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Mitochondria molecular marker primer of enteromorpha population and application thereof

A molecular marker, mitochondrial technology, applied in the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., can solve problems such as unstable operation, inability to distinguish prolifera species, and inability to accurately discriminate prolifera species. , to achieve the effect of high resolution and strong stability

Active Publication Date: 2018-07-24
INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At the molecular level, the resolution of the molecular markers ITS, rbcL and 18S commonly used in Ulva research cannot effectively distinguish Enteromorpha populations, and the 5S spacer molecular marker used for in-species analysis, due to the presence in the genome Multiple copies are often not stable enough in operation to accurately distinguish different Enteromorpha populations

Method used

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  • Mitochondria molecular marker primer of enteromorpha population and application thereof
  • Mitochondria molecular marker primer of enteromorpha population and application thereof
  • Mitochondria molecular marker primer of enteromorpha population and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0028] 1) Acquisition of the standard reference sequence of Enteromorpha DNA rps2-trnL intergenic region

[0029] ①Analysis of hypervariable regions in Enteromorpha mitochondrial genome: .

[0030] The mitochondrial genome sequences of two different Enteromorpha strains were obtained from the National Center for Biotechnology Information (NCBI, http: / / www.ncbi.nlm.nih.gov / ) (GeneGenbank Accession numbers: KT428794.1 and KT428794.1) , using the Clustal W module in the MEGA 7.0 software for genome comparison, and according to the results of the mitochondrial genome comparison, first find the segment with high sequence variation between the two strains, and further screen out the segment belonging to the intergenic region based on the genome annotation information , and finally determined that the candidate segment was the intergenic region of rps2-trnL.

[0031] ② Design of rps2-trnL sequence primer pair:

[0032] Using the upstream and downstream sequences of the intergenic r...

Embodiment 2

[0056]The Enteromorpha sample (sample1) collected from Rushan, Shandong and the Enteromorpha sample (sample2) from Haiquan Bay, Qingdao, could not confirm the population type. Carry out comparative identification with the inventive method:

[0057] Specific steps are as follows:

[0058] 1) Utilize conventional method to extract the DNA of the sample to be tested for use;

[0059] 2) PCR amplification:

[0060] The sample obtained in step 1) was used as a template, and the primers obtained in the above examples were used for PCR amplification. The PCR conditions were: pre-denaturation at 94°C for 5 minutes; denaturation at 94°C for 30s, annealing at 52°C for 40s, extension at 72°C for 40s, and 35 cycles; Finally, extend at 72°C for 10 min.

[0061] The PCR system is:

[0062]

[0063] 3) The amplified product is detected by 1.5 times agarose gel electrophoresis. The results showed that the lengths of the DNA fragments amplified by the primers (SEQ ID P01 and SEQ ID P02...

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Abstract

The invention belongs to the technical field of algae molecular markers, and particularly relates to a mitochondria molecular marker primer of enteromorpha population and application thereof. The mitochondria molecular marker primer is applied to identify a high-resolution mitochondria molecular marker (the sequence of a rps2-trnL intergenic region) of the enteromorpha population. The method can detect an algae sample and a microcosmic propagule sample, and is more reliable and practical compared with a conventional method, and accordingly the application of the mitochondria molecular marker primer of the enteromorpha population has significant meaning on the aspects of green tide disaster research, enteromorpha germplasm identification, phylogeny, systematic geography research and protection and utilization of enteromorpha resources.

Description

technical field [0001] The invention belongs to the technical field of algae molecular markers, and in particular relates to a mitochondrial molecular marker primer of Enteromorpha population and its application. Background technique [0002] Enteromorpha (Ulva prolifera) belongs to Ulva family and Ulva genus, is a common green algae in the intertidal zone, widely distributed in my country. In recent years, the explosive proliferation of Enteromorpha has caused a "green tide" disaster caused by large-scale floating in the Yellow Sea of ​​my country, which has caused serious negative impacts on the coastal economy and ecology, and has attracted widespread attention from society, the media, the government and relevant scholars. Current research has shown that floating Enteromorpha is a single and unique "floating ecotype", which is different from shore-bound Enteromorpha. Therefore, the relationship between floating Enteromorpha and settled Enteromorpha The key to tracing the...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/156
Inventor 高大海孙忠民黄超华
Owner INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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