effect of saccharomyces cerevisiae extract on …...abstract—the aim of this study was important...

Abstract—The aim of this study was important to know

ability of using yeast Saccharomyces cerevisiae extract to

amendment mycelium growth and primordia (pins) formation of Pleurotus salmoneostramineus and Pleurotus

cornucopiae var. citrinopileatus. This study was achieved by

preparation yeast extract by ice crystal formation then

dissolved it and used with PDA medium in 1, 5, 10 and 15 g

L-1. Mycelial growth rate (MGR) of Pleurotus

salmoneostramineus was 12.90 mm day-1 in PDA with 1 g L-1

of extract of yeast Saccharomyces cerevisiae, then decreased

when the concentration of yeast extract higher more than 1

g L-1. Pleurotus salmoneostramineus primordia were

appeared over PDA dish with 1 g L-1 of yeast extract in

more number, compared as less number on PDA alone.

Pleurotus cornucopiae showed less growth rate and no

appear primordia compared with of Pleurotus

salmoneostramineus.

Index Terms—pleurotus spp. saccharomyces cerevisiae.

primordia. mycelial growth rate. yeast extract.

I. INTRODUCTION

Genus Pleurotus spp. belongs to Basidiomycetes, order:

Agaricales, family: Pleurotaceae [1]. It called oyster

mushroom is an edible mushroom, can be cultivated on a

wide variety of substrates containing lignin and cellulose.

It has nutritional and medicinal properties [2]. Pleurotus

ostreatus used as biosorbent for cadmium (II) removal in

industrial wastewater [3]. Pleurotus ostreatus and

Saccharomyces cerevisiae were used in combined

mixture to upgrade the nutritional value of maize stalks

using solid state fermentation technique, which lead to

increase protein content of maize stalks in vitro [4].

Abdulhadi et al. [5] was used concentration 10% of

boiled yeast extract to increase yield and biological

efficiency of Pleurotus ostreatus that increased dry

weight and protein content in fruiting bodies. Zone

inhibition of Saccharomyces cerevisiae was 12.33-9.67

mm (40 mg ml-1

of P. ostreatus extract) [6].

Manuscript received February 21, 2014; 2014.

Yeasts fermented the produced free sugar [7]. Yeast

Extract is the water-soluble portion of autolyzed yeast.

The autolysis is carefully controlled to preserve naturally

occurring B-complex vitamins. Yeast Extract is prepared

and standardized for bacteriological use and cell cultures,

and is an excellent stimulator of bacterial growth. Yeast

Extract is typically prepared by growing Baker’s yeast,

Saccharomyces spp., in a carbohydrate-rich plant medium.

The yeast is harvested, washed, and resuspended in water,

where it undergoes autolysis, i.e., self-digestion using the

yeast’s enzymes. Yeast Extract is the total soluble portion

of this autolytic action. The autolytic activity is stopped

by a heating step, then Yeast Extract result [8]. Baker’s

yeast (Saccharomyces cerevisiae) is defined as primary

yeast because the yeast is grown for the specific purpose

of being used as a substrate in a bioprocess or as a food

product/flavoring. Yeast extract is used by the health food

industry as an inexpensive source for many of their

vitamins, and has long been recognized as a major source

of B-complex vitamins. Yeast extract, as a substrate in a

media formulation, supplies not only vitamins, but also

proteins, carbohydrates and some micronutrients [9].

The yeast extract 2 g L-1

induced mycelium growth of

Pleurotus tuber-regium (Fr.) Singer [10]. Fathy and Farid

[11] were referring to the yeast helps to activate binary

division for production the nucleic acids and proteins. El-

Din and Hendawy [12] were used dry yeast on increased

rates of growth Borago officinalis plants. Dry yeast is

producing growth regulators [13].

This study is important to know ability of using

Saccharomyces cerevisiae extract to amendment

mycelium growth and primordia formation of Pleurotus

salmoneostramineus and Pleurotus cornucopiae var.

citrinopileatus.

II. MATERIAL AND METHODS

A. Strains

Fungi strains included mushrooms and yeast. Two

oyster mushrooms species were investigated. Pleurotus

282014 Engineering and Technology Publishingdoi: 10.12720/jolst.2.1.28-31

Mustafa N. Owaid1,2

, Behar M. A. Al-Ani2, and Mowafak M. Muslat

3

1AL-Athar School, Heet Education, Ministry of Education, Heet, Anbar 31007, Iraq

2Department of Biology, College of Science, University of Anbar, Ramadi, Anbar 31001, Iraq

3 Department of Horticulture, College of Agriculture, University of Anbar, Ramadi, Anbar 31001, Iraq

Email: [email protected]

revised September 11,

Journal of Life Sciences and Technologies Vol. 2, No. 1, June 2014

Effect of Saccharomyces cerevisiae Extract on

Mycelium Growth and Primordia Formation of

Pleurotus salmoneostramineus and Pleurotus

cornucopiae (In Vitro)

cornucopiae var. citrinopileatus (bright yellow) and

Pleurotus salmoneostramineus (pink) are obtained from

Mushroom Box Company, Monmouth, UK, in form

spawn and sub cultured it on Potato Dextrose Agar (PDA)

medium at 25 C° for this experiment. Yeast was

Saccharomyces cerevisiae is obtained in form dry powder

from baker’s yeast, China.

