SNP (single nucleotide polymorphism) marker related to hevea brasiliensis stem girth and application thereof

A rubber tree and labeling technology, which is applied in the direction of recombinant DNA technology, microbial measurement/inspection, DNA/RNA fragments, etc., can solve the problems that cannot meet the needs of molecular breeding of rubber trees, achieve the effect of convenient operation and promote the breeding process

Active Publication Date: 2016-09-21
RUBBER RES INST CHINESE ACADEMY OF TROPICAL AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In recent years, some progress has been made in the development and utilization of rubber tree molecular markers, but it is far from meeting the needs of rubber tree molecular breeding

Method used

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  • SNP (single nucleotide polymorphism) marker related to hevea brasiliensis stem girth and application thereof
  • SNP (single nucleotide polymorphism) marker related to hevea brasiliensis stem girth and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0030] Embodiment 1: Obtaining of SNP markers related to rubber tree stem girth

[0031] 1.1 Acquisition of rubber tree germplasm materials

[0032] The population used is the rubber tree 1981'IRRDB germplasm, which was planted in the National Rubber Tree Germplasm Resource Garden of the Rubber Research Institute of the Chinese Academy of Tropical Agricultural Sciences. Mato Grosso and Rondonia three states. In June 2014, 34 germplasm young leaves were collected and frozen in liquid nitrogen and brought back to the laboratory for future use.

[0033] 1.2 DNA extraction of rubber tree germplasm material

[0034] Take the frozen rubber tree leaves, and use the CTAB method to extract genomic DNA: (1) Weigh 1g of rubber tree leaves, freeze them in liquid nitrogen, grind them into fine powder, and transfer them to a 50ml centrifuge tube. (2) Add 10ml of 65°C preheated 2×CTAB extraction buffer (2% β-mercaptoethanol has been added in advance), gently rotate the centrifuge tube t...

Embodiment 2

[0037] Example 2: Sequencing verification and application of SNP markers related to rubber tree stem circumference

[0038] 2.1 Amplification of nucleotide fragments containing SNP sites

[0039] Using the genomic DNA of each rubber tree to be tested obtained by the aforementioned extraction as a template, use forward primer F: 5'-GAA GCA CTA AGA CGG TCC ATA G-3' (SEQ ID NO: 2) and reverse primer R: 5' - GGA GAC AGA TTA CAA TCT GAC AGC-3' (SEQ ID NO: 3), amplifying the nucleotide fragment where the SNP to be detected is located. Among them, the PCR reaction system is 25 μl: 1 μl of 100-200ng / μl template DNA, 1 μl of 10 μM primers F and R, 2.5 μl of 10×PCR reaction buffer, 2.0 μl of 2.5mM dNTP, 0.2 μl of 5U / μl Tap DNA polymerase, Water balance; PCR reaction conditions: 95°C pre-denaturation for 5 minutes, 30 cycles at 94°C for 30s, 60°C for 30s, 72°C for 1min, and extension at 72°C for 5min.

[0040] 2.2 Sequencing to identify the genotype of the SNP site

[0041] The PCR pr...

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Abstract

The invention discloses an SNP (single nucleotide polymorphism) marker related to hevea brasiliensis stem girth and application thereof. The SNP marker is shown as in SEQID NO:1, and the base at the 132bpth site from the 5' end of the nucleotide sequence is G or T. The SNP marker is closely related to the stem girth of hevea brasiliensis, and can be effectively used for molecular marker assisted breeding of hevea brasiliensis.

Description

technical field [0001] The invention relates to a SNP marker and its application, in particular to a SNP marker related to the rubber tree stem girth and its application. Background technique [0002] Rubber tree is a tropical tree species that plays an important role in the world's economic and military development. The natural rubber tree it produces is an important industrial raw material and strategic material. At present, the area suitable for planting rubber trees in China is limited, and the actual planting area has reached the limit. There is no room to expand the planting area to increase production. A substantial increase in the production of rubber trees per unit area is a suitable way for the development of China's rubber industry and to ease the pressure of the contradiction between supply and demand of natural rubber. Feasible way. [0003] Rubber tree hybrid breeding has always been a conventional method for rubber tree breeding. However, the long cycle of co...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 龙翔宇方永军唐朝荣何斌赵庆洲
Owner RUBBER RES INST CHINESE ACADEMY OF TROPICAL AGRI SCI
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