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InDel molecular marker tightly linked with tea purple buds and application of InDel molecular marker

A molecular marker, molecular marker sequence technology, applied in the field of molecular biology, can solve the problem of not carrying out molecular marker research and so on

Active Publication Date: 2019-08-09
TEA RES INST CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the specific sequence variation of this site is still unclear, and no research on related molecular markers has been carried out.

Method used

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  • InDel molecular marker tightly linked with tea purple buds and application of InDel molecular marker
  • InDel molecular marker tightly linked with tea purple buds and application of InDel molecular marker
  • InDel molecular marker tightly linked with tea purple buds and application of InDel molecular marker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1: Development of tightly linked InDel molecular markers for tea purple buds

[0021] 1. Test materials

[0022] Plant material: 6 individual plants (4 spring green buds and 2 spring purple buds) of tea tree varieties Zijuan, Longjing 43 and Zijuan progeny planted by the Tea Research Institute of the Chinese Academy of Agricultural Sciences were used as test materials .

[0023] 2. Genomic DNA extraction:

[0024] Take 1g of fresh young shoots, add liquid nitrogen and grind to powder. Genomic DNA was extracted using a plant genomic DNA extraction kit. After the extraction is completed, use 1.1% agarose gel electrophoresis to detect the integrity of the extraction, and then use the Nanodrop 2000 micro-nucleic acid quantifier to measure the concentration and purity of the DNA. After the qualified DNA is identified, the DNA concentration of the sample is uniformly diluted to 100ng / μl. Store in -20°C refrigerator.

[0025] 3. Cloning of the tea tree CsMYB75 pro...

Embodiment 2

[0029] Example 2: Application of InDel Molecular Markers Tightly Linked to Tea Purple Buds

[0030]Pick 10g of fresh leaves of Zijuan and Longjing 43 varieties, add liquid nitrogen and grind to powder. Genomic DNA was extracted using a plant genomic DNA extraction kit. After the extraction is completed, use 1.1% agarose gel electrophoresis to detect the integrity of the extraction, and then use the Nanodrop 2000 micro-nucleic acid quantifier to measure the concentration and purity of the DNA. After the qualified DNA is identified, the DNA concentration of the sample is uniformly diluted to 100ng / μl. Store in -20°C refrigerator. According to the above InDel-marked specific primer pair (Ziya F1: 5'-GTGAATCTGTCCTCCCTGTGT-3'; Ziya R1: 5'-GATGTTTTGGTCTTGGAGTTG-3'), the primer pair can amplify a 1049bp gene in Zijuan. Fragment, which cannot be amplified in Longjing 43, such as Figure 4 shown.

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Abstract

The invention discloses an InDel molecular marker tightly linked with tea purple buds and application of the InDel molecular marker, and belongs to the technical field of molecular biology. The invention discloses an insertion fragment which is closely related to purple bud characters, and provides a functional marker for identifying purple bud characters of a tea tree. The current situation thatpurple bud gene accurate positioning and genetic mechanism research at present is almost blank at home and abroad is eliminated. The InDel marker is applied to molecular marker assisted selection, thetea tree material with the purple bud characters in Zijuan offspring can be rapidly screened out, the breeding efficiency of a purple bud tea tree variety is improved, and a foundation is laid for breeding new varieties of the tea tree.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and in particular relates to a closely linked InDel molecular marker of tea purple buds and an application thereof. Background technique [0002] Zijuan tea rich in anthocyanins is an important purple bud tea tree variety cultivated in my country. Compared with ordinary green bud tea, purple bud tea helps humans to resist oxidation and eliminate free radicals, prevent cardiovascular and cerebrovascular diseases and cancer, and reduce the risk of neurodegenerative diseases. Therefore research and development utilize purple bud tea to be closely watched by people. [0003] In recent years, tea scientists have carried out a lot of research on the color formation mechanism of purple bud tea. At present, based on the QTL mapping of genetic populations, it has been found that sequence differences near CsMYB75 can explain 28.1-40.8% of the genetic variation of the purple bud trait. However,...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 韦康庞丹丹成浩王丽鸳阮丽张亚真
Owner TEA RES INST CHINESE ACAD OF AGRI SCI
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