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Tissue culturing method for regenerating somatic cells of aquatic iris

An aquatic iris and somatic cell technology, applied in the field of plant tissue culture, to achieve the effects of complete structure, reduced tissue culture cost and high speed

Inactive Publication Date: 2012-11-14
ZHEJIANG XIAOSHAN COTTON & FLAX RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are no reports at home and abroad on the tissue culture technology research on the regeneration pathway of somatic embryos in aquatic iris

Method used

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  • Tissue culturing method for regenerating somatic cells of aquatic iris

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Embodiment 1: (a kind of tissue culture method 1 of aquatic iris somatic cell regeneration)

[0036] (1) The preparation of the culture medium, including the components of the basic medium and the culture medium at each stage of tissue culture, and the weight of each component in each liter of culture medium is:

[0037] 1) Basic medium: MS medium is used for the induction of aquatic iris buds, and WPM basic medium is used for the induction of leaf embryoid bodies, subculture proliferation, strong seedlings and rooting culture; wherein, the agar is 5g / L, pH5. 8;

[0038] 2) Bud induction medium: white sugar 30g / L MS+6-BA 0.5mg / L and NAA 0.2mg / L;

[0039] 3) Leaf embryoid body induction medium: WPM+ZT 0.5mg / L+TDZ 0.25mg / L and 2,4-D 0.1mg / L of white sugar 30g / L;

[0040] 4) Subculture proliferation medium: WPM+ZT 0.5mg / L+TDZ0.25mg / L and 2,4-D 0.1mg / L with white sugar 30g / L;

[0041] 5) Strong seedling medium: white sugar is 30g / L WPM+BA1.0mg / L and NAA 0.3mg / L;

[0042...

Embodiment 2

[0051] Embodiment 2: (a kind of tissue culture method 2 of aquatic iris somatic cell regeneration)

[0052] In this embodiment, the agar in the basic medium is 7g / L, and the pH is 5.7; the bud induction medium is: white sugar 20g / L MS+6-BA 0.8mg / L and NAA 0.3mg / L; leaf embryoid bodies The induction medium is: WPM+ZT 1.0mg / L+TDZ 0.28mg / L with 20g / L white sugar; the subculture medium is: WPM+ZT 1.0mg / L+TDZ 0.28mg / L with 20g / L white sugar and 2, 4-D 0.05mg / L; strong seedling medium: white sugar 15g / L WPM+BA 0.5mg / L and NAA 0.1mg / L; rooting medium: white sugar 15g / L WPM+TBA1.0 and NAA 0.5mg / L; the young shoots were soaked in 70% alcohol for 45 seconds, and soaked in 0.1% mercuric chloride aqueous solution for 9 minutes, then the shoot tip tissue was removed and cut into 0.35cm 3 The light intensity of each stage of bud induction, leaf embryoid body induction, subculture proliferation, strong seedling and rooting culture is 1500Lx, and the culture time of each stage is 20 days, 15...

Embodiment 3

[0053] Embodiment 3: (a kind of tissue culture method 3 of aquatic iris somatic cell regeneration)

[0054] In this embodiment, the agar in the basic medium is 8g / L, and the pH is 5.6; the bud induction medium is: white sugar 60g / L MS+6-BA 1.0mg / L and NAA 0.5mg / L; leaf embryoid bodies Induction medium: white sugar 60g / L WPM+TDZ 0.3mg / L and 2,4-D0.05mg / L; subculture medium: white sugar 60g / L WPM+ZT0.75mg / L, TDZ 0.3 mg / L and 2,4-D 0.075mg / L; strong seedling medium: white sugar 30g / L WPM+BA 0.3mg / L and NAA 0.2mg / L; rooting medium: white sugar 30g / L WPM +IBA1.5 and NAA1.0mg / L; the young shoots were soaked in 70% alcohol for 60 seconds, soaked in 0.1% mercuric solution for 10 minutes, then removed the shoot tip tissue and cut into 0.5cm 3 The light intensity of each stage of bud induction, leaf embryoid body induction, subculture proliferation, strong seedling and rooting culture is 2500Lx, and the culture time of each stage is 10 days, 30 days, 10 days, 20 days and 30 days respec...

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Abstract

The invention discloses a tissue culturing method for regenerating somatic cells of an aquatic iris, belonging to the technical field of plant tissue culturing. The tissue culturing method comprises the steps of: (1) preparing a culturing medium; (2) culturing detoxified tissue culture seedlings of the aquatic iris; (3) transplanting and culturing the tissue culture seedlings; and the like. A tissue culture rapid propagation technical system established by using aseptic seedling leaves as embryoid inducing explants is characterized by high propagation coefficient, high speed, complete structure, high regeneration rate, low production cost and the like; compared with the general organ tissue culturing method, the tissue culturing method for regenerating somatic cells of the aquatic iris has the advantages of shortening the whole culturing period to 75 days, increasing the seedling proliferation coefficient by 2.4 times, reaching a culture seedling rooting rate of 100 percent and a transplanting survival rate of 100 percent, being applicable to industrial seedling culture and realizing commercialized production. The method can be popularized and applied to enterprise production of the aquatic iris.

Description

technical field [0001] The invention relates to the technical field of plant tissue culture, in particular to a tissue culture method for regeneration using aquatic iris somatic embryos. Background technique [0002] Aquatic iris is a perennial aquatic plant of the family Iridaceae, native to Louisiana, USA. This kind of iris has been widely used in some countries in North America and Europe. It has rich flower colors and a flowering period of 4 to 5 months. Adaptability, showing cold resistance, drought resistance, and extensive tolerance, and at the same time it can remain evergreen in the south of the Yangtze River in my country. At present, the market situation of this series of flower plants is optimistic, but because they are mainly propagated by ramets and seeds, the propagation speed is relatively slow, and it takes a lot of work and time, so the supply of high-quality seedlings is in short supply and cannot meet the needs of the market. [0003] In the past, the ti...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00
Inventor 朱开元刘慧春邹清成周江华金久宏
Owner ZHEJIANG XIAOSHAN COTTON & FLAX RES INST
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