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Agraarteadus
Journal of
Agricultural Science
Vol. 32 No. 1
June 2021
JOURNAL OF AGRICULTURAL SCIENCE
AGRAARTEADUS
Tartu 2021
Akadeemilise Põllumajanduse Seltsi väljaanne
––––––––––––––
Estonian Academic Agricultural Society publication
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1 • XXXII • 2021, p-ISSN 1024-0845, e-ISSN 2228-4893
Kaastööde esitamiseks ja lugemiseks külastage: https://agrt.emu.ee
For online submission and open access visit: https://agrt.emu.ee/en
Merko Vaga Estonian University of Life Sciences, Estonia
Rein Viiralt Estonian University of Life Sciences, Estonia
Väljaandmist toetavad Eesti Maaülikool ja Eesti Taimekasvatuse Instituut
Supported by Estonian University of Life Sciences and Estonian Crop Research Institute
Abstracted / indexed: AGRICOLA, AGRIS, CABI, CABI Full Text, DOAJ, EBSCO, SCOPUS
Trükk / Print: Eesti Ülikoolide Kirjastus OÜ. Kaanepilt / Cover image by 0melapics / Freepik
p-ISSN: 1024-0845, e-ISSN: 2228-4893
Priit Tammeorg University of Helsinki, Finland
Vidmantas Pileckas Lithuanian University of Health Sciences, Lithuania
Marek Gaworski Warsaw University of Life Sciences, Poland
Margareta Emanuelson Swedish University of Agricultural Sciences, Sweden
Edenio Detmann Federal University of Viçosa, Brasil
Iveta Kociņa Institute of Food Safety, Animal Health and Environment "BIOR", Latvia
Zita Kriaučiūnienė Aleksandras Stulginskis University, Lithuania
Jan Philipsson Swedish University of Life Sciences, Sweden
Baiba Rivza Latvia University of Life Sciences and Technologies, Latvia
Aldona Stalgienė Institute of Economics and Rural Development of the Lithuanian Centre
Vita Tilvikienė Lithuanian Research Centre for Agriculture and Forestry, Lithuania
Csaba Jansik Natural Resource Institute Finland, Luke, Finland
Sven Peets Harper Adams University, UK
for Social Sciences
Martti Esala Natural Resource Institute Finland, Luke, Finland
Alo Tänavots Estonian University of Life Sciences
Peatoimetaja / Editor-in-chief
Toimetuskolleegium / Editorial Board
Tegevtoimetaja / Managing Editor
Irje Nutt Estonian University of Life Sciences
Toimetajad / Editors
Maarika Alaru Estonian University of Life Sciences
Reelika Rätsep Estonian University of Life Sciences
Marko Kass Estonian University of Life Sciences
Evelin Loit Estonian University of Life Sciences
Timo Arula University of Tartu, Estonia; University of Maryland, USA
Berit Bangoura University of Wyoming, USA
Tanel Kaart Estonian University of Life Sciences
David Arney Estonian University of Life Sciences
Marten Madissoo Estonian University of Life Sciences
National Academy of Agrarian Sciences of Ukraine
Gunita Deksne Institute of Food Safety, Animal Health and Environment "BIOR", Latvia
Volodymyr Bulgakov National University of Life and Environmental Sciences of Ukraine
Ants-Hannes Viira Estonian University of Life Sciences
Nõukogu / Advisory Board
Ants Bender Estonian Crop Research Institute, Estonia
Edward H. Cabezas-Garcia National University of San Marcos, Peru
Toomas Orro Estonian University of Life Sciences
A1
AGRAARTEADUS
JOURNAL OF AGRICULTURAL SCIENCE
1 XXXII 2021
Väljaandja: Akadeemiline Põllumajanduse Selts
Peatoimetaja: pm-dr Alo Tänavots
Tehniline toimetaja: pm-mag Irje Nutt
Aadress: Fr. R. Kreutzwaldi 1, 51006 Tartu
e-post: jas@emu.ee
www: https://aps.emu.ee, https://agrt.emu.ee
Agraarteaduses avaldatud teaduspublikatsioonid on retsenseeritud
SISUKORD
TEADUSARTIKLID
A.M.A. Al-Shammari, G.J. Hamdi
Genetic diversity analysis and DNA fingerprinting of tomato breeding lines using SSR
markers .................................................................................……………................. 1
V. Bandura, B. Kotov, S. Gyrych, V. Gricshenko, R. Kalinichenko, O. Lysenko
