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Method for inhibiting endophytic bacterial contamination of broccoli tissue culture seedlings

A technology of tissue culture seedlings and endophytes, which is applied in the field of plant tissue culture, can solve the problems of harmful mammals, microbial resistance, and easy wrong use, so as to reduce the contamination rate of endophytes, inhibit growth and spread, The effect of easy operation

Inactive Publication Date: 2017-01-11
ZHENJIANG AGRI SCI INST JIANGSU HILLY AREAS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the use of antibiotics may be harmful to mammals; if plants with a longer growth period are used, microorganisms will also develop resistance; and it is easy to use them incorrectly, such as using various antibiotics without targeting microorganisms

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Proliferation medium is: MS+6-BA 2mg / L+NAA 0.05mg / L, containing 20% ​​sucrose, 0.8% agar, 0.1‰ Ebelon, pH value is 5.8; rooting medium is 1 / 2 MS+0.1 mg / L IBA+0.15 mg / L NAA, containing 20% ​​sucrose, 0.8% agar, 0.1‰ Epelon, pH 6.0.

[0025] The rooting medium is 1 / 2 MS + 0.1 mg / L IBA + 0.1 mg / L NAA, containing 20% ​​sucrose, 0.8% agar, 0.1‰ Eppelon, and the pH value is 6.0.

[0026] The MS medium, in 1L, consists of: NH 4 HO 3 1650 mg, KNO 3 1900 mg, CaCl 2 2H 2 O 440mg, KH 2 PO 4 170 mg, MgSO 4 ·7H 2 O 370 mg, FeSO 4 ·7H 2 O 27.8 mg, Na 2 EDTA 37.3 mg, H 3 BO 3 6.2 mg, MnSO 4 ·H 2 O 16.9 mg, ZnSO 4 ·7H 2 O 8.6 mg, Na 2 MoO 4 2H 2 O 0.25 mg, CuSO 4 ·5H 2 O 0.025mg, CoCl 2 ·6H 2 O 0.025 mg, KI 0.83 mg, inositol 100 mg, glycine 2 mg, niacin 0.5 mg, vitamin B10.1 mg, vitamin B6 0.5 mg and the rest sterile water.

[0027] Retrieve the growing young stalked curds (including at least one bud point) of broccoli from the field, cut them into small pieces...

Embodiment 2

[0030]Proliferation medium is: MS+6-BA 2mg / L+NAA 0.05mg / L, containing 20% ​​sucrose, 0.9% agar, 0.15‰ Ebelon, pH value is 5.8; rooting medium is 1 / 2 MS+0.1 mg / L IBA+0.15 mg / L NAA, containing 20% ​​sucrose, 0.9% agar, 0.15‰ Epelon, pH 5.9.

[0031] The rooting medium is 1 / 2 MS + 0.15 mg / L IBA + 0.2 mg / L NAA, containing 30% sucrose, 1.0% agar, 0.2‰ Ebelon, and the pH value is 5.9.

[0032] The MS medium, in 1L, consists of: NH 4 HO 3 1650 mg, KNO 3 1900 mg, CaCl 2 2H 2 O 440mg, KH 2 PO 4 170 mg, MgSO 4 ·7H 2 O 370 mg, FeSO 4 ·7H 2 O 27.8 mg, Na 2 EDTA 37.3 mg, H 3 BO 3 6.2 mg, MnSO 4 ·H 2 O 16.9 mg, ZnSO 4 ·7H 2 O 8.6 mg, Na 2 MoO 4 2H 2 O 0.25 mg, CuSO 4 ·5H 2 O 0.025mg, CoCl 2 ·6H 2 O 0.025 mg, KI 0.83 mg, inositol 100 mg, glycine 2 mg, niacin 0.5 mg, vitamin B10.1 mg, vitamin B6 0.5 mg and the rest sterile water.

