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Dicarboxylic acid transporter gene controlling citric acid content in apple fruit and its application

A citric acid and transporter technology, applied in application, genetic engineering, plant genetic improvement and other directions, can solve problems such as unclear genetic basis, and achieve the effects of improving breeding efficiency, improving apple flavor and taste, and saving breeding time.

Active Publication Date: 2022-07-29
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 controlling citric acid content in apple fruit and its application
  • Dicarboxylic acid transporter gene controlling citric acid content in apple fruit and its application
  • Dicarboxylic acid transporter gene controlling citric acid content in apple fruit and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1: Determination of Apple Fruit Citric Acid Content in Resource Population

[0038] Remove the peel and core of ripe apple fruits, cut them into small pieces, freeze them in liquid nitrogen, and store them in an ultra-low temperature refrigerator at -80 °C; take the pulp samples frozen in the ultra-low temperature refrigerator at -80 °C, grind them into powder with a mortar, and weigh 1 g. Powder samples were added to 6 mL of ddH 2 O, vortexed and mixed; the sample was sonicated for 15 min, centrifuged at 5000 r / min for 15 min at 4°C, and the supernatant was filtered through a Waters SPE-C18 solid phase extraction cartridge and a 0.22 μm filter into a new 1.5 mL centrifuge tube; save The filtrate was used for the determination of citric acid content; the chromatographic conditions for the determination of citric acid were as follows: the chromatographic column was an Aspect liquid chromatography column LAEQ-462572 (specification 4.6×250mm, 5μm), and the mobile p...

Embodiment 2

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

[0040] Take 100 mg of young apple leaves for apple genomic DNA extraction. The 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, and uses the PCR amplification primer pair of dCAPS molecular marker CA1-dCAPS to amplify the MdTDT gene fragment. The reaction system is as follows:

[0041]

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

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

[0044]

[0045] After digestion, the products of digestion were directly detected by 2.0% agarose gel electrophoresis for SNP site polymorphism.

[0046] Table 1 List of primers used for mole...

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Abstract

The invention discloses a dicarboxylic acid transporter gene for controlling the content of citric acid in apple fruit and its application, and relates to the field of apple molecular genetic breeding. Dicarboxylic acid transporter genes controlling citric acid content in apple fruit MdTDT : The gene sequence is shown in SEQ ID NO.1, and the protein sequence encoded by the gene is shown in SEQ ID NO.2. Its application is: to increase or decrease the expression level of the gene by transgenic method to obtain the character of desired fruit citric acid content. The present invention has the following advantages and positive effects: ① identified MdTDT Gene is the key gene that causes the difference of citric acid content in apple fruit; ② CA1-dCAPS molecular marker can be used to predict the level of citric acid content in apple fruit; ③ The use of CA1-dCAPS molecular marker can guide the cross-breeding of apples, which can be screened in advance, Save breeding time and improve breeding efficiency.

Description

technical field [0001] The invention relates to the field of apple molecular genetics and breeding, in particular to a dicarboxylic acid transporter gene for controlling the content of citric acid in apple fruit and its application. Background technique [0002] Apple is an important economic fruit tree in the world, and my country is the largest apple producer and consumer in the world. According to the World Food and Agriculture Organization (FAO) statistics, 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, ranking first in the world. However, apple's high heterozygosity, self-incompatibility and long childhood severely restrict conventional apple genetic breeding and selection of new varieties. Appearance quality, nutritional quality and flavor quality are the keys to determine fruit quality, among which flavor quality is the first. The flavor quality of app...

Claims

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Application Information

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Patent Type & Authority Patents(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
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