Rice glume development gene promoter p-TRI1 and application thereof
A promoter, transgenic rice technology, applied in the application, angiosperms/flowering plants, DNA/RNA fragments, etc., to achieve the effect of broad application and market prospects
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Embodiment 1
[0028] Example 1, Discovery of rice glume development gene promoter p-TRI1
[0029]1. Discovery of rice glume development gene TRI1
[0030] After 93-11 was treated with EMS (ethyl methane sulfonate) and self-crossed for multiple generations and its traits were observed, a mutant line with a pure genotype was formed. Through the screening of these mutant systems, a mutant with abnormal glume development was found. The specific performance of this mutant is: the glume width of the mutant is narrower than that of the normal wild type, that is, the glume width of 9311 is 2.84± 0.08mm, the glume width of the mutant was 2.33±0.11mm. However, there was no difference in the length of glumes between the two. The most obvious feature is that it exhibits a variation of triangular shape at its tip, hence the name tri1.
[0031] Tri1 was crossed with 93-11, Teqing, Guichao 2, Zhonghua 17, C418 and other conventional varieties respectively, and the hybrid F 1 It represents the characte...
Embodiment 2
[0043] Example 2, the acquisition and identification of p-TRI1 transgenic rice
[0044] 1. Construction of p-TRI1 plant expression vector
[0045] 1. Construction of p-LTT7 plant expression vector
[0046] 1. Using the genomic DNA of 93-11 rice as a template, use specific primers to T7EUS (T7EUSF / T7EUSR), and use the KODplus enzyme reaction kit of TOYOBO Biological Company to perform PCR amplification. After the reaction, perform PCR amplification. The amplified product was detected by 1% agarose gel electrophoresis, and the 2229bp DNA fragment (p-LTT7) was recovered and purified (the recovery kit used was the DNA product purification kit of Tiangen Biochemical Technology Co., Ltd., catalog number: DP204-02) .
[0047] 2. Digest the PCR product recovered in step 1 with restriction endonucleases EcoRI and SalI.
[0048] 3. Digest the plant expression vector pCambia1381 with restriction endonucleases EcoRI and SalI, and recover the vector skeleton.
[0049] 4. Ligate the dig...
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