Method of culturing high-yielding crops and special specific genome segment and specific primer pair thereof
A technology of specific primer pairs and specific genes, applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc. Effect
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Embodiment 1
[0058] Embodiment 1, design and synthesis of primers
[0059] Sucrose is an important transport substance in rice. In the rice genome, multiple genes have been annotated as encoding sucrose synthase (EC 2.4.1.13), such as the gene encoding sucrose synthase 2 (sucrose synthase 2, JN944363.1; SUSY2) (hereinafter referred to as the SUSY2 gene) . The SUSY2 gene is located on rice chromosome 3. 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 SUSY2 gene (one kind of allele fragment is named as the allele fragment SUSY2_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 SUSY2_b), and they are related to rice yield per plant.
[0060] A specific primer pair was designed according to the above two allelic fragments, consisting of primer 1 and primer 2.
[0061...
Embodiment 2
[0063] Embodiment 2, the establishment of the typing method based on the allele fragment in rice
[0064] The established method is as follows:
[0065] 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.
[0066] 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.
[0067] 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 kind of PCR amplification product, and as shown in sequence 1 in the sequence table, the genotype of the rice to be tested is SUSY2_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 t...
Embodiment 3
[0069] Example 3, Correlation Analysis of Genotype Based on the Allelic Fragment in Rice and Yield per Plant of Rice
[0070] 1. Statistical yield per plant of different rice varieties
[0071] In 2014, multiple rice varieties were planted in Sanya City, Hainan Province (see Table 1, Table 2 and Table 3 for details, the names of rice varieties are listed in column 2, the origins of rice varieties are listed in column 3, and the 2009 numbers of some rice varieties are listed in column 6). In the field, a completely randomized block test design was adopted, with three repetitions. After the rice is mature, the rice plants are harvested per plant, weighed, and averaged to obtain the yield per plant of the variety (results are shown in Table 1, Table 2 and Table 3, column 5).
[0072] Two, according to the typing method established in embodiment 2, detect the genotype of each rice variety (results are shown in Table 1, Table 2 and Table 3, the 4th column).
[0073] Table 1
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