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Application of physcomitrella patens ATG3 gene in plant senescence and salt tolerance stress

A Physcomitrella patens and gene technology, applied in the application field of Physcomitrella patens ATG3 gene in plant senescence and salt tolerance

Inactive Publication Date: 2019-04-12
KUNMING INST OF BOTANY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The ATG gene is also included in Physcomitrella patens, and the functional study of the ATG3 gene has not been reported. Exploring the function of this gene in bryophytes is of great significance for plant development and evolution

Method used

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  • Application of physcomitrella patens ATG3 gene in plant senescence and salt tolerance stress
  • Application of physcomitrella patens ATG3 gene in plant senescence and salt tolerance stress
  • Application of physcomitrella patens ATG3 gene in plant senescence and salt tolerance stress

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] In the Plant Genome Database (http: / / phytozome.jgi.doe.gov / pz / portal.htmL) BLASTp comparison of the reported Arabidopsis ATG3 protein sequence and download the homologous ATG3 protein in Physcomitrella patens And gene sequence, through the online structure domain analysis tool SMART (http: / / smart.embl.de / ) and InterProScan (http: / / www.ebi.ac.uk / interpro / ) to obtain the candidate gene conservative domain identification, identification results such as figure 1 As shown, the transcribed and translated protein has three structural domains, which are the Autophagy_N domain at the N-terminus, the Autophagy_act_C domain at the middle, and the Autophagy_C domain at the C-terminus. It can be used as a candidate gene, and the number of the Physcomitrella patens ATG3 gene is Pp1s62_116V6.1.

Embodiment 2

[0021] The Physcomitrella patens ATG3 gene numbering Pp1s62_116V6.1 obtained in Example 1 is input into the online analysis website http: / / bar.utoronto.ca / efp_physcomitrella / cgi-bin / efpWeb.cgi to obtain ATG3 genes different in Physcomitrella patens. The expression levels in tissues, including protoplast, chlorocelium, axoneme, gametophyte, archegonia, sporophyte S1, sporophyte S2, sporophyte S3, sporophyte M, spores and rhizoids. The expression measurement results were as figure 2 As shown, it is expressed in many tissues.

Embodiment 3

[0023] Transient expression and stable transformation of Physcomitrella patens

[0024] BCD medium was used to cultivate moss, and the protonema material was polished and subcultured for 5-6 days, and the photoperiod of 16h light / 8h dark, 500μmol·m -2 ·s -1 Light intensity, cultured at 25°C.

[0025] gene amplification

[0026] Phanta Max Super-Fidelity DNA Polymerase amplification system: Take the 50μL system as an example, 2×Phanta Max Buffer 25μL, dNTP Mix (10mM each) 1μL, DNA / cDNA 50-400ng / 1-5μL, ForwardPrimer (10μM) 2μL, Reverse Primer (10μM) 2μL, Phanta Max Super-FidelityDNAPolymerase 1μL, add ddH at the end 2 0 to 50 μL. The reaction program was as follows: pre-denaturation at 95°C for 3 min; denaturation at 95°C for 15 sec, annealing at 56-72°C for 15 sec, extension at 72°C for 60 sec / kb, 35 cycles; complete extension at 72°C for 5 min.

[0027] Restriction sites and plasmid construction

[0028] Using Physcomitrella patens cDNA as a template, using primers PpATG...

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Abstract

The present invention provides an application of a physcomitrella patens ATG3 gene in plant senescence and salt tolerance stress, and relates to the technical field of gene application. A nucleotide sequence of the physcomitrella patens ATG3 gene is shown in SEQ ID NO.1. Compared with wild type, the physcomitrella patens ATG3 gene mutant is accelerated in senescence under normal culture conditionsand reduced in salt tolerance stress, and can be used for research on physcomitrella patens in the plant senescence and salt tolerance traits.

Description

technical field [0001] The invention belongs to the technical field of gene application, and in particular relates to the application of Physcomitrella patens ATG3 gene in plant aging and salt stress tolerance. Background technique [0002] Autophagy (autophagy for short) is a ubiquitous eukaryote that can promote the vacuolar-dependent (plant and yeast) or lysosome-dependent (animal) degradation of unwanted material components in cells and use them for regeneration. mechanism for recycling. Autophagosomes are first formed with a double-membrane structure that sequesters and encapsulates unwanted cytoplasmic components such as damaged organelles or protein aggregates that are not degraded in the autophagosome but require autophagy The body fuses with the lytic vacuole or lysosome, and finally the contents are degraded and recycled by the hydrolytic enzymes or proteases therein. Autophagy is involved in the recycling not only of proteins or nitrogen sources, but also of any...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C12N15/82A01H5/00
CPCC07K14/415C12N15/8261C12N15/8273
Inventor 刘莉陈则希李萍杨红李飞
Owner KUNMING INST OF BOTANY - CHINESE ACAD OF SCI
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