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Chloroplast genome capable of identifying populus species as well as PCR amplification primer and application of chloroplast genome

A technology of chloroplast genome and tree species, which is applied in the field of plant species identification and molecular biology, and can solve the problems of difficult amplification, sequencing and assembly, high identification rate, and cumbersome methods

Active Publication Date: 2021-06-08
SOUTHWEST FORESTRY UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the length of the whole chloroplast genome sequence of Populus species is about 150Kb, and its amplification, sequencing and assembly are difficult, and the method is cumbersome, which is not conducive to popularization
In addition, some chloroplast fragments such as atpF-H, matK, psbK-I, ​​rbcL, rpoC1, trnL-trnF, and trnH-psbA were also used as DNA barcodes for identification of Populus species, but the single fragment sequences were different at the genus or group level. The identification rate is high, but at the species level, especially the closely related Populus species are difficult to distinguish

Method used

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  • Chloroplast genome capable of identifying populus species as well as PCR amplification primer and application of chloroplast genome
  • Chloroplast genome capable of identifying populus species as well as PCR amplification primer and application of chloroplast genome
  • Chloroplast genome capable of identifying populus species as well as PCR amplification primer and application of chloroplast genome

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Embodiment Construction

[0020] The present invention will be further described below in conjunction with the accompanying drawings and specific embodiments.

[0021] 1. Materials

[0022] The young leaves of different species of Populus were collected, stored in a ziplock bag filled with silica gel for rapid drying, and brought back to the laboratory for later use. The materials used are shown in Table 1, including:

[0023] Table 1 Sample Source Table

[0024]

[0025] 2. Genomic DNA extraction

[0026] Weigh 100 mg of poplar leaves dried with color-changing silica gel into a 2 mL centrifuge tube, add an appropriate amount of PVP powder, and grind with a freeze-mixing ball mill. The ground samples are extracted from the total genomic DNA according to the Ezup Column Plant Genomic DNA Extraction Kit.

[0027] 3. PCR amplification

[0028] The 9 hypervariable regions were amplified by PCR. The total volume of the PCR reaction was 25 μL, the DNA template was 1.0 μL, the 10×buffer Mix was 12.5 μL...

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Abstract

The invention belongs to the technical field of molecular biology, relates to the field of plant species identification, and discloses a chloroplast genome sequence capable of specifically identifying populus species and a specific PCR amplification primer of the chloroplast genome sequence, the specific primer comprises 9 hypervariable regions in the populus chloroplast genome, including ccsA-ndhD, ndhC-trnV, ndhF-ccsA, psbM-trnD, psbZ-trnfM, rpoB-petN, rps15-ycf1, ycf1 and trnK-psbK respectively, and the nucleotide sequences of the 9 hypervariable regions are shown in SEQ ID NO.1-9. The invention aims to provide the 9 hypervariable regions of the chloroplast genome suitable for identifying the populus species and the specific primer of the chloroplast genome, the 9 hypervariable region sequences are combined, the populus species can be rapidly and accurately identified, and an effective DNA bar code is provided for identifying the populus species.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, relates to the field of plant species identification, in particular to a specific primer capable of specifically identifying the chloroplast genome sequence of Populus species and PCR amplification thereof. Background technique [0002] Poplar belongs to the species of Populus salicaceae. Because of its characteristics of easy cultivation, rapid growth, strong adaptability, wide distribution and wide application, it has become one of the most important tree species for afforestation. However, because poplar is easy to hybridize between species and within species, introgression phenomenon is also common, and intraspecific variation is relatively rich, which makes it difficult to identify Populus species only by phenotypic traits. [0003] Chloroplast genomes are widely used in the study of phylogenetic relationships due to their simple structure, low recombination rate, and uniparental i...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12Q1/6806C12N15/11
CPCC12Q1/6895C12Q1/6806C12Q2600/156C12Q2531/113C12Q2565/125
Inventor 纵丹何承忠江涛张晓琳王静敏
Owner SOUTHWEST FORESTRY UNIVERSITY
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