Nucleotide sequence of chorismate mutase related to rice high yield and its application
A rice and sequence technology, applied in the biological field, can solve problems such as the difficulty of affecting yield, and achieve the effect of high accuracy and simple operation
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Embodiment 1
[0062] Embodiment 1, design and synthesis of primers
[0063] Chorisnic acid is an important intermediate substance in rice. In the rice genome, several genes have been annotated as encoding chorismate mutase (EC5.4.99.5). Such as the gene encoding chorismate mutase 3 (chorismate mutase 3, CT829390.1; OSCM3) (hereinafter referred to as OSCM3 gene). Located on chromosome 1 of rice. Through a large number of preliminary experiments and sequence comparisons, the inventors of the present invention have found that there are two kinds of allele fragments in the OSCM3 gene (one kind of allele fragment is named as allele fragment OSCM3_a as shown in sequence 1 of the sequence table; the other An allele fragment is shown in Sequence 2 of the Sequence Listing, named allele fragment OSCM3_b), and they are related to rice yield per plant.
[0064] A specific primer pair was designed according to the above two allelic fragments, consisting of primer 1 and primer 2.
[0065] Primer 1 (s...
Embodiment 2
[0067] Embodiment 2, the establishment of the typing method based on the allele fragment in rice
[0068] The established method is as follows:
[0069] 1. Using the genomic DNA (about 10-100 ng) of the rice to be tested as a template, a specific primer pair composed of primer 1 and primer 2 is used to perform PCR amplification to obtain a PCR amplification product.
[0070] The reaction program of PCR amplification: 95°C for 5 minutes; 35 cycles of 95°C for 30 seconds, 60°C for 1 minute, and 72°C for 1 minute; 72°C for 8 minutes.
[0071] 2. After step 1 is completed, the PCR amplification product is sequenced, and the following judgment is made according to the sequencing result: if there is only one PCR amplification product, and as shown in sequence 1 in the sequence table, the genotype of the rice to be tested is OSCM3_a Homozygous; if there is only one PCR amplification product, and as shown in sequence 2 in the sequence listing, the genotype of the rice to be tested is...
Embodiment 3
[0073] Example 3, Correlation Analysis of Genotype Based on the Allelic Fragment in Rice and Yield per Plant of Rice
[0074] 1. According to the typing method established in Example 2, detect the genotype of each trial rice variety (the results are shown in Table 1, Table 2 and Table 3, column 4).
[0075] Table 1
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[0077]
[0078] 2. Statistical yield per plant of different rice varieties
[0079] In 2017, multiple rice varieties were planted in the Xiangshan area of Beijing (see Table 1 for details, see the second column for the name of the rice variety, and see the third column for the origin of the rice variety). After the rice is mature, the rice plants are harvested per plant, repeated at least three times, weighed, and averaged to obtain the yield per plant of the variety (results are shown in Table 1, column 5).
[0080] The results in Table 1 show that the genotypes of 17 of the 37 rice varieties are OSCM3_a homozygous, and the average yield per pl...
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