B. Preparation of Yeast Extracts

Stock solution (40 g L-1

) of Saccharomyces cerevisiae

was dissolved in distilled water containing 1% sucrose

[5], left at 25 C° for 4 h then froze for 2 days. Yeast

extract ice was dissolved at room temperature 25 C°,

prepared 1, 5, 10 and 15 g L-1

with potato dextrose agar

(PDA), autoclaved for 15 min. at 120 C° with pressure

1.5 psi and poured in dishes plates 85 mm. PDA was used

as control [14].

C. Effect of Extracts of Yeast on Mycelium Growth in

Solid Medium

When cells are returned to complete glucose medium

they quickly resume proliferation [15]. Mycelial growth

rate of Pleurotus spp. was determined after five days.

And also time of over growth of these mycelia on whole

dish was calculated.

D. Statistical Analysis

Experimental values are given as means. Statistical

significance was determined by Completely Randomized

Design (CRD) in (two ways) analysis (ANOVA) by using

GenStat Discovery Edition computer program version 7

DE3 (VSN International Ltd., UK). Differences at P>

0.05 were considered to be significant. The experiments

used three replicates.

III. RESULTS

A. Mycelial Growth of Pleurotus spp. on PDA with and

Without Extract of Yeast Saccharomyces Cerevisiae

Table I showed that Pleurotus salmoneostramineus

gave maximum diameter of mycelial colony (64.00) mm

on (1 g L-1

of yeast extract) with significant difference

(P> 0.05) compared with other concentrations (5, 10 and

15 g L-1

), while was 48.75 mm for Pleurotus cornucopiae

var. citrinopileatus on the same medium. Growth of

mycelia decreased when the yeast (Saccharomyces

cerevisiae) extract was found with PDA medium.

Minimum diameter of colony was 26.00 mm on PDA

with 1 g L-1

of yeast extract for P. cornucopiae.

Mycelial growth rate (MGR) of Pleurotus

salmoneostramineus was 12.90 mm day-1

in PDA with 1

g L-1

of extract of yeast Saccharomyces cerevisiae, then

decreased when the concentration of yeast extract higher

more than 1 g L-1

. Pleurotus cornucopiae showed less

growth rate compared with of Pleurotus

salmoneostramineus (Table II).

TABLE I. DIAMETER OF MYCELIAL GROWTH OF PLEUROTUS SPP. ON SOLID MEDIA AFTER FIVE DAYS (MM)

TABLE II. MYCELIAL GROWTH RATE OF PLEUROTUS SPP. ON SOLID MEDIA AFTER FIVE DAYS (MM DAY-1)

B. The Time of Over Growth of Pleurotus spp. on PDA

with and Without Extract of Yeast Saccharomyces

Cerevisiae

The time of over growth was 8 days for concentrations

5, 10, 15 g L-1

of yeast extract compared as 6 and 7 days

for media of PDA and PDA with 1 g L-1

respectively. Pleurotus cornucopiae take more time when yeast extract

was found in medium which covered PDA with 9.33 days,

whereas PDA with extract of Saccharomyces cerevisiae

was showed 12.33, 14.33, 15.00 and 16.00 days for

concentrations 1, 5, 10 and 15 g L-1

(Table III).

TABLE III. THE TIME OF OVER GROWTH OF PLEUROTUS SPP. IN PETRI DISHES 85 MM

Oyster mushrooms (PDA)

0 g L-1 Yeast (Saccharomyces cerevisiae ) extract with PDA

Mean of oyster

mushrooms 1 g L-1 5 g L-1 10 g L-1 15 g L-1

P. salmoneostramineus 84.00 64.50 55.00 55.50 50.83 61.97

P. cornucopiae 48.75 26.00 28.50 30.50 31.83 33.12

Yeast extract mean 66.37 45.25 41.75 43.00 41.33 -

LSD P> 0.05 Oyster M.=2.594 , Yeast=4.101 , Oyster M.* Yeast =5.800

Oyster mushrooms (PDA)

