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The main gene of watermelon umbilical spot trait and the molecular marker and application of the main gene detection

A technology of molecular markers and major genes, which is applied in biochemical equipment and methods, microbiological determination/inspection, DNA/RNA fragments, etc., can solve the problems of low efficiency in genetic research, achieve low cost, stable variation, and detection convenient effect

Active Publication Date: 2022-04-29
ZHENGZHOU FRUIT RES INST CHINESE ACADEMY OF AGRI SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Conventional genetic research is not efficient. Molecular markers such as Indel and SNP have attracted much attention today with the rapid development of high-throughput sequencing technology. They can be widely used in the construction of genetic maps of horticultural crops, QTL analysis, molecular marker-assisted selection breeding, etc.

Method used

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  • The main gene of watermelon umbilical spot trait and the molecular marker and application of the main gene detection
  • The main gene of watermelon umbilical spot trait and the molecular marker and application of the main gene detection
  • The main gene of watermelon umbilical spot trait and the molecular marker and application of the main gene detection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1 Location of gene loci for watermelon umbilical spot traits and development of molecular markers

[0030] Extraction of B176 (BC 1 ) Genome sequencing information of the paternal and maternal genomes of the population, conduct preliminary QTL mapping, and design and develop Indel and SNP molecular markers in the QTL region. Specific steps are as follows:

[0031] (1) F 1 Generation, F 1 Backcross with the mother parent to obtain BC 1 group. Identification Statistics BC 1 Population umbilical plaque phenotype. The B132 has no umbilical plaques, B135 has umbilical plaques, F 1 There is an umbilical plaque.

[0032] (2) Using whole-genome simplified sequencing technology and MSTmap software to analyze watermelon BC 1 Population development of high-density molecular signatures, construction of high-density genetic linkage maps, such as figure 1 shown.

[0033] (3) Combining genetic map and BC 1 Statistical results of population hilum spot traits, using ...

Embodiment 2

[0039] Example 2 primer pair Zshpr101 to watermelon BC 1 Identification of traits of umbilical plaques in populations

[0040] With the primer pair Zshpr101 of the dCAPS molecular label of embodiment 1 pair BC 1 Populations were analyzed for molecular markers.

[0041] Specific steps are as follows:

[0042] 2.1 Extraction of BC by CTAB method 1 Population watermelon genomic DNA

[0043] (1) Put 1 g of fresh watermelon leaves into a mortar, add liquid nitrogen to grind them into powder, then transfer them to a centrifuge tube with 1 mL of CTAB extraction solution preheated at 65°C, turn them upside down to fully Mix well, place in a constant temperature water bath at 65°C for 60 minutes, and invert and mix several times during the process to prevent precipitation;

[0044] (2) After the sample is taken out from the water bath, centrifuge at 8000rpm for 1min;

[0045] (3) Take the sample and put the supernatant into another centrifuge tube, add a mixture of chloroform and...

Embodiment 3

[0079] Example 3 Identification of the primer pair Zshpr101 on the umbilical spot traits of watermelon natural populations

[0080] Utilize the primer pair Zshpr101 according to the method for embodiment 2 to identify the umbilical spot character of watermelon natural population, the maternal genotype (no umbilical spot) is A, the male parent's genotype (with umbilical spot) is recorded as B, heterozygous The genotype (with umbilical plaque) is H, and the statistical results are shown in Table 2 below. The results showed that the concordance rate of genotype and phenotype identified by Zshpr101 in natural populations reached 69%.

[0081] The identification result of the watermelon hilum spot character in the natural population of table 2

[0082]

[0083]

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Abstract

The invention discloses the main effect gene of watermelon umbilical spot traits Cla019481 The gene and the dCAPS molecular marker of the main effect gene, the 21st base in the nucleotide sequence of the molecular marker is mutated from G to A, and the mutation site is located at 5706652 bp on chromosome 3 of the watermelon genome. The invention also discloses the application of the dCAPS molecular marker in identification of umbilical spot traits of watermelon germplasm resources and / or molecular marker assisted breeding and genotype purity identification of umbilical spot traits. through the BC 1 Validation in populations and natural populations shows that dCAPS molecular markers can effectively identify the trait of watermelon hilum spot, which can be determined on chromosome 3 Cla019481 The gene is the main gene controlling the traits of umbilical spot in watermelon. The method of the invention can quickly and accurately locate the trait gene of the watermelon umbilical spot. The invention provides new means for the identification of umbilical spot traits of watermelon germplasm resources and the research of molecular marker-assisted breeding.

Description

technical field [0001] The invention relates to the technical field of biological genes, in particular to a main gene for watermelon umbilical spot traits and a molecular marker for detecting the main gene and its application. Background technique [0002] Watermelon (Citrullus lanatus) is an annual plant of Cucurbitaceae Cucurbitaceae. The flesh is delicious, juicy, and rich in nutritional value. It is deeply loved by consumers and is known as the "king of melons and fruits". During its cultivation process, watermelon has undergone long-term natural selection and artificial selection to form many characters and varieties. Hilum spot refers to the inconsistent color between the hilum of watermelon and the surface of the seed. This trait can be inherited stably, with obvious differences between varieties, and this trait does not exist in wild watermelon. Therefore, hilum spot can become a watermelon germplasm resource , The main basis for variety identification and genetic c...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12N15/11
Inventor 马双武陈继峰张晓雨王吉明尚建立李娜周丹闫劝劝李楠楠
Owner ZHENGZHOU FRUIT RES INST CHINESE ACADEMY OF AGRI SCI