Heat-resistance related protein of plant maintaining stability of thylakoid membrane and application thereof

A plant and thylakoid technology, applied in the fields of biotechnology and botany, can solve the problem of undiscovered thylakoid membrane thermostable protein and other problems

Inactive Publication Date: 2013-09-11
SHANGHAI INST OF BIOLOGICAL SCI CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] So far, no heat-resistant protein that can maintain the stability of thylakoid membranes has been found in the art

Method used

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  • Heat-resistance related protein of plant maintaining stability of thylakoid membrane and application thereof
  • Heat-resistance related protein of plant maintaining stability of thylakoid membrane and application thereof
  • Heat-resistance related protein of plant maintaining stability of thylakoid membrane and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0085] Construction of p35S:HsfA2 vector

[0086] The cDNA coding sequence of HsfA2 was amplified by RT-PCR. Specific steps are as follows:

[0087] Using the RNeasy Plant Mini kit (purchased from Qiagen), the total RNA of col-O type Arabidopsis was isolated according to the instructions, and the isolated total RNA was reverse-transcribed using M-MLV reverse transcriptase (purchased from TOYOBO) , to obtain cDNA.

[0088] Using cDNA as a template, PCR reaction was performed using Primer star DNA polymerase (purchased from Takara Company) to amplify the cDNA coding region sequence of HsfA2. The primers used are as follows:

[0089] Upstream 121A2-F -UP 5′-AA TCTAGA CTCTGAGCTTATGGATTTGAG-3' (SEQ ID NO.: 2), wherein the underlined part ( TCTAGA ) part is restriction site XbaI;

[0090] Downstream 121A2-R-DO 5'-AAGAGCTCGACCGCAAACAAGTAGATGTG-3' (SEQ ID NO.: 3), wherein the underlined part (GAGCTC) Part of it is the enzyme cutting site SacI.

[0091] The resulting DNA amp...

Embodiment 2

[0116] get transgenic plants

[0117] The Agrobacterium obtained in Example 1 was transformed into col-O type of wild-type Arabidopsis thaliana by spraying method to produce transgenic plants. As a result, 35 transgenic lines were obtained, all of which could grow normally on the PNS medium containing kanamycin.

[0118] The overexpression strain of HsfA2 is screened by the method of RT-PCR, and the primers used are as follows:

[0119] HsfA2+5'-ATGGAAGAACTGAAAGTGGAAATG-3' (SEQ ID NO.: 4);

[0120] HsfA2-5'-GATCAAATCTTTCTGAATCCATAAG-3' (SEQ ID NO.: 5).

[0121] The results showed that the screened lines were all HsfA2 transgenic plants.

Embodiment 3

[0123] Construction of rps1 transgenic plants constitutively expressing HsfA2 gene (35S:HsfA2rps1)

[0124] The rps1 mutant (purchased from salk) obtained in Example 1 was transformed with Agrobacterium, and the rps1 transgenic plant (35S:HsfA2rps1) constitutively expressing HsfA2 was obtained. The seeds of homozygous transgenic plants of the T2 generation with a Kan resistance segregation ratio of 3:1 were used for subsequent experiments.

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Abstract

The invention relates to a heat-resistance related protein of a plant maintaining stability of a thylakoid membrane and an application thereof. Specifically, through massive screen to plant genes, the inventor accidentally obtain a heat-resisting gene of a plant related to maintaining the stability of the thylakoid membrane. The gene is a heat shock transcription factor A2 (HfA2) of the plant. The gene and protein provided by the invention can effectively maintain the thermal stability of the thylakoid membrane, improve the viability and the photosynthetic efficiency of plants under heat stress, improve the crop yield, and avoid or reduce adverse effect of high temperature to the plants.

Description

technical field [0001] The invention belongs to the field of biotechnology and botany, in particular, the invention relates to a plant heat resistance-related protein for maintaining the stability of thylakoid membrane and its application. Background technique [0002] Due to the increase of carbon dioxide emissions, the earth's greenhouse effect continues to intensify, leading to global warming. Plants grown in a natural environment are very sensitive to the rise in temperature and are prone to growth disorders, especially some spring crops (such as rice, corn, etc.) are easily affected by high temperature weather during the earing and filling period, resulting in crop yield reduction . According to the data of the International Rice Research Institute, from 1998 to 2003, every 1°C increase in temperature will reduce the global crop production by 10%. According to experts' prediction, the global temperature may rise by 5-6°C in the next 100 years. In my country, by 2050, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/84A01H5/00
Inventor 郭房庆于海东刘逊亮陈思婷
Owner SHANGHAI INST OF BIOLOGICAL SCI CHINESE ACAD OF SCI
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