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Elytrigia elongata external rectification potassium channel protein and coding gene and application thereof

A technology of E. elongatum and its coding gene, applied in the field of external rectifier potassium channel protein and its coding gene and application of E. elongatum, which can solve the problems of unknown biological function of SKOR

Active Publication Date: 2020-07-28
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the research on SKOR of E. elongatum has not been reported in the prior art. Whether SKOR exists in E. elongatum and what biological functions of SKOR are unknown

Method used

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  • Elytrigia elongata external rectification potassium channel protein and coding gene and application thereof
  • Elytrigia elongata external rectification potassium channel protein and coding gene and application thereof
  • Elytrigia elongata external rectification potassium channel protein and coding gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1 Cloning and sequence analysis of EeSKOR outer rectifier potassium channel protein gene EeSKOR of Echinopsis elongatum

[0047] Degenerate primers (P1, P2) were designed, and the cDNA synthesized by reverse transcription of the total RNA of the young roots of E. elongatum was used as a template to amplify the conserved core cDNA fragment of E. elongatum EeSKOR by PCR. After sequencing, the core fragment was found to be 555bp ( figure 1 a).

[0048] According to the 5'-RACE and 3'-RACE methods of the Clontech SMARTer RACE kit manual, 5'-cDNA and 3'-cDNA were synthesized respectively; based on the core sequence of the E. elongatum EeSKOR gene, DNAMAN 8.0 and Primer5.0 were used to The software designed the 5'-RACE outer primer P3, the Z-type primer P4 and the 3'-RACE outer primer P5, and the Z-type primer P6; the 3'-RACE amplification product was sequenced to 1033bp ( figure 1 b), the 5'-RACE amplification product is sequenced to be 1136bp ( figure 1 c).

[00...

Embodiment 2

[0062] Example 2 Effects of different concentrations of salt treatment on the expression level of EeSKOR

[0063] In order to ascertain the expression level and changes of EeSKOR gene in the roots, sheaths and leaves of E. elongatum under salt stress, qRT-PCR forward primer P7 (5'-TACGGAGGCTGCTCAGGTTT-3') and reverse primer P8 (5' -CGCATCTCCTCGCTTCATC-3'), the PCR product length is 189bp; internal reference gene Actin real-time fluorescent quantitative PCR forward primer P9 (5'-CTTGACTATGAACAAGAGCTGGAAA-3') and reverse primer P10 (5'-TGAAAGATGGCTGGAAAAGGA-3'), PCR product The length is 139bp, and the expression levels of EeSKOR gene in roots, leaf sheaths and leaves of 4-week-old Etypyrum elongatum were treated with different concentrations of NaCl (0, 25, 50, 100, 150 and 200mM) for 24h.

[0064] The results showed that with the increase of NaCl treatment concentration (25-100mM), the expression level of EeSKOR gene in roots showed an increasing trend, while the expression le...

Embodiment 3

[0065] Example 3 Construction of Etoxypyrum elongatum EeSKOR gene plant expression vector

[0066] According to the requirements of Clontech Infusion seamless connection technology, Nco was introduced at both ends of the primers upstream (P11: 5'-ACTCTTGACCATGGTATGGAGAGGGAGATTGTAGCAGAG-3') and downstream (P12: 5'-TTTACCCTCAGATCTCTACTGATCGGCTGCAACAGCAG-3') of the EeSKOR gene ORF frame. I and Bgl II restriction site, through RT-PCR amplification, by 1.2% gel electrophoresis detection target fragment PCR product ( Figure 5 ). Then, the restriction enzyme site on the pCAMBIA1301 vector was double-digested with Nco I and Bgl II, and the large fragment was recovered, named pCAMBIA1301-A; according to the corresponding procedure of Clontech Infusion seamless linking technology, the target gene EeSKOR was inserted into the linear The recombined plasmid pCAMBIA1301-35S-EeSKOR-Nos was obtained from the surface vector (pCAMBIA1301-A) of the plant, and then the specific band of about 29...

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Abstract

The invention relates to the technical field of biology, in particular to elytrigia elongata external rectification potassium channel protein and a coding gene and application thereof. The elytrigia elongata external rectification potassium channel protein EeSKOR is cloned, and the amino acid sequence of the elytrigia elongata external rectification potassium channel protein EeSKOR is shown as SEQID NO. 1. It is found that the biomass and height of plants in a high-salt environment can be increased through EeSKOR, the content of H2O2 and MDA of transgenic tobacco plants is remarkably reducedthrough the overexpression EeSKOR gene under salt treatment, the SOD activity and chlorophyll content are increased, the Na + concentration of the transgenic plants is remarkably reduced, and the in-vivo K+ concentration of the transgenic plants is increased. EeSKOR can be used for improving the growth performance of the plants under high-salt stress, the salt tolerance of the plants is improved,and the elytrigia elongata external rectification potassium channel protein and the coding gene and application thereof have great significance in cultivating new salt-resistant plant varieties.

Description

technical field [0001] The invention relates to the field of biological technology, in particular to the outer rectifier potassium channel protein of Echinopsis elongatum and its encoding gene and application. Background technique [0002] When plants are faced with abiotic stresses such as salt stress, the response involves self hypersensitivity that culminates in programmed cell death. K + / Na + If the ratio is too low, the plant degrades its contents by autophagy, or autophagy in aleurone cells, or degradative hydrolases released during the formation of tubular molecules in cell protoplasts. Therefore, high K in plants + / Na + Ratio, related to plant salt tolerance, and related to plant growth and development. K + channel and K + The transporter is the plant body for K + An important part of the absorption, transport and distribution of Gaymard et al. found that ABA treatment and acidic conditions could reduce the expression of SKOR. AtSKOR isolated from Arabidop...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/00A01H6/46
CPCC07K14/415C12N15/8273C12N15/8261
Inventor 郭强孟林周妍彤毛培春田小霞郑明利
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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