OsNRT2.3b Nitrate Transporter Overexpression for Crop Yield

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Solution Overview

Problem

Current crop production relies heavily on chemical fertilizers, with nitrogen (N) use efficiency being only 30-50%, leading to environmental degradation and economic losses, and there is a need for more nutrient-efficient crop genotypes to ensure sustainable food security and reduce environmental impacts.

Innovation Solution

Overexpression of the rice nitrate transporter OsNRT2.3b, which has a pH-sensitive regulatory site, is used in transgenic plants to improve nitrogen use efficiency and growth by enhancing both nitrate and ammonium uptake, and regulating pH homeostasis, demonstrating improved growth, yield, and nitrogen use in rice and other species.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If chemical fertilizers are used to increase crop productivity, then grain yield is improved, but nitrogen use efficiency deteriorates (only 30-50% efficiency)

Engineering Contradiction:
Improvegrain yieldVSAvoidnitrogen use efficiency
Core Design Contradiction:
ProductivityVSLoss of energy

Solution Approach 1:

The patent modifies the expression level and activity of nitrate transporter proteins (NRT1 and NRT2 families) in plant roots through genetic engineering. By overexpressing specific nitrate transporter genes, the patent increases nitrate uptake capacity and efficiency, allowing plants to acquire nitrogen more effectively from the soil, thereby improving nitrogen use efficiency while maintaining or increasing grain yield.

Inventive Principle:
Principle #35Parameter changes

2Productivity

If chemical fertilizers are applied to achieve maximum yield, then crop production is improved, but environmental degradation occurs

Engineering Contradiction:
Improvecrop productionVSAvoidenvironmental degradation
Core Design Contradiction:
ProductivityVSObject-affected harmful factors

Solution Approach 1:

The patent alters the nitrate uptake parameters by engineering enhanced expression of nitrate transporter genes. This enables plants to achieve maximum yield with reduced fertilizer input, as the improved transporter efficiency allows for better nitrogen acquisition from available soil nutrients, thereby reducing the need for excessive chemical fertilizer application and associated environmental harm.

Inventive Principle:
Principle #35Parameter changes

3Quantity of substance

If nitrate uptake systems are increased to improve nitrogen acquisition, then nitrogen use efficiency is improved, but plant complexity increases

Engineering Contradiction:
Improvenitrogen acquisitionVSAvoidtransporter system complexity
Core Design Contradiction:
Quantity of substanceVSDevice complexity

Solution Approach 1:

The patent utilizes existing nitrate transporter protein families (NRT1 and NRT2) that serve multiple functions in nitrate acquisition across different soil conditions. By overexpressing specific members of these families, the patent enhances nitrogen uptake capacity without introducing entirely new complex systems, leveraging the universal functionality of these transporter proteins to improve nitrogen acquisition efficiently.

Inventive Principle:
Principle #6Universality (Multi-functionality)

Applied Scientific Principles

This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.

Function Achieved in This Case

Transgenic plants overexpressing OsNRT2b show increased grain yield, improved nitrogen use efficiency, and better pH regulation, leading to enhanced growth and nitrogen acquisition, even in plants that typically use different nitrogen sources, thus addressing the inefficiencies of traditional fertilizer use.

Implementation Method 1

Both electrophysiological and molecular studies have shown that nitrate uptake through both HATS and LATS is an active process mediated by proton/nitrate co-transporters

Methodology Applied
Scientific EffectProton/nitrate co-transport:

Implementation Method 2

Only OsNRT2.3b had a pH-sensitive regulatory site on the cytoplasmic face of the protein. This pH sensing site was confirmed by site-directed mutagenesis of a histidine amino acid residue (H167R) in the pH sensing motif.

Methodology Applied
Scientific EffectpH sensing:

Implementation Method 3

The specialized aerenchyma cells in rice roots can transfer oxygen from the shoots to the roots and release it to the rhizosphere, where bacterial conversion of ammonium to nitrate (nitrification) can take place

Methodology Applied
Scientific EffectOxygen transfer through aerenchyma:

Data Source

PatentEP2954059B1Transgenic plants
Publication Date: 2019.04.10 PLANT BIOSCIENCE LIMITED
  • EP2954059B1 patent drawingFigure 1a~1c
  • EP2954059B1 patent drawingFigure 1d
  • EP2954059B1 patent drawingFigure 1e

AI summary

The invention relates to transgenic plants with improved growth and nitrogen use efficiency expressing nitrate transporter gene,methods of making such plants and methods for improving growth and nitrogen use efficiency.