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Method for establishing efficient plant regeneration system by using hevea brasiliensis embryonic cell suspension system

A technology of Hevea brasiliensis and embryogenic cells, applied in plant regeneration, botanical equipment and methods, gardening methods, etc., to achieve the effects of low production cost, high induction rate, and high plant regeneration rate

Inactive Publication Date: 2012-11-07
RUBBER RES INST CHINESE ACADEMY OF TROPICAL AGRI SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the limitations of many factors, there are still many difficulties in establishing a high-efficiency plant regeneration system using the suspension line of Hevea brasiliensis embryogenic cells. How to solve these difficulties and finally establish a stable and efficient regeneration system has become the most urgent task at present.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Induction of fragile embryogenic callus by embryogenic suspension cells: use a pipette gun to suck up the culture medium in the embryogenic cell suspension of Hevea brasiliensis, and select small cell clusters to inoculate into the embryogenic cell suspension to induce fragile embryogenic callus. On the culture medium of wounded tissue, cultured in dark for 20 days, and the culture temperature was 28°C. The medium for inducing fragile embryogenic callus is MS medium and additional components. The content of MS medium is 500 mg / L of magnesium sulfate heptahydrate, 400 mg / L of potassium dihydrogen phosphate, and 250 mg / L of anhydrous calcium chloride. and manganese sulfate tetrahydrate 35mg / L (the same below), the additional components are 2,4-D 1.5mg / L, 6-BA 0.5mg / L, NAA 1.0mg / L, Phytagel 2.0g / L, sucrose 70g / L And coconut water 50ml / L.

[0022] Fragile embryogenic callus subculture: transfer the fragile embryogenic callus to the medium for inducing fragile embryogenic c...

Embodiment 2

[0027] Preparation of explants: Select Reyan 7-33-97 flower buds at the single-nucleus stage as explants, disinfect the surface with 75% alcohol for 60 seconds, and then use 1g / L HgCl 2 Disinfect for 10 minutes, and finally rinse with sterile water 3 times, 5 minutes each time.

[0028]Embryogenic callus induction: the stamens from the sterilized flower buds were inoculated on the embryogenic callus induction medium and cultured in the dark at 28°C for 40 days. The active ingredients of the embryogenic callus induction medium are improved MS medium and additional components, and the modified MS medium is: magnesium sulfate heptahydrate 500mg / L, potassium dihydrogen phosphate 400mg / L, anhydrous chlorine Calcium chloride 250mg / L, manganese sulfate tetrahydrate 35mg / L (the following steps are the same), additional ingredients are 2,4-D 2.0mg / L, KT1.0mg / L, NAA 1.0mg / L, Phytagel 2.0g / L, Sucrose 70g / L and coconut water 50ml / L.

[0029] Subculture of embryogenic callus: the induced...

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Abstract

The invention discloses a method for establishing an efficient plant regeneration system by using a hevea brasiliensis embryonic cell suspension system. The method comprises the steps of friable embryonic callus induction with embryonic suspension cells, friable embryonic callus subculture, differentiation of body cell embryoid, plant regeneration and the like. The method is simple in process flow, low in production cost, high in embryoid inductivity and high in plant regeneration rate, and has great application value.

Description

technical field [0001] The invention relates to a method for establishing a high-efficiency plant regeneration system using a suspension system of embryogenic cells of Hevea brasiliensis, belonging to the field of plant tissue and cell engineering. Background technique [0002] Hevea brasiliensis Muell.Arg is a perennial cross-pollinated tree belonging to the Euphorbiaceae Hevea genus. As the main source of natural rubber, it is widely planted in tropical regions and has always played an important role in the world's economic development. [0003] Breeding of Hevea brasiliensis mainly relies on conventional breeding, but the breeding cycle is long and the materials used for conventional breeding are all derived from the strains of the Wei Kehan ​​system and its derived clones, and the genetic basis is narrow. In order to broaden the breeding methods of Brasiliensis and improve its quality, many countries have invested a lot of human, material and financial resources in exten...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00
Inventor 张全琪倪燕妹黄志彭远明王力前张能李孝云黄春华叶青
Owner RUBBER RES INST CHINESE ACADEMY OF TROPICAL AGRI SCI
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