A method for callus induction and proliferation of water lily mature embryos

A callus induction and callus technology, applied in the field of plant tissue culture, can solve the problems of unresolved callus proliferation and inapplicable seed disinfection, etc., and achieve the effect of fast callus proliferation

Active Publication Date: 2022-07-26
三亚南京农业大学研究院 +1
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, we conducted experiments and found that the aseptic seedlings of different subgenus water lilies need to use different disinfection methods. For example, it is easier to obtain aseptic seedlings from the seeds of tropical water lily Nine Pin Perfume by conventional disinfection methods, but it is not suitable for snow white water lilies,' Manla' and other hardy water lily seed disinfection, therefore, need to further explore its effective disinfection method
[0005] At present, there are no successful reports on the in vitro regeneration of water lilies at home and abroad. Sun Chunqing et al. used the immature embryos of three hybrid combinations of water lilies as explants, and studied the immature embryos 13-18 days after pollination at 1 / 2MS + 2,4-D Water lily callus was induced in 2.0 mg / L + 6-BA 2.0 mg / L medium, but related technical problems in callus proliferation have not yet been overcome

Method used

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  • A method for callus induction and proliferation of water lily mature embryos
  • A method for callus induction and proliferation of water lily mature embryos
  • A method for callus induction and proliferation of water lily mature embryos

Examples

Experimental program
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Effect test

Embodiment 1

[0053] A method for inducing and proliferating mature embryo callus of water lily, comprising the following steps:

[0054] (1) Pretreatment of mature embryos: Take plump and healthy mature embryos of snow-white water lilies for testing, rinse them under running water for 2 hours, then place them in 10g / L carbendazim solution at room temperature for vortex disinfection for 20 minutes, and rinse them with distilled water for 3~5 hours. Repeat until no carbendazim solution remains.

[0055] (2) Preparation of induction medium: prepare callus induction medium, the medium composition is 1 / 2MS + 1.5mg / L 2,4-D + 0.5 mg / L TDZ + 50g / L sucrose + 8g / L agar , the preparation method is to weigh 2.47g of 1 / 2MS medium (commercially available 1 / 2MS solid medium, without agar and sucrose), 8g of agar and 50g of sucrose in an enamel tank, add 1L of distilled water to heat and dissolve, and then add auxin 2,4-D 1.5mg, cytokinin TDZ 0.5mg, adjust pH to 5.8 with 1 mol / L NaOH, sterilize at 121°C ...

Embodiment 2

[0066] Others are the same as in Example 1, except that the addition amount of TDZ in the induction medium is 0.2 mg / L, and the addition amount of 2,4-D is 1.5 mg / L.

Embodiment 3

[0068] Others are the same as in Example 1, except that the addition amount of TDZ in the induction medium is 0.2 mg / L, and the addition amount of 2,4-D is 2.0 mg / L.

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Abstract

The invention discloses a method for inducing and multiplying callus of mature embryos of water lily. The specific steps include: material preparation, pretreatment and disinfection of mature embryos, induction of embryogenic callus and proliferation of callus. The invention uses the mature embryo of the water lily as the explant, the material is convenient to obtain, the pollution rate is low after the disinfection treatment, the callus induction rate is high, and the proliferation effect is good, and the invention can be used for genetic transformation, germplasm resource preservation and rapid propagation production, and regeneration of water lilies. System and transgenic research to establish a foundation.

Description

technical field [0001] The invention relates to a plant tissue culture technology, in particular to a method for inducing and multiplying callus of mature embryos of water lily. Background technique [0002] Nymphaea (Nymphaea) is a perennial floating-leaf aquatic herb of Nymphaeaceae (Nymphaeaceae), mainly distributed in temperate and tropical zones. In the garden waterscape (Liao Weiwei, 2016), the market demand is broad. At present, the vast majority of water lily varieties are produced by artificial hybridization and natural hybridization. The breeding cycle is long and the randomness is strong (Li Shujuan, 2019). Seedlings are produced by tissue culture, which has a fast production speed and can shorten the breeding cycle. , Effectively maintain the excellent traits of seedlings and preserve rare water lily germplasm resources. Therefore, rapid propagation through tissue culture is an effective way to meet application requirements as soon as possible. It is imminent to...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00
CPCA01H4/005A01H4/002A01H4/00
Inventor 徐迎春赵莹姜慧杨志娟金奇江王彦杰周平
Owner 三亚南京农业大学研究院
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