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Primers for identifying tapisicia sinensis gender and application thereof

A technology of the sex of the pepper tree, which is applied in the field of primers for identifying the sex of the pepper tree, can solve the problems of difficult early identification of the sex of the pepper tree, and achieve the effect of clear band pattern, good repeatability and strong reliability

Inactive Publication Date: 2016-01-06
NORTHWEST UNIV(CN)
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Aiming at the defects and deficiencies in the prior art, the present invention provides a primer for identifying the sex of the pepper tree and its application, which solves the technical problem that it is not easy to identify the sex of the pepper tree early in the prior art

Method used

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  • Primers for identifying tapisicia sinensis gender and application thereof
  • Primers for identifying tapisicia sinensis gender and application thereof
  • Primers for identifying tapisicia sinensis gender and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0027] The SSR primers used in this study came from the Unigenes obtained by transcriptional sequencing of the pepper tree, and the Unigenes sequences containing microsatellite molecular markers were selected and primers were designed. Use primer3 software for primer design, the specific parameters are: (1) GC content ranges from 40% to 60%, (2) primer length is between 18-27bp, (3) annealing temperature is 55-63°C, (4) The amplification length is 100-280bp.

[0028] The 150 included microsatellite molecular marker sequences and the designed primer sequences are shown in Table 1.

[0029] The 10 individuals of the pepper tree were collected from the Taibai Campus of Northwest University in Xi'an City, Shaanxi Province, including 5 male plants and 5 hermaphrodite plants;

[0030] 150 pairs of primers were used to screen the genomic DNA of 10 P. japonica individuals. The primers TSMHF (5'-TTGTCCCCTTCAACTTCGCT-3') and TSMHR (5'-AAAATCAACCAGCCAGTTCG-3') can be used to identify di...

Embodiment 2

[0032] The present embodiment selects 28 individuals of the pepper tree, and the 28 individuals of the pepper tree are collected from the southern slope of the Qinling Mountains, Xunyangba Town, Ningshan County, Ankang City, Shaanxi Province, wherein there are 13 male plants and 15 hermaphrodite plants;

[0033] The plant genome extraction kit selected in this embodiment is the DP305-02 plant genome DNA extraction kit of Beijing Tiangen Biochemical Technology Company;

[0034] 1. Use the plant genome extraction kit to extract genomic DNA from 28 individuals of the pepper tree respectively. The extraction method is as follows:

[0035] (1) Take about 30 mg of the dry weight of the leaves of the pepper tree, add liquid nitrogen and fully grind them;

[0036] (2) Quickly transfer the ground powder to a centrifuge tube pre-installed with 700 μl of 65°C preheating buffer GP1 (add mercaptoethanol to the preheated GP1 before the experiment, so that the final concentration is 0.1% by ...

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Abstract

The invention discloses primers for identifying tapisicia sinensis gender and application thereof. The primers include TSMHF and TSMHR, wherein the sequence of TSMHF is 5'-TTGTCCCTCTCAACTTCGCT-3', and the sequence of TSMHR is 5'-AAAATCAACCAGCCAGTTCG-3'. The primers are used for identifying male and bisexual tapisicia sinensis. The primers have the advantages that one pair of specific microsatellite marker primers for identifying tapisicia sinensis gender is obtained by accident from 150 pairs of microsatellite primers, the primers are clear in banding pattern, good in repeatability, capable of identifying tapisicia sinensis gender with an identification rate of 100%, high in reliability, and capable of being used for identifying tapisicia sinensis gender; the primers are extremely high in specificity and stability, only the steps of conventional PCR, polyacrylamide gel electrophoresis and dye development are needed, fragment separation is not needed, tapisicia sinensis gender can be identified by naked eye observation, and theoretical and technical guidance is provided to the directive breeding of tapisicia sinensis.

Description

technical field [0001] The invention belongs to the technical field of plant molecular biology, and relates to a primer for identifying the sex of the pepper tree and its application. Background technique [0002] Gall pepper tree (Tapisciasinensis), also known as silver magpie tree, belongs to the Tapisciaceae family (Tapisciaceae), and is a rare and ancient tree species unique to my country. Due to its long seed development cycle, poor natural regeneration ability, and human destruction, it is already in an endangered state and is listed as a national third-level protected plant. [0003] Gall pepper tree is an androgenic dioecious breeding system, and there are two sexes in the population: male plant and hermaphroditic plant. Male plants have male flowers and no fruit, and hermaphrodite plants have full flowers and fruit. Androgyny is extremely rare in nature, and it has unique research value in the evolution and maintenance of plant breeding systems. The leaves of the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q2600/156
Inventor 周晓君刘文哲
Owner NORTHWEST UNIV(CN)
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