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Molecular marker and application thereof

A technology of molecular markers and melons, applied in the direction of DNA/RNA fragments, measurement/inspection of microorganisms, recombinant DNA technology, etc., to achieve the effect of saving time, high accuracy and low cost

Active Publication Date: 2016-07-27
SHENZHEN BGI AGRI & CYCLE ECONOMIC TECH +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] However, at this stage, the molecular markers closely linked to the melon peel color gene still need to be discovered.

Method used

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  • Molecular marker and application thereof
  • Molecular marker and application thereof
  • Molecular marker and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1F2

[0042] Example 1 F2 Generation Segregation Population Construction and Character Investigation

[0043] (1) Material

[0044] Using the male parent "K3-92-1" and female parent "K1-7" of melon "Jinmi No. 6" provided by Xinjiang Baofeng Seed Industry Co., Ltd. as materials, the F2 generation segregation population was constructed.

[0045] The male parent "K3-92-1" is an inbred line with an average leaf length of 21.6cm and an average leaf width of 29.3cm. It has a thick-skinned muskmelon, light yellow skin, fine-grained texture, no covering pattern, orange-red flesh, and long stalks. The average weight of a single melon is 2.4kg, the fruit shape index is 1.55, the soluble solid content in the edge is 7.4%, and the soluble solid content in the center is 11.9%.

[0046] The female parent "K1-7" is an excellent single-melon line bred by self-crossing of a farm variety "Yangtou" in Kazakhstan. The average leaf length is 24.1cm, and the average leaf width is 34.5cm. Yellow-green s...

Embodiment 2

[0055] Example 2: SNP marker analysis and genetic map drawing

[0056] 1. SNP marker analysis and genetic map construction

[0057] 1.1 SNP marker analysis

[0058] Get the tender leaves of each individual plant of the parents "K1-7" (P1), "K3-92-1" (P2) and F2 populations in Example 1, use improved CTAB (cetyl trimethyl bromide Ammonium) method to extract genomic DNA, specifically as follows:

[0059] Genomic DNA of parents, F1 generation, and F2 generation individuals were extracted by CTAB method, the specific method is as follows:

[0060] (1) Weigh 1.0g of fresh leaves, cut them into pieces and put them into a mortar, grind them with liquid nitrogen, add 3mL1.5×CTAB, grind them into a homogenate and transfer them to a 15mL centrifuge tube, then add 1mL1.5×CTAB into the mortar Wash with ×CTAB and transfer to a centrifuge tube. After mixing, place in a water bath at 65°C for 30 minutes, and shake slowly from time to time during this period.

[0061] Among them, the for...

Embodiment 3

[0085] Example 3: Correlation Verification of Peel Color Correlation of Molecular Markers

[0086] The color-related molecular marker M022377 (nucleotide sequence shown in SEQIDNO: 1) of muskmelon peel determined in Example 2 is verified, specifically as follows:

[0087] Primers were designed for the above molecular markers, and the primer sequences are as follows:

[0088] Forward primer: 5'-GCCACGCTGGTAAGAAGATT-3' (SEQ ID NO: 2);

[0089] Reverse primer: 5'-ATGCACTCATTGCGTCGTTT-3' (SEQ ID NO: 3).

[0090] The polymorphism and amplification stability of the marker were verified by PCR amplification and agarose gel electrophoresis detection using the above primer pair.

[0091] Specifically, using the genomic DNA of the male parent, the female parent, the F1 generation, and the F2 generation extracted in Example 2 as templates, the above-mentioned primer pair was used to perform PCR amplification to obtain an amplified fragment, wherein,

[0092] The PCR reaction system is...

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Abstract

The invention discloses a molecular marker and application thereof. The molecular marker shows insertion / deletion length polymorphism of a nucleotide sequence as shown in SEQ ID No. 1. The molecular marker is closely related to the property of the pericarp line color of muskmelon with yellow-green pericarp and can be effectively used for molecular marker-aided breeding of the muskmelon.

Description

technical field [0001] The present invention relates to molecular markers and their applications. Specifically, the present invention relates to molecular markers related to the color of muskmelon peel, primer pairs and kits for detecting the molecular markers, the use of the aforementioned molecular markers, primer pairs, and kits in melon breeding, and detection of muskmelon peel color Methods for color traits and methods for assisted breeding of melons. Background technique [0002] Melon (Cucumismelo L.) is an important horticultural crop with high commercial and economic value. my country has a history of more than 3,000 years of melon cultivation. It is one of the earliest countries in the world to cultivate melons, and it is also the country with the largest melon cultivation area and the highest yield in the world. The fruit of muskmelon is sweet and delicious, and it is a traditional fresh fruit that is generally liked by the vast urban and rural people in my coun...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/68
Inventor 张耕耘全志武倪雪梅邹洪锋夏秋菊范荣
Owner SHENZHEN BGI AGRI & CYCLE ECONOMIC TECH
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