Application of OsGBP1 gene to regulation and control on rice flowering and grain types

A gene and rice technology, applied in the application field of OsGBP1 gene in regulating the flowering and grain shape of rice, can solve the problems of affecting yield potential, affecting rice plant height and yield, etc., and achieve the effect of reducing grain length and promoting plant growth.

Active Publication Date: 2018-05-15
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although many genes related to heading date have been cloned in rice, such as Ehd1, Ghd7, DTH8 / Ghd8 (Doi et al., 2004; Xue et al., 2008; Wei et al., 2010; Yan et al., 2010), etc., Some genes, such as Ghd7 and Ghd8, negatively regulate rice flowering and affect rice plant height and yield at the same time. They are pleiotropic genes. However, when these two genes are aggregated, rice will always be in the vegetative growth stage without flowering. Seriously affect the production potential

Method used

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  • Application of OsGBP1 gene to regulation and control on rice flowering and grain types
  • Application of OsGBP1 gene to regulation and control on rice flowering and grain types
  • Application of OsGBP1 gene to regulation and control on rice flowering and grain types

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0131] Cloning of Example 1 Gene OsGBP1

[0132] Extract the DNA of rice variety Nipponbare (primer sequence: upstream primer 5′-AGCCTCTGGATCATTCGC-3′ and downstream primer 5′-GGCAAATGGCCTCTTCGA-3′) for polymerase chain reaction (PCR), and sequence the obtained PCR product to obtain the gene The gene sequence of OsGBP1 consists of 6040 bases, and the nucleotide sequence is shown in SEQ ID NO:1. PCR program: pre-denaturation at 94°C for 5 minutes; 35 cycles (denaturation at 94°C for 30 seconds; annealing at 55°C for 30 seconds; extension at 72°C for 6 minutes), and extension at 72°C for 10 minutes. RNA was extracted from the leaves of rice variety Nipponbare, reverse-transcribed into cDNA, and polymerase chain reaction (PCR) was performed with primers (primer sequence: upstream primer 5′-GGTACCCCTCTCCACCCCCAACA-3′ and downstream primer 5′-GGATCCCTACCGGATGGTGATGTACCG-3′) , the size of the amplified product is 1095bp (containing 99bp 5' non-coding sequence), and the obtained PCR...

Embodiment 2

[0134] The construction of embodiment 2 recombinant vector and the establishment of transformed agrobacterium

[0135] (1) The sequence containing the OsGBP1 gene CDS (SEQ ID NO: 2) amplified in Example 1 was double-digested with Kpn I and BamH I, and the target product was separated and recovered, and then digested with Kpn I and BamH I. The pCAMBIA1301S vector was ligated with T4 ligase to form an overexpression vector. The above primers were synthesized by Shanghai Sangong, and the restriction enzymes BamH I, Kpn I and T4 ligase were purchased from Takara Company.

[0136] (2) According to figure 1 The vector construction technical route of using the Nipponbare leaf cDNA obtained in Example 1 as a template with primers (primer sequence: upstream primer 5'-AAAGAGCTCGGATCCGCTTCGCCAATGCTACAA-3' and downstream primer 5'-AAAACTAGTGGTACCTGATCTTCCCCATCTTTCC-3') to carry out polymerase chain reaction (PCR) to isolate a 303-base cDNA fragment of the gene OsGBP1, the sequence of wh...

Embodiment 3

[0139] Example 3 Agrobacterium-mediated genetic transformation

[0140] (1) Induction: Shell the seeds of mature rice varieties (Zhonghua 11 and Nipponbare), and then treat them with 75% volume ratio of ethanol for 1 minute, and 0.15% concentration of mercury chloride (HgCl 2 ) Disinfect the surface of the seeds for 18 minutes; wash the seeds 4-5 times with sterilized water; place the seeds on the japonica rice induction medium; place the inoculated medium in a dark place for 4 weeks at a temperature of 25±1°C.

[0141] (2) Subculture: select bright yellow, compact and relatively dry embryogenic calli, put them on the japonica rice subculture medium and culture them in the dark for 2-3 weeks at a temperature of 25±1°C.

[0142] (3) Agrobacterium culture:

[0143] In the LA medium with kanamycin resistance (product of Shanghai Shenggong Company) selected (the preparation of LA medium can refer to J. Science Press, 2002) to pre-culture Agrobacterium strains FOX and FR for two ...

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Abstract

The invention provides application of an OsGBP1 gene to regulation and control on rice flowering and grain types. The gene controls the rice flowering time; through the over-expression of the gene, the rice heading period is delayed, the grain length is reduced, but the grain weight is not affected. The gene negatively regulates and controls the seed size; through inhibiting the expression of thegene, the plant growth at the seedling stage is promoted, and later grains become longer. The application of the OsGBP1 gene, provided by the invention, has the beneficial effects that valuable genetic resources are provided for crop breeding, and the rice OsGBP1 gene can be widely applied to crop hybrid breeding and hybrid seed production.

Description

technical field [0001] The invention belongs to the technical field of plant genetic engineering, and specifically relates to the application of OsGBP1 gene in regulating rice flowering and grain type. Background technique [0002] Rice is one of the most important food crops in the world. In order to meet the food demand of the growing population, breeders have worked to address the demand by increasing rice production. However, in the past more than half a century, although the dwarf breeding brought about by the first green revolution and the successful application and large-scale promotion of cross-breeding later resulted in two qualitative leaps in rice yield, however, in the past two decades , the effect of traditional methods on increasing yield is significantly reduced, coupled with the deteriorating natural ecological environment. New functional genes to cultivate new varieties with high yield and high quality. [0003] The size of the grain shape determines the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/82A01H5/10A01H5/02A01H6/46
CPCC07K14/415C12N15/8218C12N15/8261C12N15/827
Inventor 余四斌龚蓉
Owner HUAZHONG AGRI UNIV
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