Ornamental onion planting method

Abstract

The invention discloses an ornamental shallot planting method, and belongs to the technical field of plant rapid propagation, and the method comprises the following steps: (1) explant disinfection; (2) callus induction; (3) differentiation culture; (4) multiplication culture; (5) rooting. According to the ornamental shallot planting method, the reproduction rate of ornamental shallots in each link from explant treatment to callus induction and adventitious bud induction to adventitious bud proliferation and finally to rooting is very high, the characters are stable, the excellent characters of female parents are kept to the maximum extent, and the ornamental shallots have good economic benefits. The invention establishes a good rapid propagation mode of the ornamental green onion, lays a foundation for research work such as new variety cultivation and genetic improvement on the variety by using a plant biotechnology in future, and also provides technical guidance and reference for establishment of a tissue culture rapid propagation technology of other plant varieties.

Description

Technical Field

[0001] The invention belongs to the technical field of rapid plant propagation, and in particular relates to a method for planting ornamental onions. Background Art

[0002] Ornamental onions are perennial plants of the Amaryllidaceae family and the Allium genus, such as Allium grandiflorum, Allium maslennifolium, Allium slender-stemmed, Allium long-stalked, Allium truncatum, Allium scabra, Allium narcissus-shaped, and Allium splendidum. They involve many Allium species and are a general term for Allium species with high ornamental value. There are more than 800 species of Allium plants in the world, mainly distributed in the northern hemisphere, from the Caspian Sea to southwestern Siberia, Central Asia to the arid areas of northwest China. There are 163 species (including variants) distributed in my country. Except for green onions, onions, garlic, chives, scallions, and leeks, which have been widely cultivated as vegetables for a long time, most of the others are wild species. Domestic research mainly focuses on the edible (nectar source, vegetable) and medicinal value of Allium plants, and there are few reports from the perspective of ornamental application. So far, only more than 10 species of ornamental onions have been introduced and cultivated in my country.

[0003] Many species of Allium plants have beautiful plant shapes, different flower shapes, bright colors, and high ornamental value. Some species are suitable for fresh cut flowers, some are dried flowers, and some are materials for decoration and beautification of gardens, flower borders, and rock gardens. Like bulbous flowers such as tulips, daffodils, hyacinths, and Dutch irises, ornamental onion bulbs rely on imports every year. The high cost of introduction and quarantine seriously restricts their application in gardens. The germplasm resources of the Allium genus are numerous and their characteristics vary greatly. Research on the biological characteristics, growth phenology, and bulb breeding of Allium plants is of great significance for their development and utilization.

[0004] At present, the common propagation methods of ornamental onions are planting or bulb division. However, although there are many seeds, the seed germination time is long and the technical requirements are high. Once the conditions are not suitable, the germination rate will be very low, even less than 1%, and it usually takes 5 years for the onion to bloom normally after sowing. In addition, the propagation of bulb regeneration division requires a very high size of the mother bulb, and the reproduction rate is low, only 2 to 4 can be reproduced each year, and the traits of the propagated ornamental onions are unstable, affecting the ornamental value. Summary of the invention

[0005] The purpose of the invention is to provide an ornamental onion planting method in view of the existing problems.

[0006] The present invention is achieved through the following technical solutions:

[0007] A method for planting ornamental onions comprises the following steps:

[0008] (1) Explant disinfection:

[0009] Take the bulb of ornamental onion as the explant, remove the leaves on the top and the roots at the base of the fresh bulb, rinse it under running water for 40-60 minutes, disinfect it with 75% alcohol for 30-50 seconds, disinfect it in disinfectant for 1-3 minutes, then wash it with sterile water for 5-7 times, and finally put it on sterile gauze to absorb the surface moisture before inoculation;

[0010] (2) Callus induction:

[0011] Cut the sterilized bulbs into small pieces of 0.3-0.7 cm × 0.3-0.7 cm, inoculate them into the induction medium, and place them in a light incubator to induce callus tissue;

[0012] (3) Differentiation culture:

[0013] The induced callus tissue is cut and inoculated into an adventitious bud differentiation medium, and then placed in a light incubator for adventitious bud differentiation;

[0014] (4) Proliferation culture:

[0015] The adventitious buds produced by differentiation are inoculated into a proliferation culture, and then placed in a light incubator for proliferation culture;

[0016] (5) Rooting:

[0017] The proliferated clustered shoots were inoculated into rooting medium for rooting culture.

[0018] Furthermore, the ornamental onion described in step (1) is one of Allium foldedii, Allium foldedii, Allium foldedii, and Allium foldedii.

[0019] Furthermore, the components and corresponding weight percentages of the disinfectant described in step (1) are: 1-3% α-bisabolol, 1-3% allicin, 1-3% artemisinin, 0.7-0.9% fulvic acid, 2-4% sodium hypochlorite, and the balance is sterile water.

