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A kind of molecular marker primer and method for differentiating large flat apricot, Siberian apricot and fresh apricot

A molecular marker technology, applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., can solve the problems of complex structure and composition, low copy number, multiple copies of genes, etc. The effect of improving detection speed and efficiency

Active Publication Date: 2021-02-19
INNER MONGOLIA AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Among the plant DNA barcodes, available genomic DNAs include nuclear genomic DNA (nrDNA), chloroplast genomic DNA (cpDNA) and mitochondrial genomic DNA (mtDNA). Among them, nrDNA has the fastest evolution rate and is a fully autonomous genetic system in the nucleus. The structure and composition are extremely complex, each gene family forms a functional regulatory network, and many genes have multiple copies, and there are problems such as orthologs and paralogs
However, mtDNA is prone to gene rearrangement, duplication and loss, and the copy number of plant mtDNA is low, so it is difficult to extract and purify. Therefore, the application range of mtDNA in plant identification is limited.

Method used

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  • A kind of molecular marker primer and method for differentiating large flat apricot, Siberian apricot and fresh apricot
  • A kind of molecular marker primer and method for differentiating large flat apricot, Siberian apricot and fresh apricot
  • A kind of molecular marker primer and method for differentiating large flat apricot, Siberian apricot and fresh apricot

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056]1. Collection of blade samples

[0057]The sample was identified and collected by the Inner Mongolia forest-raising center, and the vicinity, healthy and diseased blade samples were selected from the different origins of the largelactic apricots, Siberian apricots and fresh food apricots. As described in Table 1.

[0058]Table 1 Leaves samples and sources of large flatpric apricot, Siberian apricot and fresh food

[0059]

[0060]2. Extraction of the total genomic DNA of the blade sample

[0061]Through the plant DNA rapid extraction kit, the total genomic DNA of the blade sample was extracted, and the quality was detected based on 1% agarose gel electrophoresis.figure 1 The brightness, uniformity and point-like mouth spot of the electrophoresis strip are good, indicating that the purity of the sample is high, no pollution, and can be directly used for subsequent molecular biology experiments.

[0062]3. PCR Amplifies the target gene sequence in the total genomic DNA of the blade sample

[0063]Us...

Embodiment 2

[0078]Example 2 results analysis

[0079]After purification of the PCR product, two-way sequencing, and comparison of the sequence of the largelastic apricot blades, Siberian apricot blade samples and fresh food apricot leaves sample, Siberia apricot blade sample And Fresh Food Apricot The comparison of the comparison of the sequence of the chloroplast gene trylnk-uuu-rps16 is:

[0080]Big flat apricot:

[0081]AgattgaattgcacagcgcaatcaaaaaataaatttaaaaAtttctaattgattatttctgaacataataaaaatTAacggataaaAtAmaGgcatttaaataTttaaataTtt (aaatatt)+AaggaccgccccTTGTCTTTATGTTATCCATTAATGA (Attaga)+AttagtacatatttttttattaattttaatcttaatcaaaataatAAAAAAAAGACTTCTTGTATCACAGAAAATCCAAGAACCCATTATTTGAATGAAATAAAATCTATGGGACAGAGATATAAATAGATCCTTATTATCCACGATCGGATTATATTTATTTGATACACTGTTGTCAATATGAATGTTGAGAAAAAAAA-TACAAAAGAGAAAACAAATTATAAATCTAACTAACAATTTT AattcaatcaattttaataaaaaaaaaActaagtactgtgtggattgcactaataatattttatattatttggtgaagaataaattaAaggaagataaaggaaaaagtaTAaaaaaaa - TgaggttttgataAgtaaaaaaaaacaagtttataatccttgtgttttttttgtc...

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Abstract

A molecular marker primer and method for identifying apricot, Siberian apricot and fresh apricot, the molecular marker primer includes a forward primer and a reverse primer; the sequence of the forward primer trnK-UUU-rps16F is: AGATTGAATTGCACAGCG, The sequence of the reverse primer trnK-UUU-rps16R is: TGAAGTAATCGGGTCTAT. The identification method comprises the following steps: (1) collection of leaf samples; (2) extraction of the total genomic DNA of leaf samples; (3) utilizing molecular marker primers for identifying large flat apricot, Siberia apricot and fresh apricot, to step (2) The target gene in the total genomic DNA of the leaf sample is amplified to obtain the amplified product; (4) the amplified product in step (3) is purified and sequenced; (5) the sequenced result in step (4) is carried out comparative analysis. Based on the base difference in the intergenic region of trnK‑UUU‑rps16, the forward primer and reverse primer can be used as molecular marker primers to identify the germplasm of Apricot apricot, Siberian apricot and fresh apricot, and the identification method is simple and easy to operate .

Description

Technical field[0001]The invention relates to the field of biotechnology. Specifically, it is a molecular marker primer and method for identifying large flat apricot, Siberian apricot and fresh apricot.Background technique[0002]Kernel-using apricot is a plant of the Rosaceae family (Prunoideae) and apricot genus (Armeniaca). It is a unique eco-economic tree species in my country and one of the six woody grain and oil tree species in China. Mainly distributed in my country's central and western ecologically fragile areas such as Hebei, Liaoning, Gansu, Inner Mongolia, Shanxi and Shaanxi provinces. Da Bian Xing is full of treasures and is often hailed as the "hardcore dealer". Compared with other apricots, Apricot kernels are sweet and without bitter taste. They are rich in protein, fat, sugar, vitamin B17 and trace amygdalin. They can be processed into dried fruits to produce almond protein, almond oil, cosmetics, functional foods, etc. It is the raw material of high-quality food, co...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/156
Inventor 包文泉敖敦
Owner INNER MONGOLIA AGRICULTURAL UNIVERSITY
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