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Low-temperature resistant plant protein TCF1 (transcription factor1) and encoding genes and application thereof

A technology of LC-35S-GFP-TCF1 and gene, which is applied in the field of plant low temperature resistance protein TCF1 and its coding gene and related transgenic applications, can solve the problems of long period, lagging research on plant cold resistance mechanism, low efficiency, etc., and achieve improvement The effect of cold resistance

Inactive Publication Date: 2011-08-17
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

However, compared with the research on the molecular mechanism of plant drought resistance and salt-alkali resistance, the research on the mechanism of plant cold resistance is relatively lagging behind, and the work of using biotechnology to breed new varieties of cold-resistant crops is also relatively backward
For a long time, people have mainly used traditional genetic breeding methods to improve the stress resistance of crops. However, this method has the disadvantages of low efficiency and long cycle, and its effect becomes less and less obvious with long-term use

Method used

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  • Low-temperature resistant plant protein TCF1 (transcription factor1) and encoding genes and application thereof
  • Low-temperature resistant plant protein TCF1 (transcription factor1) and encoding genes and application thereof
  • Low-temperature resistant plant protein TCF1 (transcription factor1) and encoding genes and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Embodiment 1, the expression characteristic of TCF1 gene and the cloning of TCF1 gene

[0042] 1. Analysis of the expression characteristics of TCF1 gene under stress

[0043] Arabidopsis Col-0 seedlings grown at room temperature for about 12 days were subjected to the following stress treatments:

[0044] (1) Low temperature treatment: put Arabidopsis thaliana Col-0 seedlings in a 4°C incubator, light culture for 0 hours, 1 hour, 3 hours, 6 hours, 12 hours, 24 hours, take them out and freeze them with liquid nitrogen, -80°C Save for later.

[0045] (2) Abscisic acid treatment: Arabidopsis thaliana Col-0 seedlings were placed in 100 μM abscisic acid (ABA) solution, cultured in light for 3 hours, taken out and quick-frozen in liquid nitrogen, and stored at -80°C for later use.

[0046] (3) Salt treatment: Arabidopsis Col-0 seedlings were placed in 150 mM NaCl solution, cultured under light for 5 hours, the materials were taken out, quick-frozen in liquid nitrogen, and ...

Embodiment 2

[0057] Example 2, TCF1 gene deletion mutant tcf1-1 and overexpression TCF1 transgenic plant TCF1OX all improve cold resistance

[0058] 1. Obtained tcf1-1 and TCF1OX transgenic plants

[0059] SAIL Lines seeds from ABRC (Arabidopsis Biological Resource Center) were homozygously identified to obtain a homozygous mutant tcf1-1 with TCF1 gene insertion. Semi-quantitative RT-PCR results showed that tcf1-1 was a nonsense mutant of the TCF1 gene ( image 3 ); the pCAMBIA1300-35S-TCF1 vector was used to transform Arabidopsis thaliana into Col-0 to obtain pure transgenic plants, and the RT-PCR results showed that the TCF1 gene was overexpressed in TCF1OX ( Figure 4 ).

[0060] 2. Both tcf1-1 and TCF1OX transgenic plants showed cold resistance

[0061] In the case of cold treatment without cold acclimation, the survival rates of tcf1-1 after freezing treatment at -6°C, -8°C and -10°C for 2 hours were 91.3%, 52.2% and 17.4%, respectively, while Col-0 (WT ) at the corresponding trea...

Embodiment 3

[0063] Example 3, Analysis of biochemical characteristics of TCF1 protein

[0064] 1. Analysis of subcellular localization of TCF1 protein

[0065] Cut off the leaves of seedlings (cold-acclimated or non-cold-acclimated transgenic homozygous plants) grown in a petri dish for 12 days, put them in a disposable syringe (without a needle), mix the leaves with water, and then block them by hand Needle, pulled several times to make the leaves translucent, put the air hole side up on the glass slide, add a drop of water, cover the cover glass, and put it upside down on the Confocal microscope stage, Mobilize the bright field, and then scan the laser, the excitation light is 488nm; the microscope inspection is the same as above. The results showed that the TCF1 protein was nuclear localized ( Figure 8 ).

[0066] 2. Analysis of GEF Activities

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Abstract

The invention discloses a related low-temperature resistant plant protein TCF1 (transcription factor1) and encoding genes and application thereof. The protein disclosed by the invention is the protein (a) formed by amino acid sequences 1 in an amino acid sequence table or the protein (b) formed by amino acid sequences deviated from the amino acid sequences 1 by carrying out the substitution and / or deletion and / or addition by one or more amino acid residues on the amino acid sequences 1, wherein the protein (b) is relevant to the plant adversity tolerance. The protein TCF1 of which the genes are specifically cold-induced and expressed can be positioned on a cell nucleus and can be specifically combined with the histone so that the transcription and the expression of downstream genes of theprotein TCF1 can be regulated. Both the deleted protein TCF1 genes and the overexpressed protein TCF1 genes can improve the low-temperature resistance of the plant, lays a foundation for the artificial control on the expression of the related adversity-resistant genes and plays an important role in breeding the adversity-resistant plant and the adversity-resistance enhanced plant.

Description

technical field [0001] The invention relates to a plant low-temperature-resistant protein TCF1, its coding gene and related transgenic applications. Background technique [0002] Temperature is an environmental factor necessary for plant growth and development, affects all processes of plant growth and development, and is also a major factor limiting crop yield and distribution. It is a common problem in the world that chilling injury causes a substantial reduction in the yield of food crops. It is estimated that hundreds of millions of dollars are spent each year worldwide just to mitigate the damage from frost (Pearce 1999). Therefore, elucidating the mechanism of plant cold resistance not only has important scientific and theoretical significance, but also has important application value. In the past 20 years, due to the utilization of model plants and the discovery of related functional genes, we have made many breakthroughs in the study of the molecular mechanism of p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/63C12N5/10C12N15/82C07H21/04A01H5/00
Inventor 李霞纪洪涛王幼宁
Owner INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI