May 26, 2026
Protocol for Short-Term UV-C Irradiation (254 nm) of Duckweed (_Lemna_ sp.) to Study Morphogenesis and Survival
- Evhenii Hordiienko1,
- Evhenii Hordiienko2,3
- 1Independent Researcher, Kyiv, Ukraine;
- 2American Society for Microbiology;
- 3Association for the Sciences of Limnology and Oceanography
- Microbiology
Protocol Citation: Evhenii Hordiienko, Evhenii Hordiienko 2026. Protocol for Short-Term UV-C Irradiation (254 nm) of Duckweed (_Lemna_ sp.) to Study Morphogenesis and Survival. protocols.io https://dx.doi.org/10.17504/protocols.io.4r3l2dxk3g1y/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: May 25, 2026
Last Modified: May 26, 2026
Protocol Integer ID: 317867
Keywords: Lemna sp., UV-C radiation, morphogenesis, photodegradation, survival, duckweed, surviving plant, subsequent tissue degradation, duckweed, uv, term uv, morphogenesi, mother plant
Abstract
The effect of a single short-term UV-C irradiation (254 nm, 60 s, distance 26 cm) on morphogenesis and survival of young Lemna sp. plants was studied. Four individuals were used in the experiment. By the 4th day, only one individual survived (survival rate 25%). In the surviving plant, significant morphogenetic abnormalities were observed: failure of daughter fronds to separate from the mother plant, formation of a single multi-lobed star-shaped colony, and subsequent tissue degradation. The results indicate high sensitivity of Lemna sp. to UV-C radiation.
Guidelines
Safety and Operational Guidelines
1. UV-C Safety: This protocol involves hazardous short-wave UV-C radiation (254 nm). Operators must utilize a dedicated technical shielding box or wear UV-blocking safety goggles and protective clothing to prevent photokeratitis and skin burns.
2. Experimental Uniformity: Use young Lemna sp. fronds separated from the maternal colony exactly 3 days prior to irradiation to ensure consistent physiological and morphogenetic conditions.
3.Contamination Control: Because the nutrient medium contains 5.56% sucrose, it is highly susceptible to rapid bacterial and fungal contamination. Maintain clean handling conditions and use sterile, BPA-free containers.
Materials
**Object of study: Young Lemna sp. plants separated from the maternal colony 3 days prior to the experiment. A total of 4 individuals were used.
**Nutrient medium: Distilled water supplemented with 5.56% sucrose.
**Radiation source: UV lamp 254 nm (ozone-free).
**Exposure Container: Transparent plastic container (BPA-free, 100 ml capacity).
STEP-BY-STEP PROCEDURE
Isolate young Lemna sp. plants by separating them from the maternal colony exactly 3 days prior to the start of the irradiation procedure. Select a total of 4 healthy individuals for the experiment.
Prepare the nutrient medium consisting of distilled water supplemented with 5.56% sucrose. Measure and transfer a volume of 50 ml of this medium into a transparent, BPA-free plastic container with a total capacity of 100 ml.
Place the 4 selected individuals into the prepared medium container. Set up the radiation source (ozone-free UV lamp 254 nm) at a strict distance of 26 cm from the object. Expose the plants to the radiation continuously for 60 seconds.
Immediately following the 60-second exposure period, carefully transfer the irradiated plants into a fresh nutrient medium (prepared with the identical composition). Maintain the cultivation at room temperature under dim light (~6500 K).
Perform systematic observations of the culture on days 4, 6, 8, and 9 post-exposure. Upon reaching day 9, terminate the live observation and fix the experimental material in 70% ethanol for preservation.
Short-term UV-C irradiation (254 nm) for 60 seconds at a distance of 26 cm causes significant morphogenetic disturbances and high lethality in Lemna sp. The observed changes (failure of daughter fronds to separate and formation of a fused colony) are most likely caused by DNA damage, primarily the formation of pyrimidine dimers.
The experiment confirms the high sensitivity of duckweed Lemna sp. to UV-C radiation and demonstrates the possibility of inducing stable morphogenetic abnormalities under short-term exposure.