[0020] Furthermore, the induction medium formula described in step (2) is: 1 / 2MS+6-BA 0.3-0.6mg / L+2,4-D 1-2mg / L+white sugar 26-32g / L+agar 5-7g / L+special additive 0.6-0.8mL / L, pH 5.8-6;

[0021] The induction conditions were as follows: the temperature in the incubator was controlled at 25-29°C, the photoperiod was 10-12 h / d, and the light intensity was 20-35 μ·mol·m -2 ·s -1, ultrasonic treatment is carried out every 2 to 3 days, the frequency of the ultrasonic wave is 20 to 40 kHz, and each treatment time is 5 to 8 hours.

[0022] Furthermore, the formula of the adventitious bud differentiation medium described in step (3) is: 1 / 2MS + 6-BA 3-5 mg / L + NAA 3-5 mg / L + white sugar 26-32 g / L + agar 5-7 g / L + special additives 0.5-0.7 mL / L, pH 5.8-6;

[0023] The conditions for differentiation of adventitious buds are as follows: the temperature in the incubator is controlled at 25-29°C, the photoperiod is 15-17 h / d, and the light intensity is 20-35 μ·mol·m -2 ·s -1 , ultrasonic treatment is carried out every 2 to 3 days, the frequency of the ultrasonic wave is 20 to 40 kHz, and each treatment time is 5 to 8 hours.

[0024] Furthermore, the formula of the proliferation medium described in step (4) is: 1 / 2MS+6-BA 0.4-1 mg / L+NAA 0.05-0.25 mg / L+white sugar 26-32 g / L+agar 5-7 g / L+special additive 1-1.6 mL / L, pH 5.8-6;

[0025] The conditions for proliferation culture are: the temperature in the incubator is controlled at 25-29°C, the photoperiod is 14-16 h / d, and the light intensity is 20-35 μ·mol·m -2 ·s -1 .

[0026] Furthermore, the formula of the rooting medium described in step (5) is: 1 / 2MS + IBA 0.4-1 mg / L + NAA 0.4-1 mg / L + white sugar 26-32 g / L + agar 5-7 g / L + special additive 0.4-0.6 mL / L, pH 5.8-6;

[0027] The rooting culture conditions were as follows: the temperature in the incubator was controlled at 27-31°C, the photoperiod was 12-16 h / d, and the light intensity was 20-35 μ·mol·m -2 ·s -1 .

[0028] Furthermore, the components and corresponding weight percentages in the special additive are: L-lysine 2-3%, citric acid 2-4%, lactoferrin 1-2%, arginine 2-3%, and the balance is sterile water.

[0029] Furthermore, the above disinfection and inoculation operations are all carried out on a clean bench.

[0030] Compared with the prior art, the present invention has the following advantages:

[0031] 1. The present invention provides a method for planting ornamental onions, which adopts tissue culture technology for large-scale production of ornamental onions. It not only has the advantages of traditional tissue culture without regional restrictions and is more conducive to factory breeding, but also has the characteristics of high reproduction rate and stable traits of cultivated ornamental onions.

[0032] 2. Compared with mercuric chloride, the disinfectant of the present application has better disinfection effect, the sterility rate is as high as 96%, and almost no damage is caused to the explant, and the survival rate of the explant is as high as 93%. The callus formed by the present application is dense and grows fast, and the formed callus will form granular callus in the later stage, which is more conducive to the differentiation of adventitious buds. By adding the special additive of the present application to the culture medium, the induction rate of adventitious buds is as high as 94%, and the adventitious buds differentiated are dark green in color, without vitrification, and grow fast. In addition, the improvement of disinfectant and culture conditions also significantly promotes the differentiation of adventitious buds. Callus with better texture will produce dark green adventitious buds one after another after inoculation, and single bud inoculation can produce a good proliferation effect, and the bud proliferation rate is as high as 3.56 times, which is significantly better than the traditional tissue culture method. The planting method of the present application has a rooting rate of up to 95.2%, and the rooting effect is very good.

[0033] In summary, from the treatment of explants to the induction of callus tissue, the induction of adventitious buds, the proliferation of adventitious buds, and finally to rooting, the reproduction rate of ornamental onions in each link is very high, and the traits are stable, which maximizes the maintenance of the excellent traits of the mother plant.

[0034] 3. The present invention has established a good rapid propagation method for ornamental onions, which lays a foundation for future research work such as breeding new varieties and genetic improvement of this variety using plant biotechnology, and also provides technical guidance and reference for the establishment of tissue culture rapid propagation technology for other plant varieties of this genus. DETAILED DESCRIPTION

[0035] In order to further explain the present invention, it is described below in conjunction with the following specific embodiments.

