SSR (Simple Sequence Repeats) primer composition and method for identifying high-quality germplasm of picria fe-tearrae Lour.
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The technology of primer composition and identification method is applied in the field of molecular identification of new plant varieties, and achieves the effects of uniform quality of medicinal materials, promotion of development and convenient material selection.
Inactive Publication Date: 2019-04-26
GUANGXI UNIV OF CHINESE MEDICINE
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However, SSR markers have not been reported in the identification of excellent germplasm of Sophora scrophulariae, so it is necessary to establish a molecular method for quickly and accurately identifying the good germplasm of Sophora scrophulariae. Based on a series of experiments, the present invention screened out 9 For SSR core primers, it can quickly and accurately identify excellent germplasm
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Embodiment 1
[0028] The instrument, reagent and implementation process of embodiment 1 and embodiment 2 are as follows:
[0029] 1. Instruments and Reagents
[0030] 1) Main instruments: Bio-Rad PCR amplification instrument, 3730XL sequencer.
[0031] 2) Main reagents: CTBA method DNA extraction kit, PCR kit.
[0032] 2. Experimental process
[0033] 1) extract the DNA of the sample to be tested;
[0034] 2) Synthesis of primers: The primers were synthesized by Sangon Bioengineering (Shanghai) Co., Ltd., and the list of primers is shown in Table 1.
[0035] Table 1 Information of 9 pairs of primers
[0036]
[0037] 3) Reaction conditions
[0038] The volume of the PCR amplification reaction system is 20 μL, including: ddH2O 14.8 μL, dNTP 0.4 μL, Buffer 2 μL, F0.3 μL (20 μM), R0.3 μL (20 μM), DNA template 2 μL, Taq 0.2 μL. The PCR reaction program was: pre-denaturation at 94°C for 5 min; denaturation at 94°C for 30 s, annealing at 54°C (annealing temperature fluctuated around 54°C...
Embodiment 2
[0052] 1. Material: the strain No. zgb04 of Scrophulariaceae collected from Wuzhou, Guangxi, which has the second highest content of Scrophulariae IA among the tested samples, and is an excellent germplasm.
[0053] 2. Results: 9 pairs of primers were used to perform PCR amplification on the DNA extracted from zgb04, the results are shown in Table 4, and the amplification results of 6 pairs of primers are shown in the attached figure 2 .
[0054] Table 4 The results of PCR amplification of DNA extracted from zgb04 by 9 pairs of primers
[0055]
[0056] It can be seen from Table 4 that 9 pairs of primers were used to carry out PCR amplification on the DNA extracted by zgb04, and 7 pairs of primers had amplification products in the length of the fragments that should be amplified, and there was no amplification in the length of the fragments that should not be amplified. The test result meets the identification requirements.
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Abstract
The invention discloses an SSR (Simple Sequence Repeats) primer composition and method for identifying high-quality germplasm of picria fe-tearrae Lour., is used for screening and identifying the high-quality germplasm of the picria fe-tearrae Lour., and belongs to the field of molecular identification of new varieties of plants. According to the invention, 9 pairs of core SSR primers are designed, and by utilizing the 9 pairs of core SSR primers, a feature SSR molecular marker identifying method of the high-quality germplasm of the picria fe-tearrae Lour. is successfully constructed, so thatthe high-quality germplasm of the picria fe-tearrae Lour. can be rapidly and accurately identified. According to the invention, based on the 9 pairs of SSR primers, a PCR (Polymerase Chain Reaction) amplified reaction is performed; and a capillary electrophoresis fluorescence detection method is adopted, so that the high-quality germplasm of the picria fe-tearrae Lour. can be directly judged according to the presence and size of an amplified product fragment. The SSR primer composition and method disclosed by the invention can be directly applied to molecular identification of the high-qualitygermplasm of the picria fe-tearrae Lour., and has good practicability.
Description
technical field [0001] The invention discloses an SSR primer composition for identifying the excellent germplasm of Sophora scrophulariae and an identification method for the excellent germplasm, which are used for screening and identifying the excellent germplasm of Sophora scrophulariae, and belong to the field of molecular identification of new plant varieties. Background technique [0002] Picria fel-tearrae Lour. is the only plant of the genus Scrophulariaceae in the family Scrophulariaceae. It has been used as medicine in Guangxi for more than 200 years, and it is an authentic medicinal material in Guangxi. Its main pharmacological effects are swelling and pain relief, antibacterial, anti-inflammatory and analgesic, etc. One of the main raw materials of products such as Wuzhou Guiling ointment. Its main chemical components are triterpenoids, flavonoids and glycosides, and its main medicinal active ingredients are tetracyclic triterpene glycosides, of which glucoside I...
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