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STR primer for identifying eucalyptus clones and application thereof

A clone and eucalyptus technology, applied in the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., to achieve the effects of accurate detection, convenient operation, and high experimental efficiency

Inactive Publication Date: 2017-11-24
RES INST OF TROPICAL FORESTRY CHINESE ACAD OF FORESTRY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, DNA molecular markers have been widely used in the identification of germplasm resources of many crops, and certain DNA fingerprint analysis work has been carried out in different clones of eucalyptus, but there is still no simple and easy STR detection system and common eucalyptus clones. Department of detection method

Method used

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  • STR primer for identifying eucalyptus clones and application thereof
  • STR primer for identifying eucalyptus clones and application thereof
  • STR primer for identifying eucalyptus clones and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0032] (1) Extraction of Eucalyptus Clonal DNA

[0033] Leaves of Eucalyptus clones (Table 1) were collected, Genomic DNA of Eucalyptus was extracted by CTAB method, and placed in a refrigerator for use.

[0034] Table 1 58 Eucalyptus clones

[0035]

[0036]

[0037]

[0038] Weigh about 0.3g of 58 Eucalyptus clones (Table 1) leaf samples, grind them in liquid nitrogen, transfer to a 2mL centrifuge tube, add 1mL CTAB extract, keep warm at 60-65°C for about 45-60min, and shake once every 10min . Take out the sample tube, centrifuge at 12000rpm at 4°C for 10min, and take the supernatant. Add an equal volume of chloroform:isoamyl alcohol (24:1), seal, shake well; centrifuge at 12000rpm at 4°C for 10min, and take the supernatant. Add an equal volume of chloroform:isoamyl alcohol (24:1), centrifuge at 12 000 rpm for 10 min, and take the supernatant. Add refrigerated, 2 / 3 volume isopropanol to the supernatant, shake gently to mix, and let stand at -20°C for more than 1...

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Abstract

The invention discloses an STR primer for identifying eucalyptus clones and application thereof. Eight pairs of eucalyptus STR primer with high polymorphism and clear amplification bands are screened out through genotyping of a large quantity of STR markers, and are used for identifying eucalyptus clones, and building a multiplex fluorescence detection system. The STR primer group for identifying eucalyptus clones has the advantages of high efficiency, accuracy in detection, convenient operation and the like. The STR primer can distinguish fake and disordered clones effectively so as to protect the rights and interests of breeders of improved varieties and forest growers, and can provide important technological supports for rapid genetic evaluation of eucalyptus germplasm resources, genetic map establishment, molecular marker assisted selection, and clone fingerprint establishment and verification in the future.

Description

technical field [0001] The invention belongs to the technical field of molecular markers, in particular to STR primers for identifying eucalyptus clones and applications thereof. Background technique [0002] Eucalyptus is the general term for Eucalyptus tree species of Myrtaceae (Myrtaceae), and it is one of the three largest afforestation tree species in the world. Eucalyptus is widely planted in southern China because of its rich genetic diversity, rapid growth, high wood yield and strong adaptability, and has played an important role in alleviating the tense situation of wood supply and demand in my country. [0003] In production, Eucalyptus is mainly propagated by clones. With the increase of the generation of clones, the expansion of the scope of promotion and the expansion of the distribution market, the problem of confusion of clones is becoming more and more serious. Some clone names are used in tissue culture rooms and nurseries. Misrecording, there is also a phe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q2600/156C12Q2600/16
Inventor 周长品李昌荣李发根翁启杰陈升侃王莉甘四明
Owner RES INST OF TROPICAL FORESTRY CHINESE ACAD OF FORESTRY