[0036] Example 1

[0037] A method for planting ornamental onions comprises the following steps:

[0038] (1) Explant disinfection:

[0039] The bulb of Allium grandiflorum is used as an explant, and the leaves on the top and the roots at the base of the fresh bulb are removed, and then rinsed under running water for 40 minutes, and then disinfected with 75% alcohol for 30 seconds, and then disinfected in a disinfectant for 1 minute, and then washed with sterile water for 5 times, and finally placed on sterile gauze to absorb the surface moisture before inoculation; the components and corresponding weight percentages in the disinfectant are: 1% of α-bisabolol, 1% of allicin, 1% of artemisinin, 0.7% of fulvic acid, 2% of sodium hypochlorite, and the balance is sterile water;

[0040] (2) Callus induction:

[0041] The sterilized bulbs were cut into small pieces of 0.3 cm × 0.3 cm, then inoculated into the induction medium, and then placed in a light incubator for callus induction;

[0042] The formula of the induction medium is: 1 / 2MS+6-BA 0.3mg / L+2,4-D 1mg / L+white sugar 26g / L+agar 5g / L+special additive 0.6mL / L, pH 5.8;

[0043] The induction conditions were as follows: the temperature in the incubator was controlled at 25°C, the photoperiod was 10 h / d, and the light intensity was 20 μ·mol·m -2 ·s -1 , sonication was performed every 2 days, the frequency of the sonication was 20 kHz, and each treatment lasted for 5 hours;

[0044] (3) Differentiation culture:

[0045] The induced callus tissue is cut and inoculated into an adventitious bud differentiation medium, and then placed in a light incubator for adventitious bud differentiation;

[0046] The formula of adventitious bud differentiation medium is: 1 / 2MS+6-BA 3mg / L+NAA 3mg / L+white sugar 26g / L+agar 5g / L+special additive 0.5mL / L, pH 5.8;

[0047] The conditions for differentiation of adventitious buds were as follows: the temperature in the incubator was controlled at 25°C, the photoperiod was 15 h / d, and the light intensity was 20 μ·mol·m -2 ·s -1 , sonication was performed every 2 days, the frequency of the sonication was 20 kHz, and each treatment lasted for 5 hours;

[0048] (4) Proliferation culture:

[0049] The adventitious buds produced by differentiation are inoculated into a proliferation culture, and then placed in a light incubator for proliferation culture;

[0050] The formula of proliferation medium is: 1 / 2MS+6-BA 0.4mg / L+NAA 0.05mg / L+white sugar 26g / L+agar 5g / L+special additive 1mL / L, pH 5.8;

[0051] The conditions for proliferation culture were as follows: the temperature in the incubator was controlled at 25°C, the photoperiod was 14 h / d, and the light intensity was 20 μ·mol·m -2 ·s -1 ;

[0052] (5) Rooting:

[0053] The proliferated clustered buds are inoculated into a rooting medium for rooting culture;

[0054] The rooting medium formula is: 1 / 2MS + IBA 0.4mg / L + NAA 0.4mg / L + white sugar 26g / L + agar 5g / L + special additive 0.4mL / L, pH 5.8;

[0055] The rooting culture conditions were as follows: the temperature in the incubator was controlled at 27°C, the photoperiod was 12 h / d, and the light intensity was 20 μ·mol·m -2 ·s -1 .

[0056] The components and corresponding weight percentages in the special additive are: L-lysine 2%, citric acid 2%, lactoferrin 1%, arginine 2%, and the balance is sterile water.

[0057] The above disinfection and inoculation operations are all carried out on a clean bench.

[0058] Example 2

[0059] A method for planting ornamental onions comprises the following steps:

[0060] (1) Explant disinfection:

[0061] The bulb of Allium grandiflorum is used as an explant, and the leaves on the top and the roots at the base of the fresh bulb are removed, and then rinsed under running water for 50 minutes, and then disinfected with 75% alcohol for 40 seconds, and then disinfected in a disinfectant for 2 minutes, and then washed with sterile water for 6 times, and finally placed on sterile gauze to absorb surface moisture before inoculation; the components and corresponding weight percentages in the disinfectant are: 2% of α-bisabolol, 2% of allicin, 2% of artemisinin, 0.8% of fulvic acid, 3% of sodium hypochlorite, and the balance is sterile water;

[0062] (2) Callus induction:

[0063] The sterilized bulbs were cut into small pieces of 0.5 cm × 0.5 cm, then inoculated into the induction medium, and then placed in a light incubator for callus induction;

[0064] The formula of the induction medium is: 1 / 2MS+6-BA 0.45mg / L+2,4-D 1.5mg / L+white sugar 29g / L+agar 6g / L+special additive 0.7mL / L, pH 5.9;

[0065] The induction conditions were as follows: the temperature in the incubator was controlled at 27°C, the photoperiod was 11 h / d, and the light intensity was 27.5 μ·mol·m -2 ·s -1 , sonication was performed every 2.5 days, the frequency of the sonication was 30kHz, and each treatment lasted for 6 hours;

