Application of NsNHX1 protein and related biomaterials in breeding of reverse-resistant poplar

A protein and plant stress tolerance technology, applied in the biological field, can solve the problems of lack of development and research of antiporter genes

Active Publication Date: 2019-09-20
INNER MONGOLIA UNIVERSITY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

So far, the developed and applied Na + / H + Most of the antiporter genes come from Arabidopsis, tomato and other sweet soil plants, but for woody plants, especially for halophytic woody plants Na + / H + The development and research of antiporter genes are relatively scarce

Method used

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  • Application of NsNHX1 protein and related biomaterials in breeding of reverse-resistant poplar
  • Application of NsNHX1 protein and related biomaterials in breeding of reverse-resistant poplar
  • Application of NsNHX1 protein and related biomaterials in breeding of reverse-resistant poplar

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Experimental program
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Effect test

Embodiment 1

[0113] Example 1, the acquisition of NsNHX1 Populus tomentosa and its stress tolerance analysis

[0114] 1. Acquisition of NsNHX1 Populus tomentosa

[0115] 1. Construction of recombinant vector

[0116] (1) Amplification of the target gene

[0117] According to Siberian Nitraria + / H + A pair of primers (NHX-pBI-F: 5'-GC TCTAGA ATGGATCAATTAAGTT-3', 5' includes XbaI recognition sequence; NHX-pBI-R: 5'-C GAGCTC TCACTGCCATTGGGGGAT-3', including the SacI recognition sequence at the 5' end), using the RNA of Nitraria sibirica Pall as a template, using NHX-pBI-F and NHX-pBI-R primers, using TaKaRa RNA PCR Kit (AMV) Ver. The 3.0 kit amplifies the NsNHX1 ORF region to obtain a PCR product. The reaction conditions are: 94°C for 3min; 94°C for 30s, 52°C for 30s, 72°C for 1min, 35cycles; 72°C for 5min. The electrophoresis results of the PCR products were as follows: figure 1 shown.

[0118] (2) The target gene is connected to the expression vector

[0119] The binary expres...

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Abstract

The invention discloses an application of NsNHX1 protein and related biomaterials in breeding of reverse-resistant poplar. In the present invention, the nitraria sibirica Na<+> / H<+> reverse transport protein NsNHX1 is used as a research object, and is transferred into Populus tomentosa 84K to obtain transgenic NsNHX1 Populus tomentosa. Three transformants, N1, N2 and N3, are selected for identification of reverse resistance. The wild-type Populus tomentosa is used as a control to study the salt-tolerance and antioxidant capacity of transgenic NsNHX1 populus tomentosa under adversity stress. The results show that under the conditions of adversity stress treatment, the survival rate, biomass, leaf chlorophyll content, water content, plant height and antioxidant capacity of transgenic NsNHX1 are significantly higher than those of wild type populus tomentosa. The application indicates that NsNHX1 can significantly improve the reverse resistance and antioxidant capacity of the transgenic poplar, and NsNHX1 can be used as a salt tolerance gene to breed the reverse-resistant poplar varieties.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to the application of NsNHX1 protein and related biological materials in cultivating stress-tolerant poplars. Background technique [0002] Plant roots are in direct contact with Na in the soil + organs, so the Na in the root + Na + Concentration of compartmentalization is an important mechanism in determining plant salt tolerance. Na on the tonoplast + / H + The antiporter (NHX1) transports Na in the cytoplasm + Transport against the concentration gradient to compartmentalize the vacuole; Na on the plasma membrane + / H + The antiporter protein (SOS1) can transport Na in the cytoplasm + High Na + Environment. So far, the developed and applied Na + / H + Most of the antiporter genes come from Arabidopsis, tomato and other sweet soil plants, but for woody plants, especially for halophytic woody plants Na + / H + The development and research of antiporter genes are r...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N5/10C12N15/82A01H5/00A01H6/00
CPCC07K14/415C12N15/825C12N15/8271C12N15/8273C12N15/8261C12N15/8251C12N15/8243
Inventor 林晓飞耿新张文波陈首业
Owner INNER MONGOLIA UNIVERSITY
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