 100 percent, and the number of the axillary buds of the pedicel can be increased.

After the seedling is induced, the temperature is set to be the conventional temperature suitable for the growth of the butterfly orchid, the seedling grows faster when the temperature is high (24-30 ℃), and the temperature is higher (32 ℃) to lead to the high Wen Xiumian so as to stop the growth; low temperatures (18-22 ℃) lead to slow growth and reproductive growth, and excessively low temperatures (< 15 ℃) lead to cold stress of the seedlings.

Example 3

The method of this example differs from example 1 in that: the 2.5 inch or 2.8 inch flowering strain butterfly orchid 'large capsicum' is selected as a test object, and the preparation of the hormone composition and the rapid induction process of the butterfly orchid seedlings are the same as those in example 1. And finally, the experimental result shows that the seedling yield of each mother plant is 600%.

Example 4

Example 4 provides a method of formulating a phytohormone composition, comprising:

the plant hormone composition comprises DA-6, CPPU, NAA and 6-BA.

The specific preparation method comprises the following steps: 45mg of DA-6 was dissolved in 50mL of pure water to obtain solution A;

9mg of CPPU and 1.8mg of NAA are dissolved in 50mL of absolute ethyl alcohol to obtain solution B;

18mg of 6-BA was dissolved in 100mL of a 0.1N NaOH solution to give solution C;

heating and melting 1.6kg of lanolin in a water bath mode, adding the three solutions after melting, fully and uniformly stirring, and cooling to obtain a brown yellow pasty product, namely the plant hormone composition.

The rapid induction method of the phalaenopsis seedlings in example 4 was the same as in example 1.

Example 5

Example 5 provides a method of formulating a phytohormone composition, comprising:

the plant hormone composition comprises DA-6, CPPU, NAA and 6-BA.

The specific preparation method comprises the following steps: 15mg of DA-6 was dissolved in 20mL of pure water to obtain solution A;

3mg of CPPU and 0.8mg of NAA are dissolved in 20mL of absolute ethyl alcohol to obtain solution B;

7mg of 6-BA was dissolved in 60mL of a 0.1N NaOH solution to give solution C;

heating and melting 0.9kg of lanolin in a water bath mode, adding the three solutions after melting, fully and uniformly stirring, and cooling to obtain a brown yellow pasty product, namely the plant hormone composition.

The rapid induction method of the phalaenopsis seedlings in example 5 was the same as in example 1.

The plant hormone compositions of examples 4 and 5 are adopted to rapidly induce the phalaenopsis seedlings, so that the same technical effects as those of example 1 can be obtained, and the method disclosed by the invention is not only capable of damaging a mother plant, but also capable of obtaining a large number of high-quality seedlings, and is simple and feasible, and has excellent application value.

Comparative example 1

The method of this comparative example was identical to the induction conditions and method of example 1, but did not contain the above phytohormone composition, and the induction results are shown in Table 3.

TABLE 3 Phalaenopsis Capsicum annuum flower stalk axillary bud direct induced seedling condition

Number of plants/plant	Number of axillary buds of pedicel/number of axillary buds of pedicel	Total number of differentiated shoots/number of differentiated shoots	Differentiation coefficient	Yield of single plant
					50	150	0	0	0

As can be seen from Table 3, after the inflorescence was cut, the peduncle axillary buds of this comparative example were not germinated and the induced plant was 0. The comparative example shows that the purpose of inducing the seedling of the butterfly orchid plant can be achieved by combining the plant hormone composition with the induction conditions and the method.

Comparative example 2

The method of comparative example 2 differs from example 1 in that the ingredients of the phytohormone composition are adjusted. The method of induction was the same as in example 1.

Formulation of phytohormone compositions

The phytohormone composition includes NAA and 6-BA.

The specific preparation method comprises the following steps: dissolving 1mg of NAA in 50mL of absolute ethanol to obtain a solution A;

10mg of 6-BA was dissolved in 100mL of a 0.1N NaOH solution to give solution B;

heating and melting 1.8kg of lanolin in a water bath mode, adding the two solutions after melting, fully and uniformly stirring, and cooling to obtain a brown yellow pasty product, namely the plant hormone composition.

TABLE 4 Phalaenopsis Capsicum annuum flower stalk axillary bud direct induced seedling condition

Number of plants/plant	Number of axillary buds of pedicel/number of axillary buds of pedicel	Total number of differentiated shoots/number of differentiated shoots	Differentiation coefficient	Yield of single plant
					50	150	150	1	100％

The induction results are shown in Table 4, and after 6 months of inflorescence pruning, the method described in comparative example 2 can induce seedlings after the opening of butterfly orchid "big capsicum", and after induction, each axillary bud differentiated 1 tender bud.

Comparative example 3

The method of comparative example 3 is different from example 1 in that the specific procedure of seedling induction is different. The ingredients of the induced phytohormone composition were the same as in example 1.

