SNP molecular marker for grape fruit seedless major QTL site SDL and application

A molecular marker, SDL2 technology, applied in the field of grape molecular genetics and breeding, to achieve the effect of good repeatability, high identification accuracy, and accelerated improvement

Active Publication Date: 2018-02-23
ZHENGZHOU FRUIT RES INST CHINESE ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no research on QTL mapping of seedless traits of grape fruit using SNP molecular markers at home and abroad

Method used

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  • SNP molecular marker for grape fruit seedless major QTL site SDL and application
  • SNP molecular marker for grape fruit seedless major QTL site SDL and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1 Seedless main effect QTL loci in grape fruit SDL and the acquisition of its molecular markers SDL1 and SDL2

[0031] a. Using the female parent 'Red Earth' and the male parent 'Moritani Seedless' to obtain 129 F1 populations;

[0032] b. Use RAD-seq (restriction site-associated DNA sequencing) method to sequence the genomes of 131 individuals including 2 parents; first extract the genomic DNA of 131 individuals, and use Nanodrop, Gel-Electrophotometric, etc. Sample DNA is tested, qualified samples are detected, interrupted by ultrasound, and then the ends are repaired with T4 DNA polymerase and Klenow polymerase, blunt ends are prepared, A bases are added to the 3' end of the DNA fragment, and T4 DNA ligase reaction is configured System, react in Thermomixer at a suitable temperature for a certain period of time to connect the adapter and the "A" product, purify the 260-450bp DNA fragment on agarose gel, and configure the PCR reaction system to amplify the ge...

Embodiment 2

[0039] Example 2 Main-effect QTL loci with grape fruit seedless SDL Detection of linked molecular markers SDL1 and SDL2

[0040] Using the grape genome DNA as a template, PCR amplification was carried out with the primer pair of molecular markers SDL1 and SDL2, and the amplified product was subjected to TA cloning and sequencing. At the same time, the base at the 75th bp of the 150 bp DNA sequence amplified by the primer pair of SDL2 is C, which means that it is related to the main effect QTL locus of grape fruit seedless. SDL Linked molecular markers SDL1 and SDL2 are present.

[0041] The above primers are designed using primer5, the design principle is: the length of the primer is 15-30 bases, the length of the amplified fragment is 100-600 base pairs, the annealing temperature is 55-60°C, and there is no primer-dimer; the primer of the above molecular marker SDL1 The sequence of the forward primer SEQ ID NO.3 of the pair is: 5'-GGCTTCTGGTTGGGTTTCATTCT-3'; the sequence of...

Embodiment 3

[0044] Example 3 Application of Molecular Markers SDL1 and SDL2 in Grape Seedless Molecular-Assisted Breeding

[0045] a. Using 'Red Earth' as ​​the female parent and 'Moritani Seedless' as the male parent to construct the F1 generation hybrid population, and obtained 129 individual plants of the F1 generation population;

[0046] B. carry out the detection of SDL1 and SDL2 marker to the obtained 129 strains F1 generation single plant, concrete method is: take grape genome DNA as template, carry out PCR amplification with the primer pair of molecular marker SDL1 and SDL2, amplified product is used 8 % Polyacrylamide gel electrophoresis separation, TA sequencing; if the base at the 73bp of the 146bp long DNA sequence amplified by the SDL1 primer pair is A, and the base at the 75bp of the 150bp long DNA sequence amplified by the SDL2 primer pair is C, indicating that the single plant has molecular markers SDL1 and SDL2, and it is predicted that the fruit is seedless; the sequenc...

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Abstract

The invention belongs to the field of grape molecular breeding, and particularly provides a molecular marker for a grape fruit seedless major QTL site and an application. An RAD-seq method is utilizedfor sequencing and constructing a joint genetic linkage map of a female parent 'Red Globe' and a male parent 'Centenial Seedless', identification is carried out in combination with a grape fruit phenotype, and an interval mapping method is utilized for QTL analysis, then the presence of the major QTL site (i)SDL(/i) is detected in combination with a 18th linkage group in the genetic linkage map,a contribution rate of the major QTL site is 77.9%, and two SNP markers closely linked to the site are SDL1 and SDL2, with a genetic distance of 124kb. The SNP molecular marker for the grape fruit seedless major QTL site obtained in the invention can be used for early screening of grape fruit seedless traits, and has important theoretical and practical guiding significance for improving the breeding efficiency.

Description

technical field [0001] The invention belongs to the field of grape molecular genetics and breeding, in particular to a seedless main-effect QTL locus of grape fruit SDL SNP molecular markers and applications. Background technique [0002] Grape( Vitis vinifera L.) is an important economic crop in my country, and its cultivation area and output both rank first in the world. Grape fruit has high nutritional value, high sugar content, rich in anthocyanins and resveratrol, and has certain health care functions, so it is loved by consumers. Seedless is an important factor in the character of grape fruit. Seedless grapes are convenient to eat, especially suitable for the elderly, children and women. In addition, the yield and economic benefits of seedless grape varieties are relatively high. Therefore, cultivating seedless varieties has become one of the important goals of grape breeding. [0003] Grapes are perennial vines with a long generation cycle and late fruiting. In...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 张颖刘崇怀樊秀彩姜建福
Owner ZHENGZHOU FRUIT RES INST CHINESE ACADEMY OF AGRI SCI
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