Rice phosphorus absorption and transfer regulator gene OsPHF1 and application thereof

A technology for regulating genes and phosphorus absorption, which is used in applications, genetic engineering, plant genetic improvement, etc.

Active Publication Date: 2011-04-13
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0008] There are no relevant reports on the related gen...

Method used

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  • Rice phosphorus absorption and transfer regulator gene OsPHF1 and application thereof
  • Rice phosphorus absorption and transfer regulator gene OsPHF1 and application thereof
  • Rice phosphorus absorption and transfer regulator gene OsPHF1 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Embodiment 1, screening and phenotype of mutants

[0054] A large number of single-copy homozygous strains of OsPHR2 overexpression material (Oryza sativa Nipponbare, which can be purchased from Zhejiang University) were propagated, soaked in EMS (ethyl methanesulfonate) with a concentration of 1% (volume concentration) at room temperature for 12 hours Afterwards, a large number of species were propagated in Hainan. The seeds (M2) after two generations of mutant selfing were built into a mutant library and used as mutant screening objects. m 2 The seeds were rinsed with distilled water and washed with 0.6% (volume concentration) dilute HNO 3 Dormancy-breaking treatment for 16 hours, germination in the dark at 37°C until white. Sow the dew-white seeds on the nylon gauze floating on the rice culture solution with a sodium arsenate concentration of 50 μM (the formula of the culture solution is the standard formula of the International Rice Institute), at a temperature o...

Embodiment 2

[0056] Embodiment 2, gene localization

[0057] f 2 The mapping population was obtained by crossing the homozygous (Osphf1-1) with the indica rice Kasalath, and a total of 28 F 2 Individual, available phosphorus (phosphate) was measured according to a known method (Nanamori et al., 2004). Genomic DNA for gene localization was extracted from rice leaves by a rapid extraction method of rice trace DNA. Take about 0.1 g of young rice leaves, freeze them with liquid nitrogen, grind the leaves into powder in a 1.5 ml centrifuge tube, extract the total DNA, and dissolve the obtained DNA in 200 μl sterile water. 2 μl of DNA sample was used for each SSR and STS reaction.

[0058] In the preliminary mapping experiment of OsPHF1 gene, 28 F 2 Individuals were subjected to SSR analysis. According to the published molecular genetic maps created by japonica and indica rice, select SSR primers that are approximately evenly distributed on each chromosome, and SSR markers are derived from ...

Embodiment 3

[0100] Example 3, gene prediction and sequence analysis

[0101] According to the results of fine mapping, the OsPHF1 gene is located within 70kb of the STS07g479 marker at the junction of BAC clone P0506C07 and OSJNBb0084L07 ( figure 2 A). According to the rice gene annotation information of TIGR (http: / / www.tigr.org / tdb / e2k1 / osal / ) and the EST database (http: / / www.ncbi.nlm.nih.gov / ), the mapped chromosomal regions In-segment gene prediction analysis, there is a gene near this marker, and the protein encoded by it with WD40 domain is homologous to AtPHF1 in Arabidopsis, so this gene is named OsPHF1, and the cDNA sequence of OsPHF1 gene is SEQ ID NO: the cDNA sequence shown in 1. The amino acid sequence encoded by the cDNA sequence of the OsPHF1 gene is the amino acid sequence shown in SEQ ID NO:2. According to the rice genome sequence information submitted by NCBI (http: / / www.ncbi.nlm.nih.gov / ), the upstream sequence of the OsPHF1 gene on the chromosome is the promoter se...

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Abstract

The invention discloses a protein coded by a rice phosphorus absorption and transfer regulator gene OsPHF1, which has an amino acid sequence shown in SEQ ID NO:2. The invention also discloses the gene OsPHF1 coding the protein. The gene comprises a cDNA sequence and a promoter sequence, wherein the cDNA sequence is a nucleotide sequence shown in SEQ ID NO:1; and the promoter sequence is a nucleotide sequence shown in SEQ ID NO:3. The invention also discloses application of the gene OsPHF1, namely the gene OsPHF1 is used for constructing transgenic rice. Through identification of transgenic experiments and plant tissue effective phosphorus concentration determination experiments, the gene regulates phosphorus absorption and transfer of the rice.

Description

technical field [0001] The invention belongs to the field of plant genetic engineering. Specifically, the present invention relates to a cDNA sequence of the rice OsPHF1 gene cloned by map-based cloning technology. The protein encoded by the cDNA belongs to a chaperone protein similar in structure to Sec12, and is identified through transgenic experiments and plant tissue available phosphorus concentration determination experiments. , which regulates the uptake and transport of phosphorus in rice. Background technique [0002] Phosphorus is an important component of nucleic acids, phospholipids and ATP in plants. As a material for energy transfer in plants, phosphorus can activate proteins in the body and regulate the entire metabolic process of plants (Marschner, 1995). Among the various inorganic nutrients needed by plants, phosphorus is one of the most important elements, but it is extremely difficult to obtain from the soil. Although phosphorus is abundant in ecosystem...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N5/10A01H5/00
Inventor 吴平陈婕妤王以锋张帆刘于张为民陈明秀吴忠长
Owner ZHEJIANG UNIV
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