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Screening method for fast and effectively reducing false positive rate of peanut genetic transformation plant

A screening method and genetic transformation technology, applied in the field of crop seedling raising, can solve the problems of false positives of transformed plants, reduce the false positive rate of genetically transformed plants of peanuts, etc., and achieve the effect of reducing workload and saving time.

Inactive Publication Date: 2014-09-03
BIOTECH RES CENT SHANDONG ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Aiming at the problem that there are many false positives in transformed plants in the current peanut genetic transformation, the present invention provides a screening method for quickly and effectively reducing the false positive rate of peanut genetic transformation plants

Method used

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  • Screening method for fast and effectively reducing false positive rate of peanut genetic transformation plant
  • Screening method for fast and effectively reducing false positive rate of peanut genetic transformation plant
  • Screening method for fast and effectively reducing false positive rate of peanut genetic transformation plant

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Agrobacterium strain: LBA4404, available from Beijing Dingguo Changsheng Biotechnology Co., Ltd.

[0053] Plant expression vector: pROK II, which contains nptII gene, which can impart kana resistance to transgenic plants. Available from Beijing Dingguo Changsheng Biotechnology Co., Ltd.

[0054] The peanut variety "Luhua 14" is available from Qingdao Jiahua Seed Industry Co., Ltd.

[0055] MS basic medium, the components are as follows:

[0056] NH 4 NO 3 , 1.65g / L; KNO 3 , 1.9g / L; CaCl 2 2H 2 O, 0.44g / L; MgSO 4 ·7H 2 O, 0.37g / L; KH 2 PO 4 , 0.17g / L; KI, 0.83mg / L; H 3 BO 3 , 6.2mg / L; MnSO 4 4H 2 O, 22.3mg / L; ZnSO 4 ·7H 2 O, 8.6 mg / L; Na 2 MoO 4 2H 2 O, 0.25mg / L; CuSO 4 ·5H 2 O, 0.025mg / L; CoCl 2 ·6H 2 O, 0.025mg / L; FeSO 4 ·7H 2 O, 27.8mg / L; Inositol, 100mg / L; Glycine, 2mg / L; Thiamine Hydrochloride, 0.1mg / L; Pyridoxine Hydrochloride, 0.5mg / L; Niacin, 0.5mg / L; Sucrose, 30g / L; agar, 7.2g / L.

[0057] The liquid MS basal medium is obtained by remov...

Embodiment 2

[0083] Agrobacterium strain: LBA4404, available from Beijing Dingguo Changsheng Biotechnology Co., Ltd.

[0084] Plant expression vector: pCAMBIA2301, which contains nptII gene, which can impart kana resistance to transgenic plants. Also contains the reporter gene GUS. Available from Beijing Dingguo Changsheng Biotechnology Co., Ltd.

[0085]The peanut variety "Fenghua No. 1" is available from Shandong Xiangfeng Seed Industry Co., Ltd.

[0086] MS basic medium, the components are as follows:

[0087] NH 4 NO 3 , 1.65g / L; KNO 3 , 1.9g / L; CaCl 2 2H 2 O, 0.44g / L; MgSO 4 ·7H 2 O, 0.37g / L; KH 2 PO 4 , 0.17g / L; KI, 0.83mg / L; H 3 BO 3 , 6.2mg / L; MnSO 4 4H 2 O, 22.3mg / L; ZnSO 4 ·7H 2 O, 8.6 mg / L; Na 2 MoO 4 2H 2 O, 0.25mg / L; CuSO 4 ·5H 2 O, 0.025mg / L; CoCl 2 ·6H 2 O, 0.025mg / L; FeSO 4 ·7H 2 O, 27.8mg / L; Inositol, 100mg / L; Glycine, 2mg / L; Thiamine Hydrochloride, 0.1mg / L; Pyridoxine Hydrochloride, 0.5mg / L; Niacin, 0.5mg / L; Sucrose, 30g / L; agar, 7.2g / L.

[008...

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Abstract

The invention relates to a screening method for fast and effectively reducing the false positive rate of a peanut genetic transformation plant. The screening method comprises the following steps of: (1) preparing a bacteria solution to be transformed; (2) preparing a transformed hypocotyledonary axis; (3) initially screening a plant bud; (4) moderately screening the plant bud; (5) deeply screening the plant bud; and (6) culturing the deeply screened plant bud to prepare a transformation plant. The screening method disclosed by the invention can be used for removing a majority of cluster buds which are not successfully transformed at the initial stage of cluster bud induction in a screening mode of sequentially increasing screening pressure, and a part of albescent false positive plants can be removed in each subculture, thereby saving the time, greatly reducing the later workload and ensuring that the majority of finally obtained green plants are real positive plants.

Description

technical field [0001] The invention relates to a screening method for quickly and effectively reducing the false positive rate of peanut genetically transformed plants, and belongs to the technical field of crop seedling cultivation. Background technique [0002] Peanut is an important oil crop and economic crop in my country, and has very important economic value. Since the 1940s, research on peanut tissue culture has been gradually started, and a variety of peanut tissue culture regeneration systems have been established. Since Eapen and George first reported obtaining peanut transgenic plants in 1994, there have been many successful reports on peanut genetic transformation. [0003] Although some progress has been made in the genetic transformation of peanut, transgenic peanut regenerated plants can be obtained through Agrobacterium-mediated methods, but the transformation and regeneration frequency of peanut is low, which cannot meet the needs of large-scale peanut gen...

Claims

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Application Information

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IPC IPC(8): C12N15/84A01H4/00A01H5/00
Inventor 彭振英毕玉平万书波边斐王兴军李新国单雷郑玲
Owner BIOTECH RES CENT SHANDONG ACADEMY OF AGRI SCI
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