ISSR-SCAR (inter simple sequence repeat-sequence characterized amplified region) marker capable of identifying Wujiang brassica chinensis and identification method of marker

Abstract

The invention discloses an ISSR-SCAR (inter simple sequence repeat-sequence characterized amplified region) marker capable of identifying Wujiang brassica chinensis and an identification method of the marker. The method includes: performing total DNA (deoxyribose nucleic acid) extraction, PCR (polymerase chain reaction) amplification, primer selection and ISSR-SCAR detection on Wujiang brassica chinensis and other 10 brassica chinensis varieties, and finally determining an ISSR-SCAR molecular marker and a molecular identification method of the Wujiang brassica chinensis. A primer shown in the sequence such as SEQ ID NO:1 is used for amplification to obtain a specific fragment shown in the sequence such as SEQ ID NO:2. According to the sequence, a pair of primers in the sequences such as SEQ ID NO:3 and SEQ ID NO:4 is designed. The marker is simple to operate, stable and reliable, is the first molecular identification marker applied to the variety of Wujiang brassica chinensis, can be used for quickly identifying the variety and has significance of protecting germplasm resources of geographical indication products.

Description

technical field

[0001] The invention belongs to the field of molecular marker identification, in particular to an ISSR-SCAR marker capable of identifying Wujiang fragrant green vegetables and an identification method thereof. Background technique

[0002] Green vegetables( Brassica chinensis L.) is also called Chinese cabbage and non-heading Chinese cabbage. It belongs to the genus Brassicaceae. It is an annual or biennial herb and is an important vegetable and oil crop. Green vegetables are originally produced in China, but in recent years, some countries such as Southeast Asia, Europe and the United States have gradually introduced and cultivated them, and they have become a worldwide vegetable. Among them, Xiangqingcai is not only a unique green vegetable variety in my country, but also a rare vegetable variety with local traditional characteristics in Suzhou. It has been cultivated for more than 100 years. It is mainly planted along the East Taihu Lake and the south...

Examples

Embodiment Construction

[0019] Embodiments of the present invention will be described in detail below in conjunction with the accompanying drawings.

[0020] 1. Extraction of DNA from samples to be tested

[0021] Select the plants with good growth conditions of the plants to be tested (Table 1), collect 100 mg of healthy young leaves, and use the Easy Pure Plant Genomic DNA Kit (Beijing Quanshijin Biotechnology Co., Ltd.) to extract the total DNA of the samples according to the instructions. The concentration and purity of the DNA was detected with a nucleic acid protein detector (eppendorf, USA), the DNA concentration was adjusted to 20 ng / μl, and the DNA samples were stored at -20°C for later use.

[0022] Table 1 Experimental material table

[0023]

[0024] 2. Screening of ISSR primers

[0025] The PCR reaction was carried out on a BioMetra T1 PCR instrument, the reaction system was 20 μl, and its components and final concentrations were: 20 ng DNA template (20 ng / μl) 1.0 μl, 2×Reaction M...

technical field

[0001] The invention belongs to the field of molecular marker identification, in particular to an ISSR-SCAR marker capable of identifying Wujiang fragrant green vegetables and an identification method thereof. Background technique

[0002] Green vegetables( Brassica chinensis L.) is also called Chinese cabbage and non-heading Chinese cabbage. It belongs to the genus Brassicaceae. It is an annual or biennial herb and is an important vegetable and oil crop. Green vegetables are originally produced in China, but in recent years, some countries such as Southeast Asia, Europe and the United States have gradually introduced and cultivated them, and they have become a worldwide vegetable. Among them, Xiangqingcai is not only a unique green vegetable variety in my country, but also a rare vegetable variety with local traditional characteristics in Suzhou. It has been cultivated for more than 100 years. It is mainly planted along the East Taihu Lake and the south...

Examples

Embodiment Construction

[0019] Embodiments of the present invention will be described in detail below in conjunction with the accompanying drawings.

[0020] 1. Extraction of DNA from samples to be tested

[0021] Select the plants with good growth conditions of the plants to be tested (Table 1), collect 100 mg of healthy young leaves, and use the Easy Pure Plant Genomic DNA Kit (Beijing Quanshijin Biotechnology Co., Ltd.) to extract the total DNA of the samples according to the instructions. The concentration and purity of the DNA was detected with a nucleic acid protein detector (eppendorf, USA), the DNA concentration was adjusted to 20 ng / μl, and the DNA samples were stored at -20°C for later use.

[0022] Table 1 Experimental material table

[0023]

[0024] 2. Screening of ISSR primers

[0025] The PCR reaction was carried out on a BioMetra T1 PCR instrument, the reaction system was 20 μl, and its components and final concentrations were: 20 ng DNA template (20 ng / μl) 1.0 μl, 2×Reaction M...

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