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Dicarboxylic acid transporter gene for controlling content of citric acid in apple fruits, and application thereof

A technology of citric acid and transporter, which is applied in application, genetic engineering, plant genetic improvement, etc., can solve problems such as unclear genetic basis, and achieve the effects of directional improvement of apple flavor and taste, shortening cycle and improving breeding efficiency

Active Publication Date: 2021-04-16
WUHAN BOTANICAL GARDEN CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, little research has been done on citric acid accumulation in apples, and its genetic basis remains unclear, requiring further in-depth research

Method used

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  • Dicarboxylic acid transporter gene for controlling content of citric acid in apple fruits, and application thereof
  • Dicarboxylic acid transporter gene for controlling content of citric acid in apple fruits, and application thereof
  • Dicarboxylic acid transporter gene for controlling content of citric acid in apple fruits, and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Embodiment 1: the mensuration of apple fruit citric acid content in the resource group

[0038] Ripe apples are removed from the peel and core, cut into small pieces, quick-frozen in liquid nitrogen, and stored in a -80°C ultra-low temperature refrigerator; take a sample of pulp frozen in a -80°C ultra-low temperature freezer, grind it into powder with a mortar, and weigh 1g Powder samples were added to 6 mL of ddH 2 In O, vortex and mix well; the sample is ultrasonicated for 15min, centrifuged at 5000r / min at 4°C for 15min, and the supernatant is filtered through a WatersSPE-C18 solid phase extraction cartridge and a 0.22μm filter membrane into a new 1.5mL centrifuge tube; save The filtrate is used for the determination of citric acid content; the chromatographic conditions for the determination of citric acid are: the chromatographic column is Ampel liquid chromatography column LAEQ-462572 (specification 4.6 × 250mm, 5 μm), and the mobile phase is KH at pH=2.4, 0.22mo...

Embodiment 2

[0039] Example 2: Typing detection of dCAPS molecular markers

[0040] Get 100mg of young apple leaves for apple genome DNA extraction, detailed DNA extraction steps are carried out according to the plant genome DNA extraction kit of Tiangen Biochemical Technology (Beijing) Co., Ltd.; TaKaRa Ex The kit uses the diluted genomic DNA as a template to amplify the MdTDT gene fragment using the dCAPS molecular marker CA1-dCAPS PCR amplification primer pair. The PCR primer sequence is shown in Attached Table 1; 10 μL for PCR amplification The reaction system is as follows:

[0041]

[0042] The reaction program was: 95°C, 5min; [95°C, 30s; 60°C, 30s; 72°C, 3min] 35 cycles; finally, 72°C extension for 10min.

[0043] The PCR amplification product was digested by the restriction endonuclease HindIII, and the reaction system was as follows:

[0044]

[0045] After digestion, the digested product was directly subjected to SNP site polymorphism detection by 2.0% agarose gel elec...

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Abstract

The invention discloses a dicarboxylic acid transporter gene for controlling the content of citric acid in apple fruits, and application thereof, and relates to the field of apple molecular genetic breeding. The dicarboxylic acid transporter gene MdTDT for controlling the citric acid content of apple fruits has a gene sequence as shown in SEQ ID NO.1, and a protein sequence coded by the gene is as shown in SEQ ID NO.2. The application is as follows: the expression level of the gene is improved or reduced through a transgenic method so as to obtain the character of expected fruit citric acid content. The gene has the following advantages and positive effects: (1) the identified MdTDT gene is a key gene causing citric acid content difference in apple fruits; (2) a CA1-dCAPS molecular marker can be used for predicting the content of citric acid in apple fruits; and (3) the CA1-dCAPS molecular marker can be used for guiding crossbreeding of apples, screening can be carried out in advance, the breeding time is saved, and the breeding efficiency is improved.

Description

technical field [0001] The invention relates to the field of apple molecular genetic breeding, in particular to a dicarboxylic acid transporter gene for controlling citric acid content in apple fruit and application thereof. Background technique [0002] Apple is an important economic fruit tree in the world, and my country is the largest apple producing and consuming country in the world. According to the statistics of the World Food and Agriculture Organization (FAO), in 2018, my country's apple cultivation area was 2.07 million hectares, with a total output of 39.24 million tons, accounting for 42% and 45% of the global total respectively, ranking first in the world. However, the characteristics of high heterozygosity, self-incompatibility and long childhood of apple seriously restrict the conventional apple genetic breeding and the selection of new varieties. Appearance quality, nutritional quality and flavor quality are the key to determine fruit quality, among which f...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C07K14/415C12Q1/6895
Inventor 韩月彭廖燎张巍瀚王鲁郑蓓蓓张若细
Owner WUHAN BOTANICAL GARDEN CHINESE ACAD OF SCI