Populus alba sex-determining genes and application thereof for sex identification of Populus alba

A technology of silver white poplar and gender, applied in the direction of microbial measurement/testing, DNA/RNA fragments, recombinant DNA technology, etc., can solve the problems of time and profit loss, and the difficulty of distinguishing male and female white poplar, and achieve reliable technical and theoretical support, Save time and expand economic benefits

Pending Publication Date: 2019-12-31
SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, it is difficult to distinguish male and female silver poplars in the seedling stage, and it often takes 4 to 5 y

Method used

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  • Populus alba sex-determining genes and application thereof for sex identification of Populus alba
  • Populus alba sex-determining genes and application thereof for sex identification of Populus alba
  • Populus alba sex-determining genes and application thereof for sex identification of Populus alba

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1. Identification of female-specific genes

[0021] According to the genome sequence of Populus alba, the unique genes of female Populus alba were identified, named M1 and M3 respectively, and the nucleotide sequences are shown in SEQ ID NO.1 and SEQ ID NO.2, respectively.

Embodiment 2

[0022] Embodiment 2, CTAB method extracts Populus alba DNA

[0023] Take about 0.5g leaves of 19 female and male Populus alba, wash them thoroughly, add liquid nitrogen to the mortar, add 1mL CTAB, and transfer to a centrifuge tube; bathe in water at 65°C for 30 minutes, then add an equal volume of chloroform to precipitate impurities. Take the supernatant and add an equal volume of isopropanol to precipitate the DNA at -20°C, then wash it twice with 75% ethanol to collect the precipitate, air dry and add 50 μL Elution buffer; take 5 μL for electrophoresis detection.

Embodiment 3

[0024] Embodiment 3, identification DNA quality

[0025] The extracted DNA is used as a template, and the specific primers designed according to the sequence of Populus alba ACTIN are amplified:

[0026] ACTIN-F: 5'-cactcgtttgcgataatgga-3' (SEQ ID NO.3);

[0027] ACTIN-R: 5'-ctcgttgtagaaggtatgat-3' (SEQ ID NO.4);

[0028] Then use TaKaRa’s Primer start high-fidelity DNA polymerase to amplify. The PCR amplification conditions are: 95°C pre-denaturation for 5 minutes; Extend for 5 minutes, and detect the amplification quality of the target band by nucleic acid electrophoresis. The results are as follows: figure 1 shown. The results showed that the extracted DNA could amplify the target band of ACTIN with good quality, which can be used for the next step of gender identification.

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PUM

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Abstract

The invention discloses Populus alba sex-determining genes and an application thereof for sex identification of the Populus alba. The Populus alba sex-determining gene is at least one of M1 and M3, with the nucleotide sequences shown in SEQ ID NO. 1 and SEQ ID NO. 2, respectively. The M1 and the M3 are female-specific genes of the Populus alba, so that female Populus alba can be quickly and conveniently identified at any period of growth by detecting whether the genome of the Populus alba contains the M1 and the M3, thus avoiding unnecessary loss, and the sex-determining genes have huge potential application and economic value.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a sex-determining gene of Populus alba, and to the application of the sex-determining gene of Populus alba in identifying the sex of Populus alba. Background technique [0002] Populus and Salix are tree species of ecological and economic importance, both in North America and Asia. Among them, poplar is the key species in the temperate zone of the two continents and the northern forest ecosystem, providing an important place for the survival of other animals, plants and microorganisms, and is the basis for the stable development of biodiversity in the ecosystem. Poplar is also an important source of paper and timber industries in the two continents, and more than half of China's plantations are poplar. In addition, the insect defense chemicals of poplar and willow trees are of great medicinal value, such as aspirin, a well-known and commonly used modern important drug, which has bee...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12Q1/6879C12N15/11
CPCC12Q1/6879C12Q1/6895C12Q2600/13
Inventor 刘建全马涛同少飞杨雯露
Owner SICHUAN UNIV
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