This document describes an experiment on UV treatment of the fungus Monuscus sp. to produce mutants. UV treatment at distances of 20cm and 40cm for 40-60 minutes produced albino and pigment-less mutant varieties. When sub-cultured repeatedly in light, the mutants regained pigmentation. The document also provides background on Monuscus sp., its uses, and growth conditions. The objective was to observe morphological changes from UV-induced mutations.

1. UV TREATMENT IN
MONUSCUS SP.
Presented by Steve Irwin

2. INTRODUCTION
Monuscus sp is a fungus that belong to class
ascomycota and family Monasaceae. Under
microscope Monuscus purpureus appear
filamentous fungi. Can be obtained from tofu and
rice.
Some special characteristics of this fungi are :
This fungus bears asexual reproducing spores
called conidia and bears sexual reproducing
ascus. Monuscus purpurea produces red and
yellow pigment.
Monuscus sp have many commercial and medicinal
use.

3. COMMERCIAL AND MEDICINAL
USAGE
1. Pigment is used as food dyes, preservative, and
food condiments.
2. Ingredient in making rice wine, tofu, etc.
3. Red rice is used as medicine to treat dysentery,
external and internal trauma
4. Fermentate of this fungus is used to bring down
cholesterol level and triglyceride level.
5. This fungus also contain antibiotic property.

4. OBJECTIVE OF THE EXPERIMENT
 The main objective of the lab was to produce
colored mutant and albino variety of Monuscus
sp. by UV treatment and observe the morphology
of the various mutants.
 Ultraviolet light causes DNA mutation in some
cases by forming pyrimidine dimers ‘T-T’ (Klug et
al. 2007).
 Prediction was to observe change in colony
morphology, or reduced pigmentation in the
Monuscus sp.

5. GROWTH MEDIUMS FOR THE
FUNGUS
PDA – Potato Dextrose Agar. 39 grams of PDA
powder in 1000 ml of distilled water and
autoclave.
MYS- Malt Yeast Starch. 30 grams cassava starch,
3 gram malt extract, 3 gram yeast extract, 5
gram peptone, 15 gram agar in 1000 ml distilled
water and autoclave.
White Rice.

6. FUNGAL CULTURE
 Thai
 Vietnam

7. Isolate fungus
PDA
Store
Grow on PDA. 2
Weeks incubation.
Glycerol
Suspension
Isolate single
spore conidia.
3 days
incubation on
PDA
Single spore
clones
UV treatment
Grow on MYS to
see Mutation
Store

8. UV TREATMENT
 In order to do UV treatment fungus must be
cloned from conidia and grown in PDA.
 Glycerol suspension was made with monoclonal
fungus with 1% glycerol.
 The UV treatment can be done by taking 1 ml
glycerol suspension into empty Petri dishes and
treat it in 20 cm, 40 cm, 60 cm in different times
such as 20 min, 40 min, 60 min.

9. Sample Time Distance 1 Distance 2
Vietnam 10 single spore
# 1 (V10 SS 1)
20 minutes 20 cm 40 cm
40 minutes 20 cm 40 cm
60 minutes 20 cm 40 cm
Sample Time Distance 1 Distance 2
Vietnam 6 single spore #
2 (V10 SS2)
20 minutes 20 cm 40 cm
40 minutes 20 cm 40 cm
60 minutes 20 cm 40 cm

10. RESULTS OF ULTRAVIOLET
TREATMENT
Sample
(Monuscus
purpurea.)
UV
Treatment
Time
Distance Observations
Vietnam 10
(V10 SS 1 )
20 Minutes
20 cm
Negative
40 Minutes Negative
60 Minutes Negative
Vietnam 10
(V10 SS 1)
20 Minutes
40 cm
Negative
40 Minutes Negative
60 Minutes Negative

11. Sample
(Monuscus
purpurea.)
UV Treatment
Time
Distance Observations
Vietnam 6 Single
Spore 2 (V6 SS 2 )
20 Minutes
20 cm
Negative
40 Minutes
White sector was
observed in UV
treated colonies
compared to
normal.
60 Minutes
White colonies
compared to
Normal colony.
Sectors of
different
morphology.
Vietnam 6 single
spore 2 (V6 SS 2)
20 Minutes
40 cm
Negative
40 Minutes
Mutation was
observed and
white colony of
fungus was seen
without pigments.
60 Minutes Negative

12. SUB CULTURING MUTANT
V6 SS 2 mutant was sub-cultured into PDA and
MYS if more than 2 times in presence of light
will reverse pigmentation in the Monuscus sp.
Wong, C., and Lin, Y., et al. reported that slightly
colored and albino mutants of Monuscus
purpureus produced pigments after having been
exposed to UV for 5 minutes.

