SNP sites related to African rice grain length and its application
An African rice and site technology, applied in the field of plant genetic engineering, can solve the problems of poor knowledge of population genetic structure and evolutionary history, lack of research on African cultivated rice, etc.
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Embodiment 1
[0036] Example 1. Mapping of genes regulating African rice grain size and determination of SNP sites
[0037] 1. Localization of genes regulating grain size in African rice
[0038] African rice variety IRGC102305 was crossed with African wild rice W1411, and then IRGC102305 was used as the recurrent parent, and a line GIL25 ( figure 1 ), crossed it with IRGC102305 to construct a F2 segregation population containing 186 individuals. In this F2 population, kernel length was closely linked to markers RM3335 and RM5608, and the GL4 gene was located between these two genes. Then the isolated population was expanded to 6500 individual plants, and primers were designed according to the sequence between the markers RM3335 and RM5608, and finally the GL4 gene was located on the 5.9kb between the markers M3 and M4 ( figure 2 ). Based on the sequence prediction of African cultivated rice IRGC96717 (CG14) on the website http: / / ensembl.gramene.org / Oryza_glaberrima, this interval conta...
Embodiment 2
[0060] Example 2, the application of SNP site C766T in identifying the grain length of existing African rice varieties
[0061] 1. Experimental materials
[0062] Using 67 African rice varieties distributed in many countries in West Africa as experimental materials (see Table 1 for details), the application of SNP site C766T in the selection of African rice grain length was identified.
[0063] 2. Experimental method
[0064] After sampling the single plant of each test plant at the seedling stage, extract its genomic DNA and use it as a template to perform PCR amplification with primer F and primer R, and perform sequencing after obtaining the PCR product. Genotype of individual plant: if the obtained PCR product contains the nucleotide sequence shown in sequence 1, and the 64th position of sequence 1 is C homozygous type, then the individual plant to be tested is C:C genotype; if the obtained PCR product If the product contains the nucleotide sequence shown in sequence 1, ...
Embodiment 3
[0074] Example 3, the application of SNP site C766T in identifying the grain length of African rice parent hybrid offspring
[0075] 1. Experimental materials
[0076] F using PI450352 (father, C:C genotype) and PI450489 (female, T:T genotype) crossed in Table 1 2 Substitute experimental materials to identify the application of SNP site C766T in the selection of African rice grain length. At the same time, two parental controls were set.
[0077] 2. Experimental method
[0078] After each test plant was listed and sampled at the seedling stage, its genomic DNA was extracted and used as a template, PCR amplification was carried out with primer F and primer R (sequence 2 and sequence 3), and sequencing was carried out after obtaining the PCR product. As a result, the genotype of the individual plant to be tested is determined according to the following method: if the obtained PCR product contains the nucleotide sequence shown in sequence 1, and the 64th position of sequence 1...
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