Dried Maize Explant Storage for 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 availability of storable tissues competent for various transformation methods, which complicates phenotypic selection and transformation efficiency.
Innovation Solution
A method for preparing dried maize explants by drying maize embryos on a substrate at controlled temperature and humidity levels, followed by freezing, allowing for storage and subsequent rehydration and germination, enabling efficient transformation through Agrobacterium-mediated transformation and force treatment.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If transformation competent germplasm is prepared immediately prior to transformation, then transformation efficiency is maintained, but the ability to store tissues is lost and tissue culture-induced mutations occur
Solution Approach 1:
The patent applies preliminary action by preparing and drying embryo tissues in advance before transformation. Embryos are dried to specific moisture content ranges (2-20%) and stored at controlled temperatures (4°C to 30°C) with controlled humidity (20%-60% RH), enabling long-term storage while maintaining transformation competence when rehydrated and used for Agrobacterium-mediated transformation.
Solution Approach 2:
The patent utilizes parameter changes by controlling the moisture content of embryos during drying to specific ranges (2%-20%) and maintaining storage conditions at specific temperatures (4°C to 30°C) and humidity levels (20%-60% RH). These parameter changes enable the embryos to be stored long-term while preserving their transformation competence.
2Productivity
If embryo drying is performed at high temperature and low humidity, then drying speed increases, but embryo viability and germination rate decrease
Solution Approach 1:
The patent applies parameter changes by optimizing drying conditions to specific temperature (4°C to 30°C) and humidity (20%-60% RH) ranges, avoiding excessive heat and extreme dryness that would kill embryos. This balanced parameter setting enables controlled moisture removal while preserving embryo viability and germination rates of at least 65%.
Solution Approach 2:
The patent replaces aggressive mechanical drying methods with a more gentle controlled desiccation process using substrate-based drying and controlled environmental conditions, substituting high-temperature forced air drying with a milder, viability-preserving drying approach.
3Reliability
If controlled drying conditions are used, then embryo viability is maintained, but drying time increases
Solution Approach 1:
The patent optimizes the drying process by adjusting temperature (4°C to 30°C) and humidity (20%-60% RH) parameters to achieve the right balance between drying speed and viability maintenance. This optimized parameter setting reduces unnecessary drying time while ensuring embryos reach the target moisture content (2%-20%) without excessive time loss.
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
This method provides a storable and competent germplasm for plant transformation, reducing tissue culture-induced mutations and increasing transformation efficiency, while minimizing contamination risks and storage limitations.
Implementation Method 1
drying a maize embryo on a substrate at a temperature between about 4° C. and about 30° C. and at a relative humidity between about 20% and 60%
Implementation Method 2
the embryos is dried in the presence of desiccation medium comprising water, a tissue culture medium, or a sugar solution
Implementation Method 3
the method additionally comprises the step of freezing the embryo at −20° C.
Data Source
AI summary
Disclosed herein are methods for sterile extraction, drying, storage, and transformation of immature and mature maize and monocot embryos. The present disclosure also describes a dried, storable, freezable value added maize and monocot explants.