[0066] (3) Differentiation culture:

[0067] The induced callus tissue is cut and inoculated into an adventitious bud differentiation medium, and then placed in a light incubator for adventitious bud differentiation;

[0068] The formula of adventitious bud differentiation medium is: 1 / 2MS+6-BA4mg / L+NAA4mg / L+white sugar 29g / L+agar 6g / L+special additive 0.6mL / L, pH 5.9;

[0069] The conditions for differentiation of adventitious buds were as follows: the temperature in the incubator was controlled at 27°C, the photoperiod was 16 h / d, and the light intensity was 27.5 μ·mol·m -2 ·s -1 , sonication was performed every 2.5 days, the frequency of the sonication was 30kHz, and each treatment lasted for 6 hours;

[0070] (4) Proliferation culture:

[0071] The adventitious buds produced by differentiation are inoculated into a proliferation culture, and then placed in a light incubator for proliferation culture;

[0072] The formula of proliferation medium is: 1 / 2MS+6-BA 0.7mg / L+NAA 0.15mg / L+white sugar 29g / L+agar 6g / L+special additive 1.3mL / L, pH 5.9;

[0073] The conditions for proliferation culture were as follows: the temperature in the incubator was controlled at 27°C, the photoperiod was 15 h / d, and the light intensity was 27.5 μ·mol·m -2 ·s -1 ;

[0074] (5) Rooting:

[0075] The proliferated clustered buds are inoculated into a rooting medium for rooting culture;

[0076] The rooting medium formula is: 1 / 2MS + IBA 0.7mg / L + NAA 0.7mg / L + white sugar 29g / L + agar 6g / L + special additive 0.5mL / L, pH 5.9;

[0077] The rooting culture conditions were as follows: the temperature in the incubator was controlled at 29°C, the photoperiod was 14 h / d, and the light intensity was 27.5 μ·mol·m -2 ·s -1 .

[0078] The components and corresponding weight percentages in the special additive are: L-lysine 2.5%, citric acid 3%, lactoferrin 1.5%, arginine 2.5%, and the balance is sterile water.

[0079] The above disinfection and inoculation operations are all carried out on a clean bench.

[0080] Example 3

[0081] A method for planting ornamental onions comprises the following steps:

[0082] (1) Explant disinfection:

[0083] The bulb of Allium grandiflorum is used as an explant, and the leaves on the top and the roots at the base of the fresh bulb are removed, and then rinsed under running water for 60 minutes, and then disinfected with 75% alcohol for 50 seconds, and then disinfected in a disinfectant for 3 minutes, and then washed with sterile water for 7 times, and finally placed on sterile gauze to absorb surface moisture before inoculation; the components and corresponding weight percentages in the disinfectant are: 3% of α-bisabolol, 3% of allicin, 3% of artemisinin, 0.9% of fulvic acid, 4% of sodium hypochlorite, and the balance is sterile water;

[0084] (2) Callus induction:

[0085] The sterilized bulbs were cut into small pieces of 0.7 cm × 0.7 cm, then inoculated into the induction medium, and then placed in a light incubator for callus induction;

[0086] The formula of the induction medium is: 1 / 2MS+6-BA 0.6mg / L+2,4-D 2mg / L+white sugar 32g / L+agar 7g / L+special additive 0.8mL / L, pH 6;

[0087] The induction conditions were as follows: the temperature in the incubator was controlled at 29°C, the photoperiod was 12 h / d, and the light intensity was 35 μ·mol·m -2 ·s -1, sonication was performed every 3 days, the frequency of the sonication was 40 kHz, and each treatment lasted 8 hours;

[0088] (3) Differentiation culture:

[0089] The induced callus tissue is cut and inoculated into an adventitious bud differentiation medium, and then placed in a light incubator for adventitious bud differentiation;

[0090] The formula of adventitious bud differentiation medium is: 1 / 2MS+6-BA 5mg / L+NAA 5mg / L+white sugar 32g / L+agar 7g / L+special additive 0.7mL / L, pH 6;

[0091] The conditions for differentiation of adventitious buds were as follows: the temperature in the incubator was controlled at 29°C, the photoperiod was 17 h / d, and the light intensity was 35 μ·mol·m -2 ·s -1 , sonication was performed every 3 days, the frequency of the sonication was 40 kHz, and each treatment lasted 8 hours;

[0092] (4) Proliferation culture:

[0093] The adventitious buds produced by differentiation are inoculated into a proliferation culture, and then placed in a light incubator for proliferation culture;

[0094] The formula of proliferation medium is: 1 / 2MS+6-BA 1mg / L+NAA 0.25mg / L+white sugar 32g / L+agar 7g / L+special additive 1.6mL / L, pH 6;

[0095] The conditions for proliferation culture were as follows: the temperature in the incubator was controlled at 29°C, the photoperiod was 16 h / d, and the light intensity was 35 μ·mol·m -2 ·s -1 ;

[0096] (5) Rooting:

[0097] The proliferated clustered buds are inoculated into a rooting medium for rooting culture;

[0098] The rooting medium formula is: 1 / 2MS + IBA 1mg / L + NAA 1mg / L + white sugar 32g / L + agar 7g / L + special additive 0.6mL / L, pH 6;

[0099] The rooting culture conditions were as follows: the temperature in the incubator was controlled at 31°C, the photoperiod was 16 h / d, and the light intensity was 35 μ·mol·m -2 ·s -1 .

