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Populus euphratica PeMIPS1 gene and application thereof

A gene, Populus euphratica technology, applied in the field of plant molecular biology, can solve the problems of MIPS1 gene limitation and no reports yet

Active Publication Date: 2018-03-30
LUDONG UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But so far, the reported MIPS1 gene is limited to herbaceous plants, and the research on the involvement of MIPS1 gene in plant resistance to heavy metal stress has not been reported yet.

Method used

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  • Populus euphratica PeMIPS1 gene and application thereof
  • Populus euphratica PeMIPS1 gene and application thereof
  • Populus euphratica PeMIPS1 gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0020] Example 1 detects the expression level of Populus euphratica PeMIPS1 gene

[0021] The annual Populus euphratica seedlings with consistent growth were selected for various stress treatments. 30% PEG simulated drought treatment: soak the roots of Populus euphratica seedlings in 30% PEG solution, and collect leaves at 0hr, 12hr, 24hr, 48hr and 72hr respectively; 300mM NaCl treatment: soak the roots of Populus euphratica seedlings in 30% PEG solution, respectively Leaves were collected at 0hr, 12hr, 24hr, 48hr and 72hr; 1 mM CuSO 4 Treatment: Soak the roots of Populus euphratica seedlings in 30% PEG solution, and collect leaves at 0 hr, 12 hr, 24 hr, 48 hr and 72 hr, respectively. Total RNA was extracted using PureLinkTM RNA Mini kit (purchased from Invitrogen, USA), and the specific operation was performed according to the instructions of the above kit. For reverse transcription, PrimeSriptTM reagent kit (purchased from Takara, China) was used, and specific operations w...

Embodiment 2

[0022] Construction of embodiment 2 PeMIPS1 overexpression vector

[0023] In order to analyze the function of the PeMIPS1 gene, the applicant overexpressed it in poplar, and evaluated its application potential in genetic engineering improvement of plant stress resistance from the phenotype of the transgenic plants.

[0024] The method for constructing the overexpression vector is as follows: First, use the method of homologous comparison in the GenBank database (https: / / www.ncbi.nlm.nih.gov / ) of the National Center for Biotechnology Information (NCBI) A gene sequence was searched in , with annotation number XM_01103265, which was predicted to be the inositol-1-phosphate synthase gene in Populus euphratica. Taking the Populus euphratica leaf cDNA prepared in Example 1 as a template, use primer PeMIPS1-Xbal

[0025] (5′-CGC TCTAGA ATGTTTATTGAGAAGTTTAA-3′, sequence-specific primer excision Xba I site) and PeMIPS1-Sal (5′-ACG GTC GAC TCACTTGTATTCCAAAAATCA-3′, the sequence-sp...

Embodiment 3

[0026] Embodiment 3 Genetic Transformation of Populus japonicus

[0027] First, the recombinant expression vector PeMIPS1-PBI121 obtained in Example 2 was transformed into Agrobacterium EHA105. The preparation method of EHA105 competent cells is as follows:

[0028] (1) Pick a single colony of Agrobacterium EHA105, inoculate it in 5 mL of liquid YEB medium supplemented with 50 mg / L rifampicin, and culture overnight at 28°C;

[0029] (2) Take 2mL culture into liquid YEB and continue to culture to OD 800 is about 0.5;

[0030] (3) Place the culture in an ice bath for 30 minutes, centrifuge at 5000 rpm at 4°C for 5 minutes;

[0031] (4) Discard the supernatant, and suspend the bacteria in 10 mL of cold NaCl (0.1 mol / L);

[0032] (5) 4°C, 5000rpm, centrifuge for 5min;

[0033] (6) discard the supernatant, 1mL cold CaCl 2 (20mmol / L) suspension, aliquoted into 200μL / tube, frozen in liquid nitrogen and stored at -70°C.

[0034] Agrobacterium EHA105 transformation, screening me...

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Abstract

The invention relates to a populus euphratica PeMIPS1 gene and application thereof, and belongs to the field of plant molecular biology. A DNA (Deoxyribonucleic Acid) fragment containingthe populus euphratica PeMIPS1 gene is separated and applied by the invention, the length of a nucleotide sequence is 1533bp, and the sequence number is SEQ. NO. 1; the number of corresponding amino acid sequencesis 510, and the sequence number is SEQ. NO. 2. The DNA fragment is capable of enhancing the resistance to stress given to high salinity and heavy metal copper by populus tremula; a salt-resistant andheavy metal copper-resistant popular tremula new variety cultivated by utilizing the populus euphratica PeMIPS1 gene will has wide application value in plant restoration.

Description

technical field [0001] The invention belongs to the field of plant molecular biology, and in particular relates to the Populus euphratica PeMIPS1 gene and its application in improving poplar resistance to salt and heavy metal copper. Background technique [0002] Soil salinization and heavy metal pollution are the main limiting factors restricting agricultural production. In particular, heavy metal pollution not only leads to soil degradation, crop yield and quality decline, but also pollutes surface water and groundwater through runoff and leaching, seriously threatening human health and the ecological environment. According to statistics, an average of 15,000 tons of mercury, 3.4 million tons of copper, 5 million tons of lead, 15 million tons of manganese, and 1 million tons of nickel are discharged each year in the world. Heavy metal pollution in the soil is not only large in area, but also lasts for a long time. Therefore, reducing the negative effects of heavy metal po...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/90C12N15/61C12N15/82A01H5/00A01H6/00
CPCC12N9/90C12N15/8271C12N15/8273C12Y505/01004
Inventor 宿红艳王磊张静李媛媛朱世东
Owner LUDONG UNIVERSITY
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