A transposon insertion fragment of Dali tea and its application as a molecular marker to distinguish Dali tea from other tea plants

By PCR amplifying the 1082 bp transposon insertion fragment in the promoter of the CsCAD14 gene of Dali tea, special primers P1 and P2 were designed, which solved the problem of distinguishing Dali tea from other tea plants and achieved a fast, simple and low-cost identification effect.

CN118028526BActive Publication Date: 2025-09-16ANHUI AGRICULTURAL UNIVERSITY
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Patent Information

Application Number
CN202410336925.7
Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
Filing Date
2024-03-22
Publication Date
2025-09-16
Estimated Expiration
2044-03-22

AI Technical Summary

Technical Problem

Existing technologies are unable to quickly and easily distinguish Dali tea from other tea group plants, resulting in difficulties in protecting and identifying wild Dali tea resources.

Method used

PCR amplification technology was used to utilize the unique 1082 bp transposon insertion fragment in the promoter of the CsCAD14 gene of Dali tea. Special PCR primers P1 and P2 were designed for amplification, and agarose gel electrophoresis analysis was used to distinguish Dali tea from other tea plants.

Benefits of technology

The method achieved a rapid, simple and low-cost method of distinguishing Dali tea from other tea plants at the DNA level. The results had good repeatability and specificity and were suitable for the identification of common Dali tea and other tea plants.

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Abstract

The present invention provides a transposon insert of Dali tea and its use as a molecular marker to distinguish Dali tea from other tea plants, belonging to the technical field of DNA molecular marker identification of tea trees. A comparison of the CsCAD14 gene promoter fragments of Dali tea and other tea varieties within the whole genome revealed a 1082-bp transposon insert fragment, the sequence of which is shown in SEQ ID No. 1, only present in the CsCAD14 gene promoter of Dali tea. This insert fragment is not present in other tea plants. Therefore, by using two pairs of PCR primers to identify the CsCAD14 gene promoter and insert fragment, respectively, Dali tea can be effectively distinguished from other tea plants. Furthermore, compared with traditional tea variety identification methods, the present invention provides a simpler, faster, and more accurate method for distinguishing Dali tea from other tea plants on the market.
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Claims

1. Use of primers for amplifying transposon inserts of Dali tea in distinguishing Dali tea from other tea plants, characterized in that: The species of the other tea group plants are one or more of 'Foxiang No. 1', 'Fuding Dabai', 'Fuhao', 'Huangguanyin', 'Jinxuan', 'Longjing Changye', 'Puyan No. 2', 'Qingxin Damo', 'Xinxuan 801', 'Yaoshan Xiulu', 'Shucha Zao', 'Tieguanyin', 'Cinnamon', 'Yiqibao Black Tea', 'Yuncha No. 1', 'Yunkang 10', 'Yunkang 15', 'Yunkang 50', 'Zijuan', 'Purple Peony', 'Mengku Daye', 'Yinghong 9', 'Longye Baihao', 'Dachang Tea', 'Guangxi Tea', 'Jinyunshan Tea', 'Rucheng Maoye Tea', 'Xi'e Tea', 'Dashu Tea', 'Laohei Tea', 'Narrow Leaf Tea', 'Moye Tea', 'Siqiu Tea', and 'Tufang Tea'; the nucleotide sequence of the transposon insertion fragment is as shown in SEQ ID No .1; the amplification primers include an upstream primer P3 with a nucleotide sequence as shown in SEQ ID No. 6 and a downstream primer P4 with a nucleotide sequence as shown in SEQ ID No. 7; if the target band amplified by the upstream primer P3 and the downstream primer P4 is 646 bp, it is Dali tea; if there is no 646 bp target band, it is other tea group plants.

2. The use according to claim 1, characterized in that The species of the other tea group plant is 'Foxiang No. 1'.

3. The use according to claim 1, characterized in that The species of the other tea group plant is 'Fuding Dabai'.

4. The use according to claim 1, characterized in that The species of the other tea group plant is 'Fuhao'.

5. The use according to claim 1, characterized in that The species of the other tea group plant is 'Huang Guanyin'.

6. The use according to claim 1, characterized in that The species of the other tea group plant is 'Jinxuan'.

7. The use according to claim 1, characterized in that The species of the other tea group plant is 'Longjing Longye'.

8. Use of primers for amplifying the left and right conserved sequences of the transposon insert of claim 1 in distinguishing Dali tea from other tea plants, characterized in that: The amplification primers include an upstream primer P1 having a nucleotide sequence as shown in SEQ ID No. 4 and a downstream primer P2 having a nucleotide sequence as shown in SEQ ID No. 5; the species of the other tea plants are 'Foxiang No. 1', 'Fuding Dabai', 'Fuhao', 'Huangguanyin', 'Jinxuan', 'Longjing Changye', 'Puyan No. 2', 'Qingxin Damo', 'Xinxuan 801', 'Yaoshan Xiulu', 'Shucha Zao', 'Tieguanyin', 'Cinnamon', 'Yiqibao Black Tea', 'Yuncha No. 1', 'Yunkang 10', 'Yunkang 15', 'Yunkang 50', 'Zijuan', 'Zi Mudan', 'Mengku Daye', 'Yinghong 9', 'Changye Baihao', 'Dachang Tea', 'Guangxi Tea', 'Jinyunshan Tea', 'Rucheng Maoye Tea', 'Xi'e Tea', 'Dashu Tea', 'Laohei Tea', 'Narrow Leaf Tea', 'Moye Tea', 'Siqiu Tea', and 'Tufang Tea'; if the target band amplified by the upstream primer P1 and the downstream primer P2 is 1421 bp, it is Dali tea; if the target band is 339 bp, it is other tea plants.

9. The use according to claim 8, characterized in that The upstream primer P1 corresponds to the conserved sequence on the left side of the inserted fragment in Dali tea and other tea plants, and the downstream primer P2 corresponds to the conserved sequence on the right side of the inserted fragment in Dali tea and other tea plants, wherein the conserved sequence on the left side of the inserted fragment is shown as SEQ ID No. 2, and the conserved sequence on the right side of the inserted fragment is shown as SEQ ID No.

3.

10. A method for distinguishing Dali tea from other tea plants using the amplification primers according to claim 8, characterized in that: The following steps are involved: (1) Extract DNA from different varieties of tea trees; (2) using the upstream primer P1 and the downstream primer P2 to perform PCR amplification on the DNA of different varieties of tea trees obtained in step (1); (3) Analyze the PCR amplified bands by agarose gel electrophoresis. If the target band is 1421 bp, it is Dali tea. If the target band is 339 bp, then it is other tea group plants; the species of the other tea group plants are one or more of 'Foxiang No. 1', 'Fuding Dabai', 'Fuhao', 'Huangguanyin', 'Jinxuan', 'Longjing Changye', 'Puyan No. 2', 'Qingxin Damo', 'Xinxuan 801', 'Yaoshan Xiulv', 'Shucha Zao', 'Tieguanyin', 'Cinnamon', 'Yiqibao Black Tea', 'Yuncha No. 1', 'Yunkang 10', 'Yunkang 15', 'Yunkang 50', 'Zijuan', 'Purple Peony', 'Mengku Daye', 'Yinghong 9', 'Longye Baihao', 'Dachang Tea', 'Guangxi Tea', 'Jinyunshan Tea', 'Rucheng Maoye Tea', 'Xi'e Tea', 'Dashu Tea', 'Laohei Tea', 'Narrow Leaf Tea', 'Moye Tea', 'Siqiu Tea' and 'Tufang Tea'.

Citation Information

Patent Citations

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