Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

SNP (Single Nucleotide Polymorphism) marker related to bitter character of cucumber and application of SNP marker

A technology of cucumber and bitter taste, applied in the fields of genetic engineering and molecular biology, can solve problems such as difficult to ensure accurate results, unscientific, and inoperable, achieve accurate and reliable results, eliminate human factors, and speed up the breeding process

Active Publication Date: 2013-04-03
INST OF VEGETABLE & FLOWERS CHINESE ACAD OF AGRI SCI
View PDF0 Cites 21 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] It is very unscientific to rely on subjective evaluation methods such as experience or tasting to judge whether cucumbers have bitter taste. It is greatly affected by individual differences. Different individuals have different reactions to bitter taste, so it is difficult to ensure accurate results.
However, although the method of measuring cucurbitacin C by HPLC is accurate, the measurement cost is high and the sample preparation is cumbersome.
Breeders need to identify tens of thousands of materials when breeding non-bitter breeding materials, which is not feasible, so this method has not been adopted, and is only used for small-scale scientific research

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • SNP (Single Nucleotide Polymorphism) marker related to bitter character of cucumber and application of SNP marker
  • SNP (Single Nucleotide Polymorphism) marker related to bitter character of cucumber and application of SNP marker
  • SNP (Single Nucleotide Polymorphism) marker related to bitter character of cucumber and application of SNP marker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1 Acquisition of SNP markers related to cucumber bitterness traits

[0036] More than 3,000 cucumber resources were collected from all over the world, and 114 of them were selected as cucumber core germplasm resources, which represented 80% of the genetic diversity. Each resource was resequenced to generate a data volume of about 5G, covering an average of 10x cucumber genome. At the same time, these 114 germplasm resources were also identified for bitter taste phenotype. Using the method described in Yu J (Yu J, et al. (2005). Nature genetics 38(2):203-208) and Zhang Z (Zhang Z, et al. (2010). Nature genetics 42(4):355-360) , according to the Mixed Liner model, using TASSEL software for GWAS analysis, found a SNP site associated with the bitter phenotype from 1 million SNPs ( figure 1 ). The SNP site is located at the 1178th bp of the gene encoding the Csa6G088690 protein, G (leaves are bitter) is mutated to A (leaves are not bitter), and this mutation cause...

Embodiment 2

[0046] Example 2 Correlation Analysis and Detection Application of Cucumber Different Genotypes and Bitterness Traits

[0047] Using cucumber 9930 (BiBi) with bitter leaves and cucumber 9110GT (bibi) with non-bitter leaves as parents, a F2 population consisting of 1800 individuals was constructed. According to the method for embodiment 2, 1800 F2 cucumber populations have been carried out PCR-dCAPS identification, PCR amplified sequence is as shown in SEQ ID NO.2, and the analysis result of amplified sequence 26bp site is as table 1 and figure 2 shown.

[0048] Table 1 Segregation of SNP loci in 1800 F2 cucumber populations

[0049]

[0050] It can be seen from Table 1 that the ratio of bitter cucumbers to non-bitter cucumbers is about 3, and bitterness is dominant relative to non-bitterness. F2 offspring meet the segregation expectation of 3:1, indicating that the SNP is linked to cucumber bitterness traits.

Embodiment 3

[0051] Example 3 Acquisition of Cucumber Csa6G088690 Gene

[0052] Firstly, a cucumber cDNA library was prepared, and then PCR amplification was performed using the forward primer 5'-AGATTAAAAGTGGGAAAAG-3' and the reverse primer 5'-CAGTTTTGAGCTACCC-3'.

[0053] The PCR reaction system is calculated as 20μl: 1μl of 10-20ng / μl template, 1μl of 10pmol / μl forward and reverse primers, 0.4μl of 10mmol / L dNTP mix, 1μl of 0.5U / μL high-fidelity Taq DNA polymerase, 10× 2 μl of PCR reaction buffer, the balance is water.

[0054] The PCR reaction conditions were: 94°C for 5 minutes; 35 cycles of 94°C for 20 seconds, 55°C for 20 seconds, 72°C for 2 minutes and 30 seconds; 72°C for 10 minutes.

[0055] The amplified fragment with a size of 2358bp (SEQ ID No.2) was connected to the T-vector (TAKARA), and no mutation was confirmed by sequencing.

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention provides an SNP (Single Nucleotide Polymorphism) marker related to the bitter character of cucumber and application of the SNP marker. The SNP marker is located at the NO.1178bp of the gene sequence for coding the Csa6G088690 protein of cucumber; cucumber with basic groups G at the NO.1178bp is bitter; and cucumber with basic groups A at the NO.1178bp is not bitter. According to the invention, a genome wide association study (GWAS) method is adopted to screen out the SNP marker related to the bitter taste character of cucumber, a derived cleaved amplified polymorphic sequences (dCAPS) method is adopted to detect the gene types of cucumber to be detected, and bitter or non-bitter cucumber variety breeding is carried out according to the gene types, so as to speed up the breeding process of non-bitter cucumber. The SNP marker is more accurate and reliable in result than the traditional method of subjectively judging whether cucumber is bitter or not, is simpler and more convenient than HPLC (High Performance Liquid Chromatography) for identifying bitter substances, and can be used for large-scale screening of breeding materials. Besides, the invention discovers the committed step for the Csa6G088690 protein of cucumber to control the synthesis pathway of bitter of cucumber for the first time.

Description

technical field [0001] The invention relates to the fields of genetic engineering and molecular biology, in particular to a SNP marker related to cucumber bitterness and its application. Background technique [0002] The bitterness of cucumbers is caused by a class of triterpenoids called cucurbitacin C. The accumulation of cucurbitacin C in cucumber fruit will seriously affect the commercial quality of cucumber. Early classic genetic experiments found that the bitter taste of cucumber was controlled by two genes, Bi and Bt. The Bi gene mainly controls leaf bitterness, and the Bt gene mainly controls cucumber fruit bitterness. The recessive bi gene has an epistatic effect on the Bt gene. But so far, neither Bi nor Bt genes have been cloned. In order to avoid the interference of bitterness on cucumber quality, breeders often rely on subjective evaluation methods such as tasting or experience to identify the bitterness of cucumber varieties in the breeding process. In add...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C12N9/90C12N15/61C12N15/63C12N1/21C12N1/19C12N5/10C12P33/00C12N15/11C12Q1/68A01H1/04
Inventor 黄三文尚轶张忠华李颖马永硕齐建建王深浩王晔
Owner INST OF VEGETABLE & FLOWERS CHINESE ACAD OF AGRI SCI
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com