Transcription factor derived from corn and new use of coding gene thereof

A transgenic plant, gene technology, applied in genetic engineering, plant genetic improvement, application and other directions, can solve problems such as low nitrogen utilization rate

Inactive Publication Date: 2014-06-04
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Planting nitrogen-efficient corn varieties is an effective way to solve the low utilization rate of nitrogen fertilizers, reduce production costs, and reduce environmental pollution. It is also a basic requirement for sustainable agricultural development and enhancing the international competitiveness of products. The process of genetic engineering, especially the maturity of corn transgenic technology, makes it possible for us to quickly obtain nitrogen-efficient varieties through genetic engineering

Method used

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  • Transcription factor derived from corn and new use of coding gene thereof
  • Transcription factor derived from corn and new use of coding gene thereof
  • Transcription factor derived from corn and new use of coding gene thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] Example 1. Real-time quantitative PCR (Real-Time PCR) analysis of the expression characteristics of maize ZmNLP1;1 gene tissues

[0068] The maize inbred line B73 (Germplasm Resource Center, Crop Institute, Chinese Academy of Agricultural Sciences) was selected. Extract seedling corn roots, seedling corn shoots, new leaves before pollination, old leaves before pollination, panicle leaves before pollination, ear ears before pollination, tassels before pollination, new leaves 15 days after pollination, and 15 days after pollination Total RNA was extracted from old leaves, 15-day post-pollination panicle leaves, 15-day post-pollination cobs, and 15-day-pollination grain samples. The obtained total RNA was decontaminated by DNase I enzyme (TaKaRa Company) to remove the DNA in the total RNA, and M-MLV reverse transcriptase (Promega Company) and oligo d(T) 18 Reverse transcribe into cDNA first strand. Using the Real-time PCR method, the maize ZmNLP1;1 gene was amplified wit...

Embodiment 2

[0091] Embodiment 2, cultivating trans ZmNLP1; 1 gene Arabidopsis

[0092] 1. Amplification of full-length ZmNLP1;1 gene cDNA

[0093] Design a pair of primers P1-F (5'-ATGGAGCAGGCGCCGCAGAC-3') and P1-R (5'-TTACTGCATACAAACCAATCC-3'), where P1-F just contains the translation start codon ATG, and P1-R contains the translation stop codon The sub-TAA; using the cDNA library of reverse transcription of the total RNA of the maize inbred line B73 as a template, the high-fidelity Primer STAR enzyme was used to amplify the ZmNLP1;1 gene fragment containing the complete open reading frame (its nucleotide sequence is sequence 1 from the 355th to the 3216th nucleotide of the 5' end). Wherein, the reaction system is as follows:

[0094]

[0095] PCR reaction conditions: 98°C pre-denaturation for 2 minutes; then 98°C for 10 seconds, 65°C for 10 seconds, 72°C for 3 minutes and 30 seconds, 30 cycles; then add 1 μl Taq enzyme, 72°C for extension and tailing for 20 minutes.

[0096]Take 8...

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Abstract

The invention discloses a transcription factor derived from corn and new use of a coding gene thereof. The new use provided by the invention is a method using a coding nucleic acid molecule of ZmNLP1;1 to cultivate a transgenic plant. The method is a method for cultivating the transgenic plant with at least one characters of the following 1)-4): 1) aboveground biomass is higher than that of a receptor plant; 2) after induction of a nitrate, the nitrate transport protein gene expression is higher than that of the receptor plant; 3) after induction of the nitrate, nitrate reductase gene expression is higher than that of the receptor plant; and 4) after induction of the nitrate, nitrite reductase gene expression is higher than that of the receptor plant; and the method comprises a step for introducing a ZmNLP1;1 gene into the receptor plant; and the ZmNLP1;1 gene codes a protein shown in SEQ ID No.2. Experiments show that, in the condition of using nitric nitrogen (NO<3->) as a sole nitrogen source, the aboveground biomass and the total biomass of arabidopsis transgenic with the ZmNLP1;1 gene are increased significantly compared with that of receptor arabidopsis.

Description

technical field [0001] The present invention relates to a new application of a corn-derived protein transcription factor and its encoding gene, in particular to a new application of a corn-derived transcription factor related to nitrate absorption and utilization and its encoding gene. Background technique [0002] Nitrogen is one of the essential mineral nutrients for plants. It is an essential component for the synthesis of proteins, nucleic acids and many biologically active substances. It is closely related to crop yield and quality. The use of nitrogen fertilizers in agricultural production has greatly increased crop yields. Currently, about 80 to 90 million tons of nitrogen fertilizer are applied globally every year, and it is predicted that it will increase to 240 million tons by 2050 (Tilman et al., 1999, Proc Natl Acad Sci USA 96:5995-6000). The increase in energy costs has led to a continuous increase in the price of nitrogen fertilizers, thereby increasing the co...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82C12N15/29C12N1/15C12N1/19C12N1/21C07K14/415A01H5/00
Inventor 袁力行王章奎张蕾顾日良米国华张福锁
Owner CHINA AGRI UNIV
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