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Huahuachai plasma membrane na + /H + Antiporter and its coding gene and application

A technology of antiporter and variegated protein, applied in the field of genetic engineering, can solve problems such as unclear functions

Active Publication Date: 2020-12-18
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, currently available information on SOS1-mediated Na + The role of efflux in salinity tolerance in Huachai still remains unclear

Method used

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  • Huahuachai plasma membrane na <sup>+</sup> /H <sup>+</sup> Antiporter and its coding gene and application
  • Huahuachai plasma membrane na <sup>+</sup> /H <sup>+</sup> Antiporter and its coding gene and application
  • Huahuachai plasma membrane na <sup>+</sup> /H <sup>+</sup> Antiporter and its coding gene and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Embodiment 1 Huahuachai Na + / H + Cloning of Antiporter Gene KcSOS1

[0053] The 2-week-old tissue culture seedlings of Huachai with the same growth vigor were transferred to a culture box (20cm×15cm×10cm), cultured with Hoagland nutrient solution for 4 weeks, added 150mM NaCl for stress treatment for 24h, and the leaves were cut and washed with sterile water. Put it on sterile filter paper to absorb water, weigh 100 mg, fully grind it in liquid nitrogen, extract total RNA according to the instructions of TaKaRa MiniBEST Plant RNAExtraction Kit, measure the concentration by Quawell5000 nucleic acid protein analyzer, and use TaKaRa PrimeScriptRTase cDNA first-strand Synthesis kit instructions Perform reverse transcription to synthesize cDNA.

[0054] Search the cloned plant plasma membrane Na according to NCBI database + / H + The nucleotide sequence of the antiporter gene SOS1 was found by DNAMAN 8.0 software to find the highly conserved region, and following the pri...

Embodiment 2

[0070] Example 2 Plasma membrane Na + / H + Predictive analysis of the structural features of the antiporter KcSOS1

[0071] The amino acid sequence of KcSOS1 of Huahuachai and other plants in GenBank were analyzed by DNAMAN8.0 software and Blast (https: / / blast.ncbi.nlm.nih.gov), and it was found that KcSOS1 was closely related to dicotyledonous plants Artemisia annua, chrysanthemum and Arabidopsis SOS1 protein homology of 88%, 87% and 55%, respectively ( Figure 2A ). Analysis of KcSOS1 protein using TMHMMServer v.2.0 (http: / / www.cbs.dtu.dk / services / TMHMM / ) and HMMTOP2.0 (http: / / www.enzim.hu / hmmtop / html / submit.html) Transmembrane regions and topology are predicted. The results showed that its N-terminus contained 12 transmembrane regions, its C-terminus contained a long hydrophilic tail, and it contained a cyclic nucleotide (cNMP) binding domain at 738-828aa ( Figure 2B ). Therefore, Huahuachai KcSOS1 belongs to the plasma membrane Na + / H + Antiporter.

Embodiment 3

[0072] Example 3 Plasma membrane Na + / H + Evolutionary tree analysis of the antiporter gene KcSOS1

[0073] In order to analyze the genetic relationship between the KcSOS1 gene of Huahuachai and other plant SOS1 genes, MEGA 6.0 software was used for phylogenetic tree analysis. The results showed that KcSOS1 was closely related to the dicotyledonous plants AjSOS1 and CcSOS1 of chrysanthemum, but was relatively distantly related to the monocotyledonous plants such as wheat TaSOS1 and rice OsSOS1 ( image 3 ). This implies that KcSOS1 encodes plasma membrane Na + / H + Antiporter.

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Abstract

The invention discloses a karelinia caspica(Pall.)Less. plasma membrane Na+ / H+ reversal transport protein and an encoding gent and application thereof. An amino acid residue sequence of the kareliniacaspica(Pall.)Less. plasma membrane Na+ / H+ reversal transport protein KcSOS1 is shown in SEQ ID No.2 in a sequence table; a sequence of the gene encoding the karelinia caspica(Pall.)Less. plasma membrane Na+ / H+ reversal transport protein is shown in SEQ ID No.1. It is proved through an experiment that when a karelinia caspica(Pall.)Less. KcSOS1 gene is transferred into tobacco, the saltresistanceof the transgenic tobacco can be significantly improved. The provided karelinia caspica(Pall.)Less. KcSOS1 gene can be used for improving the salt stress resistance performance of crops, and has a wide application prospect in the field of salt tolerance inheritance improvement for the crops and fine forage grass.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to a Huahuachai plasma membrane Na + / H + Antiporter KcSOS1 and its coding gene and application. Background technique [0002] Drought and soil salinization are the main environmental factors limiting plant growth and development. There are about 1 billion hectares of saline-alkali land in the world, of which my country's saline-alkali land accounts for about 10% of the world's total area (Wang Zunqin et al., 1993; Rozema and Flowers, 2008); especially in recent years, frequent drought and flood climate changes have led to increasing soil salinization in my country. Aggravating, seriously restricting my country's agricultural and animal husbandry production and ecological environment construction (Guo et al., 2013, 2015). Therefore, how to effectively control and alleviate soil salinization has become an urgent problem to be solved under the background of the continuou...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/00
CPCC07K14/415C12N15/8273
Inventor 郭强孟林张晨毛培春田小霞张国芳
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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