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Chorispora bungeana ferric reductase gene CbFRO and amino acid sequence of encoding product of Chorispora bungeana ferric reductase gene CbFRO

A technology of alpine ion mustard and ferric iron, applied in the field of plant genetic engineering, can solve the problems of iron deficiency and chlorosis of plants, loss of agricultural production and the like

Inactive Publication Date: 2013-10-23
LANZHOU UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

But in alkaline soil (pH7.4~8.5), the concentration of soluble iron is only 10 -10 mol / L (Guerinot and Yi 1994), thus plants exhibit iron deficiency chlorosis, causing great losses to agricultural production

Method used

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  • Chorispora bungeana ferric reductase gene CbFRO and amino acid sequence of encoding product of Chorispora bungeana ferric reductase gene CbFRO
  • Chorispora bungeana ferric reductase gene CbFRO and amino acid sequence of encoding product of Chorispora bungeana ferric reductase gene CbFRO
  • Chorispora bungeana ferric reductase gene CbFRO and amino acid sequence of encoding product of Chorispora bungeana ferric reductase gene CbFRO

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Embodiment 1

[0015] Example 1. Cloning method

[0016] The Trizol method was used to obtain high-quality total RNA from Alpine ion mustard that meets the requirements of a reverse transcription template. According to the sequence of plant ferroportin gene published in GeneBank, degenerate primers were designed to amplify the conserved region of CDS (B=T+G+C; Y=T+C; D=A+T+G; N=A+C+G +T; R =A +G), get the target fragment. According to the sequencing results and sequence comparison results of the first cloned fragments, the trivalent iron reductase gene of Arabidopsis thaliana FRO Designed 5`end RACE primer and 3`end RACE primer for template. Amplify with P3 and P4 as primers to obtain an obvious target band. The other end was amplified with P5 and P6 primers to obtain the target band. The sequencing results of the three PCR products are spliced ​​together to obtain a complete CDS sequence. Ferric iron reductase gene CbFRO It has the nucleotide sequence shown in SEQ ID NO: 1, the gene has a...

Embodiment 2

[0025] Example 2, CbFRO Identification of genes

[0026] The invention induces different Fe concentration CbFRO The genes were identified. Treat Alpine ion mustard under normal (100μM), iron deficiency (0μM), and iron-rich (500μM) environments, and determine at 12 h, 24 h, and 36 h CbFRO The amount of expression. The results show that( figure 2 ), under the iron deficiency treatment, there will be a significant increase. After 12 h of treatment, the expression level even increased to 33 times that of the control; the expression level of the iron-rich treatment also increased to 4.7 times that of the control; with the extension of time, two The expression of CbFRO decreased significantly under the concentration treatment. When the treatment reached 36 hours, the expression level suddenly increased, and the iron deficiency treatment was significantly higher than the iron-rich treatment. The experimental results show that the alpine ion mustard CbFRO Genes are highly expressed in ...

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Abstract

The invention provides a Chorispora bungeana ferric reductase gene CbFRO which has a nucleotide sequence shown as SEQ ID NO:1. The invention also provides an encoding product CbFRO of the Chorispora bungeana ferric reductase gene CbFRO, wherein the encoding product CbFRO has an amino acid sequence shown as SEQ ID NO:2.

Description

Technical field [0001] The present invention relates to the field of plant genetic engineering, in particular to a ferric reductase gene of alpine ion mustard CbFRO And the amino acid sequence of the encoded product. Background technique [0002] Alpine ion mustard (Chorispora bungeana) is a perennial plant of the cruciferous ion mustard genus, distributed in high-altitude subalpine meadows and gravel hillsides. The environmental conditions are characterized by cold climate, high salinity, thin air, strong radiation, and strong wind. Alpine ion mustard is mainly distributed in Liushidu in the headwater area of ​​the Urumqi River in my country, with an altitude of 3600-3900m. The annual average temperature is between -5°C and -7°C, and the average temperature from June to August is 3.6°C. The daily temperature difference is relatively large. It is a freezing environment accompanied by repeated freezing and thawing. And on a windward slope, the water retention of gravel in the fl...

Claims

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Application Information

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IPC IPC(8): C12N15/53C12N9/02
Inventor 安黎哲张丽静徐世健宋渊张华向云盛红梅陈书燕何文亮李田
Owner LANZHOU UNIVERSITY
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