Bryophyte reversal-resistant protein PpLEA3-20 and encoding gene and application thereof

A technology encoding genes and plant stress resistance, applied in the fields of bryophyte stress tolerance protein PpLEA3-20 and its encoding genes and applications, to achieve the effect of improving drought tolerance

Inactive Publication Date: 2012-07-18
CAPITAL NORMAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The most critical technical bottleneck in this method is the sc...

Method used

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  • Bryophyte reversal-resistant protein PpLEA3-20 and encoding gene and application thereof
  • Bryophyte reversal-resistant protein PpLEA3-20 and encoding gene and application thereof
  • Bryophyte reversal-resistant protein PpLEA3-20 and encoding gene and application thereof

Examples

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Embodiment 1

[0044] Example 1. Screening of moss stress tolerance-related protein PpLEA3-20 encoding gene PpLEA3-20 and its cDNA clone.

[0045] Physcomitrella patens drought treatment: Physcomitrella patens (Physcomitrella patens) grown on BCD medium (Geng Xuke et al., 2008 Plant Gene Targeting——Application of Model Plant Physcomitrella patens. Biological Bulletin, 43 (4) : 13-15; the public can obtain from Capital Normal University) the stem and leaf bodies are transferred to the soaked filter paper, put into a 500ml beaker, and seal with absorbent paper. The drought treatment is carried out in a cultivation room with a temperature of 24±1° C. and a relative humidity of 30±5%. After 3 days, the desiccated stem and leaf bodies were taken out for protein extraction. Take the stem and leaf body without drought treatment as the control experiment.

[0046] The protein extraction of Physcomitrella patens stems and leaves adopts the method of combining step-by-step extraction and phenol extr...

Embodiment 2

[0051] Example 2, Transgenic PpLEA3-20 gene cultivation of drought-tolerant plants and its detection and analysis

[0052] 1) Construction of PpLEA3-20 expression vector pCU-PpLEA3-20

[0053] Insert the maize Ubiquitin promoter shown in Sequence 3 in the sequence listing (amplified from maize genomic DNA using 5'-CCAAGCTTAAGCTTTCTAGTGCAGTGCAGCGTGAC-3' and 5'-GGGGTACCCTGCAGCCTCTAGTGCAGAAGTAACACCA-3' as primers) into the plant binary expression vector pC1390 (purchased from Cambia, Queenslan d.Australia) in the HindIII and KpnI sites constitute the intermediate vector pC1390-Ubi. The cDNA fragment of the PpLEA3-20 gene amplified in the above-mentioned Example 1 was double-digested with Spe I and Kpn I, and inserted forward into the Ubiquitin promoter of the intermediate vector pC1390-Ubi to obtain a recombinant vector. The recombinant expression vector of the PpLEA3-20 gene containing the 1-829th deoxyribonucleotide from the 5' end of the sequence 1 in the sequence listing is ...

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Abstract

The invention discloses a bryophyte reversal-resistant protein PpLEA3-20 and an encoding gene and an application thereof. A plant resistance-related protein provided by the invention is named PpLEA3-20, is derived from physcomitrellapatens, and is a protein shown as (1) or (2), wherein (1) is constituted by an amino acid sequence shown as a sequence 2 in a sequence table, or (2) is obtained by performing substitution and/or deletion and/or addition of one or more amino acid residues on the amino acid residue sequence shown as the sequence 2 in the sequence table, is relevant to plant resistance and is derived from (1). As proved by an experiment for introducing the encoding gene of PpLEA3-20 into oryza sativassp.japonica, the survival rates (44 percent, 36 percent, 60 percent and 72 percent) and growth states of four obtained T2-generation PpLEA3-20 transgenic plants (20-1, 20-2, 20-3 and 20-4) in a drought stress experiment are remarkably better than those of a check plant.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a bryophyte stress-tolerant protein PpLEA3-20 and 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. The most critical technical bottleneck in this method is the screening and function discovery of effective stress resistance 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 l...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N5/10C12N1/15C12N1/19C12N1/21C12N15/63C12N15/11A01H5/00
Inventor 崔素霞何奕騉路铁刚张治国曹保久宋晓璇宋旸蔺平亮任桂林陈希郭士磊
Owner CAPITAL NORMAL UNIVERSITY
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