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Application of the transcription factor hinge1 in the regulation of nitrogen-phosphorus homeostasis in plants

A technology for phosphorus absorption and nucleic acid molecules, applied in angiosperm/flowering plants, applications, plant peptides, etc., can solve problems such as incomplete understanding of signaling pathways

Active Publication Date: 2022-03-22
INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It has been found that an increase in nitrogen can promote phosphorus uptake, but the signaling pathway is not fully understood

Method used

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  • Application of the transcription factor hinge1 in the regulation of nitrogen-phosphorus homeostasis in plants
  • Application of the transcription factor hinge1 in the regulation of nitrogen-phosphorus homeostasis in plants
  • Application of the transcription factor hinge1 in the regulation of nitrogen-phosphorus homeostasis in plants

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0078] Example 1 Analysis and cloning of HINGE1 gene

[0079] 1. Analysis of HINGE1 gene

[0080] The inventors first performed transcriptome analysis (RNA-seq) on the seedlings of rice variety ZH11 supplied with different nitrogen sources, and the results are shown in figure 1 B (pure ammonium treatment labeled NH 4 Cl, pure nitrate treatment is marked as KNO 3 , ammonium nitrate 1:1 (calculated as nitrogen, mass ratio) mixed treatment marked as NH 4 NO 3 ); The inventors performed transcriptome analysis (RNA-seq) on rice seedlings induced by short-term nitrate, and the results are shown in figure 1 A (short-term nitrate-inducing treatment labeled KNO 3 , KCl as the control).

[0081] According to the above results, by screening nitrate-induced transcription factors, the inventors obtained a gene (LOC_Os04g56990) that was induced the strongest by nitrate in roots, which encoded the GARP class MYB family transcription factors, and named it HINGE1 (Highly Induced by Nitr...

Embodiment 2

[0110] Embodiment 2, expression analysis of HINGE1 gene

[0111] 1. Expression analysis of rice HINGE1 supplied with different nitrogen sources

[0112] The inventors used qRT-PCR (primers are qHINGE1-1F and qHINGE1-1R) to analyze the expression of HINGE1 in rice ZH11 seedlings supplied with different nitrogen sources, and the results are shown in figure 1 At the same time, the expression of HINGE1 in rice seedlings induced by different concentrations (0mM, 0.2mM, 5mM) of nitrate in a short period (0.5, 1, 2, 4, 8, 16, 24 hours) was analyzed, and the results are shown in figure 1 D. The above results all proved that the expression of HINGE1 was induced by nitrate.

[0113] 2. Analysis of HINGE1 expression in different rice tissues

[0114] 2.1qRT-PCR analysis

[0115] The expression of HINGE1 in different tissues of rice was further analyzed by qRT-PCR, the primers used were qHINGE1-1F and qHINGE1-1R, and it was found that the expression of HINGE1 was the highest in roots ...

Embodiment 3

[0127] Example 3, HINGE1 transcription factor characteristic analysis

[0128] 1. Subcellular localization analysis

[0129] Transcription factors generally perform their transcriptional activation or repression functions in the nucleus, and the inventors analyzed the subcellular localization of HINGE1. Therefore, the inventor constructed a plasmid in which eGFP (enchanced GFP) was fused to the C-terminus of the HINGE1 protein driven by the 35S promoter, and transformed rice protoplasts. The specific steps were as follows:

[0130] 1.1 Construction of HINGE1-eGFP vector

[0131] Primers containing In-Fusion adapters were designed for the pCAMBIA2300-35S-eGFP(C) vector, and the primer sequences are as follows:

[0132] HINGE1-GFP-F:5' (The sequence indicated by the underline is the KpnI enzyme recognition site sequence, and the sequence indicated by the bold font is the In-Fusion linker sequence);

[0133] HINGE1-GFP-R:5' (The sequence indicated by the double underline i...

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Abstract

The invention discloses a transcription factor HINGE1. External nitrate induces the expression of HINGE1, and the abundantly expressed HINGE1 can activate downstream phosphorus starvation-induced genes, increase phosphorus uptake, and maintain nitrogen-phosphorus homeostasis in rice. Compared with the recipient rice, the transgenic rice expressing the HINGE1 gene has a significantly enhanced phosphorus absorption capacity, indicating that the HINGE1 gene is a gene related to the regulation of plant nitrogen-phosphorus homeostasis, and the HINGE1 gene can be used to coordinate plant nitrogen-phosphorus uptake.

Description

technical field [0001] The invention relates to the application of a transcription factor HINGE1 in regulating plant nitrogen-phosphorus homeostasis in the field of biotechnology. Background technique [0002] Nitrogen and phosphorus are two of the most important plant mineral nutrients. The input of chemical fertilizers represented by nitrogen and phosphorus fertilizers is one of the important factors contributing to the "green revolution" of modern agriculture. However, a large amount of chemical fertilizer input also brings many environmental problems such as eutrophication of water bodies. In addition, phosphate rock is a non-renewable resource, and its reserves are also limited. As the global population continues to increase, how to reduce the input of chemical fertilizers while ensuring an increase in food production is an urgent problem to be solved. As an important food crop, rice (Oryzasativa L.) faces a particularly severe problem of "weight loss and efficiency en...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/00A01H6/46
CPCC07K14/415C12N15/8261
Inventor 储成才张志华胡斌李钊王威蒋志敏
Owner INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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