6 Ways to Save Money on Culture Medias & Containers
Summary
The cost of production is directly proportional to the cost of culture media and containers. The type of culture vessels influences the efficiency of transfer during subculture and production of propagules per unit area.
Proper choice of media and containers can reduce the cost of micropropagation. The replacement of expensive imported vessels with reusable glass jars and lids, alternatives to gelling agents, use of household sucrose, and some medium components can reduce costs of production.
Bulk making of media and storage as deep frozen stocks also reduces labor costs
1. Gelling Agent
The growth of plant tissues and organs is strongly influenced by the physical consistency of the culture medium. Gelling agents are usually added to culture medium to increase its viscosity, which results in plant tissues remaining above the surface of nutrient medium.
Media chemicals cost less than 15% of micro-plant production. In some cases the cost may be as low as 5%. Of the medium components, the gelling agents such as agar contribute 70% of the costs.
Agar is the most commonly used gelling agent for preparation of solid and semi-solid media.
Gelling Agent Concentration
Agar is usually used at 0.6–0.8% (w/v). It is advisable to prepare sample media in small quantities using various concentrations of agar, e.g., 0.7, 0.75, 0.8, 0.85 and 0.9%.The appropriate concentration should then be used for large-scale production purposes.
A semi-solid medium is often preferred over solid medium
Why ? Because …
A. Semi-solid medium ensures adequate contact between the plant tissue and the medium.
B. It is beneficial to growth as it allows better diffusion of medium constituents, and is easily removed from plantlets before their transfer to in vitro conditions.
Low Cost Alternative to Agar
Cheaper alternatives to agar include various types of starches and plant gums such as :
A. 8.0% tapioca starch to the MS medium was found to be a good substitute for ‘Bacto-agar’ for potato shoot-culture
B. Barley starch (60.0 g/l) has also been used for culturing potato-tuber discs, and for anther culture of barley
The approx. cost of sago is $0.5/kg.
The approx. cost of ‘Isubgol’ is about $4/kg.
The cost of Corn Starch was $1.8/kg compared with $200/kg of agar.
C. Sago (obtained from the stem pith of Metroxylon) at 13% concentration. The number of shoots and leaves, and root length were significantly higher on sago than on agar.
D. ‘Isubgol’, a husk derived from the seeds of Plantago ovata at 3% in MS medium has been used for the propagation of chrysanthemum.
E. Corn starch as a gelling agent has been used along with low concentration of ‘Gelrite’ (0.5 g ‘Gelrite’ + 50.0 g CS /l) for the propagation of fruit trees, such as apple, pear and raspberry, banana, and sugarcane, ginger and turmeric
The shoot proliferation was better on corn starch-medium than on agar.
2. Suspension Culture
Suspension cultures without gelling agents are commonly used for culturing callus, cell clusters, buds and somatic embryos. Suspension systems allow greater contact between the explant and the medium.
Sterilized, non-chlorine bleached, rolled, pure cotton fiber
Support to cultures in liquid media has been reported in the commercial propagation of orchids, banana, chrysanthemum, and potato in Pakistan and Bangladesh.
Advantages
A. Cotton fiber and MS liquid medium produced rapid growth of banana shoots, which could be sub-cultured after two instead of six weeks
B. Protocorm initiation and shoot development of orchids was much faster on cotton than on agar-based medium.
The cost of cotton fiber is about $2/kg, and of agar $100- 200/kg
Other alternative culture supports include foam, plastic, filter paper bridges, glass beads ‘Viscose’ sponge, glass wool and rock wool in liquid media.
3. Medium Carbon Source
Sucrose is the most commonly used carbon source in the micropropagation of plants which adds significantly to the media cost.
Replace Sucrose from Household Sugar as common sugar reduces the cost of the medium between 78 to 87%
Why ?
A. The cost of the local sugar was US$ 0.55/kg against the $40.0/kg for the imported sucrose
B. Sugar sold in grocery stores is sufficiently pure for micro propagation
4. Source of Water
Water is the main component of all plant tissue culture media. Usually in tissue culture research, distilled or doubled distilled and de-ionized water is used. Distilled water produced through electrical distillation is expensive
In Bangladesh, the changeover of water distillation from electrical to gas operated unit reduced the cost from US$260 to $5/month for producing 50–60 liter water per day ( Approx.)
Replace distilled or doubled distilled and de-ionized water from
A. Rain water Harvested water
B. Clean drinkable tap water — if free from heavy metals & containments
5. Save Time ( & Money ) While Making Media
Readymade Medias (either in liquid or powder form) can be considered when:
1. Helps to save time
2. Errors in media preparation is less frequent
3. Affordable if required in small quantities
Large scale lab require at least 200 to 500 Lts of medium per day.
Make sure to:
1. Media should be made in small batches, so that even if the mistake occurs , there is not much loss
2. Non-sterile media should not be stored at room temperature for more than 24 hr, especially when it contains high amounts of sucrose.
Usually stocks are prepared in 10x to 100x concentrations frozen in plastic bags sufficient to create 1 to 5 liters of solution at a time.
Low Cost Options for Containers
Plastic bags (approximately 10x15cm) have been used for large-scale micropropagation and are very cost effective.
Why ?
A. Disposable plastic bags eliminate the cost of washing and of lids
B. Plastic bags are sterile due to high temperature during manufacture
A private company in India has been producing more than five million plants of various ornamentals (foliage and flowering), plantation, medicinal, farm and forestry plants using low cost media ingredients and containers.
How ?
After pouring pre sterilized medium under the laminar flow:
A. Top 2 to 5 cm of the bags is folded and several bags are held together with either large paper clips or plastic cloth-hanging pegs.
B. After transfer of the explants and cuttings, the bags are closed with a heat-sealing machine or by knotting if the bags are 18–20 cm long.
C. Plastic bags being lightweight can also be hung by thread and do not need elaborate shelving.
The production cost of tissue-cultured plants can be reduced by 50–90% by using low cost media-ingredients and containers as described
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