Common wild rice grain shape related coding gene and application thereof
An encoding gene and wild rice technology, which is applied to the common wild rice grain type-related encoding gene and its application field, can solve the problems of small selection range of excellent genes in rice and narrow gene source range.
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Embodiment 1
[0055] Construction of overexpression vector of common wild rice grain type related gene GL12-1
[0056] 1. Acquisition of GL12-1 gene
[0057] Using the DNA of common wild rice Y12 (Oryzarufipogon Griff.) as a template, use the following primers primer1 and primer2 to perform PCR amplification to obtain the target gene:
[0058] primer1: 5'ATGAACATCGCCACTGGG 3';
[0059] primer2: 5' CTATAGTTGTTCCGAAAC 3'.
[0060] After recovering and purifying the PCR product, it was connected to Zero (purchased from Beijing Quanshijin Company) sequencing vector, transformed into DH5α competent cells, and the positive clones were selected for sequencing.
[0061] Sequencing results show that the amplified PCR product sequence is the nucleotide sequence shown in SEQ ID No.1, with a length of 2460bp, named GL12-1 gene, and the amino acid sequence of the protein encoded by the GL12-1 gene is shown in SEQ ID No. .2 shown.
[0062] 2. Construction of the overexpression vector (recombinant exp...
Embodiment 2
[0067] Breeding and identification of transgenic plants overexpressing GL12-1 with increased expression level of common wild rice grain type-related gene GL12-1
[0068] 1. Cultivate overexpressed GL12-1 transgenic plants with increased expression level of the common wild rice grain type-related gene GL12-1, and transform the recombinant vector OE-GL12-1 through Agrobacterium tumefaciens EHA105 to transform indica rice 253 japonica rice, the specific method as follows:
[0069] 1. The recombinant vector OE-GL12-1 obtained in Example 1 was introduced into Agrobacterium tumefaciens EHA105 by heat shock method to obtain recombinant Agrobacterium tumefaciens EHA105 containing recombinant vector OE-GL12-1; The recombinant Agrobacterium tumefaciens EHA105 of 1 was cultured at 28°C for 16 hours, and the bacterial cells were collected; the bacterial cells were diluted with N6 liquid medium (Sigma, catalog number C1416) containing 100 μM acetosyringone to obtain the diluted bacterial l...
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