[0100] The components and corresponding weight percentages in the special additive are: 3% L-lysine, 4% citric acid, 2% lactoferrin, 3% arginine, and the balance is sterile water.

[0101] The above disinfection and inoculation operations are all carried out on a clean bench.

[0102] Comparative Example 1

[0103] A method for planting ornamental onions comprises the following steps:

[0104] (1) Explant disinfection:

[0105] The bulb of Allium grandiflorum is used as an explant, and the leaves on the top and the roots at the base of the fresh bulb are removed, and then rinsed under running water for 50 minutes, and then disinfected with 75% alcohol for 40 seconds, and then disinfected in a disinfectant for 2 minutes, and then washed with sterile water for 6 times, and finally placed on sterile gauze to absorb surface moisture before inoculation; the components and corresponding weight percentages in the disinfectant are: 2% of α-bisabolol, 2% of allicin, 2% of artemisinin, 0.8% of fulvic acid, 3% of sodium hypochlorite, and the balance is sterile water;

[0106] (2) Callus induction:

[0107] The sterilized bulbs were cut into small pieces of 0.5 cm × 0.5 cm, then inoculated into the induction medium, and then placed in a light incubator for callus induction;

[0108] The formula of the induction medium is: 1 / 2MS+6-BA 0.45mg / L+2,4-D 1.5mg / L+white sugar 29g / L+agar 6g / L+special additive 0.7mL / L, pH 5.9;

[0109] The induction conditions were as follows: the temperature in the incubator was controlled at 27°C, the photoperiod was 11 h / d, and the light intensity was 27.5 μ·mol·m -2 ·s -1 , sonication was performed every 2.5 days, the frequency of the sonication was 30kHz, and each treatment lasted for 6 hours;

[0110] (3) Differentiation culture:

[0111] The induced callus tissue is cut and inoculated into an adventitious bud differentiation medium, and then placed in a light incubator for adventitious bud differentiation;

[0112] The formula of adventitious bud differentiation medium is: 1 / 2MS+6-BA4mg / L+NAA4mg / L+white sugar 29g / L+agar 6g / L+special additive 0.6mL / L, pH 5.9;

[0113] The conditions for differentiation of adventitious buds were as follows: the temperature in the incubator was controlled at 27°C, the photoperiod was 16 h / d, and the light intensity was 27.5 μ·mol·m -2 ·s -1 , sonication was performed every 2.5 days, the frequency of the sonication was 30kHz, and each treatment lasted for 6 hours;

[0114] (4) Proliferation culture:

[0115] The adventitious buds produced by differentiation are inoculated into a proliferation culture, and then placed in a light incubator for proliferation culture;

[0116] The formula of proliferation medium is: 1 / 2MS+6-BA 0.7mg / L+NAA 0.15mg / L+white sugar 29g / L+agar 6g / L+special additive 1.3mL / L, pH 5.9;

[0117] The conditions for proliferation culture were as follows: the temperature in the incubator was controlled at 27°C, the photoperiod was 15 h / d, and the light intensity was 27.5 μ·mol·m -2 ·s -1 ;

[0118] (5) Rooting:

[0119] The proliferated clustered buds are inoculated into a rooting medium for rooting culture;

[0120] The rooting medium formula is: 1 / 2MS + IBA 0.7mg / L + NAA 0.7mg / L + white sugar 29g / L + agar 6g / L + special additive 0.5mL / L, pH 5.9;

[0121] The rooting culture conditions were as follows: the temperature in the incubator was controlled at 29°C, the photoperiod was 14 h / d, and the light intensity was 27.5 μ·mol·m -2 ·s -1 .

[0122] The components and corresponding weight percentages in the special additive are: L-lysine 2.5%, citric acid 3%, lactoferrin 1.5%, arginine 2.5%, and the balance is sterile water.

[0123] The above disinfection and inoculation operations are all carried out on a clean bench.