Identification of mathematical description of the dynamics of extraction of oil materials
in the electric field of high frequency........................................................................... 8
A. Bender
Kasvuregulaatori ja kevadise täiendava lämmastik-väetise mõju põldtimuti (Phleum
pratense L.) seemnesaagile ja seemnete kvaliteedile …………..……………..…………. 17
O.O. Borshch, S. Ruban, O.V. Borshch
Review: The influence of genotypic and phenotypic factors on the comfort and welfare
rates of cows during the period of global climate changes .......................….......……. 25
L.F. Gar, B.R. Bader, J.S.E. Al-Esawi, M.A. Abood, M. O. Sallume, G.J. Hamdi
Response of onion growth and yield grown in soils of semi-arid regions to foliar
application of iron under water stress conditions ..................................................... 35
Y. Ihnatiev, V. Bulgakov, V. Bonchik, Z. Ruzhylo, A. Zaryshnyak, V. Volskiy, V. Melnik, J. Olt
Experimental research into operation of potato harvester with rotary tool ................... 41
P. Karis, H. Jaakson, K.i Ling, M. Runin, M. Henno, A. Waldmann, M. Ots
Body condition effects on dry matter intake and metabolic status during the transition
period in Holstein dairy cows ................................................................................... 49
M. Kvitko, N. Getman, A. Butenko, G. Demydas, V. Moisiienko, S. Stotska, L. Burko,
V. Onychko
Factors of increasing alfalfa yield capacity under conditions of the forest-steppe ........ 59
R. Kõlli, T. Tõnutare
Rähkmuldade seisund ja levik eesti põllumajandus- ning metsamaastikes ................... 67
I. Mubarak, M. Janat
Bottle gourd (Lagenaria siceraria L.) crop response to different planting densities
under both drip and wide-spaced furrow irrigation methods .....................…………… 79
A2
E. Nugis, J. Kuht, A. Komarov
Potato yield forecast by using guttation test method in household laboratory conditions ... 86
J. Olt, V. Adamchuk, V. Kornuchin, V. Melnik, H. Kaletnik, Y. Ihnatiev, R. Ilves
Research into the parameters of a potato harvester's potato heap distributor, and the
justification of those parameters ...................................................…………………… 92
Z. Pacanoski, A. Mehmeti
Efficacy and selectivity of pre-em herbicide on dependance of soil types and
precipitation in sunflower crop ..........................................................................…… 100
A. Panfilova, V. Gamayunova, N. Potryvaieva
The impact of nutrition optimization on crop yield and grain quality of spring barley
varieties (Hordeum vulgare L.) .....................................................…………………… 111
P. Prakash, B. Karthikeyan, M.M. Joe
Chemotactic responses of sweet flag (Acorus calamus L.) root exudates and evaluation
of inoculation effects on its growth ........................................................................... 117
S. Shrestha, S. Bhattarai, R.K. Shrestha, J. Shrestha
Effect of different substrate sterilization methods on performance of oyster mushroom
(Pleurotus ostreatus) ..........………..................................................................……… 127
S. Subedi, S. Neupane
Antifungal assessment of plant extracts, biocontrol agents and fungicides against
Fusarium verticillioides (Sacc.) causing ear rot of maize ................................……… 133
O. Ulyanych, S. Poltoretskyi, V. Liubych, A. Yatsenko, V. Yatsenko, O. Lazariev,
V. Kravchenko
Effect of surface drip irrigation and cultivars on physiological state and productivity
of faba bean crop ..........……….......................................................................……… 139
B. Winarto, I.G. Cempaka, R.K. Jatuningtyas, S.C. Budisetyaningrum, B. Hartoyo
In vitro shoot growth performances and responses of potato (Solanum tuberosum L.)
