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Molecular marker d4.3 closely linked to spinach sex gene y and its application

A technology of molecular markers and spinach, which is applied in the field of plant molecular genetics and breeding, can solve the problems of insufficient stability and non-linkage of amplification effects, and achieve the effect of improving breeding efficiency

Active Publication Date: 2017-03-29
中蔬种业科技(北京)有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are 4 linkage markers published at home and abroad. The first one is the co-segregation of T11A, V20A and spinach sex gene Y, but the amplification effect of V20A is not stable enough in large population verification, and the co-segregation of T11A and Y; the second is SO4, which is 4.3cM away from spinach sex gene Y, is not linked when verified in the population, and another SSR is developed on this gene fragment for analysis, and the genetic distance is about 11cM; the third is Yang Jinhua (2009) through ISSR analysis, A female-associated marker was screened, with a full length of 1176bp, and converted to a SCAR marker that was not linked when validated in a population

Method used

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  • Molecular marker d4.3 closely linked to spinach sex gene y and its application
  • Molecular marker d4.3 closely linked to spinach sex gene y and its application
  • Molecular marker d4.3 closely linked to spinach sex gene y and its application

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Effect test

Embodiment 1

[0036] Example 1 Acquisition of Molecular Marker D4.3 Closely Linked to Spinach Sex Gene

[0037] 1. Construction of spinach genetic population

[0038] The F2 population was constructed by using female plants×male plants of the first-generation commercial spinach hybrid variety Donglu 1 (F1). The traits of the F2 population were investigated until the flowering stage. There were 343 plants in the F2 population, including 189 female plants and 154 male plants. The chi-square test was consistent with 1:1, and the sex of spinach was controlled by the sex gene X / Y.

[0039] 2. Genomic DNA extraction and construction of male and female pools

[0040] Genomic DNA of 343 F2 individual plants was extracted by CTAB method.

[0041] Take 10 female plants and 10 male plants in F2 to construct female and male pools for Bulk Segregating Analysis (BSA), and measure the concentration of DNA with a DNA detector (NanoDrop Spectrophotometer). Dilute to the same concentration of 200ng / μL, ta...

Embodiment 2

[0050] Example 2 Application of Molecular Marker D4.3

[0051] Using the successfully transformed dCAPS marker D4.3, F2 populations were constructed in the other 7 populations of spinach [Population description: female plants × male plants in the first-generation spinach hybrid commercial variety Bonfit (F1), and the female plants in the F2 population × Male plants were used to construct F3 populations, and 4 F3 populations were selected for molecular marker gender identification (that is, the experimental population numbers were 48, 50, 52, and 53 respectively); F2 population was constructed by plant x male plant, and F3 population was constructed by female plant x male plant in F2 population, and three F3 populations were selected for molecular marker sex identification (that is, the experimental population numbers were 54, 58, 59)]. Gender identification, molecular marker identification results are consistent with phenotype identification results, see image 3 , the result...

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Abstract

The invention provides a molecular marker D4.3 closely-linked to spinach gender gene Y and application of the molecular marker D4.3 and belongs to the field of molecular genetic breeding of plants. The molecular marker D4.3 can amplify bands of 179bp in both male spinach plants and female spinach plants. After the female plant (XX) amplified bands are subjected to enzyme digestion with restriction enzymes AluI, only one band exists. After the male plant (XY) amplified bands are subjected to enzyme digestion with the restriction enzymes AluI, two bands exist. The primer sequence used for amplifying the molecular marker D4.3 is as shown in SEQ ID NO. 2-3. The molecular marker D4.3 is closely-linked to the spinach gender gene, and the genetic distance is about 0.3cM. The molecular marker D4.3 can identify the spinach gender in the seedling stage, the female and male spinach plants can be screened in advance, and the breeding efficiency is greatly improved. The molecular marker D4.3 has an important value in the spinach production practice and breeding.

Description

technical field [0001] The invention belongs to the field of plant molecular genetics and breeding, in particular to molecular marker D4.3 closely linked with spinach sex gene Y and application thereof. Background technique [0002] Spinach (Spinacia oleracea L.), also known as Persian grass and red root vegetable, belongs to the genus Spinacia, has a long history of cultivation and is one of the important vegetable crops in my country. Spinach is rich in nutrition, strong cold resistance, wide adaptability, short production cycle, high multiple cropping index, high yield and output value, and is very popular among producers and consumers. [0003] Spinach is generally a dioecious crop, and a small number of plants are monoecious. Spinach sex type is affected by genetic factors and environmental factors. Rosa et al. (1925) believed that the sex type of spinach is mainly controlled by genetic factors, and is not affected by external factors such as soil fertility, distance ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q2600/13
Inventor 钱伟徐兆生刘丹张合龙王晓武武剑刘伟
Owner 中蔬种业科技(北京)有限公司