Dendrobium DNA (Deoxyribonucleic Acid) fingerprint spectrum and special primer and establishment method for dendrobium DNA fingerprint spectrum

A technology of DNA fingerprinting and fingerprinting, which is applied in the field of plant DNA fingerprinting to achieve the effect of facilitating computer identification and analysis

Inactive Publication Date: 2015-09-09
上海孙桥现代农业联合发展有限公司
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  • Claims
  • Application Information

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  • Dendrobium DNA (Deoxyribonucleic Acid) fingerprint spectrum and special primer and establishment method for dendrobium DNA fingerprint spectrum

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Embodiment 1

[0029] In this example, 20 widely representative Dendrobium strains collected were used as test materials, DNA was extracted from tissue cultured seedlings, SRAP analysis was performed, and corresponding fingerprints were established. The obtained fingerprints were used to identify these 20 Dendrobium species, and provided an effective method for reference for germplasm identification of other Dendrobium species. The concrete operation of setting up the dendrobium DNA fingerprint is as follows:

[0030] 1. Selection of representative Dendrobium species

[0031] The present embodiment adopts 20 parts of Dendrobium species shown in Table 2, including 1 part of Dendrobium nobile Lindl., 1 part of Dendrobium huoshanense, and 18 parts of Dendrobium officinale. The materials are selected to be widely planted in production. Dendrobium strains, see Table 1. In this example, DNA was extracted from 20 dendrobium leaves, and the DNA fingerprints of the 20 dendrobium strains were establ...

Embodiment 2

[0053] Dendrobium dendrobium DNA fingerprint of the present invention is used for germplasm identification of dendrobium

[0054] Assuming that SH in the test material is an excellent variety used in production, it cannot be determined whether the existing batch of Dendrobium is the real Dendrobium SH-15. Its DNA fingerprint can be identified as follows.

[0055] After the dendrobium standard DNA fingerprint is established, when a sample is to be authenticated, extract the DNA of the sample to be tested, use the extracted DNA as a template, use ME1 and EM7; ME1 and EM14; ME3 and EM3; ME3 and EM5; The 6 pairs of primers ME8 and EM14; ME9 and EM3 were amplified by SRAP-PCR, and the amplified products were electrophoresed on a 4% denatured polyacrylamide gel, and silver-stained to draw the DNA fingerprint of the sample to be tested. By comparing the standard DNA fingerprints, it is possible to know whether the variety is the real Dendrobium SH-15.

[0056] The results showed th...

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Abstract

The invention discloses a dendrobium DNA (Deoxyribonucleic Acid) fingerprint spectrum and a special primer and an establishment method for the dendrobium DNA fingerprint spectrum. A nucleotide sequence of the special primer is as shown in a sequence table. The special primer is selected from one or more pairs in the following primer pairs: SEQ ID NO.1 and SEQ ID NO.7; SEQ ID NO.1 and SEQ ID NO.8; SEQ ID NO.2 and SEQ ID NO.5; SEQ ID NO.2 and SEQ ID NO.6; SEQ ID NO.3 and SEQ ID NO.8; SEQ ID NO.4 and SEQ ID NO.5. A genomic DNA of a dendrobium material serves as a template, and any special primer is used for SRAP-PCR (Sequence Related Amplified Polymorphism-Polymerase Chain Reaction) amplification; an obtained SRAP-PCR amplification product is subjected to modified polyacrylamide gel electrophoresis, and a DNA strip is displayed by silver staining to obtain the dendrobium DNA fingerprint spectrum. The dendrobium DNA fingerprint spectrum is created, and dendrobium strains can be subjected to germplasm identification in genetic essence accurately and reliably.

Description

technical field [0001] The invention belongs to the technical field of plant DNA fingerprints, in particular to a dendrobium DNA fingerprint, special primers and a method for establishing the same. Background technique [0002] Dendrobium is the largest genus in the family Orchidaceae. The Dendrobium plants widely planted in my country at present include Dendrobium nobile, Dendrobium Huoshanense, Dendrobium officinale and so on. In particular, Dendrobium officinale Kimura et Migo has the effects of nourishing the stomach and promoting body fluid, nourishing yin and clearing heat (Chinese Pharmacopoeia (2010)), and the planting area has developed rapidly in my country. In production, the quality of Dendrobium officinale is mostly evaluated by appearances such as sweet taste, heavy quality, flexibility, and strong stickiness. At present, Dendrobium candidum is expensive, and there are many varieties of Dendrobium candidum in the market. There are differences in the color of ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q1/6858C12Q2600/13C12Q2600/156C12Q2531/113C12Q2565/125
Inventor 李怀志刘士辉姚永康孙文华金静郭飞孙洪助刘青海
Owner 上海孙桥现代农业联合发展有限公司
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