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Use of histone acetylation enzyme gene OsELP3 in rice anthesis regulation

An acetylase, flowering stage technology, applied in application, genetic engineering, plant genetic improvement and other directions, can solve problems such as unclear

Inactive Publication Date: 2011-08-31
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But how plant flowering is controlled at the level of chromatin modification is still not well understood

Method used

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  • Use of histone acetylation enzyme gene OsELP3 in rice anthesis regulation
  • Use of histone acetylation enzyme gene OsELP3 in rice anthesis regulation
  • Use of histone acetylation enzyme gene OsELP3 in rice anthesis regulation

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1 Cloning and sequence analysis of OsELP3 gene:

[0029] For the genes needed for the invention, by RT-PCR method (see: J. Sambrook, EF Fritsch, T Mani Atis, translated by Huang Peitang, Wang Jiaxi, etc., Molecular Cloning Experiment Guide (Third Edition), Science Press, 2002 Edition) to amplify to obtain its full-length coding sequence, the specific steps are: according to the public database (http: / / www.ncbi.nih.gov / , http: / / cdna01.dna.affrc.go. The full-length cDNA sequence of the rice OsELP3 gene published in jp / cDNA / ) (see Table 1 for details) was used to design primers for PCR amplification. The amplified product was connected to pGEM T-vector (Promega) by T / A cloning for sequencing verification. The primers used to clone the full-length gene are: FLELP-F and FLELP-R, and the specific sequences of the primers are shown in Table 4 (see the end of the instructions)

[0030]The same method was used to obtain RNAi inhibitory fragments. The primers used to c...

Embodiment 2

[0033] The construction of embodiment 2 binary Ti plasmid vector and the establishment of transforming Agrobacterium:

[0034] Specific steps are as follows:

[0035] 1) The TA clone with the full-length cDNA of OsELP3 was digested with KpnI and BamHI, and the target band was recovered, and the expression vector plasmid pU1301 digested with KpnI and BamHI (see attached figure 1 A, refer to Huang et al., Down-regulation of a Silent InformationRegulator2-related gene, OsSRT1, induces DNA fragmentation and cell death in rice. Plant Physiol, 2007, 144: 1508-1519.) connection, that is, to construct excess Expression vector. The T / A clone with the full-length interference fragment of OsELP3 was digested with KpnI and BamHI, the target band was recovered, and the expression vector plasmid pDS1301 digested with KpnI and BamHI (see attached figure 1 B, see: Chuet al., Promoter mutations of an essential gene for pollen development result in disease resistance in rice. Genes Dev, 2006,...

Embodiment 3

[0040] Example 3 Transformation of binary Ti plasmid vector and positive detection of transgenic plants:

[0041] 1) Transform TU-ELP3 and TR-ELP3 into the rice recipient variety "Zhonghua 11" (from the Institute of Crop Science, Chinese Academy of Agricultural Sciences), and the transformation method refers to the method reported by Hiei et al. (Hiei et al, Efficient transformation of rice (Oryza sativa L.) mediated by Agrobacterium and sequence analysis of the boundaries of the T-DNA. Plant J, 1994, 6: 271-282.). The obtained transgenic plants of the T0 generation were named EU-n and ER-n, wherein n=1, 2, 3... represent different transgenic families.

[0042] 2) Total DNA was extracted from the leaves of transformed plants of the T0 generation. The DNA extraction method was the CTAB method (Murray et al, Rapid isolation of high molecular weight plant DNA. Nucleic Acids Res, 1980, 8: 4321-4325.). Then use common PCR method to carry out positive detection on T0 generation tra...

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Abstract

The invention belongs to the technical field of plant genetic engineering, and particularly relates to a rice histone acetyltransferase gene, encoding protein functions and applications thereof. The invention discloses the rice histone acetyltransferase gene OsELP3, which is related to plant florescence. Under long-day sunlight, overexpression of the gene can bring rice blooming forward, and inhibition of the gene expression can delay rice blooming. After being processed under short-day sunlight, mutant phenotype of an inhibiting material is recovered to be wild. Analysis discovers that the OsELP3 gene can influence the expression levels of three blooming genes Hd3a, Hd1 and OsGI, and take rhythmic variation of self expression under short-day sunlight. The Western Blot experiment analysisshows that the gene has the histone acetylation function and the functionary locus on H3K9.

Description

technical field [0001] The invention belongs to the technical field of plant genetic engineering. It specifically relates to the isolation, cloning, functional verification and application of a histone acetylase gene OsELP3 (Elongation Protein 3) that regulates the flowering period of rice. Said genes are related to the flowering period of plants. Background technique [0002] The complete life cycle of higher plants from seed germination to new seed production can be divided into two stages: vegetative growth and reproductive growth. The process of transition from vegetative growth to reproductive growth is called floral induction. This process is affected by both the plant's own genetic factors and the external environment. The floral induction process of different higher plants has certain similarities and regularities: after a certain period of vegetative growth, the plants reach the mature state of flowers, and the internal flower-forming substances will feel the infl...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/54A01H1/00A01H5/00C12Q1/68G01N33/53
Inventor 周道绣李晨
Owner HUAZHONG AGRI UNIV
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