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Plant Stress Tolerance Related Transcription Factors and Their Encoding Genes and Applications

A plant stress tolerance and transcription factor technology, which is applied to plant stress tolerance-related transcription factors and their encoded genes and applications, can solve the problems of lack of systematic theoretical research on stress resistance mechanism of pear rootstocks, inability to fully develop and utilize them, and improve the The effect of salt tolerance and drought tolerance

Active Publication Date: 2019-04-12
JIANGSU ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the lack of theoretical research on the stress-resistant mechanism system of pear rootstock, it has been unable to fully develop and utilize the rootstock to breed excellent stress-resistant strains.

Method used

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  • Plant Stress Tolerance Related Transcription Factors and Their Encoding Genes and Applications
  • Plant Stress Tolerance Related Transcription Factors and Their Encoding Genes and Applications
  • Plant Stress Tolerance Related Transcription Factors and Their Encoding Genes and Applications

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1 Screening of PbHB12 gene encoding gene PbHB12 related to stress tolerance in Du pear and its cDNA clone

[0034] Using Arabidopsis thaliana ATHB7 (NM_130233.4) as a probe, NCBI homology alignment was carried out, and a primer pair (upstream primer PbHB12_F1 and downstream primer PbHB12_F1 and downstream Primer PbHB12_F2) was used to amplify stress tolerance-related transcription factor PbHB12 gene from Du pear.

[0035] The rooted seedlings of Du pear were treated with 150mM NaCl aqueous solution, and the leaves of Du pear were cut after 24 hours, and the total RNA of the stress-treated leaves of Du pear was extracted by conventional methods. The cDNA first-strand synthesis kit (M-MuLV First Strand cDNASynthesis Kit, Shanghai Sangon Bioengineering Co., Ltd.) was used to synthesize the first-strand cDNA, and then using the cDNA as a template, the upstream primer PbHB12_F1, the downstream primer PbHB12_F2 and the linker primer pair ( OUTER and INNER) for nested...

Embodiment 2

[0046] Example 2, PbHB12 gene tissue expression characteristics and stress expression research

[0047] In order to confirm whether the PbHB12 gene is involved in the regulation of plant stress tolerance, the present invention uses semi-quantitative RT-PCR and real-time fluorescent quantitative RT-PCR techniques to analyze the expression patterns of the PbHB12 gene in different tissues of Duli pear and under different stress conditions.

[0048] 1. Analysis of tissue expression of PbHB12 gene under normal growth conditions

[0049] Under normal growth conditions, the roots, stems, leaves, flowers, and fruits of Duli pear were collected and quickly frozen in liquid nitrogen. About 200 mg of each part was taken, and ground into powder in liquid nitrogen. RNA was extracted using TRIzon kit (Shanghai Life Technologies Corporation). Using the RNA extracted from each part as a template, reverse transcription kit ( PrimeScript TM RTreagent Kit with gDNA Eraser, Treasure Bioengin...

Embodiment 3

[0066] Embodiment 3, transformation PbHB12 escherichia coli growth curve and gene expression analysis

[0067] The prokaryotic expression vector pET-22b(+) and the recombinant plasmid p-MD18-PbHB12 obtained in Example 1 were digested with EcoR Ⅰ and HindIII (Thermo Scientific, USA) respectively, and TaKaRa MiniBESTAgarose Gel DNAExtraction Kit Ver4.0 was used to (Treasure Bioengineering (Dalian) Co., Ltd.) for gel recovery, and the recovered product was ligated with T4 DNA Ligase (Thermo Scientific, USA) and then transformed into Escherichia coli E.coli DH5α competent cells (Treasure Bioengineering (Dalian) Co., Ltd.). The recombinant plasmid was picked on the ampicillin (Amp) resistant LB plate, identified by enzyme digestion and sequenced, and the expression vector successfully fused with the target gene was named pET-22b(+)-PbHB12.

[0068] The empty vectors pET-22b(+) and pET-22b(+)-PbHB12 were respectively transformed into BL21(DE3) to obtain recombinant bacteria BL21-pET...

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Abstract

The invention provides a plant stress tolerance associated transcription factor, an encoding gene of the plant stress tolerance associated transcription factor, and application of the encoding gene. The transcription factor is protein consisting of an amino acid sequence shown in SEQ ID NO: 2 or protein which is obtained by adding or missing of an amino acid sequence shown in the SEQ ID NO:2 and related with stress tolerance. The encoding gene of the transcription factor is the 77th-772th nucleotides from 5' end of a sequence shown in the SEQ ID NO: 1 or the sequence shown in the SEQ ID NO: 1. An experiment of transferring a PbHB12 gene into tomatoes testifies that in a salt stress and drought stress experiment, the survival rate, the water content and the chlorophyll content of three PbHB12 transgenic plants of T2 generation are remarkably better than those of wild plants. That is, the PbHB12 gene and protein encoded thereby can remarkably improve the salt tolerance and drought tolerance of plants.

Description

technical field [0001] The invention relates to the field of plant genetic engineering, in particular to transcription factors related to plant stress tolerance, coding genes and applications thereof. Background technique [0002] Drought stress and salt stress are important limiting factors for crop growth and yield. The water shortage caused by drought stress and salt stress severely inhibits leaf cell swelling and expansion, thereby affecting leaf growth and the normal development of aboveground parts, and ultimately limiting plant flowering and fruiting. Because of the perennial immobility of woody plants, the study of stress tolerance mechanism and the selection of stress-tolerant varieties are particularly important. [0003] Homeobox (homeobox) refers to the encoding of 60 conserved amino acid homeodomain (homeodomain, HD). 60 amino acids are folded into a triple helix structure and specifically bind to DNA. Homeoboxes are ubiquitous in plants, animals, humans, fru...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/00A01H6/82
CPCC07K14/415C12N15/8273
Inventor 王宏蔺经常有宏李晓刚
Owner JIANGSU ACAD OF AGRI SCI
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