Mitra Ayu's Tissue Culture: Stages for Innovative Essential Oil Production
Mitra Ayu is embracing the tissue culture method to expedite the propagation of various essential oil plant species, such as Massoia, Tea Tree, Eucalyptus plants and many others in our Solok Plantation. By carefully manipulating the culture medium, we can stimulate the rapid development of new roots, and the entire plants.
The growing global demand for economically valuable natural products derived from aromatic plants has led to a shortage of raw materials . Plant tissue cultures offer a solution by enabling the rapid multiplication of a single explant (a small piece of tissue) into thousands of plants within a short timeframe and limited space, independent of the seasonal variations and weather patterns. The other benefits of tissue culture are numerous, including accelerated multiplication, disease-free plantlets, genetic stability and the conservation of rare and endangered species.
Figure 1: Mitra Ayu's Tissue Culture
Tissue culture, a sophisticated and systematic method of cultivating and propagating cells, typically encompasses a series of well-defined stages. Each stage plays a crucial role in the overall success of the process, ensuring the proper growth and development of cells or tissues in a controlled environment.
These stages include preparation of nutrient medium, sterilization, inoculation, controlled multiplication and ultimately the harvest of the cultivated tissue. This systematic approach ensures precision and reproducibility, making tissue culture a versatile tool in aromatic plant propagation for essential oils production.
The stages of tissue cultures are explained below.
Preparation of nutrient medium.
The medium is prepared as semi-solid medium containing macro and micro elements, amino acids, vitamins, iron source, carbon source and phytohormones . The medium contains all the nutrients that required for the normal growth and development of plants. The pH of the medium is an important factor that affects both the growth of plants and activity of plant growth regulators (PGR). In addition, PGR such as auxins, cytokinin, and gibberellins, also play an important role in determining the development pathway of plant cells and tissues. The type of hormones and its concentrations will depend on the species of the plant, the tissue or organ cultured and the objective of the experiment .
Establishment of aseptic culture.
An actively growing shoot tip of axillary or terminal bud or shoot tip of a plant is usually the starting material for the process and will undergo a sterilization process. This crucial step is implemented to eliminate any potential contaminants, such as microorganisms or fungi, that might interfere with the intended cellular development. The sterilization procedure ensures aseptic conditions within the culture environment, promoting the successful initiation of the tissue culture process and minimizing the risk of unintended biological interactions .
During this phase, the explants or micro shoots—carefully prepared sections of plant tissue—undergo a strategic transition onto the sterilized nutrient medium. The transfer process is conducted under sterile conditions to prevent any external contaminants from compromising the integrity of the culture .
Following inoculation, the bottles containing the cultured material are sealed and relocated to a growth room. This controlled environment serves as a crucial setting for initiating developmental processes. Under diffused light, typically provided by fluorescent bulbs with an intensity ranging from 1000 to 2000 lux, the plant cultures are exposed to a temperature of 25℃ and a relative humidity maintained at 50 to 60% .
Given the delicate nature of the plantlets during their growth within the laboratory setup, they are particularly susceptible to potential harm from abrupt shifts in environmental conditions. Recognizing this vulnerability, a thoughtful acclimatization period becomes essential for ensuring the robust performance of the plantlets. This period allows the cultured plants to gradually adjust to external conditions, reducing the risk of stress-induced damage and promoting their successful transition to more varied and fluctuating environments 
Once the cultured tissues or plantlets have reached an optimum stage of development, they are ready for harvest. The precise timing of this stage is often determined by specific growth indicators, such as the formation of well-defined roots, the emergence of healthy shoots, or the attainment of desired size and maturity.
Harvesting involves carefully removing the cultured tissues or plantlets from the growth medium. This delicate process is executed with precision to avoid damage to the developed structures and to maintain the integrity of the harvested material. The harvested tissues may then undergo additional steps, such as washing or rinsing, to eliminate any residual medium or contaminants.
Sustainable Essential Oil Production through Tissue Culture