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Application of rice nitrate transporter nrt1.1b in improving plant nitrogen use efficiency

A technology of NRT1.1B, nitrate transport, applied in application, plant peptides, plant products, etc., to achieve the effect of increasing nitrate content

Active Publication Date: 2017-04-26
INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, the excessive use of nitrogen fertilizer poses a strategic challenge to the sustainable development of agriculture

Method used

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  • Application of rice nitrate transporter nrt1.1b in improving plant nitrogen use efficiency
  • Application of rice nitrate transporter nrt1.1b in improving plant nitrogen use efficiency
  • Application of rice nitrate transporter nrt1.1b in improving plant nitrogen use efficiency

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1, Nucleotide variation in the CDS region of indica and japonica NRT1.1B

[0036] Through the evolutionary analysis of large-scale rice varieties, the results showed that the nitrate transporter gene NRT1.1B (LOC_Os10g40600) showed significant differentiation between indica and japonica varieties ( figure 1 a). The results of sequence analysis showed that there were two single nucleotide variations (SNP1, SNP2) in the CDS region of NRT1.1B in indica and japonica rice, such as figure 1 b and figure 1 As shown in c, and the SNP1 (T<C) at the 980th position in the CDS region leads to amino acid variation (Met<Thr) between indica and japonica rice subspecies.

Embodiment 2

[0037] Embodiment 2, indica rice and japonica rice 15 NO 3 - Content and Chlorate Sensitivity Assays

[0038] The composition of the improved Kimura nutrient solution includes: 2mM KNO 3 , 0.36mM Ca(Cl) 2 4H 2 O, 0.54mMMgSO 4 ·7H 2 O, 0.18mM KH 2 PO 4 , 40μM Fe(II)-EDTA, 18.8μM H 3 BO 3 , 13.4μMnCl 2 4H 2 O, 0.32 μM CuSO 4 ·5H 2 O, 0.3 μM ZnSO 4 4H 2 O and 0.03 μM Na 2 MoO 4 4H 2 O.

[0039] The seedlings of indica and japonica rice were cultured for 2 weeks and then transferred to the seedling containing 5mM 15 N-KNO 3The improved Kimura nutrient solution was absorbed for 24 hours, then treated with 0.1mm CaSO4 for 1 minute, rinsed with deionized water for 3 times, and the aerial part was dried at 70°C. The dried samples were ground to powder, and the 15N content was determined using an isotope ratio mass spectrometer (Thermo Finnigan Delta plus XP; Flash EA1112). The results showed that indica cultivars exhibited higher nitrate content than japonica cu...

Embodiment 3

[0041] Example 3, Determination of Nitrate Transport Ability of Indica and Japonica NRT1.1B in Xenopus Oocytes

[0042] The RNAs of the indica rice IR24 and the japonica rice Nipponbare were extracted and reverse transcribed into cDNA. The cDNAs obtained respectively were used as templates, and the NRT1.1B CDS of indica rice and japonica rice were respectively amplified by PCR using the following primer sequences. The two ends of the primers used for amplification were respectively introduced into the recognition sites of restriction endonucleases BamH I and EcoR I (as shown in the underline), primer sequence: F: 5'- GGATCC ATGGCGATGGTGTTGCCG-3';R:5'- GAATTC TTAGTGGCCGACGGCGATGGT-3'.

[0043] The amplified fragments of interest were ligated into the Xenopus oocyte expression vector pCS2+, and were transcribed in vitro using the mMESSAGEmMACHINE (Ambion; AM1340) kit. Injection and culture of cRNA were performed according to the reported method (Zhang et al., 1998). The cu...

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Abstract

The invention discloses the application of rice nitrate transporter NRT1.1B in improving plant nitrogen utilization efficiency. The protein NRT1.1B is the protein shown in Sequence 2 of the Sequence Listing, which can be used to increase the plant nitrate content or improve the transport capacity of plant nitrate, thereby improving the nitrogen utilization efficiency of plants. The experimental results proved that the nitrate content of the transgenic lines obtained by overexpressing the rice nitrate transporter NRT1.1B was significantly increased, indicating that the nitrate transporter NRT1.1B has important application potential in improving plant nitrogen use efficiency. The invention provides a new approach for cultivating crops with high nitrogen utilization rate.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the application of rice nitrate transporter NRT1.1B in improving plant nitrogen utilization efficiency. Background technique [0002] Among various nutrient elements, nitrogen occupies the primary position in the process of plant life activities. Nitrogen is many important compounds in plants, such as nucleic acids (DNA, RNA), proteins (including enzymes), phospholipids, chlorophyll, photosensitizers, vitamins (B1, B2, B6, etc.), plant hormones (IAA, CTK), alkaloids, etc. Therefore, nitrogen plays a multifaceted role in maintaining and regulating plant physiological functions. However, in the natural soil environment, the content of nitrogen sources that can be directly absorbed and utilized by plants is low, so nitrogen is an important limiting factor affecting plant growth and development. Nitrogen fertilizers are widely used in agriculture to increase crop yields. Since the 198...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/84A01H5/00A01H1/02
CPCA01H1/02C07K14/415C12N15/8222C12N15/8261Y02A40/146C12N15/8241C12N15/8243
Inventor 储成才胡斌王威张志华李华梁成真车荣会
Owner INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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