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A rice long-grain related gene and its application

A kind of use and grain technology, applied in the field of agronomy, can solve the problems of rice long-grain-related gene cloning and research without breakthroughs, few genes for cloning and in-depth research, etc.

Active Publication Date: 2016-08-17
CAS CENT FOR EXCELLENCE IN MOLECULAR PLANT SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Even though some quantitative trait loci have been mapped using molecular marker technology in recent years, there are still very few genes that have been successfully cloned and studied in depth
[0004] So far, no breakthroughs have been made in the cloning and research of rice long-grain-related genes in this field

Method used

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  • A rice long-grain related gene and its application
  • A rice long-grain related gene and its application
  • A rice long-grain related gene and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0114] Molecular marker-assisted selection breeding of embodiment 1GL3

[0115] Extract the DNA of the long / large-grain variety WY3 and the small-grain variety FAZ1, and design PCR oligonucleotide primers in the GL3 gene. The primer sequences are as follows:

[0116] 5' end primer: 5'-GGTCGTTGTTGTGTAAGG-3' (SEQ ID NO.: 11);

[0117] 3' end primer: 5'-TAGTGAAGATGGGTGCTG-3' (SEQ ID NO.: 12).

[0118] PCR amplification was carried out with Taq enzyme; the amplified product was digested with restriction endonuclease DraI, and the DNA polymorphism between the long / large grain variety and the small grain variety was detected by 1.5% agarose gel electrophoresis. The amplified molecular weight of the small-grain variety is about 1kb, and the amplified molecular weight of the long / large-grain variety is about 500bp, so the primers can be used as molecular markers to specifically identify the long-grain GL3 gene and the small-grain GL3 gene. The molecular marker can be used to quickly...

Embodiment 2

[0159] Embodiment 2 GL3 rice transgenic experiment

[0160] In this example, the commercially available binary expression vector pHB was used as the rice transgenic vector. The vector has a bacterial origin of replication (ori), a kanamycin resistance gene (Kan r ), hygromycin resistance gene (Hyg r ), herbicide resistance gene (Bar), CaMV35S promoter, termination signal sequence of NOS gene, and restriction enzyme cloning site (MCS). The cDNA of GL3 can be inserted forward or reverse into the restriction endonuclease cloning site to construct a transgenic plasmid.

[0161] Construction of transgenic plasmids for positive sense overexpression of GL3

[0162] In this embodiment, the RNA of the constructed long / large-grain and small-grain varieties is used as a template to synthesize the first strand cDNA, and the PCR oligonucleotides at the 5' and 3' ends of the DNA sequence are used as primers (SEQ ID NO.: 3 and SEQ ID NO.: 4), amplified with the high-fidelity Taq enzyme K...

Embodiment 3G

[0168] Embodiment 3 GL3 transforms rice

[0169] 1.1 Four kinds of recombinant plasmids prepared in Example 2 were introduced into Agrobacterium strain EHA105 (purchased from Biovector Company) by freeze-thaw method, the method is as follows:

[0170] Add 0.5-1 μg (about 10 μl) plasmid DNA to every 200 μl of EHA105 competent cells and mix well, place on ice, liquid nitrogen, and 37°C water bath for 5 minutes each; dilute to 1ml with fresh YEB liquid medium, and store at 28°C Incubate with shaking for 2-4 hours; take 200 μl and spread it on a YEB plate containing antibiotic Kan (50 μg / ml), and culture at 28° C. for 2-3 days. The grown colonies were marked on the YEB plate containing antibiotics for 3 consecutive times. Referring to the method of Hiei et al. (1994), pick a single colony of Agrobacterium from the YEB plate and inoculate it into 3ml of YEB liquid medium containing antibiotics, culture it overnight at 28°C with shaking, and transfer it to 50ml on the second day wi...

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Abstract

The invention relate to a rice long-grain related gene and an application for the same, and in particular discloses a rice long-grain related gene GL3 for the first time via researches on lots of rice quantitative trait loca, which is used for encoding serine / threonine phosphatase. Compared with a GL3 protein in a small-grain parent, a GL3 protein in a long-grain / large-grain parent has the mutations of amino acids, thus being declined in the phosphatase activity. The invention further relates to a GL3 mutant protein with a declined enzymatic activity, the encoding sequence thereof, and an application for the GL3 mutant protein in the aspects of modifying crops to increase grain length and crop yield, and the like.

Description

technical field [0001] The invention relates to the field of agronomy, in particular, the invention relates to a rice long-grain related gene and application thereof. Background technique [0002] The food problem is a worldwide problem. With the rapid increase of population and the decrease of available arable land, it has become an important strategic measure for all countries to increase food production as soon as possible. Rice (Oryza sativa) is the main food variety and economic crop for Asian populations, and the improvement of rice yield is of great significance to solve the food problem. [0003] Rice yield is mainly determined by three factors: grain weight, number of grains per panicle and number of panicles. Most of these traits are quantitative traits, and the genes controlling these traits are mainly quantitative trait loci (QTL). Therefore, identifying and understanding the functions of these genes is of great significance for breeding and explaining the mech...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/16C12N15/55C12N15/63C12N1/15C12N1/19C12N1/21C12N5/10C12Q1/68A01H5/00
Inventor 林鸿宣祁澎宋献军高继平单军祥朱美珍施敏
Owner CAS CENT FOR EXCELLENCE IN MOLECULAR PLANT SCI
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