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Corn ZmNST3 gene and application

A gene and corn technology, applied in the field of corn ZmNST3 gene and its application, to achieve the effect of increasing accumulation and facilitating development and utilization

Inactive Publication Date: 2017-06-06
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, there are no reports on the function of other maize secondary wall-associated NAC transcription factors in regulating secondary wall development

Method used

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  • Corn ZmNST3 gene and application
  • Corn ZmNST3 gene and application
  • Corn ZmNST3 gene and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1 Cloning of ZmNST3

[0029] Total RNA was extracted from maize seedlings at the V3 stage, cDNA was synthesized by reverse transcription using the cDNA Synthesis Kit of Promega Company, and amplified with specific primers to obtain the ZmNST3 CDS sequence, with a full length of 1104bp (SEQ ID No.1). The nucleotide sequence of the primer pair is:

[0030] Upstream primer: 5'-TGATGAGCATCTCGGTGAACGG-3',

[0031] Downstream primer: 5'-GACTAGGCGTTGTTCATGGCCAA-3'.

Embodiment 2

[0032] Embodiment 2 is used for the construction of the expression vector expressing ZmNST3 gene in Arabidopsis and Agrobacterium transformation

[0033] The expression cassette is obtained from the Super promoter of Agrobacterium tumefaciens (that is, the mas promoter is fused with 3 ocs enhancers and 1 mas enhancer, see Leisner SM, Gelvin SB (1988) Structure of the octopinesynthse upstream activator SEQuence. Proc Natl Acad Sci USA 85, 2553-2557) and the 3' transcription termination region from the nopaline synthase (nos) gene (Depicker A, Stachel S., Dhaese P, Zambryski P, Goodman HM (1982). Nopaline synthase: transcript mapping and DNASEQuence.J Mol.Appl Genet.1,561-573), the selection marker gene is hygromycin phosphotransferase gene (hpt). Its T-DNA region such as figure 1 shown.

[0034] Using the plasmid containing the ZmNST3 gene as a template, use the PCR method to obtain a ZmNST3 gene fragment with XbaI and SmaI restriction sites at both ends, connect it to the pM...

Embodiment 3

[0035] Example 3 Construction and Genetic Transformation of Expression Vectors for Expressing ZmNST3 in Maize

[0036] The expression cassette is composed of the CaMV35s promoter and the 3' transcription termination region from the nopaline synthase (nos) gene, and the selectable marker gene is the hygromycin phosphotransferase gene (hpt). Its expression box is as figure 2 shown.

[0037] Using the plasmid containing the ZmNST3 gene as a template, use the PCR method to obtain a ZmNST3 gene fragment with SmaI and KpnI restriction sites at both ends, connect it to the pMD19T vector, perform double digestion with SmaI and KpnI after the sequence is correct, and recover about 1100bp fragments. The expression vector pROK219 (containing the CaMV35s promoter) was double digested with SmaI and KpnI, and the large fragment of the vector was recovered. The vector and the target fragment were connected, and the E. coli competent cells were transformed to obtain the recombinant plasmi...

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Abstract

A CDS sequence (SEQ ID No.1) of a ZmNST3 gene related to development of a secondary cell wall of corn is firstly obtained through cloning, and the gene is transformed into the plant, so that accumulation of the secondary cell wall in plant stems can be improved, and the content of cellulose and lignin in the cell wall is improved. The invention provides a novel method for improving accumulation of the secondary cell wall of a plant by using the ZmNST3 gene. The corn ZmNST3 gene can be used for cultivation of a novel high-biomass plant variety, and is beneficial to development and utilization of biomass energy.

Description

technical field [0001] The invention relates to the field of plant genetic engineering, in particular to the corn ZmNST3 gene and its application. Background technique [0002] Energy is an important material basis for national economic and social development. In today's energy system, fossil energy still accounts for nearly 90% of the total, but fossil energy is non-renewable and has limited reserves. Extensive use of fossil energy has caused a series of increasingly serious environmental problems while causing a shortage of energy resources, which has negative impacts on ecosystems and human health. It is imminent to seek and develop new energy sources, especially environmentally friendly renewable energy sources. Biomass energy is a new type of renewable energy, and the development of biomass advanced liquid fuels that can replace internal combustion engine fuels will help alleviate oil shortages. Lignocellulosic biomass is the main raw material for the preparation of ...

Claims

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Application Information

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IPC IPC(8): C12N15/29C12N15/82A01H5/00
CPCC07K14/415C12N15/8205C12N15/8255
Inventor 于静娟肖文翰赵倩朱登云
Owner CHINA AGRI UNIV
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