SSR molecular marker for screening content and phenotype of apple fruit sugar and application thereof

A technology of molecular markers and sugar content, applied in the determination/inspection of microorganisms, DNA/RNA fragments, recombinant DNA technology, etc., can solve the problems of high heterozygosity, restriction of breeding process, and long childhood in apples, and achieve efficient screening Effect

Active Publication Date: 2019-10-01
WUHAN BOTANICAL GARDEN CHINESE ACAD OF SCI
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Problems solved by technology

However, the characteristics of high heterozygosity, self-incompatibili

Method used

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  • SSR molecular marker for screening content and phenotype of apple fruit sugar and application thereof
  • SSR molecular marker for screening content and phenotype of apple fruit sugar and application thereof
  • SSR molecular marker for screening content and phenotype of apple fruit sugar and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1: Detection of sugar accumulation-related genes by SSR molecular marker typing

[0027] (1) Determination of population phenotype data

[0028] Ripe apple fruits were picked and their skins and cores were removed, and the apple pulp cut into small pieces was quick-frozen in liquid nitrogen and stored in a -80°C ultra-low temperature refrigerator. Grind the pulp sample frozen in a -80°C ultra-low temperature refrigerator into a powder, weigh 1g of the pulp powder sample into a 15mL centrifuge tube and add 6mL ultrapure water to fully mix the sample. Centrifuge in the machine for 15min at a speed of 5000rpm. The supernatant produced by centrifugation was filtered through a 0.22 μm Sep-Pak microporous membrane, and the filtrate was saved for sugar content determination. The chromatographic conditions for sugar determination are: CarboSep CHO-620 calcium sugar column (300×6.5mm, pore size 10μm) and CarboSep CHO-620 guard column core, the operating temperature is ...

Embodiment 2

[0044] Example 2: Verification of Candidate Gene Functions in Strawberry Fruit and Apple Callus Systems

[0045] The coding sequence of the MdSUT4.1 gene corresponding to the SSR molecular marker SugarA3 was inserted into the multiple cloning site of the pSAK277 binary vector to construct the pSAK277-MdSUT4.1 expression vector and transformed into EHA105 Agrobacterium competent cells by heat shock method. The strains were cultured in LB liquid medium containing corresponding antibiotics in a shaker at 28° C. to OD600 = 1.0, and the bacterial cells were collected to prepare an Agrobacterium infection solution. Agrobacterium infection solution A (5mg / mL glucose, 50mM MES, 2mM Na 3 PO 4 12H 2 0 and 100 μM acetosyringone) to resuspend the bacterial body to OD600=0.2, inject the bacterial solution from the base of the green fruit of "Hongyan" strawberry through a 1mL syringe, place the transformed strawberry plants in a 16°C greenhouse for 9 days, and pick the strawberries The t...

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Abstract

The invention discloses an SSR molecular marker for screening the content and phenotype of apple fruit sugar and application thereof. The SSR molecular marker SugarA3 is successfully developed by determining candidate molecular markers through content and phenotype data of genetic typing related sugar. PCR amplification is conducted through the SugarA3, a simple sequence repetitive unit (AG)n is determined in combination with sequencing, and accordingly the content of the fruit sugar is estimated. Three types of genes can be generated through the SugarA3 and include (AG)9/9, (AG)6/9 and (AG)6/6 or 11, wherein one or two (AG)9 sequence motifs indicate that the corresponding content of fruit sugar and glucose in apple fruits is remarkably lower than that of the apple species without (AG)9 sequence motifs. Thus, the sweetness screening of applies in the seedling period can be easily realized by applying the molecular marker SugarA3, the breeding age limit of the excellent species meetingthe market demands is shortened, and the apple quality and breeding efficiency are improved.

Description

technical field [0001] The invention relates to the technical field of molecular markers, in particular to an SSR molecular marker for screening apple fruit sugar content phenotypes and an application thereof. Background technique [0002] The application of molecular genetic methods to crop breeding has broken the limitations of natural evolution and traditional physical and chemical methods, and has broad prospects for development. However, the early studies related to breeding focused on the construction of genetic linkage maps. In addition, due to the lack of high-precision genome reference information, the gene mapping of important agronomic traits is difficult to apply to production practice. With the rapid development of modern genomics, a number of high-quality genome sequences and annotation information have emerged, and a powerful platform based on genome-wide association analysis and molecular marker-assisted breeding has been established to promote the future bre...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 韩月彭彭倩马百全
Owner WUHAN BOTANICAL GARDEN CHINESE ACAD OF SCI
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