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Pyrophosphatase of ammopiptanthus mongolicus, and coding gene and application thereof

A technology of pyrophosphatase encoding and pyrophosphatase of holly holly, which is applied in the fields of application, genetic engineering, plant gene improvement, etc., and can solve the problems of staying in the physiological and biochemical stage

Inactive Publication Date: 2011-04-06
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the research on the stress tolerance of Ilex japonica only stays at the physiological and biochemical stage, and it is a blank to explain the stress resistance mechanism of Ilex japonica genetically.

Method used

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  • Pyrophosphatase of ammopiptanthus mongolicus, and coding gene and application thereof
  • Pyrophosphatase of ammopiptanthus mongolicus, and coding gene and application thereof
  • Pyrophosphatase of ammopiptanthus mongolicus, and coding gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Example 1. Acquisition of the pyrophosphatase gene of Ilex japonica.

[0020] 1.1 Salt stress treatment of Ilex japonica

[0021] Sand holly seeds with 0.01% HgCl 2 The surface was sterilized for 15 minutes, rinsed with sterile water 5 times, soaked in sterile water at 37°C overnight, and transferred to 1 / 2 MS + 1.5% sucrose + 1.0% agar + 200mmNaCl salt stress medium. Cultivate for one week at 24°C under 16h L / 8h D light.

[0022] 1.2 RNA extraction

[0023] Take about 0.1 g of the young root material of Ilex japonica treated with salt stress. After the liquid nitrogen is fully ground, transfer to a 1.5ml centrifuge tube, add 1ml (Invitrogen Company), after mixing, place at room temperature for 15 min, add 0.2ml chloroform:isoamyl alcohol (24:1), shake vigorously for 15 s, place at room temperature for 5 min, centrifuge at 13000rpm, 4°C for 15 min. Take the supernatant and add an equal volume of isopropanol, mix carefully, place at room temperature for 15 min, cen...

Embodiment 2

[0052] Example 2: Analysis of the function of the pyrophosphatase gene of Ilex japonicus.

[0053] 2.1 Sequence comparison analysis

[0054] Multiple sequence alignment was performed using Genedoc software, and ORF (Opening Reading Frame) search was performed using molecular analysis software Lasergene6.0. ORF translation was performed using Primer5 software. Multiple sequence alignment shows that AmVP1 has high homology with pyrophosphatase protein sequences from Arabidopsis thaliana, tobacco, rice, corn, barley, sorghum and other plant sources, as shown in the attached figure 2 shown. The accession numbers of each protein gene are as follows: AtVP1, Arabidopsis tonoplast membrane pyrophosphatase (NP_173021); NtVP1, tobacco tonoplast membrane pyrophosphatase (CAA58701); OsVP1, rice tonoplast membrane pyrophosphatase (BAA31523); ZmVP1, maize tonoplast membrane pyrophosphatase Pyrophosphatase (CAG29370); HvVP1, barley tonoplast pyrophosphatase (Q06572); TaVP1, sorghum tonop...

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Abstract

The invention belongs to the technical field of plant genetic engineering and in particular relates to pyrophosphatase of ammopiptanthus mongolicus, and a coding gene and application thereof. The pyrophosphatase provided by the invention comes from the ammopiptanthus mongolicus, named AmVP1, wherein the amino acid sequence of the pyrophosphatase is shown as SEQ ID NO.2. The coding gene of the pyrophosphatase of the ammopiptanthus mongolicus is one of the following nucleotide sequences: SEQ ID NO.1 and polynucleotide with the amino acid sequence shown as SEQ ID NO.2. The gene of the invention can be applied to the research of the salt-resisting and drought-enduring molecular mechanism of the ammopiptanthus mongolicus, and is used for improving the salt-resisting and drought-enduring property of plants.

Description

technical field [0001] The invention belongs to the technical field of plant genetic engineering, and in particular relates to a tonoplast membrane pyrophosphatase and its encoding gene and application, in particular to the pyrophosphatase, encoding gene and application of the holly. Background technique [0002] Salt damage and drought are two limiting factors that limit the growth of crops. The area of ​​China's arid region accounts for 1 / 3 of the total area of ​​the country, and saline-alkali soil is widely distributed in it. The coexistence of drought and salinity seriously limits the increase of crop yield and agricultural development in our country. Therefore, it is of great practical and strategic significance to understand and study the mechanism of salt-alkali and drought-resistant plants; to clone salt- and drought-resistant genes from unique resource organisms in my country to obtain valuable gene resources; to cultivate drought- and saline-alkali-tolerant crops. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/14C12N15/55C12N15/63C12N5/10A01H5/00
Inventor 蒯本科魏强郭玉娟梁宁菁
Owner FUDAN UNIV
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