AGR 506 AGRICULTURAL BIOTECHNOLOGY

Lab Report

Name: Nur Wajihah bt Jenal @ Zainal

Student no: 2019406202
Group: M3AT2263A
Name: Nur Wajihah bt Jenal @ Zainal id: 2019406202 Group: M3AT2262A
Title: tissue culture at home
Objective:
1. To understand procedures for growing plant in nutrient medium without soil.
2. To understand importance of sterile techniques.

Overview: Tissue culture is the growth of tissues or cells in an artificial medium separate
from the parent organism. This technique is also called micropropagation. This is typically
facilitated via use of a liquid, semi-solid, or solid growth medium, such as broth or agar. In
this experiment we used agar as the medium. Tissue culture is an important tool for the
study of the biology of cells from multicellular organisms. Using the appropriate growing
conditions for each explant type, plants can be induced to rapidly produce new shoots, and,
with the addition of suitable hormones new roots. These plantlets can also be divided,
usually at the shoot stage, to produce large numbers of new plantlets. The new plants can
then be placed in soil and grown in the normal manner.
Materials: 45g agar powder, plants, 3g 8N:8P:8K fertilizer, fan, confined space,1L filtered
water, 60% chlorox, disinfectant spray, tweezer, knife, glove, face mask, container.
Calculation:
60% chlorox=80ml filtered water+ 120ml chlorox
50% agar percentage in 200 ml= 45/90 g x 400ml = 50%
200ml 0.00008% NPK = 0.08/1000ml
Procedure:
Media preparation:
1. 3 grams of fertilizer was diluted with 1L of water
2. 200 ml of diluted fertilizer was poured into container that has been disinfected.
3. Then 45g of agar was added into the solution
4. The solution then microwaved till boiled. Then the solution is removed, and container
was wrapped with saran wrap and put aside till it solidified.
Workspace setup:
1. Tweezer and knife were sterilized in 100°C hot boil water then was put in 60%
chlorox.
2. workspace and hand were disinfected before the experiment started using
disinfectant spray.
3. fan was on which acts as laminar air flow after workspace has been disinfected.
Tissue culture:
1. Sanitized hand. Wear a mask and put-on glove before experiment started.
2. Sprayed the whole container of the media using disinfectant spray to avoid
contamination.
3. Cut desired plant part using knife and tweezer.
4. sanitized the plants by dipping it in 60% chlorox for 10 minutes.
5. After 10 minutes, take out the plants and plant were dipped into filtered water three
times.
6. Pieces of plants then were dried using paper towel that has been sprayed with
disinfectant spray.
 7. Put pieces of plant in the media.
8. Make sure the wound touches the media.
9. Used saran wrap to cover the media.
10. Avoid exposed the media in sunlight and environment as it can highly risk the
contamination.
Result:
Day 1

Figure 1 Day 1 of culture media show no contamination

Day 2

Figure 2 Day 2 of culture media show (white colonial around 1of the plants) sign of microbial or contamination.
Day 3

Figure 3 Day 3 show contamination on almost all the plants

Day 4:

Figure 4 Day 4 shows all the plant get contaminations.

Discussion: The experiment considered as fail due to contamination. No sign of any growth
of plants. Contamination is common in tissue culture. regular sanitizing throughout the tissue
culture procedure is needed to reduce contamination. Sterile conditions require the complete
absence of microorganisms including bacteria, fungi, yeast, and their spores. This is to
ensure the succeed of the tissue culture. Clear work area of items not in immediate use and
use face mask. There are common types of culture contamination such as bacterial, fungal
(including molds) and yeast contamination are usually visible to the unaided eye as rapid-
onset turbidity and color change of the culture medium. Using effective amount disinfectant
which is 70% alcohol e.g., ethanol helped against bacteria and most viruses. Use saran
wrap to reduce contamination. Store tissue culture in the dark because adverse effects of
light exposure on cells in culture are attributable to riboflavin-related production of free
radicals followed by generation of peroxides and other photoproducts. Photo effects
disappear when riboflavin is deleted from the media.
Conclusion: different techniques in plant tissue culture offer advantages such as the
production of exact copies of plants that produce particularly good flowers, fruits, or have
other desirable traits, production of plants from seeds that otherwise have very low chances
of germinating and growing and many more. A single microbe or fungi can contaminate and
failed the tissue culture.

References
Campbell, N. A., Urry, L. A., Cain, M. L., Wasserman, S. A., Minorsky, P. V., & Reece, J. B. (2017).
Biology: A Global Approach. In N. A. Campbell, L. A. Urry, M. L. Cain, S. A. Wasserman, P. V.
Minorsky, & J. B. Reece, Biology: A Global Approach (pp. 905-906). New York: Pearson
Education.

J, R. (2020, Jun 01). Plant Cell Technology. Retrieved from Common Types of Tissu Culture
Contamination: https://www.plantcelltechnology.com/pct-blog/common-types-of-tissue-
culture-contamination-/

M, Y. (2012). The Prerequisite of the Success in Plant Tissue Culture: High Frequency Shoot
Regeneration. Recent Advances in Plant in vitro Culture, 63.

Tissue Culture. (2015, July 07). Retrieved from https://www.britannica.com/science/tissue-culture