'Muhzoto' under different treatments and explant types ..................................……… 150
KROONIKA
M. Kass, H. Kiiman
Akadeemilise Põllumajanduse Seltsi 2019. aasta tegevusaruanne ............................... 159
JUUBELID
A. Sirendi
Erkki-Gennadi Hannolainen – 80 .....................………....………………………………. 161
K. Tamm
Jaanus Siim – 80 ................................................................…….………………….... 162
IN MEMORIAM
M. Kass
Jaak Samarütel – in memoriam …….......................………....…………………………. 163
MÄLESTUSPÄEVAD
A. Bender
Mart Jaagus – 100 ...............................................................…….………………….... 165
A3
AGRAARTEADUS
JOURNAL OF AGRICULTURAL SCIENCE
1 XXXII 2021
Published by: Estonian Academic Agricultural Society
Editor in Chief: Alo Tänavots DSc (agriculture)
Technical Editor: Irje Nutt MSc (animal science)
Address: Fr. R. Kreutzwaldi 1, 51006 Tartu, Estonia
e-mail: jas@emu.ee
www: https://aps.emu.ee, https://agrt.emu.ee
Research articles published in Agraarteadus are peer-reviewed
CONTENTS
RESEARCH ARTICLES
A.M.A. Al-Shammari, G.J. Hamdi
Genetic diversity analysis and DNA fingerprinting of tomato breeding lines using SSR
markers .................................................................................……………................. 1
V. Bandura, B. Kotov, S. Gyrych, V. Gricshenko, R. Kalinichenko, O. Lysenko
Identification of mathematical description of the dynamics of extraction of oil materials
in the electric field of high frequency........................................................................... 8
A. Bender
Effect of plant growth regulator and additional nitrogen fertilization in spring on the seed
yield and seed quality of timothy (Phleum pratense L.)…………..……………..…………. 17
O.O. Borshch, S. Ruban, O.V. Borshch
Review: The influence of genotypic and phenotypic factors on the comfort and welfare
rates of cows during the period of global climate changes .......................….......……. 25
L.F. Gar, B.R. Bader, J.S.E. Al-Esawi, M.A. Abood, M. O. Sallume, G.J. Hamdi
Response of onion growth and yield grown in soils of semi-arid regions to foliar
application of iron under water stress conditions ..................................................... 35
Y. Ihnatiev, V. Bulgakov, V. Bonchik, Z. Ruzhylo, A. Zaryshnyak, V. Volskiy, V. Melnik, J. Olt
Experimental research into operation of potato harvester with rotary tool ................... 41
P. Karis, H. Jaakson, K.i Ling, M. Runin, M. Henno, A. Waldmann, M. Ots
Body condition effects on dry matter intake and metabolic status during the transition
period in Holstein dairy cows ................................................................................... 49
M. Kvitko, N. Getman, A. Butenko, G. Demydas, V. Moisiienko, S. Stotska, L. Burko,
V. Onychko
Factors of increasing alfalfa yield capacity under conditions of the forest-steppe ........ 59
R. Kõlli, T. Tõnutare
Status and distribution of ryhky soils in Estonian agricultural and forest landscapes ... 67
I. Mubarak, M. Janat
Bottle gourd (Lagenaria siceraria L.) crop response to different planting densities
under both drip and wide-spaced furrow irrigation methods .....................…………… 79
A4
E. Nugis, J. Kuht, A. Komarov
Potato yield forecast by using guttation test method in household laboratory conditions ... 86
J. Olt, V. Adamchuk, V. Kornuchin, V. Melnik, H. Kaletnik, Y. Ihnatiev, R. Ilves
Research into the parameters of a potato harvester's potato heap distributor, and the
justification of those parameters ...................................................…………………… 92
Z. Pacanoski, A. Mehmeti
Efficacy and selectivity of pre-em herbicide on dependance of soil types and
precipitation in sunflower crop ..........................................................................…… 100
A. Panfilova, V. Gamayunova, N. Potryvaieva
The impact of nutrition optimization on crop yield and grain quality of spring barley
varieties (Hordeum vulgare L.) .....................................................…………………… 111
P. Prakash, B. Karthikeyan, M.M. Joe
Chemotactic responses of sweet flag (Acorus calamus L.) root exudates and evaluation
of inoculation effects on its growth ........................................................................... 117
S. Shrestha, S. Bhattarai, R.K. Shrestha, J. Shrestha
Effect of different substrate sterilization methods on performance of oyster mushroom
(Pleurotus ostreatus) ..........………..................................................................……… 127
S. Subedi, S. Neupane
Antifungal assessment of plant extracts, biocontrol agents and fungicides against
Fusarium verticillioides (Sacc.) causing ear rot of maize ................................……… 133
O. Ulyanych, S. Poltoretskyi, V. Liubych, A. Yatsenko, V. Yatsenko, O. Lazariev,
V. Kravchenko
Effect of surface drip irrigation and cultivars on physiological state and productivity
of faba bean crop ..........……….......................................................................……… 139
B. Winarto, I.G. Cempaka, R.K. Jatuningtyas, S.C. Budisetyaningrum, B. Hartoyo
In vitro shoot growth performances and responses of potato (Solanum tuberosum L.)
