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SNP molecular marker for identifying and controlling formation of low-temperature yellowing property of Chinese cabbage leaves and application thereof

A DNA molecule, low temperature technology, applied in the biological field, to achieve the effect of high-efficiency assisted breeding methods and technologies, high specificity and high throughput

Active Publication Date: 2018-06-05
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Few studies have been reported on low temperature-induced leaf color mutants in cruciferous crops

Method used

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  • SNP molecular marker for identifying and controlling formation of low-temperature yellowing property of Chinese cabbage leaves and application thereof
  • SNP molecular marker for identifying and controlling formation of low-temperature yellowing property of Chinese cabbage leaves and application thereof
  • SNP molecular marker for identifying and controlling formation of low-temperature yellowing property of Chinese cabbage leaves and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Embodiment 1, the development of SNP molecular marker

[0063] Resequencing of the low-temperature-resistant Chinese cabbage material (ST64) and the low-temperature-resistant Chinese cabbage material (ST336), after data analysis, it was found that there was a SNP site at the 13595848th nucleotide of the A05 chromosome (the 300th in the sequence listing sequence 4 nucleotide), which is a T / G polymorphism, and the SNP site is named A05-13595848T / G site.

[0064] For the A05-13595848T / G locus, based on the principle of competitive allele-specific PCR, the primers were designed as follows:

[0065] Upstream primer A05-13595848T / G-FF (sequence 1 of the sequence listing): 5'- GAAGGTGACCAAGT TCATGCTGGAGGGGTTGTTGGGACGg-3';

[0066] Upstream primer A05-13595848T / G-FV (sequence 2 of the sequence listing): 5'- GAAGGTCGGAGT CAACGGATTGGGAGGGGTTGTTGGGACGt-3':

[0067] Downstream primer A05-13595848T / G-R (sequence 3 in the sequence listing): 5'-CACTCATCGCCGTCCACGCGTA-3'.

[006...

Embodiment 2

[0069] Example 2. Application of A05-13595848T / G Loci in Identifying Yellowing and Non-yellowing Materials of Chinese Cabbage After Low Temperature Induction

[0070] 1. DNA extraction

[0071] Genomic DNA of the 1881 cabbage materials to be tested in Table 1 were extracted by conventional CTAB method.

[0072] 1881 materials are the hybrid offspring F of parents ST64 and ST336 2 For each individual plant, the specific method is as follows: cross the low-temperature non-yellowing parent material ST64 and the low-temperature yellowing parent material ST336 to obtain F 1 ;F 1 Continue to inbred to get F 2 group.

[0073] The quality of the extracted DNA was tested by agarose electrophoresis and Nanodrop2100 respectively, and it was found that the extracted genomic DNA met the relevant quality requirements, that is, agarose electrophoresis showed a single DNA band without obvious dispersion; Nanodrop2100 detected A260 / 280 between 1.8-2.0 between (DNA samples without protein ...

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Abstract

The invention discloses an SNP molecular marker for identifying and controlling formation of low-temperature yellowing property of Chinese cabbage leaves and an application thereof. An SNP site provided by the invention is the 300th nucleotide from terminal 5' of a sequence 4 in a sequence table of a Chinese cabbage genome, which is T / G polymorphic. By means of QTL positioning of a Chinese cabbageF2 group, a primary effect QTL tightly linked to low-temperature yellowing of the Chinese cabbage is obtained, the SNP marker tightly linked to low-temperature yellowing of the Chinese cabbage is developed in the region, and the accuracy of the marker can reach 95.9% by verifying 1881 single plants of the F2 group. The marker can be completely used for low-temperature-resistant molecular marker assistant breeding of Chinese cabbage. The molecular marker provided by the invention is low in cost, high in throughput and high in specificity in actual applications, thereby providing an efficient assistant breeding method and technology for accelerating low-temperature-resistant breeding of Chinese cabbage.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a SNP molecular marker for identifying and controlling the formation of low-temperature yellowing traits of cabbage leaves and an application thereof. Background technique [0002] Chlorophyll is one of the most important pigments in the process of photosynthesis, which plays an important role in the growth and development of plants, but free chlorophyll and its precursors will cause photooxidative damage to plant cells, so plants have formed a series of complex The mechanism to precisely regulate the biosynthesis of chlorophyll, so that it can adapt to the changing physiological and biochemical state and the external environment, resulting in different types of leaf color mutations such as albino, yellow, evergreen, dark green, light green, zebra and stripes body. Albinism is one of the most common leaf color mutations. Its mechanism involves many regulatory metabolic pathways and ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 汪维红张凤兰于拴仓苏同兵温常龙余阳俊张德双赵岫云李佩荣
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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