Drought resisting protein PpLEA3-25 (Pogonatherum paniceum Late Embryogenesis Abundant 3-25) derived from bryophyte as well as coding gene and application thereof

A plant and gene technology, applied to the drought-resistant protein PpLEA3-25 derived from mosses and its coding gene and application field, to achieve the effect of improving drought tolerance

Inactive Publication Date: 2014-04-09
THE INST OF BIOTECHNOLOGY OF THE CHINESE ACAD OF AGRI SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, it is an effective method to obtain stress-tolerant crop varieties through genetic engineering breeding, and the most critical technical bottleneck in this method is the screening and functional discovery of effective stress-resistant genes

Method used

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  • Drought resisting protein PpLEA3-25 (Pogonatherum paniceum Late Embryogenesis Abundant 3-25) derived from bryophyte as well as coding gene and application thereof
  • Drought resisting protein PpLEA3-25 (Pogonatherum paniceum Late Embryogenesis Abundant 3-25) derived from bryophyte as well as coding gene and application thereof
  • Drought resisting protein PpLEA3-25 (Pogonatherum paniceum Late Embryogenesis Abundant 3-25) derived from bryophyte as well as coding gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Embodiment 1, the discovery of PpLEA3-25 protein and its coding gene

[0022] 1. Cultivate Physcomitrella patens on BCD medium.

[0023] 2. Put the stem and leaf body on the soaked filter paper, put it into a 500ml beaker, seal it with absorbent paper, and incubate at 24±1°C.

[0024] 3. Drought treatment: Take the protocelium, put it into a crystallization dish covered with dry filter paper, seal it with a parafilm, and cultivate it at 24±1°C for 3 days (it can be observed that the degree of dehydration is over 90%); normal control: take Put the protocelium into a crystallization dish covered with wet filter paper, seal it with parafilm, and incubate at 24±1°C for 3 days.

[0025] 4. Take the stem and leaf bodies in step 3 and step 4 respectively, extract the total protein and perform IEF / SDS-PAGE two-dimensional electrophoresis (2DE) and fluorescence difference gel electrophoresis (DIGE), and the abundance change of separation exceeds 2 times under drought conditions...

Embodiment 2

[0028] Embodiment 2, the acquisition and identification of transgenic plants

[0029] 1. Construction of intermediate carrier

[0030] The maize Ubiquitin promoter shown in Sequence 3 in the sequence listing was inserted forwardly between the HindIII and KpnI restriction sites of the plant binary expression vector pCambia1390 to obtain the intermediate vector pCambia1390-Ubi.

[0031] 2. Construction of recombinant plasmids

[0032] 1. Extract the total RNA from the stem and leaf body of Physcomitrella patens treated with drought (without watering for 15 consecutive days) and reverse transcribe it into cDNA.

[0033] 2. Using the cDNA obtained in step 1 as a template, perform PCR amplification with a primer pair composed of F1 and R1 to obtain a PCR amplification product.

[0034] F1: 5'- GGATCC AGAGAGCAGTAGGCGAACAG-3';

[0035] R1: 5'- ACTAGT TGGGCACTCAACATAGGTTTC-3'.

[0036] 3. Digest the PCR amplified product obtained in step 2 with restriction endonucleases BamHI an...

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Abstract

The invention discloses a drought resisting protein PpLEA3-25 (Pogonatherum paniceum Late Embryogenesis Abundant 3-25) derived from bryophyte as well as a coding gene and application thereof. The protein provided by the invention is derived from Physcomitrella patens, is named as a PpLEA3-25 protein, and is (a) or (b), (a) a protein consisting of an amino acid sequence shown in a sequence 1 in a sequence table; (b) is a protein relevant to drought resisting capacity of a plant, which is derived from a sequence 1 by virtue of substitution and/or deficiency and/or addition of one or more amino acid residues on an amino acid sequence of the sequence 1. According to the drought resisting protein PpLEA3-25, as well as the coding gene and the application thereof, the drought resistance of the plant can be improved. The drought resisting protein PpLEA3-25 has an important theoretical and practical signficiance for cultivating new varieties of drought resisting plants, especially, cultivating new varieties of drought resisting rice.

Description

technical field [0001] The invention relates to a drought-resistance protein PpLEA3-25 derived from bryophytes, its coding gene and application. Background technique [0002] Stress factors such as drought, salinity, and low temperature in the natural environment have an important impact on the growth and development of plants. In severe cases, they will cause large-scale crop yield reduction. Cultivating stress-tolerant crops is one of the main goals of planting. At present, it is an effective method to obtain stress-tolerant crop varieties through genetic engineering breeding, and the most critical technical bottleneck in this method is the screening and function discovery of effective stress-resistant genes. [0003] Bryophytes are terrestrial pioneer plants that appeared in the Paleo-Ordovician period 500 million years ago. In their life history, the gametophyte of the "stem and leaf body" is the main vegetative growth stage. The "stem and leaf body" leaves are composed...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/00
CPCC07K14/415C12N15/8273
Inventor 吴金霞张治国张芊孙学辉路铁刚何奕昆崔素霞
Owner THE INST OF BIOTECHNOLOGY OF THE CHINESE ACAD OF AGRI SCI
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