Tissue culture is an in vitro method of growing a new organism from an extracted part of another under aseptic and properly sterilized conditions. (Hussain, Qarshi, Nazir, & Ullah, 2012). It is an important aspect of biotechnology as it facilitates the development of more nutritional, disease resistant and better-quality food crops to satisfy the high demands for food globally. This aids in shortening the growth/production time for crops and increase yields. The procedure is carried out in a series of stages namely: Selection of a suitable source of explant (stage 0), Conduction of surface sterilization followed by Isolation of explants and Initiation into culture (stage 1), Production of suitable propagules with shoots (stage 2), Plantlets preparation via Shoot Elongation and Rooting for transferal to natural environment (stage 3) and finally, Transfer of plantlets to soil and exposure to the natural environments (stage 4).
The media utilized for propagation is crucial as there needs to be adequate nutrients available to ensure the plant gets the correct environment to grow. The most popular and most common growth media utilized for tissue culture is Murashige and Skoog Media. (Phillips & Garda, 2019). Growth regulators are also added in some cases when there is a specific type of growth that is preferred. Growth regulators are plant hormones that influence the forms of growth that they undergo. (Hill & Schaller, 2013). PGRs include auxins, gibberellins and cytokinin that in their own way impact and influence the growth forms of plants.
Sterilization is also an integral part of tissue culture as this allows for the destruction of microorganisms and pathogens. The introduction of contaminants via microorganisms into culture is detrimental to the process as they can cause depletion to the nutrients within the media or cause damage to the explant itself by feeding on it for its own benefit. Bleach is most times utilized to clean the plant materials before they are inoculated into media to ensure bacterial and fungal spores’ growth are not facilitated. (Bello Oluwakemi, Esan Edward B, & Obembe Olawole., 2018). A laminar flow hood is the ideal apparatus for use to maintain the aseptic conditions when initiating explants into culture. 70% ethanol should be used to wipe down the surfaces of the flow hood, but not the back/top, depending on where the HEPA filter is. The HEPA (high efficiency particulate air filter) is the main filter used in the laminar flow as this is designed to keep microorganisms out of the system creating a sterile environment for culturing to take place. (A.O.Miller, M.W.Henry, & B.D.Brause, 2017). The materials that enter the laminar flow also must be sanitized with 70% ethanol along with the hands of the experimenter to ensure no form of contaminant is tracked into the environment to lead to the propagation of microbes to cause contamination.
Locally, tissue culture as aforementioned has been employed extensively in the agriculture sector to ensure demands are met for both exported good and those for internal distribution. Crop item which are grown via tissue culture in Jamaica are Bananas, Pineapples, Ginger, and Yam etc. (Davis, 2015). It has managed to cut growth times almost in half for these crops and even facilitated growth out of their normal seasons to ensure no supply chain fallout.
By Ricardo M. L. Hickling
References
A.O.Miller, M.W.Henry, & B.D.Brause. (2017). Prevention of joint infections. In J. C. Arts, & J. Geurts (Eds.), Management of Periprosthetic Joint Infections (pp. 3-23). Retrieved May 2022, from https://doi.org/10.1016/B978-0-08-100205-6.00001-X
Bello Oluwakemi, A., Esan Edward B, & Obembe Olawole., O. (2018). Establishing surface sterilization protocol for nodal culture of Solanecio biafrae. Conf. Series: Earth and Environmental Science 2. Retrieved May 2022, from doi:10.1088/1755-1315/210/1/012007
Davis, G. (2015). SRC Providing Disease-Free Plants For Agriculture Sector. Retrieved May 2022, from JAMAICA INFORMATION SERVICE: https://jis.gov.jm/src-providing-disease-free-plants-for-agriculture-sector/#:~:text=farmers%20and%20horticulturists.-,These%20include%20banana%2C%20plantain%2C%20pineapple%2C%20ginger%2C%20several%20varieties,Ann's%20Bay%20Parish%20Library%2C%20St.
Hill, K., & Schaller, G. E. (2013). Enhancing plant regeneration in tissue culture. Plant Signaling & Behavior. doi:10.4161/psb.25709
Hussain, A., Qarshi, I. A., Nazir, H., & Ullah, I. (2012). Plant Tissue Culture: Current Status and Opportunities. In A. Leva, & L. M. Rinaldi (Eds.), Recent Advances in Plant in vitro Culture. doi:10.5772/50568
Phillips, G. C., & Garda, M. (2019). Plant tissue culture media and practices: an overview. In Vitro Cellular & Developmental Biology, 242-257. Retrieved May 2022, from https://pubag.nal.usda.gov/catalog/6382024