Specifically, the method of comparative example 3 is different from example 1 in the growth management after the induction treatment:

and (3) growth management: in the first month, the illumination intensity maintains natural illumination, and the maximum light intensity is not more than 400 mu mol.m ^-2 ·s ^-1 The method comprises the steps of carrying out a first treatment on the surface of the Humidity is 70-80%; the temperature of day and night is 28/24 ℃; 3000 times of liquid of Huaduo No. 1 is used as fertilizer to be supplied for 1 week and 1 time;

from the second month, 50% carbendazim wettable powder is applied once a month, diluted 500 times for spraying, and the red spider or scale insect is prevented by adopting a 2000 times liquid of the cyromazine or the fast flutter, the white fly is prevented by adopting a 3000 times liquid of the delphagostimulant, and the normal maintenance is achieved.

The upper part of the butterfly orchid 'large capsicum' axillary bud induced by the comparative example can germinate, but can not be induced into seedlings, but germinate into inflorescences, and secondary flowering is carried out, and the induction result is shown in figure 8.

The phytohormone composition of the present application may be used in any common strain of butterfly orchid. The method is simple, easy to operate, high in seedling yield and free of special sterile environment and special technology. Compared with the method, the traditional plant tissue culture method induces the phalaenopsis seedlings, and although the induction rate is higher, the method requires special operation technology and sterile environment, and is not beneficial to large-area popularization.

The above examples are illustrative of embodiments of the present invention, and embodiments of the present invention are not limited to the above examples, and any other changes, modifications, substitutions, combinations, and simplifications that do not depart from the spirit and principles of the present invention should be made by the equivalent substitution manner, and are included in the scope of the present invention.

Claims

1. A plant hormone composition for inducing phalaenopsis seedlings, which is prepared by the following method:

1) Dissolving DA-6 in water to obtain a solution A; dissolving CPPU and NAA in an organic solvent I to obtain a solution B; dissolving 6-BA in an alkali solution to obtain a solution C;

2) Mixing the solutions A, B, C to obtain a mixed solution, wherein the concentration of DA-6 is 10.0-25.0 mg/L, the concentration of CPPU is 1.0-5.0 mg/L, the concentration of 6-BA is 5.0-10.0 mg/L, and the concentration of NAA is 0.5-1.0 mg/L;

3) Heating and melting the semi-solid plant hormone carrier, placing the semi-solid plant hormone carrier into the mixed solution in the step 2), uniformly mixing, and cooling to room temperature to obtain a solid product, namely the plant hormone composition;

the organic solvent I is absolute ethyl alcohol, acetone or methanol; the alkali solution is NaOH solution, and the concentration is 0.1N;

the semi-solid phytohormone carrier is lanolin or vaseline.

2. The phytohormone composition of claim 1 wherein the ratio of semi-solid phytohormone carrier to mixed solution in step 3) is (7-10) g:1ml.

3. Use of the phytohormone composition of claim 1 for inducing germination of phalaenopsis seedlings.

4. The use according to claim 3, wherein the phalaenopsis is a total inflorescence variety with peduncles and axillary buds, in particular: "big capsicum", "Fule sunset" or "Neishan girl".

5. A method for inducing germination of phalaenopsis seedlings by using the plant hormone composition of claim 1, comprising the steps of:

(1) Pretreatment: applying fertilizer in the flowering period of butterfly orchid for 1-2 times per week;

(2) Induction treatment: after the butterfly orchid flowers wither, cutting off the withered flowers from the basal parts of the pedicel, stripping off bracts covered by axillary buds of the pedicel, exposing bud points in internodes, applying the plant hormone composition once every 4 days, and applying for 2 times or 5 times in total;

(3) Post-induction growth management: light-shielding treatment in the first month of induced growth, the light intensity is less than 50 mu mol ∙ m ^-2 ∙s ^-1 Adjusting the ambient temperature to 28-32deg.C, applying fertilizer during the period, and 1-2 times per week;

the axillary bud point of the flower stalk of the second month expands and differentiates, and the light treatment is carried out on the 30 th day of cutting off inflorescences, and the maximum light intensity is not more than 15000Lux; adjusting the ambient temperature to 24-30 ℃; and fertilizer is applied for 1-2 times per week;

the phytohormone composition of step (2) is applied 5 times, and is specifically operated as follows: uniformly smearing the plant hormone composition on exposed internode nodes, wherein each bud node is smeared with 4-6mg, and 3 pedicel axillary buds are smeared once every 4 days, namely, the 1 st day, the 5 th day, the 9 th day, the 13 th day and the 17 th day of inflorescence pruning are respectively carried out;

the phytohormone composition of step (2) is applied 2 times by the following specific procedures: the plant hormone composition is uniformly smeared on the exposed internode nodes, each bud node is smeared with 4-6mg, each plant is smeared with 1 pedicel axillary bud, and each 4 days is respectively the 1 st day and the 5 th day of the inflorescence pruning, and the smearing is carried out 2 times.