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A SNP functional molecular marker of lodging resistance gene in japonica rice and its application
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A functional molecule, anti-lodging technology, applied in the field of plant molecular biology, can solve the problems of complex operation, high labor input, inaccurate manual identification results, etc.
Inactive Publication Date: 2020-07-07
YANGTZE NORMAL UNIVERSITY
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[0005] In view of the above-mentioned deficiencies in the prior art, the purpose of the present invention is to provide a SNP functional molecular marker of the lodging resistance gene of japonica rice, so as to solve the problems of long time-consuming and high manpower input in identification of the lodging resistance phenotype of japonica rice
[0006] The invention also provides a primer set and KASP method for detecting SNP functional molecular markers, which solves the problems of unsafe experimental operation, complicated operation and inaccurate manual identification results, etc.
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Embodiment 1
[0033] Example 1: Phenotypic verification of natural genetic material of SNP marker K_06g_SCM2-1
[0034] 23 natural genetic materials were selected for verification, including 11 lodging-resistant materials and 12 lodging-sensitive materials.
[0035] 1. Genomic DNA was extracted from japonica rice leaves by simplified CTAB method.
[0036] (1) Freeze-dry or dry the japonica rice leaves, put an appropriate amount of leaves into a 2.0mL centrifuge tube, add two steel balls, and grind them on a tissue grinder;
[0037] (2) Add 750 μL of CTAB solution, oscillate to homogenate, and incubate at 65°C for 0.5-1 h;
[0038] (3) Cool to room temperature, add 750 μL of chloroform:isoamyl alcohol (24:1) in the fume hood and mix by inverting 3-4 times;
[0039] (4) Centrifuge at 12000rmp for 10min, transfer 500μL supernatant to a new 1.5mL centrifuge tube;
[0040] (5) Add an equal volume of isopropanol solution and shake gently to mix, precipitate at -20°C for more than 1 hour, centr...
Embodiment 2
[0054] Example 2 Phenotype verification of F2 segregation population of SNP marker K_06g_SCM2-1
[0055] Detect the individual plant genotype of the F2 segregation population crossed by the donor parent Habataki and the recipient parent Sasanishiki;
[0056] 1. The genotyping method is the same as in Example 1. For the genotyping results of 80 F2 isolated populations, see figure 2 .
[0057] 2. Select two homozygous genotypes for phenotypic identification. During the period from the booting stage to the heading of rice, remove the top leaves of the plants, measure the mechanical strength and diameter of the plant stems, and comprehensively evaluate the lodging resistance of the plants.
[0058] The genotype and phenotype data of homozygous individual plants are shown in Table 3.
[0059] Table 3 The phenotype identification results of the F2 segregation population of marker K_06g_SCM2-1
[0060]
[0061]
[0062] Note: "R" indicates that the plant phenotype is resist...
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Abstract
The invention discloses an SNP functional molecular marker of a lodging-resistant gene of japonica rice and application of the marker. The lodging-resistant gene is SCM2; the SNP functional molecular marker is K_06g_SCM2-1 and is positioned at the 27481645th basic group of the No. 6 chromosome of rice; the polymorphism of the SNP molecular marker is T / G; the sequence of the marker is shown in SEQ ID No.1; the base group at the 55bp site is T or G and is positioned in a promoter region of the gene SCM2. The SNP functional molecular marker disclosed by the invention can be used for predicting the lodging resistance of a japonica subspecies material of rice in an early stage (seedling stage or embryonic stage), and can be used for performing accurate screening, thereby promoting genetic improvement and breeding of lodging resistance traits of the japonica rice. According to the invention, a KASP method is adopted to detect an SNP site, and the detection method is accurate and reliable, easy and convenient to operate, and suitable for application in high-throughput commercialized molecular breeding.
Description
technical field [0001] The invention relates to the technical field of plant molecular biology, in particular to a SNP functional molecular marker of a japonica rice lodging resistance gene and its application. Background technique [0002] Traditional pedigree selection is a method commonly used by rice breeders. Through years of efforts by breeders, a large number of new rice varieties with high yield, high resistance and high quality have been bred and created. However, the traditional breeding model combined with phenotypic identification has shortcomings such as inaccurate control of phenotypes, large demand for genetic populations, high labor costs, and long breeding cycles, making it difficult to achieve large-scale commercial integrated breeding or genetic improvement of materials. With the development of molecular biology and bioinformatics, molecular marker-assisted selection breeding (Molecular Assisted Selectoin, MAS) has shown great technical advantages, realizi...
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