Molecular marker for identifying tremella aurantialba and application of molecular marker

A technology of molecular markers and cloud ears, applied in the field of genetic engineering, can solve problems such as identification errors, unstable band patterns, and uncertain sequence of characteristic bands, and achieve highly specific effects

Active Publication Date: 2020-11-13
GUANGXI ZHUANG AUTONOMOUS REGION ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Although there are many differences between the Bose cloud fungus strains and the black fungus commercial strains imported from Guangxi, the appearance of fruiting bodies is greatly affected by environmental conditions, and there is a lack of quantitative indicators. Misidentification; although ISSR analysis technology can identify and distinguish two types of black fungus species, it also has disadvantages such as unstable band pattern and uncertain characteristic band sequence, and often requires multiple primers, and the fingerprint of ISSR is also complicated. Need to rely on software analysis, the process is more cumbersome

Method used

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  • Molecular marker for identifying tremella aurantialba and application of molecular marker
  • Molecular marker for identifying tremella aurantialba and application of molecular marker
  • Molecular marker for identifying tremella aurantialba and application of molecular marker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Genetic difference analysis of the commercial strains of black fungus from Baise and foreign black fungus (selected samples include the fungus from Baise: 1: Guiyun No. 3, ty013, ly041, ly051, ty063, ty071, YE1 and Baiyun No. 6; source The cloud ear from Lishui, Zhejiang: 916; Yuner Xinke No. 4 from Jiangdu, Jiangsu), it was found that the germplasm resources of the cloud ear from Baise had the same specific band at about 1000bp, and the band was cut Glue recovery, fingerprint strips of glue recovery such as figure 1 as shown, figure 1 Among them, M is: DL2000, which are 2000bp, 1000bp, 750bp, 500bp, 250bp and 100bp from top to bottom; number 1-4: from left to right are Guiyun No. 3, ty013, ly041, ly051 ( Both are Bose cloud ear), from figure 1 It can be seen that samples 1-4 have an obvious common band at about 1000bp, and the common band is recovered by gel cutting; sequencing is performed using the DNA fragment of the cloud ear fungus strain ty063 as a template, an...

Embodiment 2

[0028] Multiple samples were amplified using the primer sequences of Example 1: the sources of samples amplified in this example are shown in Table 1:

[0029] Table 1 Variety names and sources of different cloud ear strains

[0030]

[0031]

[0032] The DNA of the above-mentioned samples No. 1-23 was extracted respectively, and amplified by PCR amplification method, wherein the PCR reaction system was: 2×Es TaqMasterMix 11.5 μL, ddH 2 O 10.5 μl, 1 μl of 10 μmol / L upstream primer, 1 μl of 10 μmol / L downstream primer, 1 μl of genomic DNA of black fungus strain to be tested;

[0033] The reaction program of PCR amplification was: pre-denaturation at 95°C for 5 min, denaturation at 95°C for 30 sec, annealing at 60°C for 30 sec, extension at 72°C for 60 sec, 30 cycles, extension at 72°C for 7 min, and storage at 4°C for 30 min.

[0034] The result of the reaction is as figure 2 Shown: M: DL2000 (from top to bottom are 2000bp, 1000bp, 750bp, 500bp, 250bp and 100bp), lanes...

Embodiment 3

[0036]Those skilled in the art can according to the primer pair of embodiment 1 and embodiment 2, design the test kit or probe that contains above-mentioned primer pair to be used for specific identification Bose cloud ear, therefore, another object of the present invention is also to protect a kind of A kit comprising the above-mentioned primer pair 3-TY063-5F: 5'-CAGATATGCAGCACATCACAC-3'; 3-TY063-5R: 5'-CCAACCAAACCAGAAACATCC-3'; if the above-mentioned molecular marker, kit or probe can be used in The size of 635bp was amplified in the cloud ear DNA, indicating that the cloud ear species came from Baise, if not, it indicated that the cloud ear species was not Baise cloud ear.

[0037] To sum up, the primers of the present application can amplify the common fragments of the cloud fungus, which can well distinguish the cloud fungus from other geographical areas, and can quickly identify the cloud fungus through the primers. Identification from the perspective of molecular biolo...

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Abstract

The invention relates to the technical field of gene engineering, in particular to a molecular marker for identifying tremella aurantialba and an application of the molecular marker, a primer pair ishigh in specificity, tremella aurantialba DNA derived from tremella aurantialba can be subjected to specific amplification, 635bp fragments can be amplified, and no amplification effect on other localtremella aurantialba strains exists. The molecular markers, probes and/or kits for the tremella aurantialba can be specifically designed by utilizing the primer pair, the tremella aurantialba is screened and identified, tremella aurantialba strains from the tremella aurantialba can be simply, conveniently, quickly and accurately screened by utilizing the method, and the method has important significance for genetic research and market popularization of the tremella aurantialba strains.

Description

【Technical field】 [0001] The invention relates to the technical field of genetic engineering, in particular to a molecular marker for identifying cloud fungus and its application. 【Background technique】 [0002] Cloud ear (Auricularia heimuer) is a common name for black fungus in southern my country, belonging to Basidiomycotina, Heterobasidiomycetes, Auricularia, Auriculariaceae, and Auricularia genus. Black fungus is widely distributed in my country. The main producing areas include Jilin, Heilongjiang, Liaoning, Inner Mongolia, Guangxi, Yunnan, Guizhou, Sichuan, Hubei, Shaanxi and Zhejiang provinces. One of the varieties. [0003] Baise cloud ear refers to the wild black fungus naturally distributed in Tianlin County, Tianyang, Youjiang, Lingyun, Leye, Longlin, Xilin and Tiandong counties (districts) in Baise, Guangxi. It is a typical growth suitable for the geographical conditions of subtropical vegetation. ecological species. Baise cloud fungus is a precious black fun...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/156Y02A50/30
Inventor 吴圣进陈雪凤王灿琴韦仕岩
Owner GUANGXI ZHUANG AUTONOMOUS REGION ACAD OF AGRI SCI
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