13. B) Vietnam 6 single spore 2
colonyA) Vietnam 6 single spore
2 Mutant 3 colony
Figure 1.1

14. Figure 1.3:- V6 SS2 M1a Pigment less single spore colony of Monuscus
purpureus on MYS.
Date of Sub culturing: April 2, 2009.
Date of Snap Shot: April 6, 2009.
Figure 1.2
A) B)

15. Figure 1.3:- V6 SS2 M3b comparison between albino and normal V6 SS2 pigment
production on PDA.
Date of Sub culturing: April 2, 2009.
Date of snap shot: April 6, 2009.
Figure 1.3
A) B)

16. Figure 1.4:- V6 SS2 M3b comparison between albino and normal V6 SS2 pigment
production on PDA.
Date of Sub culturing: April 2, 2009.
Date of Snap Shot: April 9, 2009.
Figure 1. 4
A) B)

17. Figure 1.5: - Comparison between normal pigment producing V6 SS2 in rice medium
and pigment less mutant V6 SS2 M3 in rice medium.
Date of inoculation: - April 13, 2009
Date of snap shot: - April 24, 2009
Figure 1. 5

18. DISCUSSION
UV treatment produced mutant variety at distance
20 cm and 40 cm, in 40 minutes and 60 minutes.
Reverse in pigment production in albino mutant
was observed after sub-culturing the mutant
more than two times in presence of light.
Albino mutant can be retained in dark in MYS
medium.

19. CONCLUSION
It was possible to produce mutant variety of
Monuscus sp with Ultra Violet light.
It can be predicted that Ultra violet light interrupt
with red pigment producing gene.
For further studies, molecular techniques can be
used to isolate red pigment gene.

20. ACKNOWLEDGEMENT
I would like to thank Dr. Kung for guiding me
through out the project. I would like to think Ms.
Sonya and Isha also for helping me when I
needed their help.

21. REFERENCE
 Kranz, C. Pantiz, C. and Kunz, B. (1992). Biotransformation of Free Fatty Acids in
Mixtures to Methyl Ketones by Monuscus Purpureus. Applied Microbiology and
Biotechnology, 36, pp. 436-439.
 Lin, Ching-Fwu. (1973). Isolation and Cultural Conditions of Monuscus sp. for the
Production of pigment in a Submerged Culture. Journal of Fermentation and
Technology, 51 (6), pp. 407-414.
 Lotong, N. and Suwanarit, P. (1989). Fermentation of ang-kak in plastic bags and
regulation of pigmentation by initial moisture content. Journal of Bacteriology, 68,
pp. 567-570.
 Omamor, B.I. Eziashi, E. I. and Adekunle, A.A. (2008). Carbon Nutrition in Relation
to Growth of Three Monuscus sp. isolated from decaying date fruits. African Journal
of Microbiological Research, 2, pp. 153-155.
 Wong, Hin-Chung and Bau, Yun-Shen. (1978). Morphology and Photo responses of
Fast-Neutron and X-Ray-Induced Strains of Monuscus Purpureus. Mycologia, 70, pp.
645-648.
 Wong, Hin-Chun and Koehler, P. (1981). Production and Isolation of an Antibiotic
from Monuscus Purpureus and its relationship to pigment production. Journal of
Food Science, 46 (2), pp. 589-592.
 Wong, H., C. Lin, Y., C. and Koehler, P. E. (1981). Regulation of Growth and
Pigmentation of Monuscus purpureus By Carbon and Nitrogen Concentrations.
Mycologia, 73, pp. 649-654.
 Klun, William, S. Cummings, Michael, R. and Spencer, Charlotte, A. (2007). Gene
Mutation, DNA Repair and Transcription. In: Klun, William, S. Cummings, Michael,
R. and Spencer, Charlotte, A. Essentials of Genetics. 5th
Edition. USA: William S.
Klug and Michael R. Cummings, pp. 314-315.
 Anonymous. (2001). Red Yeast Rice (Monuscus purpureus). [Online] Penton Media.
Available from: http://www.healthwell.com/healthnotes/Herb/Red_Rice.cfm?path=hw
[25 September 2001]