Method for efficiently inducing seedling propagation of paris polyphylla

A technology of Huazhong and seedlings, which is applied in the field of plant breeding, can solve the problems of seriousness, waste of raw materials, pollution of primary culture, etc., and achieve the effects of strong anti-adversity ability, convenient management and high reproduction coefficient.

Inactive Publication Date: 2018-11-16

BIOLOGICAL TECH INST OF FUJIAN ACADEMY OF AGRI SCI

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AI-Extracted Technical Summary

Problems solved by technology

[0005] Chinese patent CN201310529238.9 discloses a rapid propagation method of Huachonglou plants. This patent uses the rhizomes of Huachonglou as explants, which has the following disadvantages: because the rhizomes grow in the soi...

Abstract

The invention discloses a method for efficiently inducing seedling propagation of paris polyphylla. According to the method, a paris polyphylla seed of which the radicle is already differentiated is adopted as explants, callus induction and propagation culture are carried out, after a high propagation rate is met, differential culture, tuber expansion, rooting culture and transplanting are furthercarried out, and then a great amount of paris polyphylla seedlings can be propagated, and the paris polyphylla medicinal material growth period can be shortened. By adopting the method, the problemsthat in the prior art paris polyphylla seedlings are low in propagation rate and slow in growth and requirements of the market on paris polyphylla seedlings cannot be met, can be solved. By adopting the method disclosed by the invention, the propagation rate can be up to 60000 times or greater within two years, and the propagation rate is much faster than that of common seed propagation. The method is not only high in propagation rate, but also capable of shortening the field growth period of the paris polyphylla medicinal material, in addition high in fixed planting survival rate of tissue culture tubers, and applicable to on-scale production of paris polyphylla.

Application Domain

Cultivating equipmentsHorticulture methods +2

Technology Topic

Plant TubersPropagation rate +8

Examples

Experimental program(3)

Example Embodiment

[0046] Example 1

[0047] A method for efficiently inducing the reproduction of Huazhonglou seedlings, including the acquisition of sterile explants: culture through the following steps:

[0048] 1) Disinfection treatment: Select the Huazhonglou seeds that have differentiated their radicles, rinse them with running water, soak them with washing powder, brush lightly with a brush, then rinse them with tap water, and then remove the radicles and remove the remaining part. As an explant, soak the explant in 75% alcohol for 30 s on a clean bench, then sterilize it with 0.1% mercury liters for 10 min, and finally rinse with sterile water 4 times;

[0049] 2) Induction culture: Inoculate the sterilized explants on the induction medium, and cultivate them for 55 days at a temperature of 20 ℃ and dark conditions to obtain callus;

[0050] The induction medium is MS medium with pH=5.8, and the components and their concentrations in the induction medium are as follows:

[0051] 6- BA 2.5mg·L -1

[0052] NAA 0.4 mg·L -1

[0053] 2,4-D 0.2 mg·L -1

[0054] Sucrose 30000 mg·L -1

[0055] Agar 7500 mg·L -1

[0056] 3) Proliferation culture: under aseptic conditions, cut the callus obtained in step 2) into 2cm 3 After size, transfer it to a proliferation medium and culture it for 40 days at a temperature of 24°C in the dark. The callus proliferates in large quantities;

[0057] The value-added medium is MS medium with pH=5.8, and the components and their concentrations in the value-added medium are as follows:

[0058] 6- BA 1.5 mg·L -1

[0059] NAA 0.2 mg·L -1

[0060] 2,4-D 0.1 mg·L -1

[0061] Sucrose 30000 mg·L -1

[0062] Agar 7500 mg·L -1

[0063] 4) Differentiation culture: cut the callus obtained in step 3) into 2 cm 3 After size, transfer it to differentiation medium, culture it for 60 d under dark conditions at a temperature of 24 ℃, the callus can differentiate into protrusions (ie small tubers);

[0064] The differentiation medium is MS medium with pH=5.8, and the components and their concentrations in the differentiation medium are as follows:

[0065] 6- BA 1.0 mg·L -1

[0066] NAA 0.2 mg·L -1

[0067] Sucrose 30000 mg·L -1

[0068] Agar 7500 mg·L -1

[0069] 5) Swelling and rooting culture: The small tubers obtained in step 4) are cut into individual pieces and transferred to an swelling and rooting medium, cultured for 80 d under dark conditions at a temperature of 24 ℃, the small tubers quickly swell and take roots;

[0070] The swelling and rooting medium is MS medium with pH=5.8, and the components and their concentrations in the swelling and rooting culture basis are as follows:

[0071] 6- BA 1.0 mg·L -1

[0072] NAA 0.1 mg·L -1

[0073] IAA 0.1 mg·L -1

[0074] Sucrose 60000 mg·L -1

[0075] Agar 7500 mg·L -1

[0076] 6) Transplanting: When the root length of the tuber cultivated in step 5) is 1.5 cm, take out the tuber, wash away the root medium, and then transplant it into a mixed medium with a 1:1 volume ratio of peat and perlite for growth.