'Muhzoto' under different treatments and explant types ..................................……… 150
TEADUSARTIKLID / RESEARCH ARTICLES 127
Agraarteadus
1 ● XXXII ● 2021 127–132
Journal of Agricultural Science
1 ● XXXII ● 2021 127–132
EFFECT OF DIFFERENT SUBSTRATE STERILIZATION METHODS
ON PERFORMANCE OF OYSTER MUSHROOM (Pleurotus ostreatus)
Sanju Shrestha1, Samikshya Bhattarai2, Ram Kumar Shrestha1, Jiban Shrestha3
1Institute of Agriculture and Animal Science, Lamjung Campus,
Sundarbazar 07, Sundarbazar Municipality, 33600, Nepal, ramkuma.shrestha@mail.huji.ac.il
2Texas A&M AgriLife Research and Extension Center, Texas A&M University,
Uvalde TX 78801, USA, bhattaraisami01@gmail.com
3Nepal Agricultural Research Council, National Plant Breeding and Genetics Research Centre,
Khumaltar15, Lalitpur Metropolitan City, 44700, Nepal, jibshrestha@gmail.com
Saabunud:
21.01.2021
Received:
Aktsepteeritud:
16.04.2021
Accepted:
Avaldatud veebis:
16.04.2021
Published online:
Vastutav autor:
Sanju
Shrestha
Corresponding author:
E-mail: sanjustha1997@gmail.com
Keywords: biological efficiency,
oyster mushroom, spawn-run,
sterilization, yield.
DOI: 10.15159/jas.21.03
ABSTRACT. Proper sterilization of substrates is an indispensable step in
oyster mushroom cultivation. Oyster mushroom growers in Nepal usually
follow three different substrate sterilization methods; however, their
comparative effectiveness is vastly unexplored. Thus, these experiments
were carried out at the Institute of Agriculture and Animal Science
(IAAS), Lamjung Campus, Lamjung, Nepal from January to March, in the
years 2017 and 2019. The objective of these experiments was to identify
the most appropriate method of sterilization. Three different types of
sterilization methods viz chemical sterilization (formaldehyde +
carbendazim), steam sterilization, and hot-water sterilization were
evaluated for the growth parameters and productivity of oyster mushroom
cultivated on rice straw. The experiments were laid out on Completely
Randomized Design (CRD) with ten replications. The results showed that
the spawning rate was 3.2% of the wet substrate. Data were collected until
the third flush. A significantly longer duration to colonize the substrate
(29.7 days) was observed under chemical sterilization. The oyster
mushroom performed best under steam sterilization as it took the shortest
time for pinhead formation (34.30 days), fruiting body formation
(43.60 days), cropping duration (89.30 days), and produced the highest
mushroom yield (1401.9 g per 4 kg bag), and consequently, the highest
biological efficiency (101.38%). Average pileus diameter and stipe length
were statistically indifferent among the treatments suggesting the
significant effect of sterilization methods on the yield of oyster mushroom
but not on its morphological attributes.
