Application of gibberellin transport gene osnpf3.1 in improving rice yield
A gene and rice technology, which is applied in the application field of gibberellin transport gene OsNPF3.1 in improving rice yield, can solve the problem that there is no report on the biological impact of rice, and achieve the effect of increasing yield
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Embodiment 1
[0024] Construction of embodiment 1 OsNPF3.1 gene overexpression plant
[0025] Extract the RNA of rice Zhonghua 11 and reverse transcribe it into cDNA, using the primer pair:
[0026] F1: 5'- AGATCT ATGGCGGAGGAGGAGGAGGCGAAGAAGA-3' (Bgl II),
[0027] R1: 5'- CTTAAG TCAATGACTCAAAGGTGAAGCCTT-3' (Afl II);
[0028] After the cDNA of the OsNPF3.1 gene was amplified by PCR, it was digested with Bgl II and Afl II and then ligated into the pCAMBIA-1301 vector (the pCAMBIA-1301 vector was purchased from Cambia Company) to construct the overexpression vector OsNPF3 of the OsNPF3.1 gene. 1-p1301. Using the genetic transformation method mediated by Agrobacterium EHA105, the overexpression vector was introduced into the normal rice variety Zhonghua 11.
[0029] Transplant all the obtained transgenic seedlings into baskets with soil, water and fertilize them regularly, and plant them in the field when the seedlings grow about 10cm in height. After the seedlings grow up, extract genom...
Embodiment 2
[0033] The acquisition of embodiment 2 OsNPF3.1 gene mutant plant
[0034] F3: ACCACCCTCACCAACTTCGG CGG ,
[0035] F4: TCCGCCATGACCCCGCTCAT CGG .
[0036] Using the above two target sequences, the gene knockout vector OsNPF3.1-C of the OsNPF3.1 gene was constructed (for the method refer to Ma X et al, A robust CRISPR / Cas9 system for convenient, high-efficiency multiplex genome editing in monocot and dicot plants. Mol Plant. 2015, 8(8):1274-1284). The genetic transformation method mediated by Agrobacterium EHA105 was used to introduce the gene knockout expression vector into the normal japonica rice variety Zhonghua 11. The mutant plants were sequenced in the T0 generation to confirm that the gene had been knocked out, and continued to propagate until the T1 generation to obtain the mutant plants of the OsNPF3.1 gene. The number of tillers in the OsNPF3.1 mutant plants was much more than that in the control Zhonghua 11 plants, and the difference was significant, as shown...
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