[0124] Comparative Example 2

[0125] A method for planting ornamental onions comprises the following steps:

[0126] (1) Explant disinfection:

[0127] The bulb of Allium grandiflorum is used as an explant, and the leaves on the top and the roots at the base of the fresh bulb are removed, and then rinsed under running water for 50 minutes, and then disinfected with 75% alcohol for 40 seconds, and then disinfected in a disinfectant for 2 minutes, and then washed with sterile water for 6 times, and finally placed on sterile gauze to absorb surface moisture before inoculation; the components and corresponding weight percentages in the disinfectant are: 2% of α-bisabolol, 2% of allicin, 2% of artemisinin, 0.8% of fulvic acid, 3% of sodium hypochlorite, and the balance is sterile water;

[0128] (2) Callus induction:

[0129] The sterilized bulbs were cut into small pieces of 0.5 cm × 0.5 cm, then inoculated into the induction medium, and then placed in a light incubator for callus induction;

[0130] The formula of the induction medium is: 1 / 2MS+6-BA 0.45mg / L+2,4-D 1.5mg / L+white sugar 29g / L+agar 6g / L+special additive 0.7mL / L, pH 5.9;

[0131] The induction conditions were as follows: the temperature in the incubator was controlled at 27°C, the photoperiod was 11 h / d, and the light intensity was 27.5 μ·mol·m -2 ·s -1 , sonication was performed every 2.5 days, the frequency of the sonication was 30kHz, and each treatment lasted for 6 hours;

[0132] (3) Differentiation culture:

[0133] The induced callus tissue is cut and inoculated into an adventitious bud differentiation medium, and then placed in a light incubator for adventitious bud differentiation;

[0134] The formula of adventitious bud differentiation medium is: 1 / 2MS+6-BA4mg / L+NAA4mg / L+white sugar 29g / L+agar 6g / L+special additive 0.6mL / L, pH 5.9;

[0135] The conditions for differentiation of adventitious buds were as follows: the temperature in the incubator was controlled at 27°C, the photoperiod was 16 h / d, and the light intensity was 27.5 μ·mol·m -2 ·s -1 , sonication was performed every 2.5 days, the frequency of the sonication was 30kHz, and each treatment lasted for 6 hours;

[0136] (4) Proliferation culture:

[0137] The adventitious buds produced by differentiation are inoculated into a proliferation culture, and then placed in a light incubator for proliferation culture;

[0138] The formula of proliferation medium is: 1 / 2MS+6-BA 0.7mg / L+NAA 0.15mg / L+white sugar 29g / L+agar 6g / L+special additive 1.3mL / L, pH 5.9;

[0139] The conditions for proliferation culture were as follows: the temperature in the incubator was controlled at 27°C, the photoperiod was 15 h / d, and the light intensity was 27.5 μ·mol·m -2 ·s -1 ;

[0140] (5) Rooting:

[0141] The proliferated clustered buds are inoculated into a rooting medium for rooting culture;

[0142] The rooting medium formula is: 1 / 2MS + IBA 0.7mg / L + NAA 0.7mg / L + white sugar 29g / L + agar 6g / L + special additive 0.5mL / L, pH 5.9;

[0143] The rooting culture conditions were as follows: the temperature in the incubator was controlled at 29°C, the photoperiod was 14 h / d, and the light intensity was 27.5 μ·mol·m -2 ·s -1 .

[0144] The components and corresponding weight percentages in the special additive are: L-lysine 2.5%, citric acid 3%, lactoferrin 1.5%, arginine 2.5%, and the balance is sterile water.

[0145] The above disinfection and inoculation operations are all carried out on a clean bench.

[0146] Comparative Example 3

[0147] A method for planting ornamental onions comprises the following steps:

[0148] (1) Explant disinfection:

[0149] The bulb of Allium grandiflorum is used as an explant, and the leaves on the top and the roots at the base of the fresh bulb are removed, and then rinsed under running water for 50 minutes, and then disinfected with 75% alcohol for 40 seconds, and then disinfected in a disinfectant for 2 minutes, and then washed with sterile water for 6 times, and finally placed on sterile gauze to absorb surface moisture before inoculation; the components and corresponding weight percentages in the disinfectant are: 2% of α-bisabolol, 2% of allicin, 2% of artemisinin, 0.8% of fulvic acid, 3% of sodium hypochlorite, and the balance is sterile water;

[0150] (2) Callus induction:

[0151] The sterilized bulbs were cut into small pieces of 0.5 cm × 0.5 cm, then inoculated into the induction medium, and then placed in a light incubator for callus induction;

[0152] The formula of the induction medium is: 1 / 2MS+6-BA 0.45mg / L+2,4-D 1.5mg / L+white sugar 29g / L+agar 6g / L+special additive 0.7mL / L, pH 5.9;

[0153] The induction conditions were as follows: the temperature in the incubator was controlled at 27°C, the photoperiod was 11 h / d, and the light intensity was 27.5 μ·mol·m -2 ·s -1 , sonication was performed every 2.5 days, the frequency of the sonication was 30kHz, and each treatment lasted for 6 hours;

[0154] (3) Differentiation culture:

