GB/T 38484-2020 in English
VALIDDetermination of the biological activity for plant hormone-related secondary metabolites. Cytological method
- Issued on:2020-11-19
- Implemented on:2020-11-19
- File Format:PDF
- Delivery:Via email within 1~3 business days
$156.00
《GB/T 38484-2020植物激素类次生代谢产物的生物活性测定 细胞学评价法》由424-cnis（中国标准化研究院）归口，主管部门为国家市场监督管理总局。
Introduction
Analysis of the core content of the standard
This standard establishes a cytological evaluation system based on transgenic Arabidopsis protoplasts, and quantitatively determines the activity of three types of plant hormones through the GUS reporter gene expression:
| Hormone type | Reporter gene | Response element | Standard material |
|---|---|---|---|
| Auxin | AuxRE::GUS | AuxREs | NAA |
| Cytokinin | CTKRE::GUS | ARR1AT | ZT |
| Gibberellins | GARE::GUS | TATC-boxs | GA3 |
Key Technical Principles
4-MU fluorescence intensity was detected by ELISA, and GUS enzyme activity was converted according to the standard curve:
- The hormone-receptor complex activates the reporter gene to express GUS protein
- GUS hydrolyzes 4-MUG to generate the fluorescent substance 4-MU
- The fluorescence intensity change rate was measured under 37℃ water bath conditions
Implementation Points
Sample Processing Requirements
- Concentration of living cells ≥1×107 cells/mL
- Standard working solution must be prepared and used immediately, and gradient dilution shall adopt a logarithmic concentration sequence
- The sample should contain more than 3 effective concentration gradients
Key quality control indicators
| Detection link | Control parameters | Allowance deviation |
|---|---|---|
| Culture conditions | 25±1℃ | ±0.5℃ |
| Enzyme reaction | 37℃ water bath | ±0.2℃ |
| Repeatability | Three independent experiments | ≤20% |
Application value of the standard
Compared with traditional bioassay methods, this method has the advantages of high sensitivity (detection limit of 10-8 mg/mL) and good specificity, and has been successfully applied in the following fields:
- Evaluation of the activity of microbial fermentation products
- Determination of the potency of synthetic plant growth regulators
- Monitoring of endogenous hormone levels in genetically modified crops
Precautions
- Coomassie Brilliant Blue staining solution needs to be prepared fresh every month
- 4-MU standard solution must be kept away from light
- The ELISA plate needs to be accurately zeroed with stop solution before reading
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