Common wild rice grain shape related coding gene and application thereof
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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.
Active Publication Date: 2021-03-16
GUANGXI ZHUANG AUTONOMOUS REGION ACAD OF AGRI SCI
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[0004]
The main problem of high-quality rice breeding today is the narrow range of gene sources, resulting in a limited selection of good genes in rice
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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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Abstract
The invention discloses an application of a common wild rice grain shape related coding gene GL12-1 in regulation and control of a rice grain shape. The common wild rice grain shape related coding gene GL12-1 is derived from a DNA sequence of a Y12 strain of oryza common wild rice (Oryzarufipogon Griff.), and the regulation and control of the rice grain shape refers to the regulation and control of the rice grain shape to be shortened and narrowed. According to the coding gene, the common wild rice grain shape related gene GL12-1 is identified for the first time, and the grain shape related common wild rice can be obtained by the GL12-1 under the condition of function enhancement or expression quantity increase, so that the common wild rice grain shape related gene protein or the protein thereof is proved to play an important role in controlling the common wild rice grain shape; the transgenic rice with increased GL12-1 gene expression obtained by the invention is used as a new rice germplasm material, and can be used for researching the grain shape rice mechanism and discovering more genes for regulating and controlling the grain shape development of common wild rice.
Description
technical field [0001] The invention relates to the technical field of genetic engineering, in particular to a common wild rice grain type-related coding gene and its application. Background technique [0002] Rice is one of the most important food crops for human beings. More than 100 countries in the world grow rice, and 2 / 3 of the population uses rice as their staple food. The demand for rice in Asia is increasing year by year. Due to rapid economic development, increasing population pressure, sharp increase in industrial land, and decrease in arable land, rice production is facing tremendous pressure. Rice quality is closely related to grain shape, and its appearance quality is mainly measured by indicators such as grain shape, chalkiness and transparency. Therefore, it is the main research direction of scientists to provide grain type gene resources for high-quality rice breeding. [0003] The main genes regulating grain length in rice mainly include GS3, qGL3, GL4 an...
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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/16C12N15/82A01H5/10A01H6/46
CPCC12N9/16C12N15/8261C12Y301/03003
Inventor 潘英华邓国富高利军戴高兴
Owner GUANGXI ZHUANG AUTONOMOUS REGION ACAD OF AGRI SCI
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