[0155] The induced callus tissue is cut and inoculated into an adventitious bud differentiation medium, and then placed in a light incubator for adventitious bud differentiation;

[0156] The formula of adventitious bud differentiation medium is: 1 / 2MS+6-BA4mg / L+NAA4mg / L+white sugar 29g / L+agar 6g / L+special additive 0.6mL / L, pH 5.9;

[0157] The conditions for differentiation of adventitious buds were as follows: the temperature in the incubator was controlled at 27°C, the photoperiod was 16 h / d, and the light intensity was 27.5 μ·mol·m -2 ·s -1 , sonication was performed every 2.5 days, the frequency of the sonication was 30kHz, and each treatment lasted for 6 hours;

[0158] (4) Proliferation culture:

[0159] The adventitious buds produced by differentiation are inoculated into a proliferation culture, and then placed in a light incubator for proliferation culture;

[0160] The formula of proliferation medium is: 1 / 2MS+6-BA 0.7mg / L+NAA 0.15mg / L+white sugar 29g / L+agar 6g / L+special additive 1.3mL / L, pH 5.9;

[0161] The conditions for proliferation culture were as follows: the temperature in the incubator was controlled at 27°C, the photoperiod was 15 h / d, and the light intensity was 27.5 μ·mol·m -2 ·s -1 ;

[0162] (5) Rooting:

[0163] The proliferated clustered buds are inoculated into a rooting medium for rooting culture;

[0164] The rooting medium formula is: 1 / 2MS + IBA 0.7mg / L + NAA 0.7mg / L + white sugar 29g / L + agar 6g / L + special additive 0.5mL / L, pH 5.9;

[0165] The rooting culture conditions were as follows: the temperature in the incubator was controlled at 29°C, the photoperiod was 14 h / d, and the light intensity was 27.5 μ·mol·m -2 ·s -1 .

[0166] The components and corresponding weight percentages in the special additive are: L-lysine 2.5%, citric acid 3%, lactoferrin 1.5%, arginine 2.5%, and the balance is sterile water.

[0167] The above disinfection and inoculation operations are all carried out on a clean bench.

[0168] In order to compare the technical effects of the present application, fresh Allium grandiflorum bulbs of similar size and growth conditions were selected as experimental objects, and then randomly divided into 4 equal parts of equal quality and quantity, and the methods of the above-mentioned Example 2 and Comparative Examples 1 to 3 were used to carry out rapid propagation of Allium grandiflorum. During the tissue culture process, the survival rate, callus induction rate, adventitious bud induction rate, adventitious bud proliferation and rooting were measured. The specific results are as follows:

[0169] Table 1 Disinfection of Allium grandiflorum explants in each group

[0170] Explant sterility rate / % Explant survival rate / % Example 2 96 93 Comparative Example 1 89 83 Comparative Example 2 85 80 Comparative Example 3 91 87

[0171] It can be seen from Table 1 above that the sterility rate of explants using the disinfectant of the present application is the highest, reaching 96%. Compared with mercuric chloride, the disinfectant of the present application has a better disinfection effect and hardly causes damage to the explants. The survival rate of the explants is as high as 93% for rapid propagation using the method of the present application, which is significantly better than comparative examples 1 to 3.

[0172] Table 2 Callus induction of Allium grandiflorum in each group

[0173]

[0174] It can be concluded from Table 2 that the callus tissue formed in the present application is dense and grows fast. In the experiment, it was found that the callus tissue of Example 2 will form granular callus in the later stage, which is more conducive to the differentiation of adventitious buds. The induction rate of the callus tissue of Comparative Examples 1 to 3 is significantly lower than that of Example 2, and the growth rate of the callus tissue is general, and the rapid propagation effect is not significant.

[0175] Table 3 Effect of each group on the differentiation of adventitious buds from Allium grandiflorum callus

[0176]

[0177] It can be concluded from Table 3 that the induction rate of adventitious buds can reach 94% by adding the special additive of the present application to the culture medium, and the differentiated adventitious buds have dark green leaves, no vitrification phenomenon, and fast growth rate. In addition, the improvement of disinfectants and culture conditions also significantly promotes the differentiation of adventitious buds.

[0178] Table 4 Proliferation of adventitious buds of Allium grandiflorum in each group

[0179] Growth status Proliferation times / fold Example 2 Adventitious buds grow normally and proliferate quickly 3.56 Comparative Example 1 Adventitious buds grow normally and proliferate quickly 2.56 Comparative Example 2 Adventitious buds grow normally but proliferate slowly 1.75 Comparative Example 3 Adventitious buds grow normally and proliferate at a normal rate 2.89

[0180] It can be seen from Table 4 that callus tissue with better texture will gradually produce dark green adventitious buds after inoculation, and single bud inoculation can produce a good proliferation effect, with the bud proliferation rate as high as 3.56 times, which is significantly better than the control example.