0 g L-1 Yeast (Saccharomyces cerevisiae ) extract with PDA Mean of oyster

mushrooms 1 g L-1 5 g L-1 10 g L-1 15 g L-1


P. cornucopiae 9.75 5.20 5.70 6.10 6.37 6.62


LSD P> 0.05 Oyster M.=0.519 , Yeast=0.820 , Oyster M.* Yeast=1.160

Oyster mushrooms

(PDA) 0 g L-1

Yeast (Saccharomyces cerevisiae) extract with PDA Mean of oyster

mushrooms 1 g L-1 5 g L-1

10 g L-1 15 g L-1


P. cornucopiae 9.33 12.33 14.33 15.00 16.00 13.40


LSD P> 0.05 Oyster M.= 0.2409 , Yeast=0.3808, Oyster M.* Yeast=0.5386

292014 Engineering and Technology Publishing


C. Primordia Appearance of Pleurotus spp. over PDA

(In Vitro)

Pleurotus salmoneostramineus primordia were

appeared over PDA dish with 1 g L-1

of yeast extract in

more number, compared as less number on PDA without

yeast extract after 11.00 and 10.33 days respectively

(Table IV). The concentration 1 g L-1

of yeast extract did

not effect on P. salmoneostramineus that gave more of

primordia in petri dish 1 g L-1

(Fig. 1) compared with

PDA but other concentrations did not appear any

primordia.

TABLE IV. TIME OF PRIMORDIA APPEARANCE OF PLEUROTUS SPP. ON PDA WITH YEAST EXTRACT (DAYS)

No: No appear any primordia.

Figure 1. Primordia appearance of Pleurotus salmoneostramineus on

solid media which contained yeast extract after 13 days 0g L-1

(PDA): Potato dextrose agar without Saccharomyces cerevisiae extract. 1, 5, 10

and 15 g L-1: PDA with S. cervisia extract.

IV. DISCUSSION

The less concentration (1 g L-1

) of yeast

Saccharomyces cerevisiae extract was showed best

number of Pleurotus salmoneostramineus primordia compared with PDA alone while other concentrations did

not show any primordia, that due to method of

preparation the extract (freeze-thaw injury then heat

shock after sterilization) and other compounds

accumulating in heat-stressed cells: Trehalose and

Sphingolipids [16] when high concentrations of yeast

extract, which inhibit primordia formation (Fig. 1) and

mycelium growth rate (Table II). In synthetic media; low

concentration of yeast extract is generally employed in

the concentration of 0.3% - 0.5% [8]. The heat shock-

induced proteins and their functions from extract are

important to Pleurotus spp. growth in the suitable

concentration. Many of the proteins are involved directly

in the degradation or reactivation of damaged proteins

[16], which lead to speed of form primordia of P.

salmoneostramineus (in vitro) (Fig. 1). Mycelial growth

rate of Pleurotus salmoneostramineus was better than P.

cornucopiae because of genetic information between

these strains [17]. Other studies on P. ostreatus was achieved by

scientists (in field) like: Abdulhadi et al. [5] was used

concentration 10% of yeast extract to increase yield and

biological efficiency of Pleurotus ostreatus that increased

protein content in fruiting bodies, whereas Alhaje-Abod

et al. [18] used bread yeast at 1g kg-1

or 2 g kg-1

wet

substrate that lead to increase the dry weight and protein

content yield and biological efficiency of Pleurotus

ostreatus. Abdulhadi et al. [5] referred to role sucrose 1%

with yeast extract to increase the growth Pleurotus

ostreatus.

ACKNOWLEDGMENT

This work was financially supported by Department of

Biology, College of Science, University of Anbar, Iraq.

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Oyster mushrooms (PDA) 0 g L-1

Yeast (Saccharomyces cerevisiae) extract with PDA

1 g L-1 5 g L-1 10 g L-1 15 g L-1

P. salmoneostramineus 10.33 11.00 No No No

P. cornucopiae No No No No No



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Mustafa N. Owaid was born in Heet, Anbar, Iraq, on 1981. He graduated the Bachelor

Degree of Science (B.Sc.) Biology from

University of Anbar, Iraq on 2003. He obtained his Master degree of Science (M.Sc.)

in Microbiology from University of Anbar, Iraq on 2010. He was completed Ph.D. in

Mycology from the same university on 2014.

Since 2005, he is a teacher at the Ministry of Education, Iraq. His research area is

mushroom and their applications. He was study Ph.D. in mushroom field. Now, he also is member in British Mycological Society (BMS) in

UK, International Society for Mushroom Science (ISMS) in South

Africa and North American Mycological Association (NAMA) in USA. Also he had memberships in many world associations. His website

www.alheeti.com/mustafa

Behar M. A. Al-Ani graduated the Bachelor Degree of Science (B.Sc.)

Biology from University of Anbar, Iraq. He obtained here Master degree of Science (M.Sc.) in Plant Pathology from University of Anbar,

Iraq on 2010. Now, she Assist. Lecture in Department of Biology, University of Anbar.

Mowafak M. Muslat (Ph.D) is lecture in Department of Horticulture,

College of Agriculture, University of Anbar, Iraq. He is Assist. He interested in mushroom field.



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