Example Embodiment

[0077] Example 2

[0078] A method for efficiently inducing the reproduction of Huazhonglou seedlings, including the acquisition of sterile explants: culture through the following steps:

[0079] 1) Disinfection treatment: Select the Huazhonglou seeds that have differentiated their radicles, rinse them with running water, soak them with washing powder, brush lightly with a brush, then rinse them with tap water, and then remove the radicles and remove the remaining part. As an explant, soak the explant in 70% alcohol for 35 s on an ultra-clean workbench, then disinfect with 0.15% mercury liters for 8 minutes, and finally rinse with sterile water for 3 times;

[0080] 2) Induction culture: Inoculate the sterilized explants on the induction medium, and culture them for 60 days at 18 ℃ in the dark to obtain callus;

[0081] The induction medium is MS medium with pH=5.6, and the components and their concentrations in the induction medium are as follows:

[0082] 6- BA 2.0 mg·L -1

[0083] NAA 0.3 mg·L -1

[0084] 2,4-D 0.1 mg·L -1

[0085] Sucrose 28000 mg·L -1

[0086] Agar 7200 mg·L -1

[0087] 3) Proliferation culture: under aseptic conditions, cut the callus obtained in step 2) into 1 cm 3 After size, transfer it to a proliferation medium and culture it for 50 d at a temperature of 22°C in the dark. The callus proliferates in large quantities;

[0088] The proliferation medium is MS medium with pH=5.6, and the components and their concentrations in the proliferation medium are as follows:

[0089] 6- BA 1.0 mg·L -1

[0090] NAA 0.2 mg·L -1

[0091] 2,4-D 0.1 mg·L -1

[0092] Sucrose 28000 mg·L -1

[0093] Agar 7200 mg·L -1

[0094] 4) Differentiation culture: cut the callus obtained in step 3) into 1cm 3 After size, transfer to differentiation medium, culture for 70 d at 22°C in the dark, the callus can differentiate into protrusions (ie small tubers);

[0095] The differentiation medium is MS medium with pH=5.6, and the components and their concentrations in the differentiation medium are as follows:

[0096] 6- BA 1.0 mg·L -1

[0097] NAA 0.2 mg·L -1

[0098] Sucrose 28000 mg·L -1

[0099] Agar 7200 mg·L -1

[0100] 5) Swelling and rooting culture: The small tubers obtained in step 4) are cut into individual pieces and then transferred to an swelling and rooting medium, cultured for 90 d under dark conditions at a temperature of 22 ℃, the small tubers rapidly swell and take roots;

[0101] The swelling and rooting medium is MS medium with pH=5.8, and the components and their concentrations in the swelling and rooting culture basis are as follows:

[0102] 6- BA 1.0 mg·L -1

[0103] NAA 0.1 mg·L -1

[0104] IAA 0.1 mg·L -1

[0105] Sucrose 58000 mg·L -1

[0106] Agar 7200 mg·L -1

[0107] 6) Transplanting: When the root length of the tuber cultivated in step 5) is 0.5cm, take out the tuber, wash the root medium and transplant it into a mixed medium with a 1:1 volume ratio of peat and perlite for growth.

Example Embodiment

[0108] Example 3

[0109] A method for efficiently inducing the reproduction of Huazhonglou seedlings, including the acquisition of sterile explants: culture through the following steps:

[0110] 1) Disinfection treatment: Select the Huazhonglou seeds that have differentiated their radicles, rinse them with running water, soak them with washing powder, brush lightly with a brush, then rinse them with tap water, and then remove the radicles and remove the remaining part. As an explant, soak the explant in 75% alcohol for 25s on a clean bench, then disinfect with 0.1% mercury liters for 10 minutes, and finally rinse with sterile water 4 times;

[0111] 2) Induction culture: Inoculate the sterilized explants on the induction medium, and culture them for 50 days at a temperature of 22 ℃ and dark conditions to obtain callus;

[0112] The induction medium is MS medium with pH=6.0, and the components and their concentrations in the induction medium are as follows:

[0113] 6- BA 3.0 mg·L -1

[0114] NAA 0.5 mg·L -1

[0115] 2,4-D 0.3 mg·L -1

[0116] Sucrose 30000 mg·L -1

[0117] Agar 7500 mg·L -1

[0118] 3) Proliferation culture: under aseptic conditions, cut the callus obtained in step 2) into 3cm 3 After size, transfer to a multiplication medium, culture for 30 d at a temperature of 26°C in the dark, and the callus proliferates in large quantities;

[0119] The value-added medium is MS medium with pH=6.0, and the components and their concentrations in the value-added medium are as follows:

[0120] 6- BA 2.0 mg·L -1

[0121] NAA 0.2 mg·L -1

[0122] 2,4-D 0.1 mg·L -1

[0123] Sucrose 30000 mg·L -1

[0124] Agar 7500 mg·L -1

[0125] 4) Differentiation culture: cut the callus obtained in step 3) into 3cm 3 After the size is changed, it is transferred to differentiation medium and cultured for 50 d under dark conditions at a temperature of 26°C, the callus can differentiate into protrusions (ie small tubers);

[0126] The differentiation medium is MS medium with pH=6.0, and the components and their concentrations in the differentiation medium are as follows:

[0127] 6- BA 1.0 mg·L -1

[0128] NAA 0.2 mg·L -1

[0129] Sucrose 30000 mg·L -1

[0130] Agar 7500 mg·L -1

[0131] 5) Swelling and rooting culture: the small tubers obtained in step 4) are cut into individual pieces and transferred to an swelling and rooting medium, cultured for 70 d under dark conditions at a temperature of 26 ℃, the small tubers rapidly swell and take roots;

[0132] The swelling and rooting medium is MS medium with pH=6.0, and the components and their concentrations in the swelling and rooting culture basis are as follows:

[0133] 6- BA 1.0 mg·L -1

[0134] NAA 0.1 mg·L -1

[0135] IAA 0.1 mg·L -1

[0136] Sucrose 60000 mg·L -1

[0137] Agar 7500 mg·L -1

[0138] 6) Transplanting: When the root length of the tuber cultured in step 5) is 2.0 cm, take out the tuber, wash away the root medium, and then transplant it into a mixed medium with a 1:1 volume ratio of peat and perlite for growth.

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Method for efficiently inducing seedling propagation of paris polyphylla

AI-Extracted Technical Summary

Problems solved by technology

Abstract

Examples

Example Embodiment

Example Embodiment

Example Embodiment

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Description & Claims & Application Information

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