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Watermelon stick seed gene SNP (single nucleotide polymorphism) molecular marker as well as screening method and application thereof

A technology of molecular markers and watermelons, applied in biochemical equipment and methods, recombinant DNA technology, DNA/RNA fragments, etc., can solve the problems of unobtained molecular markers, improve accuracy and selection efficiency, accelerate improvement process, and scientifically The effect of practical detection and identification technology

Active Publication Date: 2018-07-24
ZHENGZHOU FRUIT RES INST CHINESE ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But so far, molecular markers associated with sticky-seed watermelon types that are common among different watermelon materials have not been obtained

Method used

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  • Watermelon stick seed gene SNP (single nucleotide polymorphism) molecular marker as well as screening method and application thereof
  • Watermelon stick seed gene SNP (single nucleotide polymorphism) molecular marker as well as screening method and application thereof
  • Watermelon stick seed gene SNP (single nucleotide polymorphism) molecular marker as well as screening method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1: Determination of the sticky seed gene of watermelon seeds and the development of molecular markers

[0035] Simplified genome sequencing of different types of watermelon materials, combined with differences in population polymorphisms to determine the selected regions between populations, and obtaining chromosomal intervals related to sticky-seeded watermelon types, using deep resequencing of sticky-seeded watermelon materials and common watermelon materials The SNP sites in the target chromosome interval were screened, and the CAPS / dCAPS molecular marker primers were designed, and then the CAPS / dCAPS molecular marker primers and watermelon genomic DNA were used for PCR amplification, and finally the amplified products were digested and verified by electrophoresis.

[0036] Specific steps are as follows:

[0037] (1) Choose different types of watermelon materials, including sticky watermelon, common watermelon and wild watermelon;

[0038] (2) Using the meth...

Embodiment 2

[0044] Example 2: Watermelon DNA Molecular Marker Analysis

[0045] 1. Select 5 parts of sticky watermelon, 5 parts of ordinary watermelon and 5 parts of wild watermelon, and use the CTAB method to extract the total DNA of watermelon leaves. Three parallel experiments are set up for each sample. The specific steps are as follows:

[0046] (1) Put 1 g of fresh leaves into a mortar, add liquid nitrogen to grind into powder, then transfer to a centrifuge tube with 1 ml of CTAB extraction solution, mix the two thoroughly, and then place in a constant temperature water bath at 65°C 60 min, during which the mixture was inverted 2 to 3 times;

[0047] (2) After taking it out from the water bath, centrifuge at 8000 rpm for 1 min;

[0048] (3) Take the supernatant and put it in another centrifuge tube, add an equal volume of chloroform: isoamyl alcohol (24:1, V / V), invert gently to mix well;

[0049] (4) Centrifuge at 10,000 rpm for 5 min, and take the supernatant;

[0050](5) Add 0...

Embodiment 3

[0071] Embodiment three: identify and assist the germplasm type of screening watermelon

[0072] Carry out Npr372 molecular marker in 5 parts of sticky watermelon, 5 parts of wild watermelon and 5 parts of common watermelon genotype identification after carrying out embodiment 2, check the distribution situation of two genotypes of Npr372 molecular marker in 93 different types of watermelon materials .

[0073] Table 1 Identification and verification of NPr372 in 93 different types of watermelon materials

[0074] material code

breed name

Germplasm type

NPr372

S001

PI 248178

Sticky Seed Watermelon

A

S002

PI 494527

Sticky Seed Watermelon

A

S003

PI 559992

Sticky Seed Watermelon

A

S004

PI 559993

Sticky Seed Watermelon

A

S005

PI 559994

Sticky Seed Watermelon

A

S006

PI 559996

Sticky Seed Watermelon

A

S007

PI 559999

Sticky Seed Watermelon

A

...

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Abstract

The invention discloses a watermelon stick seed gene SNP (single nucleotide polymorphism) molecular marker as well as a screening method and application thereof. A SNP site of the watermelon stick seed gene is defined. A dCAPs molecular marker of the SNP site is designed, and the upstream primer nucleotide sequence is shown as SEQ ID NO.1; the downstream primer nucleotide sequence is shown as SEQID NO.2. A screening method of the dCAPs molecular marker of the SNP site and an identification method of the watermelon stick seed gene are designed. The dCAPs molecular marker of the SNP site is applicable to the identification and auxiliary screening of different germplasm kinds of watermelons. The crossbreeding parent selection cam be guided, so that the directed genetic improvement of the stick seed watermelons can be fast and accurately performed; a new means is provided for identifying whether the watermelons are in a stick seed type; the improvement process of the watermelon stick seedspecific character is accelerated; the accuracy and the selection efficiency of the breeding are improved.

Description

technical field [0001] The invention relates to the technical field of watermelon breeding, in particular to a watermelon sticky seed gene SNP molecular marker and a screening method and application thereof. Background technique [0002] Sticky-seed watermelon is resistant to dryness, early barrenness and strong resistance, especially has high anti-virus traits that cultivated watermelons do not have. The seed has an extremely thin seed coat, the thickness of which is only 0.064mm, while the thickness of the seed coat of ordinary watermelon is above 0.2mm; the relative weight of the seed coat of sticky-seeded watermelon is very light, accounting for only 17.7% of the total weight of the seed, and other watermelons All are above 49%, so the edible rate of sticky watermelon seeds is 30% to 40% higher than other watermelons; sticky watermelon seeds have higher oil content, among which unsaturated fatty acid content is higher, and have higher nutritional value. [0003] However...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/6895C12Q1/6858
CPCC12Q1/6858C12Q1/6895C12Q2600/13C12Q2600/156C12Q2531/113C12Q2521/301C12Q2565/125
Inventor 李娜马双武尚建立王吉明周丹李楠楠
Owner ZHENGZHOU FRUIT RES INST CHINESE ACADEMY OF AGRI SCI
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