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A Molecular Marker for Identifying Hybrid Trees and Its Application

A technology of molecular markers and paper mulching, which is used in the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc.

Active Publication Date: 2021-06-01
中科创构(北京)科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no DNA molecular marker specifically for hybrid trees on the market

Method used

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  • A Molecular Marker for Identifying Hybrid Trees and Its Application
  • A Molecular Marker for Identifying Hybrid Trees and Its Application
  • A Molecular Marker for Identifying Hybrid Trees and Its Application

Examples

Experimental program
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Effect test

Embodiment 1

[0072] Example 1. Molecular markers for identification of hybridization trees and the design of specific amplification primer pairs

[0073] 1. Molecular markers of hybrid tree

[0074] The present invention provides a DNA barcode (DNA barcode) derived from a hybrid tree (Broussonetia kazinoki×B.papyrifera), the nucleotide sequence of which is shown in sequence 3 in the sequence listing, and it is named Bpmcbode. The DNA molecule shown in sequence 3 is the molecular marker used to identify the hybrid tree.

[0075] 2. Specific primer pairs

[0076] According to the Bpmcbode sequence unique to the hybrid tree obtained in step 1, a primer pair for specific amplification of its full length is designed, and the primer sequence is as follows:

[0077] F: 5'-AACCTGGGGCGTATTGCTCAA-3' (SEQ ID NO: 1);

[0078] R: 5'-TTACCATATATATATAACACAAAATTTC-3' (SEQ ID NO: 2).

Embodiment 2

[0079] Embodiment 2, the specific detection that is used for identifying the primer pair of hybridization tree

[0080] 1. DNA extraction from plant material

[0081] The total DNA of each plant material in Table 1 was extracted respectively, and the extracted total DNA was detected by 1% agarose gel electrophoresis. Among them, the agarose gel electrophoresis detection results of the total DNA extracted from the leaves, roots or stems of the hybrid tree are as follows: figure 1 shown. The results showed that the extracted DNA had obvious electrophoresis bands, which were larger than 20kb, indicating that a relatively high-purity and relatively complete DNA was obtained.

[0082] 2. PCR amplification

[0083] The total DNA sample extracted in step 1 was used as a template, and the primer pair designed in step 2 of Example 1 was used for PCR amplification to obtain PCR products respectively.

[0084] The PCR reaction system was as follows (total volume: 25 μL): DNA template...

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Abstract

The invention discloses a molecular marker for identifying a hybrid tree and its application. The molecule is marked as 1) or 2) as follows: 1) DNA molecule shown in sequence 3 in the sequence listing: 2) sequence 3 undergoes one or several nucleotide substitutions and / or deletions and / or additions and is combined with Sequence 3 is a DNA molecule with the same function. The molecular markers derived from hybrid mulberry provided by the present invention can be applied to distinguish hybrid mulberry from other plants such as wild mulberry and small mulberry. It has important theoretical and practical significance for the species identification of hybrid mulberry and can be used in agricultural The cultivation and identification of plant varieties required for animal husbandry and ecological environment management have high practical application value. The invention has broad application prospects in the fields of agriculture and economic energy crops.

Description

technical field [0001] The invention relates to a plant molecular marker, in particular to a molecular marker for identifying a hybrid tree and its application. Background technique [0002] DNA molecular identification can be applied to paternity identification, disease diagnosis, prenatal screening, identification of Chinese herbal medicine, identification of plant varieties, identification of genetically modified genes, etc. DNA molecular identification is a new biological identification system created by using standard, sufficiently variable, easy-to-amplify and relatively short DNA fragments to create their own specificity within species and diversity among species. Rapid automatic identification of species or samples is possible. [0003] Broussonetia papyrifera (Broussonetia papyrifera), also known as mulberry tree, mulberry tree, glaucous mulberry tree, mulberry peach, corn pulp tree, deer tree, Dangdang tree, milk tree, paper wood, belongs to nitrogen fixation in t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895
Inventor 沈世华彭献军
Owner 中科创构(北京)科技有限公司