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Molecular marker for itrus reticulata cv. chachiensis strain identification and application thereof
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A technology of molecular marker and tangerine tangerine, applied in the field of gene detection, can solve the problem of mixed tangerine tangerine strains, and achieve the effects of less SNP sites, accurate identification of strains, and avoidance of strain degradation.
Pending Publication Date: 2022-02-18
SOUTH CHINA AGRI UNIV +3
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[0007] The purpose of the present invention is to provide a set of molecular markers and its application for the identification of citrus strains to solve the problem of mixed citrus strains in the prior art
Method used
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[0025] In this example, the plantgenomic DNA rapid extraction kit (N1192), manufacturer: Guangzhou Dongsheng Biotechnology Co., Ltd., was used to extract the DNA of plant tissue samples, and the specific operations were as follows;
[0027] 2. Quickly transfer the ground powder to a centrifuge tube pre-filled with 700μl 65°C pre-warmed buffer GPL, add 1μl RNase, invert and mix quickly, place the centrifuge tube in a 65°C water bath for 20 minutes, and invert the centrifuge during the water bath. tube to mix the sample.
[0028] 3. Add 700 μl of chloroform, mix well, and centrifuge at 12,000 rpm (~13,400×g) for 5 min.
[0029] 4. Carefully transfer the upper aqueous phase obtained in the previous step into a new centrifuge tube, add an equal volume of buffer GPD, and mix well.
[0030] 5. Transfer the mixed ...
Embodiment 2
[0068] In this example, Dadong No. 5, large oil body, and fine oil body are used as standard samples, and 24 Haitou Dawei tea branch orange strain samples (HTDW17-40) are selected as samples to be tested for identification testing. The specific sampling information is as follows Table 4 shows.
[0069] Table 4
[0070]
[0071] In this embodiment, 337 SNP sites of 24 samples to be tested (HTDW17-40) were screened out with reference to the sample DNA extraction and detection, library construction and high-throughput sequencing methods, and molecular marker detection methods described in Example 1, and then The genotypes of 337 SNP sites of 24 samples to be tested (HTDW17-40) and the 8 samples of Dadong No. 5 obtained in Example 1, 8 samples of large oil bodies, and 337 samples of 8 samples of fine oil bodies The genotypes of the SNP sites are processed together with reference to the PCA analysis method in Example 1. The PCA identification results of the Haitoudawei sample ...
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Abstract
The invention discloses a set of molecular marker for citrus reticulata cv. chachiensis strain identification and application thereof. The molecular marker comprises 337 SNP sites given in tables 2-3, and the citrus reticulata cv. chachiensis is Dadong No.5, a large-seed oil body variety and a fine-seed oil body variety . The molecular marker has the advantages that few SNP sites are used, strain identification is accurate, genetic breeding application is facilitated, strain degeneration is avoided, and a basis is provided for guaranteeing the quality of the pericarpium citri reticulatae market.
Description
technical field [0001] The invention relates to the technical field of gene detection, in particular to the identification technology of plant varieties. Background technique [0002] Orange peel (Pericarpium Citri Reticulatae) is a traditional Chinese medicine in my country, also known as orange peel and orange peel; it is the dried peel of Citrus reticulata Blanco and its cultivars. According to the origin, tangerine peel can be divided into "tangerine peel" and "guang tangerine peel". Since ancient times, "Guang Chenpi" has been the top-grade medicinal material of tangerine peel. In the late Qing Dynasty medical book "Materia Medica Benefit", there is a record that "Guangdong Xinhuipi is the best, and the old one is good, hence the name Chenpi". Citrus reticulatacv. Chachiensis is the original plant of tangerine peel, belonging to the genus Citrus of the family Reticulata, mainly produced in Xinhui District, Jiangmen City, Guangdong Province. Tea branch citrus, also kn...
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