Watermelon grain size gene and its SNP molecular marker and application

A technology of molecular markers and watermelon seeds, which is applied in the field of molecular biology, can solve problems such as the inability to determine the position and effect of a single quality/quantity trait gene, difficulty in screening, and many candidate genes, so as to achieve accurate directional genetic improvement, improve accuracy and The effect of selecting efficiency and accelerating the improvement process

Active Publication Date: 2021-07-09
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

[0005] Traditional genetics uses mathematical statistics methods, which can only study one or more genes controlling quality / quantity traits as a whole, and cannot determine the position and effect of a single quality / quantity trait gene on the chromosome (Xu Yunbi, 1992)
[0006] With the development of molecular marker technology and the improvement of molecular quantitative genetics, a number of QTL mapping work has been carried out in recent years (Fan et al., 2000; Hawkins et al., 2001; Yi Ke, 2002; Prothro et al., 2012; Meru and McGregor, 2013), in order to reveal the genetic basis of watermelon seed size, but these studies have located a large range, many candidate genes and are difficult to screen, so far no watermelon seed size gene has been identified

Method used

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  • Watermelon grain size gene and its SNP molecular marker and application
  • Watermelon grain size gene and its SNP molecular marker and application
  • Watermelon grain size gene and its SNP molecular marker and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1: Determination of watermelon seed size genes and development of molecular markers

[0038] 1. Fine mapping of main effect QTL for watermelon seed size

[0039] F 2 The high-density genetic linkage map constructed by the population, the parents, F 1 and F 2 Visual identification of grain size in 93 single melons from the population; combined genetic linkage map and F 2 As a result of population phenotype identification, the Rqtl-IM-binary method was used for initial QTL mapping, and the main QTL for grain size was identified, with a peak LOD of 46.94, explaining 26.16% of the phenotypic variation, and a confidence interval (2-LOD) of 3.68 ~31.67 cM.

[0040] 2. Comparative transcriptomic analysis of kernel size correlation

[0041] (1) Construct recombinant inbred lines (RILs) by using the watermelon large-grain female parent "ZXG01478" and small-grain male parent "14CB11", and screen one of the large-grain (RIL_L) and small-grain (RIL_S) postanthesis 7, 8...

Embodiment 2

[0053] Embodiment 2: Watermelon F 2 Population molecular marker analysis

[0054] 1. Using CTAB method to extract watermelon F 2 The total DNA of the leaves of the population, the specific steps are as follows:

[0055] (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;

[0056] (2) After taking it out from the water bath, centrifuge at 8000 rpm for 1 min;

[0057] (3) Take the supernatant and put it in another centrifuge tube, add an equal volume of chloroform: isoamyl alcohol, and gently invert to mix well; the volume ratio of chloroform and isoamyl alcohol is 24:1;

[0058] (4) Centrifuge at 10,000 rpm for 5 min, and take the supernatant;

[0059] (5) Add 0.7 times the volume of pre-cooled isopropanol and mix well, then f...

Embodiment 3

[0091] Embodiment three: F 2 Population genotype analysis

[0092] Carry out dcaps9_S6 molecular marker through embodiment 2 in constructing F by " ZXG01478 " and " 14CB11 " 2 After genotype identification in the population, examine the dcaps9_S6 molecular marker for both genotypes at 93 F 2 distribution across populations, the dcaps9_S6 genotype was found to completely co-segregate with the kernel size phenotype. The results are shown in Table 2, where A represents the maternal band type, B represents the paternal band type, and H represents the heterozygous genotype.

[0093] The genotype of the molecular marker dcaps9_S6 was B (113bp) in 24 of the 69 small-grain (thousand-grain weight<37g) lines, and H in 44. In the 24 large-grain lines, all of them were A (142bp).

[0094] The 24 strains with molecular marker InDel14_S6 genotype A are all large-grained strains, while the 24 strains with genotype B are all small-grained strains. The accuracy rate of genotype identifica...

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Abstract

The invention discloses a watermelon grain size gene and its SNP molecular marker and application. The nucleotide sequence of the gene is shown in SEQ ID NO.1, and the encoded protein sequence is shown in SEQ ID NO.2; a gene having more than 90% homology with the gene and encoding the same functional protein has also been designed; A SNP site co-segregated by the watermelon seed size gene is located at 5418365bp of chromosome 6 of the watermelon genome, and its base is mutated from G to A; the dCAPs molecular marker of the SNP site is designed; a watermelon seed size genotype is designed The identification method of watermelon grain size gene or dCAPs molecular marker is applied in watermelon grain size molecular marker-assisted screening and breeding. It can provide a new method for identifying the grain size of watermelon and accelerate the improvement process of grain size traits of watermelon.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a watermelon seed size gene and its SNP molecular marker and application. Background technique [0002] The first use of watermelon is to eat fresh, and the pulp is the edible part. The large seeds affect the consumption. Consumers prefer watermelons with no seeds or small seeds. Therefore, small-grained varieties are one of the breeding goals for watermelon cultivation. Another use of watermelon is for seeds, which are the main product of watermelons for seeds. Studies have shown that watermelon seeds for seeds are high-quality protein and vegetable oil resources, and are rich in vitamin D. Therefore, large-grained varieties are the main breeding targets for watermelons for seeds. [0003] In the process of collecting a large number of resources and data collection, the inventors found that the large grains are usually wild watermelons, sticky watermelons and local va...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29C07K14/415C12N15/11C12Q1/6895C12Q1/686
CPCC07K14/415C12Q1/686C12Q1/6895C12Q2521/301C12Q2565/125
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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