Transcription factor gene OsHOX4 for controlling synthesis of rice gibberellin and use thereof

A transcription factor, gibberellin technology, applied in the genetic engineering of rice, new plant field, can solve the problem that the function needs to be further studied

Inactive Publication Date: 2007-05-23
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the classification of plant species, Arabidopsis is a dicotyledonous plant and maize is a monocotyledonous plant. Therefore, whether this gene can transform maize and regulate the synthesis of gibberellin in maize plants remains to be further studied.

Method used

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  • Transcription factor gene OsHOX4 for controlling synthesis of rice gibberellin and use thereof
  • Transcription factor gene OsHOX4 for controlling synthesis of rice gibberellin and use thereof
  • Transcription factor gene OsHOX4 for controlling synthesis of rice gibberellin and use thereof

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Experimental program
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Effect test

Embodiment 1

[0032] The cloning of embodiment 1 candidate gene OsHOX4:

[0033] Using Arabidopsis AtHB6 (Homeobox (HOX) class transcription factor, NCBI protein accession number AAD41726; in the REDB website ( http: / / redb.ncpgr.cn / ), select the Regularblast tool in the "RiceEST" database for tblastx analysis, and find the homologous clone in rice (numbered as EI#77-F09), from the cDNA library (see references: Chu Zhaohui, Peng Kaiman, etc., the whole growth period of rice is uniform The construction and identification of the cDNA library, Science Bulletin, 2002, 47 (21).1656-1662) obtained this clone, sequenced and obtained the full-length cDNA sequence (sequence determination was completed by the Shanghai National Gene Sequencing Center), and the candidate gene was named as OsHOX4, its cDNA nucleotide sequence is shown in SEQ NO: 1 in the sequence table. Use GenDoc software (version is selected from: GenDoc3.2) to the protein sequence analysis of the gene OsHOX4 of reported AtHB6 and th...

Embodiment 2

[0034] The construction of embodiment 2 binary Ti plasmid vector and the establishment of transforming Agrobacterium:

[0035] Specific steps are as follows:

[0036] 1) Cloning the cDNA with OsHOX4, loading the plasmid as pSPORT1 (purchased from Promega, a special plasmid vector for constructing cDNA library) digested with KpnI and BamHI, and then isolating the target gene fragment (plus the enzyme derived from pSPORT1 Cut the linker fragment, the size of the cloned OsHOX4 gene is 1560bp), and directly connect with the vector plasmid pDS1301 digested with KpnI and BamHI (the endonucleases used are all purchased from TAKARA company, and the specific usage and dosage refer to the product manual; The ligase was purchased from Invitrogen, and the usage and dosage refer to the instructions of the product);

[0037] 2) The connection product is introduced into DH10B (purchased from Promega Company) by electrotransformation (the electrotransformer is a product of eppendorf, and the...

Embodiment 3

[0040] The transformation of embodiment 3 binary Ti plasmid vectors and the positive detection of transgenic plants:

[0041] Specific steps are as follows:

[0042] 1) The transformed strain T-HOX4 obtained in Example 2 was introduced into the recipient rice variety "Nipponbare", and the transformation method was referred to the method reported by Hiei et al. (see: Efficient transformation of rice, Oryza sativa L., mediated by Agrobacterium and sequence analysis of the boundaries of the T-DNA, 1994, Plant Journal 6:271-282). The obtained T0 transgenic rice plants were named HOX4-n, wherein n=1, 2, 3...represent different transgenic families.

[0043] Total DNA was extracted from leaves of transformed rice plants of the T0 generation. The DNA extraction method is the CTAB method (with reference to Zhang et al., the method and steps shown in genetic diversity and differentiation of indica an japonica rice detected by RFLP analysis, 1992, Theor Appl Genet, 83, 495-499), and th...

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Abstract

The invention belongs to the field of plant gene engineering, and specifically involves the separation and clone, function identification and application of DNA fragment (gene) for regulating rice gibberellin synthesis. The said gene OsHOX4 relates to the rice gibberellin synthesis. The invention includes: combining the complete coding sequence of the said gene with Cauliflower mosaic virus promoter CaMV35S, transferring into rice directly, the transgenic plants are obviously shorter than wild control plants, the transgenic plants can recover to normal height under the effect of exogenous gibberellin (GA3), it is found by further analysis that the expression of gibberellin oxidase (OsGA3ox2) in transgenic plants is inhibited, showing that OsHOX4 can regulate the expression of OsGA3ox2 and control the synthesis of gibberellin.

Description

technical field [0001] The invention belongs to the technical field of new plants, in particular to the technical field of genetic engineering of rice. A transcription factor gene OsHOX4 that regulates gibberellin synthesis in rice was isolated, the function of the gene was verified and its application in transgenic rice. Background technique [0002] Gibberellin (GA) belongs to diterpenoids, and more than 70 species have been isolated from higher plants and microorganisms. All gibberellins have a common basic skeleton in chemical structure, that is, gibberellane, and all contain carboxyl groups, so they are acidic. The structural differences between various gibberellins mainly lie in the number and position of double bonds and hydroxyl groups. [0003] In higher plants, almost all tissues and organs contain gibberellin, but the content is generally very small, and the content is often higher in vigorously growing parts. The main parts of gibb...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C12N15/82
Inventor 代明球周道绣张启发
Owner HUAZHONG AGRI UNIV
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