© 2021 Akadeemiline Põllumajanduse Selts. | © 2021 Estonian Academic Agricultural Society.
Introduction
The use of mushrooms as food is probably as old as
civilization itself. Over 200 mushroom species have
long been used as functional foods around the world
(Kalač, 2013), but only about 35 species have been
commercially cultivated (Aida et al., 2009). Over time,
an increase in awareness about mushrooms nutritive
and medicinal value has enhanced their consumption
(Chang, 1999).
Among the cultivated mushrooms, oyster mushroom
(Pleurotus species) is flourishing in the temperate and
sub-tropical environment due to its excellent flavour
and taste (Ganeshan et al., 1989). Pleurotus ostreatus
is the second-largest commercially cultivated mush-
room species in the world (Royse, 2013). Oyster
mushroom can be grown on various substrates due to
their strong enzymatic features (Atila, 2016). Pleurotus
ostreatus is easier to cultivate, favourable to eat, and
grow economically on different kinds of organic waste
raw materials and different climatic conditions (Kong,
2004; Sitaula, 2018; Tekeste et al., 2020).
The extensive cultivation of oyster mushroom is also
because of its simplistic cultivation, high biological
efficiency, and greater nutritional significance (Singh
et al., 1990). Nutritionally, the oyster mushroom is an
ideal food for humans. It is rich in proteins (30.4%), fat
(2.2%), carbohydrates (57.6%), and fibre (8.7) with
128 Sanju Shrestha, Samikshya Bhattarai, Ram Kumar Shrestha, Jiban Shrestha
Agraarteadus | Journal of Agricultural Science 1 ● XXXII ● 2021 127–132
345 Kcal energy in 100 g of mushroom on a dry weight
basis (Pandey, Ghosh, 1996).
Oyster mushrooms, having a considerable economic,
medicinal and nutritional value, are most commonly
and commercially cultivated on the non-composted
lignocellulosic substrates (Savoie et al., 2007) due to
which the agriculture, industrial, and forest wastes can
be efficiently utilized (Sanjel et al., 2021). Various
substrates, including paddy straw, maize stalks/cobs,
vegetable plant residues, and bagasse, are extensively
used in its cultivation (Sher et al., 2011). Successful
cultivation of oyster mushroom on the various
substrates, including sawdust, chopped office papers,
cardboard, and plant fibres, are also reported from
multiple research works (Mandeel et al., 2005). Dubey
et al. (2019) found that rice straw was found most
favourable for mushroom cultivation rather than wheat
straw, sugarcane bagasse and banana leaves.
Growing P.ostreatus requires sterilization of substra-
tes, which means pre-treatment of the substrates to
eliminate pathogenic and competitive micro-orga-
nisms, and to enhance the mycelial growth of mush-
room. Sterilization of the substrates for the production
of mushrooms is carried out mainly to avoid the
presence of pathogens that appear to compete for the
nutrients available in the unsterilized substrates. It is
one of the crucial steps in oyster cultivation, which can
determine the success of the cultivation (Ali et al.,
2007). Three different treatments, viz. steam sterili-
zation, immersion in hot water, and chemical treatment,
are explained in the literature (Mejía, Albertó, 2013).
However, there are not enough works comparing the
effectiveness of these sterilization methods influencing
the quality and quantity of the mushroom produced.
Therefore, the present research was undertaken to find
out the most appropriate sterilization method, which
will be helpful for further improvement in the yield of
mushrooms.
Material and methods
Two independent experiments were conducted in
Institute of Agriculture and Animal Science (IAAS),
Lamjung Campus, Lamjung, Nepal, from January to
March, in the year 2017 and 2019, to explore the best
sterilization method, i.e., chemical, steam and hot-
water sterilization in the production of oyster mush-
room. Rice straw was taken as the substrate and
subjected to hand-chopping to the required length of
5 cm.
