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Rice gene OsPGSIP1 and application thereof

A rice and genetic technology, applied in the field of genetic engineering, can solve the problems of low production efficiency, excessive application of nitrogen fertilizer, environmental pollution, etc., and achieve the goal of increasing chlorophyll and nitrogen content, increasing the mechanical strength of stems, and increasing the biomass of seedlings Effect

Active Publication Date: 2018-12-18
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, driven by the goal of high yield, nitrogen fertilizer is easily over-applied, which not only leads to environmental pollution and low production efficiency, but also makes rice stems fragile and easy to break or lodging

Method used

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  • Rice gene OsPGSIP1 and application thereof
  • Rice gene OsPGSIP1 and application thereof
  • Rice gene OsPGSIP1 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0129] The cloning of embodiment 1 gene OsPGSIP1:

[0130] (1) Extract the DNA of the rice public variety Zhenshan 97 maintainer line (ZS97B) (purchased from the China Rice Research Institute), and polymerize with primers (primer sequences are 5'-GGGAAATTGCCATCCCTAGT-3' and 5'-AATGGGGCTCATCACTTGAG-3') Enzyme chain reaction (PCR), the obtained PCR product is sequenced to obtain the gene sequence of the rice gene OsPGSIP1, which consists of 10263 bases, and the nucleotide sequence shown is 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 10 minutes), and extension at 72°C for 5 minutes.

[0131](2) extract the RNA of rice ZS97B seedling stage leaves, reverse transcribe into cDNA, and carry out polymerase chain reaction (PCR) with primers (primer sequence: 5'-aaaGAGCTCctcgcagagaacgaaagagagtccc-3' and 5'-aaaCTGCAGatctggtatagggctatctacacat-3') , the obtained PCR...

Embodiment 2

[0134] Embodiment 2: the construction of recombinant vector and the establishment of transformed Agrobacterium:

[0135] (1) if figure 1 As shown (the cDNA of the gene OsPGSIP1 is connected to the expression vector pC1301S to form an overexpression recombinant vector), the cDNA obtained in Example 1 (shown in SEQ ID NO.2) is digested with SacI and PstI, and the target product is separated and recovered. The pC1301S vector (Mao et al., 2010) digested with SacI and PstI was ligated with T4 ligase to form an overexpression vector. The above primers were synthesized by Shanghai Sangon, and the restriction enzymes SacI, PstI and T4 ligase were purchased from Takara Company;

[0136] (2) According to figure 2 The technical route, the CDS that embodiment 1 obtains uses primer (primer sequence is:

[0137] 5'-aaaGAGCTCGGATCCtgctgaccccaccaatcctctat-3' and

[0138] 5'-aaaACTAGTGGTACCacttgtgaggctcgcggtaggt-3') by polymerase chain reaction (PCR) to obtain a 403-base cDNA fragment of ...

Embodiment 3

[0142] Example 3: Agrobacterium-mediated genetic transformation:

[0143] (1) Induction: the mature rice Zhenshan 97 maintainer line (ZS97B) (purchased from China Rice Research Institute) seeds were shelled, and then treated with 70% ethanol by volume for 1 minute, and 0.15% concentration of mercuric chloride ( HgCl 2 ) seed surface disinfection for 15-20 minutes; wash the seeds 6-8 times with sterilized water; place 8-10 seeds evenly on the indica rice induction medium; place the inoculated medium in a dark place for 5 weeks at a temperature of 25 ±1°C.

[0144] (2) Subculture: select pale yellow, compact, relatively dry, and vigorous embryogenic calli, and place them on an indica subculture medium for 20 days in the dark at a temperature of 25±1°C.

[0145] (3) Pre-cultivation: select compact and relatively dry embryogenic calli, put them on an indica rice pre-cultivation medium and culture them in the dark for 4-5 days at a temperature of 25±1°C.

[0146] (4) Agrobacteri...

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Abstract

The invention discloses a rice gene OsPGSIP1. The rice gene OsPGSIP1 has a nucleotide sequence represented by SEQ ID NO.1, or has a coding sequence represented by SEQ ID NO.2; or the rice gene OsPGSIP1 has an amino acid sequence represented by SEQ ID NO.4. The deficiencies in the prior art are overcome, the rice OsPGSIP1 gene is overexpressed in rice Zhenshan 97B, the OsPGSIP1expression level of atransgenic plant increase, and the thousand kernel weight, the seedling stage biomass, the cellulose content and the hemicellulose content increase; when the expression of the OsPGSIP1 gene in the Zhenshan 97B is suppressed, the expression level of OsPGSIP1 in the transgenic plant decrease, and the chlorophyll content and the nitrogen content in leaves increase; and introduction of alleles of OsPGSIP1 derived from wild rice ACC10 into the Zhenshan 97B can significantly increase the seedling stage biomass, the thousand kernel weight, the cellulose content, the hemicellulose content and the stem mechanical strength of the Zhenshan 97B.

Description

technical field [0001] The invention belongs to the field of genetic engineering, and in particular relates to a rice gene OsPGSIP1 and its application. Background technique [0002] Rice is one of the most important food crops. In recent years, the improvement of the yield potential of hybrid and conventional rice has been extremely limited, and rice breeding has entered a so-called "platform" stage of slow progress. The level of rice yield depends on the size of the sink capacity (the number of spikelets per unit area × grain weight), the strength of the source (leaf area × net assimilation rate) and the flow (transportation of photosynthetic products to the sink). Therefore, improving photosynthetic efficiency, breaking the "platform" of yield, and further increasing rice yield per unit area are still the basic requirements for food production safety. [0003] Chlorophyll is an important pigment and material carrier for plants to participate in photosynthesis. It captur...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C12N15/82A01H5/00A01H6/46
CPCC07K14/415C12N15/8243C12N15/825C12N15/8261
Inventor 余四斌高大伟孙文强王电文
Owner HUAZHONG AGRI UNIV
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