[0033] Use broccoli seeds as explants (the explants are full and free from diseases and insect pests), sterilize the surface with 75% etha...

Embodiment 3

[0036] Proliferation medium is: MS+6-BA 2mg / L+NAA 0.05mg / L, containing 20% ​​sucrose, 0.9% agar, 0.2‰ Ebelon, pH value is 6.0; rooting medium is 1 / 2 MS+0.1 mg / L IBA+0.15 mg / L NAA, containing 20% ​​sucrose, 0.9% agar, 0.2‰ Epelon, pH 6.0.

[0037] The rooting medium is 1 / 2 MS + 0.13 mg / L IBA + 0.15 mg / L NAA, containing 25% sucrose, 0.9% agar, 0.15‰ Epelon, and the pH value is 5.8.

[0038] The MS medium, in 1L, consists of: NH 4 HO 3 1650 mg, KNO 3 1900 mg, CaCl 2 2H 2 O 440mg, KH 2 PO 4 170 mg, MgSO 4 ·7H 2 O 370 mg, FeSO 4 ·7H 2 O 27.8 mg, Na 2 EDTA 37.3 mg, H 3 BO 3 6.2 mg, MnSO 4 ·H 2 O 16.9 mg, ZnSO 4 ·7H 2 O 8.6 mg, Na 2 MoO 4 2H 2 O 0.25 mg, CuSO 4 ·5H 2 O 0.025mg, CoCl 2 ·6H 2 O 0.025 mg, KI 0.83 mg, inositol 100 mg, glycine 2 mg, niacin 0.5 mg, vitamin B10.1 mg, vitamin B6 0.5 mg and the rest sterile water.

[0039] Retrieve the growing young stalked curds (including at least one bud point) of broccoli from the field, cut them into small pieces...

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Abstract

The invention discloses a method for inhibiting endophytic bacterial contamination of broccoli tissue culture seedlings. The method mainly comprises the following steps: take seeds or axillary buds as explants, rinsing in tap water for 30-60 minutes, absorbing water on the surfaces of the explants by using absorbent paper, putting the explants into a sterile culture flask, pertinently sterilizing for 30-60 seconds by using 75% ethanol, then disinfecting for 13-20 minutes by using a sterilizing agent, then rinsing for 3-5 times by using sterile water, and then inoculating a multiplying medium with the sterile explants, and culturing in a culture chamber at the temperature of 25+ / -2 DEG C; on the 20th-25th day in the culturing process, multiplying to form new buds, separating the new buds, and sterilely transferring the new buds to the culture medium for further culturing; transferring regrown seedlings suffering from the endophytic bacterial contamination to a rooting medium for culturing, and then transferring the regrown seedlings to a kindred rooting medium for culturing until rooting. The method has the advantages of simple and convenient operation, low pollution rate and the like.

Description

technical field [0001] The invention relates to the technical field of plant tissue culture, in particular to a method for inhibiting endophyte pollution of broccoli tissue culture seedlings. Background technique [0002] Broccoli ( Brassia oleracea L. var. varitalica Planch), also known as broccoli, green cauliflower, and tender stem broccoli, is a brassica vegetable of the genus Brassica, rich in various vitamins, minerals, and natural anti-cancer and anti-cancer substances. Very tall vegetables. Broccoli originated from the Mediterranean coast, domestic germplasm resources are scarce, seeds are expensive, and foreign excellent germplasm is difficult to obtain. These factors restrict the development of domestic broccoli variety breeding. The cultivation of high-quality, stress-resistant and disease-resistant varieties has always been one of the hot issues that breeders pay attention to. The primary key to cultivating fine varieties is to breed broccoli germplasm resourc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
CPCA01H4/001A01H4/008
Inventor 张振超姚悦梅秦文斌戴忠良肖燕吴国平孙春青潘永飞毛忠良马志虎
Owner ZHENJIANG AGRI SCI INST JIANGSU HILLY AREAS
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