Molecular marker tightly linked to kiwi fruit total acid content trait QTL locus and application of molecular marker

A technology for total acid content and kiwifruit, which is applied in the fields of molecular biology and genetic breeding, can solve the problems that the research of total acid has not yet been carried out, and achieve the effect of convenient and fast detection method, accurate and fast screening, and saving production cost

Active Publication Date: 2017-07-25
WUHAN BOTANICAL GARDEN CHINESE ACAD OF SCI
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  • Abstract
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AI Technical Summary

Problems solved by technology

[0004] At present, the QTL mapping of total acid and the discovery of molecular markers related to total acid have not been carried out in kiwifruit.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Acquisition of the main effect QTL and molecular markers linked to total acid in kiwifruit:

[0023] ① In this example, the F 1 Separate groups. In the 6th year of the group's results, 125 F 1 The individual plants of the offspring of the population were the research objects.

[0024] ②A total of 125 individual plant leaves were collected from the population, and the total DNA was extracted by CTAB method. According to the RADseq method, the libraries of Chinese kiwifruit 'Guihai 4', Shanli kiwifruit 'MT570001' and 125 offspring were constructed and sequenced.

[0025] ③ Filter the sequencing data to remove sequences with low-quality base content greater than 50%, and remove sequences with sequencing adapter contamination and a large number of repetitions. Use the software SOAP2 to compare the sequencing data that meet the filtering criteria to the reference genome, perform locus variation detection for each sample, and count the genetic types of polymorphic loci in...

Embodiment 2

[0035] Application of a molecular marker closely linked to kiwifruit total acidity QTL loci in kiwifruit breeding, the steps are:

[0036] (1) F 1 generation. choose F 1 60 individual plants in the generation population were the research objects.

[0037] (2) At the fruit maturity stage, select F 1 60 individual plants in the generation group, 10 fruits per plant were collected, and the contents of quinic acid, citric acid and malic acid in the fruit were determined by gas chromatography using an Agilent 7890 N gas chromatograph, and the contents of quinic acid, citric acid and malic acid The sum of is counted as the total acid content of the fruit.

[0038] (3) Extract F 1 Generation of total DNA of 60 individual plants, according to Premix Ex Taq TM II (Tli RNaseHPlus), ROX plus kit instruction, design 20uL reaction system: 5ng·uL -1 Kiwifruit genomic DNA template 2ul, 2×SYBR Premix 10ul, 50×ROX Dye II 0.4ul, 10uM TA-F and TA-R each 0.8ul, double distilled water to ...

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PUM

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Abstract

The invention discloses a molecular marker tightly linked to a kiwi fruit total acid content trait QTL locus and application of the molecular marker. As for a primer designed according to the molecular marker, a forward primer TA-F is 5'-GTAAATGCATGGTAAAGAGGTCC-3', and a reverse primer TA-R is 5'-AGGAGAAAGGGAAAATAAAACGC-3'. As for the molecular marker, a major QTL locus for controlling total acid in a kiwi fruit is located for the first time, 21.75 percent phenotype variance can be explained, and the total acid content of the kiwi fruit can be forecasted by detecting the molecular marker relevant to the total acid content, so that superior individual plants with higher total acid content are accurately and rapidly screened.

Description

technical field [0001] The invention belongs to the technical field of molecular biology and genetic breeding, and more specifically relates to a molecular marker closely linked to a QTL site of kiwifruit total acid content traits and its application. Background technique [0002] Kiwi fruit is rich in vitamin C, dietary fiber and various mineral nutrients, and is a healthy fruit favored by consumers. China is the original origin center of the genus Actinidia. Actinidia is extremely rich in resources and has high genetic diversity. China has about 96% of the world's total resources of Actinidia species. Relying on abundant natural resources, breeding new varieties with excellent traits plays a key role in the healthy and stable development of the kiwifruit industry. The main acid components of kiwi fruit are quinic acid and citric acid, and also contain a small amount of malic acid. The content of sugar and acid in fruit and their ratio have a very important influence on t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6895C12Q2600/156
Inventor 钟彩虹张琼黄宏文姚小洪刘义飞刘春燕
Owner WUHAN BOTANICAL GARDEN CHINESE ACAD OF SCI
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