Storable Dried Explants for Plant Transformation Efficiency

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Solution Overview

Problem

Current plant genetic transformation and gene editing methods rely on 'transformation competent' germplasm that is prone to tissue culture-induced mutations and cannot be stored, limiting the efficiency and availability of transformation processes.

Innovation Solution

A method for preparing storable dried explants from seeds, involving rehydration, excision of meristematic tissue, and drying, with optional inclusion of priming agents and transformation supplements, allowing for longer storage and increased transformation efficiency.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If transformation competent germplasm is prepared immediately prior to transformation, then transformation efficiency is maintained, but the germplasm cannot be stored and tissue culture-induced mutations occur

Engineering Contradiction:

Improvetransformation efficiencyVSAvoidstorage duration

Core Design Contradiction:

ReliabilityVSDuration of action of stationary object

Solution Approach 1:

The patent applies preliminary action by preparing and drying explants before transformation, allowing them to be stored for later use. The explants are dried to a specific moisture content (5-15%) and stored at controlled temperatures (4-10°C) to maintain their transformation competence over extended periods without undergoing tissue culture-induced mutations.

Inventive Principle:

Principle #10Preliminary action

Solution Approach 2:

The patent utilizes parameter changes by controlling the moisture content of the explants during drying and storage. By adjusting moisture content to specific ranges (5-15%) and storage temperature (4-10°C), the explants maintain metabolic stability and transformation competence while enabling long-term storage, thus resolving the contradiction between immediate transformation efficiency and storage capability.

Inventive Principle:

Principle #35Parameter changes

2Productivity

If germplasm tissues are stored for extended periods, then availability for transformation is improved, but transformation efficiency decreases and mutations occur

Engineering Contradiction:

Improveavailability for transformationVSAvoidtransformation efficiency

Core Design Contradiction:

ProductivityVSReliability

Solution Approach 1:

The patent maintains transformation efficiency during storage by precisely controlling physical parameters: moisture content is reduced to 5-15% through drying, and storage temperature is maintained at 4-10°C. These parameter changes preserve the biological activity and transformation competence of the explants, allowing extended storage without loss of functionality.

Inventive Principle:

Principle #35Parameter changes

Solution Approach 2:

The patent introduces controlled storage conditions as an intermediary state between explant preparation and transformation. By maintaining explants in a controlled dry state at specific temperatures, the system enables long-term availability while preserving transformation efficiency, acting as a stable intermediary that bridges storage and transformation requirements.

Inventive Principle:

Principle #24Intermediary (Mediator)

3Ease of manufacture

If tissue culture is used to prepare transformation competent germplasm, then transformation can be performed, but tissue culture-induced mutations complicate phenotypic selection

Engineering Contradiction:

Improvetransformation capabilityVSAvoidphenotypic selection accuracy

Core Design Contradiction:

Ease of manufactureVSManufacturing precision

Solution Approach 1:

The patent extracts and eliminates the tissue culture step from the transformation process by using dried explants prepared directly from seeds. This extraction removes the source of tissue culture-induced mutations while maintaining transformation capability, thereby improving phenotypic selection accuracy without sacrificing ease of manufacture.

Inventive Principle:

Principle #2Taking out (Extraction)

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

Enables the production of storable explants suitable for various transformation methods, enhancing transformation efficiency and allowing for longer storage periods, thus addressing the limitations of existing methods.

Implementation Method 1

rehydrating a dry seed in a hydration medium

Methodology Applied

Scientific EffectAbsorption: Absorption (physical)

Implementation Method 2

drying the explant to form a dried explant

Methodology Applied

Scientific EffectEvaporation: Evaporation

Data Source

PatentUS11266086B2Efficient plant transformation method

Publication Date: 2022.03.08 WISCONSIN ALUMNI RES FOUND

AI summary

Disclosed herein are methods for the production of value added explants from the seeds of dicotyledonous cultivars of interest. Also described are methods of transformation using the value added explants produced by the methods disclosed herein.