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.
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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.
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