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Application of ospex11 gene and its protein in improving rice salt tolerance

A technology of salt tolerance and rice, applied in the field of molecular biology, can solve problems such as plant damage, enzyme inactivation, protein degradation, membrane lipid peroxidation, etc.

Inactive Publication Date: 2019-07-09
ZHEJIANG FORESTRY UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Excessive accumulation of reactive oxygen species in cells under salt stress will lead to enzyme inactivation, protein degradation and membrane lipid peroxidation, thus causing damage to plants

Method used

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  • Application of ospex11 gene and its protein in improving rice salt tolerance
  • Application of ospex11 gene and its protein in improving rice salt tolerance
  • Application of ospex11 gene and its protein in improving rice salt tolerance

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] 1. Preparation and co-transformation of yeast Y2H competent cells

[0046] Co-transform yeast expression vector pGBKT7+OsCYP2 and rice pGADT7+cDNA library expression vector into yeast strain Y2H, preparation and transformation of yeast competent cells;

[0047] Specific steps are as follows:

[0048] 1) Take 1 μl of the yeast strain Y2H and spread it on the YPDA medium, and culture it at 30°C for 3-5 days;

[0049] 2) Pick 2-3mm single clones and inoculate them in 3ml YPDA liquid medium, culture at 30°C, 250rpm for 8-12hr;

[0050] 3) Transfer 5 μl to 50ml YPDA liquid medium, culture at 30°C, 250rpm to OD 600 Reach 0.15-0.3;

[0051] 4) Centrifuge at 700g room temperature for 5min, discard the supernatant, resuspend in 100ml YPDA liquid medium, culture at 30℃, 250rpm until OD 600 Reach 0.4-0.5;

[0052] 5) Centrifuge at room temperature at 700g for 5min, discard the supernatant, and resuspend in 30ml sterilized ddH 2 O;

[0053] 6) Centrifuge at 700g room tempera...

Embodiment 2

[0118] 1. Salt stress phenotype analysis of OsPEX11 gene overexpression and interference plants

[0119] In order to verify the salt tolerance function of the OsPEX11 gene, the OsPEX11 gene was cloned into p1300-Ubi and p1300-RNAi, and the gene overexpression and interference vectors were successfully constructed; the genetic expression vector was introduced into Agrobacterium EHA105, and the rice genetic transformation test method was referred to literature (Hiei Y , Ohta S, Komari T, et al. Efficient transformation of rice (Oryza sativa L.) mediated by Agrobacterium and sequence analysis of the boundaries of the T-DNA [J]. Plant Journal, 1994, 6:271-282), on Transform the regenerated plants with hygromycin selection markers for PCR detection verification (such as Figure 5 shown), to obtain overexpression and interference positive transformed plants.

[0120] Using 10-day-old seedlings of wild-type, OsPEX11 gene overexpression plants and interfering plants as test materials...

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Abstract

The invention discloses application of an OsPEX11 gene to improvement of rice salt tolerance. Downstream target protein OsPEX11 interacting with vital function protein OsCYP2 for adjusting and controlling rice salt tolerance is screened and verified through a yeast two-hybrid system (in vivo) and the GST pull-down (in vitro) technology, and it is proved that the gene participates in adjusting and controlling rice salt stress tolerance through the RNAi technology. The novel gene for adjusting and controlling rice salt tolerance is explored, the rice salt tolerance gene adjusting and controlling network and coherent signal transduction pathway are enriched, and a new thought is provided for the rice salt stress gene function research. Meanwhile, great significance is achieved for revealing the rice salt stress adjusting and controlling mechanism and improving rice productivity.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to the application of OsPEX11 gene and its protein in improving the salt tolerance of rice. Background technique [0002] The salinization of agricultural land is becoming more and more serious, and there are about 6.7×10 4 km 2 The saline-alkali land (Zhang Jianfeng, Shu Yumin, Xing Shangjun, etc. Saline-alkali land improvement and utilization and afforestation technology [J]. Journal of Northeast Forestry University, 2002, 30(6): 124-129), has become a sustainable agricultural development Constraints. [0003] Under high-salt environmental conditions, crop growth and development are significantly inhibited, mainly manifested in low photosynthetic rate, slow vegetative growth and reproductive growth, and low accumulation of effective dry matter, which seriously affects the seed setting rate. Salt stress is one of the main abiotic stresses that limit the growth and yield of crops...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29C12N15/82A01H5/00A01H6/46
CPCC07K14/415C12N15/8273
Inventor 刘宏波崔鹏周伟军阮松林李兰
Owner ZHEJIANG FORESTRY UNIVERSITY