SNP molecular marker for column number of secondary milk ducts of rubber trees and application of SNP molecular marker

A rubber tree and marker technology, which is applied in the determination/inspection of microorganisms, recombinant DNA technology, biochemical equipment and methods, etc., can solve problems such as not involving cultivated varieties, improve breeding efficiency, shorten selection cycle, and reduce breeding workload. Effect

Active Publication Date: 2021-12-28
RUBBER RES INST CHINESE ACADEMY OF TROPICAL AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although Tang Chaorong et al. reported a SNP related to rubber tree laticifers (Tang Chaorong, Long Xiangyu, Qi Jiyan, Yang Jianghua, He Bin. A SNP marker related to the number of rubber tree laticifers and its application. Chinese invention patent, CN 105838809 A ), but its analysis samples were only based on 34 wild germplasms, and did not involve cultivars with excellent yield and number of milk ducts in current production

Method used

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  • SNP molecular marker for column number of secondary milk ducts of rubber trees and application of SNP molecular marker
  • SNP molecular marker for column number of secondary milk ducts of rubber trees and application of SNP molecular marker
  • SNP molecular marker for column number of secondary milk ducts of rubber trees and application of SNP molecular marker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1: Acquisition of SNP markers related to the number of secondary milk duct rows

[0025] The number of secondary milk ducts in cultivars of rubber trees was significantly more than that in wild species. The materials used in this study are 208 germplasm resources of rubber trees preserved in the National Rubber Tree Germplasm Resource Garden (Danzhou, Hainan), including 93 cultivated species and 115 wild species. Fresh leaves were obtained, genomic DNA was extracted by CTAB method, and whole genome resequencing was performed. GATK software was used to obtain the variation data of all materials, and the SNP differences between wild species and cultivated species were analyzed, and a SNP site related to the number of secondary laticiferous ducts of rubber trees was identified.

[0026] The site is located at the 238bp site from the 5' end of the sequence shown in SEQ ID NO: 1, and the site is represented by "g" in the sequence of SEQ ID NO: 1, and the base of the...

Embodiment 2

[0027] Example 2: Evaluation of GG genotype and AA genotype on missed selection rate and misselection rate of germplasm with large number of duct rows

[0028] 1. Experimental materials

[0029] The laticifer differentiation ability of 286 hybrid progenies with unknown pedigree that were planted in the ninth team of the Institute of Rubber, Chinese Academy of Tropical Agricultural Sciences was evaluated, and 25 progenies with a very large number of secondary laticifer columns (more than 12 columns) were selected to obtain fresh leaves Genomic DNA was extracted according to the instructions of the Plant Genomic DNA Extraction Kit (Tiangen Biochemical Technology Co., Ltd.), and the extracted DNA was tested by 1.5% agarose gel electrophoresis and NanoDrop 2000 for later use.

[0030] 2. PCR amplification

[0031] PCR amplification was carried out using the extracted genomic DNA as a template, the sequence of the forward primer used was SEQ ID NO.2, and the sequence of the revers...

Embodiment 3

[0036] Example 3: Evaluation of AA genotype and GG genotype on missed selection rate and misselection rate of germplasm with few milk duct rows

[0037] 1. Experimental materials

[0038] The 286 progenies of 7 hybrid combination populations were evaluated for laticiferial differentiation ability (same as Example 2), and 24 progenies with a very small number of secondary laticifer columns (less than 3 columns) were selected to obtain fresh leaves. According to the plant genome DNA Genomic DNA was extracted according to the instructions of the extraction kit (Tiangen Biochemical Technology Co., Ltd).

[0039] 2. PCR amplification

[0040] PCR amplification was carried out using the extracted genomic DNA as a template, the sequence of the forward primer used was SEQ ID NO.2, and the sequence of the reverse primer was SEQ ID NO.3. The PCR amplification system is 20ul: 10 μL of 2x PCR buffer, 1 μL of forward and reverse primers (10 μmol L-1), 1 μL of DNA template, ddH 2 O is 7 ...

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Abstract

The invention discloses an SNP marker related to the column number of secondary milk ducts of a rubber tree. The SNP marker is as follows: the basic group from the 238th site from the 5'end of a nucleotide sequence shown as SEQ ID No.1 is G or A. According to the SNP marker related to the column number of the secondary milk ducts of the rubber tree, from the big data level, it is identified that the number of SNP located at the 238th site of the nucleotide sequence SEQ ID NO.1 is closely associated with the number of rows of the secondary milk ducts for the first time, detection is facilitated, the accuracy and reliability are achieved, and operation is convenient; when the SNP marker is used for identifying the column number character of a secondary milk duct of a hybridized combination population, the accuracy reaches 91% or above; the SNP marker and a detection method are not influenced by the age of the rubber tree, can be selected in a seedling stage, greatly reduce the breeding workload, remarkably shorten the yield breeding selection period of the rubber tree, improve the breeding efficiency, and can be used for yield molecular marker-assisted breeding of the rubber tree in practice.

Description

technical field [0001] The invention relates to a SNP marker and its application, in particular to a SNP molecular marker of the number of secondary laticiferous ducts of rubber trees and its application. Background technique [0002] Rubber trees are highly heterozygous, but seedlings can be propagated by budding, so that excellent hybrid offspring can be stably preserved. This is the basis of the current rubber tree cross-breeding program. For a long time, one of the main breeding goals of rubber trees is to breed high-yielding varieties, and then evaluate other secondary traits on this basis. This leads to the unsatisfactory stress resistance of the high-yield varieties currently used in production. The varieties with ideal stress resistance have lower yields. Research and development of molecular markers related to yield and stress resistance traits to assist in the selection of hybrid parents and hybrid offspring is the only way to efficiently breed high-yield and st...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12Q1/6858C12N15/11
CPCC12Q1/6895C12Q1/6858C12Q2600/13C12Q2600/156C12Q2600/172C12Q2531/113C12Q2565/125Y02A40/81
Inventor 晁金泉田维敏吴绍华张世鑫杨署光
Owner RUBBER RES INST CHINESE ACADEMY OF TROPICAL AGRI SCI
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