Preparation of substrates
Chemical sterilization was carried out as explained by
Siddhant et al. (2014). The chopped straw was dipped
into a 200 L metal drum containing 125 ml formalin
mixture and 15 g Carbendazim per 100 L of water. The
mixture was stirred with the help of a wooden pole. The
drum was made airtight, and the substrate was allowed
to soak for 18 hours in the solution. Then, the solution
was drained out and the moisture content of about 65%
was maintained in the wet substrate before spawning.
For steam sterilization, the chopped substrate was
soaked in water for 12 hours before sterilization.
Presoaked substrate was exposed to steam for about
15–20 minutes, and cooled before spawning, as
explained by (Gowda, Manvi, 2019).
We followed the Kalita (2015) method for hot water
sterilization. The presoaked substrate was kept in the
drum and boiled for 30 minutes, and the solution was
drained before spawning. The experiment was set up
using a complete randomized design (CRD) with three
treatments and ten replications.
Spawning and incubation
Spawning was carried with fresh grain spawn on a
3.2% (weight/weight) rate based on the wet substrate.
On every layer of around 4 inches of sterilized straw in
polybags, spawn was placed near the periphery forming
a circle, and bags were pressed to compact. Three layers
of spawning were done to make balls of 4 kg, and
perforation of the bag was done after completion of
packing by making 8–10 holes of size 8 mm for
aeration. Cotton was plugged into each hole to prevent
contamination as well as entry of insects. Then, the
bags were suspended on strings in a dark and pre-
fumigated room with no ventilation. Thus, prepared
each ball represented one replication. The temperature
of the room was maintained at 13–14 °C, and relative
humidity was 80%. The continuous observation was
done till the full spawn run, and then the bags were cut
from two sides for fructification. Ventilation with
diffused light was provided with two gentle irrigations
per day and other necessary crop management.
Watering was stopped 24 hours before harvest.
Data collection and analysis
Harvesting was done when the majority of caps
attained their maximum size. The experiment was
terminated after three flushes from each replication.
Data were recorded periodically on different para-
meters, including the number of days to full coloni-
zation, the number of days to first fruiting, harvesting
duration (day), and mushroom yield in every flush (g),
pileus diameter (cm), stipe length (cm), and biological
efficiency (BE). The fresh mushroom yield on each
harvest was measured, then total fresh yield and
biological efficiency were calculated (Eq. 1).
𝐵𝐸 = 𝑌𝑖𝑒𝑙𝑑 𝑜𝑓 𝑓𝑟𝑒𝑠ℎ 𝑚𝑢𝑠ℎ𝑟𝑜𝑜𝑚𝑠, 𝑔
𝑇𝑜𝑡𝑎𝑙 𝑤𝑒𝑖𝑔ℎ𝑡 𝑜𝑓 𝑑𝑟𝑦 𝑠𝑢𝑏𝑠𝑡𝑟𝑎𝑡𝑒 𝑢𝑠𝑒𝑑 , 𝑔 ×100 (1)
Statistical analysis
Data were analyzed using RStudio 1.2.5033. The
treatment means were separated using Least Significant
Difference (LSD) at the 5% level of significance.
Results and discussion
Vegetative and reproductive growth
The time required for full colonization of substrate,
also known as spawn run duration, and was compared
between three methods of sterilization. In our
experiment, a significantly prolonged duration
(29.70 days) for complete mycelial growth was
observed in the chemically sterilized substrate (Fig. 1).
Effect of different substrate sterilization methods on performance of oyster mushroom (Pleurotus ostreatus) 129
Agraarteadus | Journal of Agricultural Science 1 ● XXXII ● 2021 127–132
Hot-water sterilization took the shortest duration
(26.20 days), which was statistically at par with steam
sterilization. The pattern was similar for days taken for
the first pinhead formation and the cropping duration,
reflecting the significant effect of the method of
sterilization used. Chemical sterilization took the
longest duration of 38.20 days for the appearance of the
first pinhead, whereas steam sterilization took the
shortest duration of 34.30 days. In all the treatments,
the pinheads were formed between 8–9 days after the
full colonization of the substrate. A significantly longer
cropping duration of 96.1 days was recorded in
chemical sterilization.
