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Zinc finger protein gene OsRLZP for regulation and control of paddy rice leaf shape and use thereof

A zinc finger protein and rice technology, which is applied in application, genetic engineering, plant genetic improvement, etc., can solve the problems that the research on the relationship between genes is not deep enough, and the molecular regulatory network cannot be completely and clearly outlined, so as to achieve easy access and safety high performance effect

Inactive Publication Date: 2014-08-20
HEFEI UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But so far, there are not many genes that regulate leaf shape and the research on the relationship between genes is not deep enough, and the molecular regulatory network of rice leaf shape establishment and development cannot be fully and clearly outlined (Xu Jing et al., Acta Crops, 2013 )

Method used

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  • Zinc finger protein gene OsRLZP for regulation and control of paddy rice leaf shape and use thereof
  • Zinc finger protein gene OsRLZP for regulation and control of paddy rice leaf shape and use thereof
  • Zinc finger protein gene OsRLZP for regulation and control of paddy rice leaf shape and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Embodiment 1: Cloning of rice OsRLZP gene

[0022] 1. Reagents

[0023] RNA extraction reagent Trizol was purchased from Invitrogen; reverse transcriptase ReverTraAce was purchased from Toyobo; high-fidelity DNA polymerase PrimeStar was purchased from TaKaRa; cloning vector pEASY TM -Blunt Simple Cloning Vector was purchased from Beijing Quanshijin Biotechnology Co., Ltd.; the primers were synthesized by Shanghai Yingjun Biotechnology Co., Ltd., and the rest of the reagents were imported or domestic analytically pure products.

[0024] 2. E. coli strains and plant material

[0025] Escherichia coli (Escherichia coli) strain DH5α was purchased from Beijing Quanshijin Biotechnology Co., Ltd.; rice japonica variety Nipponbare (Oryza sative L.ssp.japonica cv.Nipponbare) seeds were provided by Sichuan Academy of Agricultural Sciences.

[0026] 3. Medium and solution

[0027] LB medium: tryptone 10g / L, yeast powder 5g / L, NaCl 10g / L. Adjust the pH to 7.0 with NaOH and auto...

Embodiment 2

[0071] Embodiment 2: Construction of rice OsRLZP gene overexpression recombinant plasmid

[0072] 1. Reagents

[0073] The plasmid extraction kit EasyPure Plasmid MiniPrep Kit was purchased from Beijing Quanshijin Biotechnology Co., Ltd.; the agarose gel recovery kit EasyPure Quick Gel Extraction Kit was purchased from Beijing Quanshijin Biotechnology Co., Ltd.; restriction enzymes Hind III, Sac I. T4 ligase was purchased from TaKaRa Company.

[0074] Other imported sub-packages and conventional reagents are the same as in Example 1.

[0075] 2. Agrobacterium strains and plant expression vectors

[0076] The Agrobacterium tumefaciens strain EHA105 used for transgenesis was purchased from Clontech; the eukaryotic expression vector pHB was constructed by the laboratory of Professor Hongquan Yang of Shanghai Jiaotong University (Mao et al., Proc Natl Acad Sci USA, 2005).

[0077] 3. Medium

[0078] YEB medium: yeast extract 1g / L, beef extract 5g / L, egg white 5g / L, sucrose 5g / ...

Embodiment 3

[0122] Embodiment 3: Obtaining of rice OsRLZP overexpression transgenic plants

[0123] 1. Reagents

[0124] RNA extraction reagent Trizol was purchased from Invitrogen; reverse transcriptase ReverTraAce was purchased from Toyobo; real-time quantitative PCR reagent TransStart TM Green qPCR SuperMix was purchased from Beijing Quanshijin Biotechnology Co., Ltd. Other reagents are imported sub-packages or domestic analytically pure products.

[0125] 2. Medium

[0126] Plant tissue culture medium:

[0127]

[0128]

[0129] (1) Induction medium: NB+2mg / L2, 4-D, pH 5.8-5.9; (2) Co-cultivation medium: NB+2mg / L2, 4-D+100μmol / L acetosyringone, pH 5.2; (3) Screening medium: NB+2mg / L2, 4-D+250mg / L carbenicillin+30~50mg / L hygromycin, pH 5.8~5.9; (4) Differentiation medium: NB+10mg / L KT+0.4mg / LNAA+250mg / L carbenicillin, pH 5.8~5.9; (5) Rooting medium: 1 / 2MS, pH 5.8~5.9.

[0130] 3. Method

[0131] 3.1 Agrobacterium-mediated transformation of rice

[0132] 3.1.1 Induction a...

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Abstract

The invention relates to a zinc finger protein gene OsRLZP for regulation and control of a paddy rice leaf shape and a use thereof. The zinc finger protein gene OsRLZP has a nucleotide sequence shown in the formula of SEQ ID NO: 1 in the sequence table. A protein coded by the zinc finger protein gene OsRLZP has an amino acid sequence shown in the formula of SEQ ID NO: 2 in the sequence table. The gene full-length coding sequence shown in the formula of SEQ ID NO: 1 is inserted into an eukaryotic vector so that an eukaryotic recombinant plasmid excessively expressing the gene OsRLZP is obtained, and the eukaryotic recombinant plasmid is transformed into paddy rice so that an OsRLZP-overexpression transgenic plant is obtained. An experiment proves that the gene shown in the formula of SEQ ID NO: 1 can control paddy rice leaf development. Compared with a wild type plant, the OsRLZP up-regulation transgenic plant leaf has thick midrib, narrow leaf, involute and vertical leaves of the whole plant and less leaf surface hair. Therefore, the zinc finger protein gene OsRLZP has regulation and control effects on paddy rice leaf development and can be used in paddy rice plant-improved genetic breeding.

Description

technical field [0001] The invention belongs to the field of plant genetic engineering, and in particular relates to a rice leaf shape control gene and its application in genetic breeding for plant type improvement of crops. Background technique [0002] Rice leaves, as the main organ for photosynthesis, are the decisive factors for the formation of rice yield. A larger leaf area can form a larger photosynthetic group, but when the leaves are too long or too wide, they are prone to drooping, which will reduce the total light-receiving area of ​​the group and affect the ventilation of the planting group, making it easy for pathogenic microorganisms to breed and multiply. Therefore, the improvement of rice leaf morphology has always been an important goal of plant type improvement. [0003] The main indicators of rice leaf morphology include: leaf curl, inclination, degree of divergence, and leaf width. The research progress of molecular genetics shows that rice leaf curling...

Claims

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Application Information

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IPC IPC(8): C12N15/29C12N15/82C07K14/415A01H5/00
Inventor 牛向丽吴海云毛云
Owner HEFEI UNIV OF TECH
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