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Cucumber fruit beta carotene content related SNP marker and its application

A technology of β-carotene and cucumber, which is applied in the field of genetic engineering and molecular biology, can solve the problems of unclear molecular mechanism and slow progress of molecular breeding work, and achieve the effect of speeding up the breeding process, accurate and reliable results, and high genetic stability

Inactive Publication Date: 2013-06-12
INST OF VEGETABLE & FLOWERS CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, because the ore gene that controls the color of the pulp has not been found, the molecular mechanism of how β-carotene accumulates in Xishuangbanna cucumber fruit is still unclear, resulting in slow progress in related molecular breeding work

Method used

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  • Cucumber fruit beta carotene content related SNP marker and its application
  • Cucumber fruit beta carotene content related SNP marker and its application
  • Cucumber fruit beta carotene content related SNP marker and its application

Examples

Experimental program
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Effect test

Embodiment 1

[0026] Example 1 Acquisition of SNP markers related to cucumber fruit β-carotene content and pulp color traits

[0027] More than 3,000 accessions of cucumber were collected from all over the world, and 114 accessions were selected as cucumber core germplasm resources, which represented 80% of the genetic diversity (Lv et al.). Each resource was resequenced to generate a data volume of about 5G, covering an average of 10x cucumber genome. According to the calculated Fst value, the cucumber resources in the world can be divided into 3 cultivated groups and 1 wild group ( figure 1 ). In order to adapt to the local natural environment and be affected by human factors, these three cultivated groups have undergone independent evolution. Among them, the content of β-carotene in the fruit of Xishuangbanna population is very high, and the color of the pulp is orange-red, which is not found in other populations. Therefore, using bioinformatics, set Fst=1, and further search for hete...

Embodiment 2

[0028] Example 2 Acquisition of Cucumber Csa3G183920 Gene

[0029] Using primers 5'-GCCTCTCCGCCGCCACTCTC-3' and 5'-TCAAGAACCATCTATTGATTTTCGAGCCGT-3', the mutant and wild-type Csa3G183920 genes were amplified from the cDNA of Xishuangbanna and East Asian cucumber fruit, respectively, and connected to the T vector (TAKARA). mutation.

[0030] The nucleotide sequence of the cucumber Csa3G183920 gene is shown in SEQ ID No.2, and the amino acid sequence of its encoded protein is shown in SEQ ID No.1.

Embodiment 3

[0031] Example 3 Verification of the function of the Csa3G183920 gene by biochemical experiments

[0032] Using primers 5'-CGGGATCCATGGCGGCCGGCCTCTCCGCCGCCACTC-3' and 5'-ACGCGTCGACTCAAGAACCATCTATTGATTTTCGAGCCGT-3', the wild-type and mutated Csa3G183920 genes were constructed into the prokaryotic expression vector pET32 (Novogen), and then sequenced and identified correctly and then transferred into engineered Escherichia coli (Schwartz et al.). This Escherichia coli can express β-carotene synthase, so that excessive β-carotene is produced in the bacteria, and the color of the bacteria is orange-yellow. The Csa3G183920 gene is induced to express in the bacteria, and the Csa3G183920 protein has the activity of β-carotene hydroxylase, which can degrade β-carotene and cause the color of the bacteria to become lighter. The chemical composition of the bacteria was further detected by HPLC, and it was found that zeaxanthin was produced, and it was determined that Csa3G183920 could d...

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Abstract

The invention provides a cucumber fruit beta carotene content related SNP marker and its application. The SNP marker is located at the 770bp site in a gene sequence of a coded cucumber Csa3G183920 protein, the cucumber fruit beta carotene content is high at the site with A as a basic group, and the flesh color is orange yellow. The cucumber fruit beta carotene content is low at the site with C as a basic group, and the flesh color is white. In the invention, variomics, population genetics and other methods are utilized to screen the SNP marker related to cucumber fruit beta carotene content and flesh color characters. And through a biochemical method, the molecular mechanism that Csa3G183920 regulates fruit color by degrading beta carotene is illuminated. The genotype of a to-be-tested cucumber can be detected through sequencing, dCAPS or other methods. And according to the genotype, the cucumber fruit beta carotene content and the flesh color can be determined, thus accelerating the cultivation process of high nutrition cucumber varieties.

Description

technical field [0001] The invention belongs to the fields of genetic engineering and molecular biology, and specifically relates to a SNP marker related to cucumber fruit beta-carotene content and pulp color properties and its application. Background technique [0002] β-carotene is a typical representative of the carotenoid family and one of the indispensable nutrients for human health. In most cucumber varieties, the content of β-carotene in the fruit is very low, but in the cucumber varieties adapted to Xishuangbanna, the content of β-carotene in the fruit is very high, and the color of the pulp is orange-yellow. The fruit color is positively correlated with the content of β-carotene, and the higher the content of β-carotene, the darker the color of the pulp. In order to improve the nutritional value of cucumber fruits, breeders have been trying to transfer the characteristic of high β-carotene content of Xishuangbanna cucumber varieties to other cucumber varieties. Tr...

Claims

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Application Information

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IPC IPC(8): C12N9/02C12N15/53C12N15/63C12N5/10C12N15/11C12Q1/68A01H1/04
Inventor 黄三文张忠华尚轶沈嫡
Owner INST OF VEGETABLE & FLOWERS CHINESE ACAD OF AGRI SCI
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