In oyster mushroom cultivation, the above parameters
reflect the speed of vegetative and reproductive growth
of the mushroom. Rapid growth and development are
desirable for the profitable business; however, multiple
factors, including ambient environmental condition,
substrate physical and chemical properties, and the
presence of competitive and pathogenic organisms, and
antigrowth substances, will influence the rate of
growth. Generally, the oyster mushroom takes up to
three weeks for a full spawn run in its optimal
environment (Shah et al., 2004; Kalita, 2015). How-
ever, we observed a delay of 6–10 days, reflecting the
suboptimal temperature (13–14 °C) for mycelia growth
in our experiments. Atila (2016) reported that 7–
12 days is taken from full spawn run to pinhead
formation which is by our result. The fruiting bodies
appeared on substrates sterilized by different methods
17–20 days later after a full spawn run. Our results are
similar to Tan (1981), who reported that P. ostreatus
took 14–21 days for fruiting body formation after the
full spawn run.
The increased cropping duration in chemical
sterilization is probably due to the anti-fungal effect of
fungicides that hindered the mycelial growth of the
mushroom, which subsequently delayed all the later
developmental stage up to the third harvest. A similar
delay in cropping duration was seen by Ali et al. (2007)
where all the species of Pleurotus treated with formalin
took long to complete the mycelial growth.
Figure 1. Bar plots showing the effect of sterilization methods on the days taken for full spawn run, pinhead formation, fruiting
body formation, and cropping duration of P. ostreatus. Values in a bar with the same letter(s) are not significantly different at
P = 0.05, according to LSD (least significant difference) test. The black bars in the plots indicate the standard error of the mean
130 Sanju Shrestha, Samikshya Bhattarai, Ram Kumar Shrestha, Jiban Shrestha
Agraarteadus | Journal of Agricultural Science 1 ● XXXII ● 2021 127–132
Mushroom morphology
There was no significant difference between the
treatments for pileus diameter and stipe length. The
stipe lengths ranged from 7.0 to 7.1, and the diameters
of pileus ranged from 9.3 to 9.6 (Table 1). It is
reasonable that sterilization methods had no impact on
the morphological properties of the mushroom. Such
parameters highly depend on the mushroom strain and
the nutrient factor of substrates. In our experiment, the
substrate was the same in all treatments; thus, no
variation was observed. The Pileus diameter of
P. ostreatus was recorded in different growing
substrates between 4.0–10 cm (Yildiz et al., 2002). The
variation in their work may be due to the variation in
nutrient factors of multiple substrates.
Table 1. Pileus diameter and stipe length of P. ostreatus ±
standard error of the mean, as affected by different sterilization
methods
Treatment
Diameter of
pileus, cm
Stipe length, cm
Chemical sterilization (T1)
9.408 ± 0.36
7.187 ± 0.19
Steam sterilization (T2)
9.638 ± 0.40
7.07 ± 0.25
Hot-water sterilization (T3)
9.303 ± 0.53
7.027 ± 0.27
F test
NS
NS
LSD (0.05)
0.943
0.711
CV (%)
10.877
10.915
Values in a column with the same letter(s) are not significantly
different at P = 0.05 according to LSD (least significant difference).
CV: coefficient of variation. NS: not significantly different at
P < 0.05
Mushroom yield
There were significant effects of the methods of
sterilization on the fresh mushroom yield of P. ostreatus.
The highest total yield of 1.40 kg ball–1 was obtained in
steam sterilization, which was statistically at par with
hot-water sterilization (Fig. 2). The lowest total yield
(1.02 kg ball–1) was obtained in chemical sterilization.
Ali et al. (2007) found that steam sterilization produced
the highest total yield than all other methods which is by
our results. The superiority of the steam sterilization
might be due to the generation of optimal physical and
chemical property of substrate for the mycelium growth.
The biological efficiency also varied significantly among
the methods of sterilization. The highest biological
efficiency (BE) of 101.38% was observed in steam
sterilization, and the least BE (79.75%) was observed in
chemical sterilization (Table 2).
