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Application of Miscanthus WRKY transcription factor in increase of biomass of plant fiber

A transcription factor, Miscanthus technology, applied in the field of plant genetic engineering, can solve the problem of ignorance of genetic and molecular mechanisms

Inactive Publication Date: 2015-10-14
QINGDAO INST OF BIOENERGY & BIOPROCESS TECH CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, the research on Miscanthus WRKY transcription factors is still blank, especially the genetic and molecular mechanisms of its regulation are still almost unknown

Method used

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  • Application of Miscanthus WRKY transcription factor in increase of biomass of plant fiber
  • Application of Miscanthus WRKY transcription factor in increase of biomass of plant fiber
  • Application of Miscanthus WRKY transcription factor in increase of biomass of plant fiber

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1 Miscanthus WRKY family genes MlWRKY12 obtained.

[0029] The clones and isolation of genes were obtained from the stems of miscanthus that had been cultivated for about 4 weeks under the conditions of 14h L / 10h D light, temperature 25°C, and relative humidity 60%. The total RNA was extracted by Trizol method, and isolated from Miscanthus using RT-PCR technology MlWRKY12 The cNDA of the gene, its coded amino acid sequence is shown in SEQ ID No.2.

[0030] The amplified miscanthus designed in this embodiment MlWRKY12 The specific primers for the core fragment of the gene are:

[0031] Forward primer: 5'-ATGGAGGGAGGCCAGTTGAG-3' (SEQ ID No.3)

[0032] Reverse primer: 5'-TCAGAAGGAGCTGAAGCAATC-3' (SEQ ID No. 4).

[0033] The PCR product was ligated with the pMD18-T vector, transformed into Escherichia coli DH5α competent cells and sequenced.

Embodiment 2

[0034] Example 2 MlWRKY12 Gene sequence analysis.

[0035] MlWRKY12 The gene CDS sequence contains 702 nucleotide sequences and 233 amino acid sequences, and its molecular weight is predicted to be 25.7kDa, which was analyzed by DNAMAN software MlWRKY12 Homology to other Arabidopsis WRKY family proteins. The results show that MlWRKY12 Zinc finger structure modification with typical conserved C2H2 ( figure 1 ). Built using MEGA software MlWRKY12 Phylogenetic tree with other WRKY family proteins, the results show that MlWRKY12 Belongs to WRKY-IIc subfamily, and sweet sorghum Sb04g033240, Brachypodium tachypodium BdWRKY14 , rice OsWRKY36 , Arabidopsis AtWRKY12 closest relatives. figure 2 exhibit MlWRKY12 Phylogenetic relationships of proteins.

Embodiment 3

[0036] Example 3 MlWRKY12 Subcellular localization of genes.

[0037] will remove the stop codon MlWRKY12 The open reading frame (ORF) of the gene was connected with the pB7FWG2 vector, and the vector fused with GFP was constructed, and Arabidopsis thaliana was transgenic, and the analysis of the transgenic plants of the T2 generation showed that the gene was mainly expressed in the nucleus, as shown in image 3 shown.

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Abstract

The invention belongs to the technical field of plant genetic engineering and particularly relates to an MIWRKY12 gene obtained from Miscanthus by separating and cloning and relevant application of the MIWRKY12 gene in parenchymal cell incrassation and blooming of plants. A nucleotide sequence of the MIWRKY12 gene is represented by SEQ ID No.1, and an amino acid sequence of the MIWRKY12 gene is represented by SEQ ID No.2. According to the application, a feasible method is provided for increasing the biomass of the plants and regulating and controlling the flowering phase of the plants.

Description

technical field [0001] The invention relates to Miscanthus WRKY transcription factors MlWRKY12 The coding gene and application thereof belong to the technical field of plant genetic engineering. Background technique [0002] Miscanthus (Miscanthus) is a kind of perennial C4 plant with rhizomes. It is mostly distributed in the tropics to the southeast of Asia. It is a fiber energy plant with great development potential. The high-yield fibrous perennial energy plant widely distributed in the Yangtze River Basin has important uses, and its genome sequence has a high sequence similarity with sweet sorghum. [0003] The secondary cell walls of plants are an important source of lignocellulosic biomass, which also provide fabric, wood and potential second-generation bioenergy for human survival. NAC and MYB transcription factors involved in cell wall (i.e., cellulose, hemicellulose, and lignin) synthesis are well-studied in different species, and the recently discovered Arabidops...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C12N15/82C07K14/415A01H5/00
Inventor 周功克贺郭于延冲胡瑞波王华美曹英萍付春祥
Owner QINGDAO INST OF BIOENERGY & BIOPROCESS TECH CHINESE ACADEMY OF SCI
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