Molecular biological differentiation method aiming at genders of asparagus dioecian plants and application of molecular biological differentiation method

A molecular biology and dioecious technology, which is applied in the field of molecular biology identification of the sex of asparagus dioecious plants, can solve the problems of inaccurate sex identification of asparagus dioecious plants and the like, and achieves simple and convenient operation. Effect

Active Publication Date: 2017-09-26
VEGETABLE & FLOWER INST JIANGXI ACADEMY OF AGRI SCI
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  • Abstract
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The present invention aims at the technical defects of the prior art, and provides a molecular biological identification method for the gender of dioecious plants of the genus Asparagus, so as to solve the inaccurate sex identification of dioecious plants of the genus Asparagus in the prior art question
[0007] Another technical problem to be solved by the present invention is to solve the technical problem of the molecular biological identification method of sexless dioecious plants of the genus Asparagus except asparagus in the prior art

Method used

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  • Molecular biological differentiation method aiming at genders of asparagus dioecian plants and application of molecular biological differentiation method
  • Molecular biological differentiation method aiming at genders of asparagus dioecian plants and application of molecular biological differentiation method
  • Molecular biological differentiation method aiming at genders of asparagus dioecian plants and application of molecular biological differentiation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Example 1 Primer Design

[0054] Based on the CDS sequence of asparagus SOFF gene, using NCBI / Primer-BLAST combined with Primer Premier5.0 software, we designed and developed four functional STS molecular markers that can identify the sex of asparagus from male to female, named Soff1, Soff2, Soff4, and Soff5 respectively. , and define the forward and reverse primer sequences of each molecular marker, and the corresponding characteristic bands. The details are shown in Table 1.

[0055] Table 1 Characteristics of STS molecular markers of female suppressor gene SOFF in asparagus

[0056]

Embodiment 2

[0057] Example 2 Polymorphism Analysis of STS Molecular Markers in Asparagus Germplasm Resources Female and Male DNA Pools

[0058] (1) Test material

[0059] Select 20 germplasm resources of Asparagus in Table 2 as test materials, among which, V1-1 and V1-2 are double haploid plants cultured from asparagus anthers, and the sex-determining loci are all homozygous; V2- V14 is a diploid cultivar of asparagus; V15-V17 is a tetraploid cultivar of asparagus; V15-V17 is a wild species related to Asparagus, which is a representative of dioecious germplasm resources.

[0060] Table 2 Germplasm resources of dioecious plants of the genus Asparagus

[0061]

[0062]

[0063] (2) Construction of female and male DNA pools

[0064] V2-17 randomly selects 5 male plants and 5 male plants according to the variety, takes their pseudo-leaf respectively, mixes them in equal amounts according to the gender of the female and male varieties, extracts DNA, and uses them as the female and male...

Embodiment 3

[0073] Example 3 Genetic linkage analysis of STS molecular markers and asparagus female and male sex phenotypes

[0074] (1) Test material

[0075] The hybrid F 1, hybrid F 1-1 (♂), F 1-2 (♀) Sisters get F 2 The population, with a total of 331 strains, was used as a genetic population for genetic linkage analysis of STS molecular markers and asparagus female and male sex phenotypes.

[0076] (2) DNA extraction

[0077] Take 1-2g of the above F 2 population, and the female parent Tainanxuan No. 3 (TN3), the male parent Xing'an Tianmendong (XA), F 1-1 (♂), F 1-2 (♀) Young pseudo-leaves were ground into powder using a high-throughput tissue grinder, and plant genomic DNA was extracted by the CTAB method.

[0078] (3) PCR amplification system

[0079] The 25mL PCR reaction system is: 10× amplification buffer 2.5μL; MgCl 2 (25mmol·L-1) 2.5μL; dNTPs (1mmol·L-1) 2.5μL; primer (5pmol·L-1) 2μL, template DNA (1.25-20ng·μL-1) 2μL; Taq DNA polymerase (1U ·μL-1) 1μL; add ddH2O t...

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Abstract

The invention provides a molecular biological differentiation method aiming at genders of asparagus dioecian plants and application of the molecular biological differentiation method. According to the technical scheme, four functional STS (Sequence-tagged Site) molecular markers of the gene are developed based on a CDS (Coding Sequence) of a female inhibitor gene SOFF on a Y chromosome of asparagus. On the basis, corresponding primers and universal PCR (Polymerase Chain Reaction) amplification conditions are designed for self-characteristics of the four STS molecular markers; furthermore, the length of a basic group of a characteristic strip of a male plant is definitely oriented. An experiment proves that the four molecular markers provided by the invention are all located in a coding region of the female inhibitor gene SOFF of the asparagus, and are co-separated from phenotypes of the female and male genders in the aspect of molecular heredity, and the differentiation accuracy reaches 100 percent; the four molecular markers can be widely used for the differentiation of the female and male genders of the asparagus and molecular marker assisted selection of all-male or all-female varieties. Moreover, the experiment shows that the method can still be used for gender differentiation of the asparagus dioecian plants including asparagus dauricus (A.dauricus), asparagus longiflorus (A.longiflorus), A.maritumus and the like.

Description

technical field [0001] The invention relates to the technical field of plant molecular genetics, and further relates to a molecular marker-based plant sex identification technology, in particular to a molecular biology identification method for the sex of dioecious plants of the genus Asparagus and an application thereof. Background technique [0002] There are about 300 species of Asparagus plants, which are distributed in temperate to tropical regions all over the world except the Americas; there are 24 species and some exotic cultivars in China, which are widely distributed all over the country. In addition to being used for viewing, some species have edible properties. or medicinal value, and therefore belong to the category of plants grown in agriculture or horticulturally. Among them, asparagus (Asparagus officinalis L.), also known as Asparagus officinalis L., is a perennial herb of Asparagus family Asparagus. Its young stems are tender and delicious, rich in nutritio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68A01G1/00
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 周劲松陈光宇罗绍春汤泳萍尹玉玲张岳平谢启鑫
Owner VEGETABLE & FLOWER INST JIANGXI ACADEMY OF AGRI SCI
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