Two different dominant contaminants viz Tricho-
derma spp. and Coprinus spp. were found during the
cultivation period in the experiments. The microbial
contamination was observed the highest in hot water
sterilization (16.00%) and the lowest in chemical
sterilization (11.75%) (Table 2). Ashraf et al. (2007)
found that the fungus associated with oyster mushroom
growing media (compost) such as Aspergillus,
Trichoderma, Mucor, Penicillium, Alternaria, Clado-
sporium, Monilia, Helminthosporium, Coccidioides
and Scedosporium. Sharma et al. (2007) in the results
of his research in India showed that the fungus
Aspergillus spp., Aspergillus niger, Fusarium spp.,
Mucor spp. and Trichoderma spp. were competitor
fungus or may cause disease in cultivated mushrooms
such as oyster mushrooms. Lopez-Arevalo et al. (1996)
also found that Penicillium sp., Aspergillus sp.,
Trichoderma sp. and Cunninghamella sp. a fungal
contaminant in the tropical country of Mexico.
Figure 2. Bar plot showing the effect of sterilization methods
on the total yield of P. ostreatus. Values in the bar with the
same letter(s) are not significantly different at P = 0.05,
according to LSD (least significant difference) test. The black
bars in the plots indicate the standard error of the mean.
BE measures the ratio of the weight of fresh
mushroom to the weight of dry substrate used in its
production. In our laboratory, we have observed the BE
for oyster mushroom up to 140%. BE in oyster
mushroom production varies from 75.5 to 128.8%
(Zhang et al., 2002). In the real ground, the BE may be
affected by several factors as substrate composition,
ambient environment, mushroom strain, disease pest
and other multiple management factors. BE of around
90% in hot water sterilization and 80% in chemical
sterilization is an acceptable level. However, with the
increase in room temperature, competitive moulds and
microbes may reduce the BE of the mushroom crop
(Biswas, 2014). Thus, we suggest all oyster mushroom
growers wisely choose the sterilization techniques to
obtain the economically profitable level of BE under
suboptimal growing conditions.
Three substrate sterilization methods were evaluated
for their effect on the growth and productivity para-
meters of oyster mushroom. There were significant
differences in all the growth and yield parameters, but
the morphological attributes were indifferent. Steam
sterilization produced the highest fresh mushroom yield
of 1409.1 g per bag of 4 kg wet substrate resulting in
the highest biological efficiency of 101.3%. Chemical
and hot water sterilization resulted in an acceptable
production level with BE of around 80 and 90%,
respectively. A Significantly longer cropping duration
of 96.1 days was seen in chemical sterilization than hot
water and steam sterilization. The same pattern was
seen for other growth parameters, including days taken
for full spawn run and fruiting body formation.
Effect of different substrate sterilization methods on performance of oyster mushroom (Pleurotus ostreatus) 131
Agraarteadus | Journal of Agricultural Science 1 ● XXXII ● 2021 127–132
Table 2. Effect of the sterilization methods on the quantity harvests and biological efficiency of P. ostreatus.
Values in a column with the same letter(s) are not significantly different at P = 0.05, according to LSD test.
CV: coefficient of variation; *** and ** significant different at P < 0.001 and P < 0.01, respectively
Conclusions
Proper sterilization of substrates is important for the
effective and smooth cultivation of mushrooms. In our
study, the different methods of sterilization have
influenced the vegetative growth, morphology,
cropping duration, mushroom yield and biological
yield of oyster mushroom, however, the steam
sterilization method was found more efficient with a
shorter cropping duration, higher yield and higher
biological efficiency than other methods. Thus, stem
sterilization was found the best method of sterilization
for the cultivation of oyster mushroom.
Funding
The financial support for this study was received from
the Institute of Agriculture and Animal Science
(IAAS), Lamjung Campus, Nepal.
Conflict of interest
The authors declare that they have no conflict of interest.
Author contributions
SS – lead investigator and responsible for literature search
and write-up.
SB, RKS, JS – responsible for the literature review and
provided critical feedback on the manuscript.
All the authors read and approved the final manuscript.
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