[0181] Table 5 Rooting situation of allium sibiricum in each group

[0182]

[0183]

[0184] It can be seen from Table 5 that the planting method of the present application has a rooting rate of up to 95.2%, and has the best rooting effect.

[0185] The embodiments of this specific implementation method are all preferred embodiments of the present invention, and are not intended to limit the protection scope of the present invention. Therefore, any equivalent changes made based on the structure, shape, and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A method for planting ornamental onions, characterized in that: The steps include: (1) Explant disinfection: Take the bulb of ornamental onion as the explant, remove the leaves on the top and the roots at the base of the fresh bulb, rinse it under running water for 40-60 minutes, disinfect it with 75% alcohol for 30-50 seconds, disinfect it in disinfectant for 1-3 minutes, then wash it with sterile water for 5-7 times, and finally put it on sterile gauze to absorb the surface moisture before inoculation; (2) Callus induction: Cut the sterilized bulbs into small pieces of 0.3-0.7 cm × 0.3-0.7 cm, inoculate them into the induction medium, and place them in a light incubator to induce callus tissue; (3) Differentiation culture: The induced callus tissue is cut and inoculated into an adventitious bud differentiation medium, and then placed in a light incubator for adventitious bud differentiation; (4) Proliferation culture: The adventitious buds produced by differentiation are inoculated into a proliferation culture, and then placed in a light incubator for proliferation culture; (5) Rooting: The proliferated clustered shoots were inoculated into rooting medium for rooting culture.

2. The method for planting ornamental onions according to claim 1, characterized in that: The ornamental onion described in step (1) is one of Allium foldedii, Allium foldedii, Allium foldedii, and Allium foldedii.

3. The method for planting ornamental onions according to claim 1, characterized in that: The components and corresponding weight percentages of the disinfectant described in step (1) are: 1-3% α-bisabolol, 1-3% allicin, 1-3% artemisinin, 0.7-0.9% fulvic acid, 2-4% sodium hypochlorite, and the balance is sterile water.

4. The method for planting ornamental onions according to claim 1, characterized in that: The induction medium formula described in step (2) is: 1 / 2MS+6-BA0.3-0.6mg / L+2,4-D1-2mg / L+white sugar 26-32g / L+agar 5-7g / L+special additive 0.6-0.8mL / L, pH 5.8-6; The induction conditions were as follows: the temperature in the incubator was controlled at 25-29°C, the photoperiod was 10-12 h / d, and the light intensity was 20-35 μ·mol·m -2 ·s -1 , ultrasonic treatment is carried out every 2 to 3 days, the frequency of the ultrasonic wave is 20 to 40 kHz, and each treatment time is 5 to 8 hours.

5. The method for planting ornamental onions according to claim 1, characterized in that: The formula of the adventitious bud differentiation medium described in step (3) is: 1 / 2MS + 6-BA 3-5 mg / L + NAA 3-5 mg / L + white sugar 26-32 g / L + agar 5-7 g / L + special additives 0.5-0.7 mL / L, pH 5.8-6; The conditions for differentiation of adventitious buds are as follows: the temperature in the incubator is controlled at 25-29°C, the photoperiod is 15-17 h / d, and the light intensity is 20-35 μ·mol·m -2 ·s -1 , ultrasonic treatment is carried out every 2 to 3 days, the frequency of the ultrasonic wave is 20 to 40 kHz, and each treatment time is 5 to 8 hours.

6. The method for planting ornamental onions according to claim 1, characterized in that: The formula of the proliferation medium described in step (4) is: 1 / 2MS + 6-BA 0.4-1 mg / L + NAA 0.05-0.25 mg / L + white sugar 26-32 g / L + agar 5-7 g / L + special additives 1-1.6 mL / L, pH 5.8-6; The conditions for proliferation culture are: the temperature in the incubator is controlled at 25-29°C, the photoperiod is 14-16h / d, and the light intensity is 20-35μ·mol·m -2 ·s -1 .

7. The method for planting ornamental onions according to claim 1, characterized in that: The rooting medium formula described in step (5) is: 1 / 2MS + IBA 0.4-1 mg / L + NAA 0.4-1 mg / L + white sugar 26-32 g / L + agar 5-7 g / L + special additive 0.4-0.6 mL / L, pH 5.8-6; The rooting culture conditions were as follows: the temperature in the incubator was controlled at 27-31°C, the photoperiod was 12-16 h / d, and the light intensity was 20-35 μ·mol·m -2 ·s -1 .

8. A method for planting ornamental onions according to any one of claims 4 to 7, characterized in that: The components and corresponding weight percentages in the special additive are: L-lysine 2-3%, citric acid 2-4%, lactoferrin 1-2%, arginine 2-3%, and the balance is sterile water.

9. The method for planting ornamental onions according to claim 1, characterized in that: The above disinfection and inoculation operations are all carried out on a clean bench.

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