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Application of trnH-psbA sequences as DNA barcodes in identifying Trapa acornis Nakano and identifying method

1. The technology of trnh-psba and barcode is applied in the direction of recombinant DNA technology, DNA/RNA fragments, biochemical equipment and methods, etc., which can solve the problems of the limitation of Nanhu water chestnut identification and achieve short identification time and high repeatability , the effect of high detection accuracy

Active Publication Date: 2015-01-21
ZHEJIANG JIAXING AGRI SCI ACADEMY INST
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Problems solved by technology

Since the leaf morphology of water chestnut and the nutrient form of pulp after removing the shell of water chestnut are basically the same, there are limitations in the identification of water chestnut by traditional morphological taxonomy.

Method used

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  • Application of trnH-psbA sequences as DNA barcodes in identifying Trapa acornis Nakano and identifying method
  • Application of trnH-psbA sequences as DNA barcodes in identifying Trapa acornis Nakano and identifying method
  • Application of trnH-psbA sequences as DNA barcodes in identifying Trapa acornis Nakano and identifying method

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Embodiment 1

[0025] The molecular identification of embodiment 1 Nanhu water chestnut

[0026] Water chestnut fruits of five populations of three types of water chestnut were collected from Zhejiang, Jiangsu and other places, among which Nanhu water chestnut was sampled from four locations in Nanhu District and Xiuzhou District of Jiaxing City, Zhejiang Province. The 5 identified samples were Zhejiang Jiaxing Nanhuling, Zhejiang Wenzhou Erjiao, Jiangsu Nanjing Erjiao, Zhejiang Jiaxing Sijiaohong and Jiangsu Nanjing Sijiaoling. Please refer to the attachment for photos of Nanhu water chestnut fruit figure 1 .

[0027] The genomic DNA of the water chestnut flesh was extracted by CTAB method, PCR amplification was carried out using Trapa-F1 and Trapa-R1 primers, and rTaq (Takara, Dalian, China) was used for PCR amplification.

[0028] Trapa-F1: 5'-TCCGCCCCTTACTTTCTTCC-3'

[0029] Trapa-R1: 5'-CGTAATGCTCACAACTTCCCT-3'

[0030] PCR amplification system: 10×PCR buffer, 2 μL; dNTP mixture, 0....

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Abstract

The invention discloses an application of trnH-psbA sequences as DNA barcodes in identifying Trapa acornis Nakano and an identifying method. The identifying method comprises the following steps: amplifying trnH-psbA sequences of to-be-detected samples by using PCR (polymerase chain reaction) and sequencing, if a 278bp base of the trnH-psbA sequence is A and a 308bp base is T, determining that the to-be-detected samples are Trapa acornis Nakano. Compared with the traditional morphological identification method and DNA fingerprint marking, in the invention, trnH-psbA sequences can be used as DNA barcodes for identifying Trapa acornis Nakano, so that through carrying out PCR amplification and sequencing on the trnH-psbA sequences, identification results can be directly obtained, and the identifying method is high in detection accuracy, high in repeatability and short in identification time.

Description

technical field [0001] The invention relates to the technical field of molecular identification, in particular to the application and identification method of the trnH-psbA sequence as a DNA barcode in identifying Nanhu water chestnut. Background technique [0002] Nanhu water chestnut (Trapa acornis Nakano) is a plant of the genus Trapa acornis Nakano, also known as non-horned water chestnut, monk water chestnut, etc. It is named after its main production in Jiaxing, Zhejiang Province. It is a special aquatic economic crop in my country. It was awarded the National Geographical Indication Protection Product in 2009. title. Nanhu water chestnut fruit has no horns, thin shell, rich in various amino acids, vitamins, minerals and anti-cancer active ingredients, and has important nutritional and medicinal value. [0003] At present, there is no literature report on the molecular identification of Nanhuling. The taxonomic studies on Rhizome plants mostly focus on conventional mo...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q2600/156
Inventor 李军李白高广春黄嬛
Owner ZHEJIANG JIAXING AGRI SCI ACADEMY INST
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