BAGUIO CITY NATIONAL SCIENCE HIGH SCHOOL
PUROK 12, IRISAN, BAGUIO CITY
Subject: RESEARCH IV
Quarter: 2ND QUARTER
Activity: LITERATURE FOR THE METHODOLOGY
Date: October 31, 2023 and November 3, 2023 (online class)
NAME OF Carl Jericho P. Garcia
THE
LEARNER:
Objectives: The learner should be able to:
a. find a literature that will support the methodology of their own study;
b. identify appropriate research design, tools, procedure, and data analysis that will
be used and adapted in their study; and
c. write down the information needed for their study
DIRECTION Each learner must search for a study or a literature that will be used as a reference for
: the methodology of their study. Give the necessary information below.
ACTIVITY PROPER
Title of Write the title of your study.
your study
Mugwort (Artemisia vulgaris) extract as an antifungal spray against
White mold (Trichoderma longibrachiatum)
Read and give the information being asked below.
Make sure that the literature you found is related and is useful in
your study.
Title of the Trichoderma species occurring on wood with decay symptoms in
literature mountain forests in Central Europe: genetic and enzymatic
your found characterization
Author and Błaszczyk, L., Strakowska, J., Chełkowski, J., Gąbka-Buszek, A., &
the date of Kaczmarek, J. (2015)
the
literature
Problem Write the problem the researcher is trying to solve in the study.
The problem the researchers are trying to solve is finding out how
many species of Trichoderma occur in the mountains of Central
Europe.
Research Find the research gap in the study. Then write below.
Gap
The research gap is that there were already some species of
Trichoderma present in the mountains but there were still some
that were undiscovered.
Objectives Write the objectives and importance of the study of the literature
and
Importanc The aim of the study was to explore the species diversity of
e Trichoderma obtained from samples of wood collected in the forests
of the Gorce Mountains (location A), Karkonosze Mountains
(location B) and Tatra Mountains (location C) in Central Europe and
to examine the cellulolytic and xylanolytic activity of these species
as an expression of their probable role in wood decay processes.
Research Write the research design used in the literature
Design:
The research design used in the literature is a qualitative design since the literature
only wanted to observe the various species of Trichoderma that were present.
Tools What are the tools used in the study? Give the details and specifications of the tools.
Incubation chamber- This would be used to keep the microbial cells of the
Trichoderma alive for its observation.
Microscope- This will be used to observe the details of the different species of
Trichoderma in more detail.
Procedure What are the procedures used in the study? Be specific. (Make sure you understand
the procedure and is related and can be used in your study).
First they collected samples of decaying wood collected in the for-ests of the Tatra
Mountains, Gorce Mountains andKarkonosze Mountains, located in Central Europe in
Poland.
They proceeded to pack the pieces of wood with Trichoderma, dried them at room
temperature if they were wet, and then stored them until Isolation. In the laboratory,
small sterile samples (0.5×0.2 cm )were taken from each piece of wood, plated on
SynthetischerNährstoffarmer Agar and incubated at 20 °C for 6 days. All colonies
resembling Trichoderma were twice subcultured on potato dextrose agar and
incubated as above. Pure cultures were subsequently transferred to tubes containing
SNA and stored at 4 °C for further study.
The next step was to identify the Trichoderma isolates obtained. In this study they
identified at the species level by a combination of morphological and molecular
analyses. The morphological characteristics were based on the key by Gams and
Bissett (1998) and Chaverriet al. (2015).
In addition they conducted a Phylogenetic analysis with the use of ITS andtef1 gene
sequences (each allele identified in ITS and tef1 locus) obtained in this work as well as
on the sequences retrieved from NCBI GenBank
They moved on with an analysis of their enzyme production. Trichoderma reseei QM
9414, sourced from the Czech Col-lection of Microorganisms (CCM), Brno, Czech
Republic was used as the reference strain. The Trichoderma collection of119 isolates
were screened for cellulase and xylanase activity under the same conditions,
specifically optimized for cellulase/xylanase production with T. reseei QM 9414 strain.
Finally they conducted an Assay of enzyme activities where they cultured in an
induction medium, mycelial pellets are removed by centrifugation (15 min at
12,000g). Total cellu-lase activity was determined using the filter paper activity(FPA)
assay (Ghose 198 7). Whatman No. 1 filter paper(50 mg, 1×6 cm strip; Whatman
International, UK) was incu-bated for 60 min in 0.5 mL of 0.05 M Na-citrate buffer
solu-tion (pH 4.8) at 50 °C with an addition of 0.5 mL enzyme solution (supernatant).
The reaction mixture was incubated at 50 °C for 15 min. Released reducing sugars
were deter-mined by dinitrosalicylic acid reagent (DNS) method ofMiller (1959) and
Eveleigh et al. (2009 ), by adding 1 mLof DNS solution and then incubating the mixture
at 95 °Cfor 5 min. Absorbance was measured at 540 nm (UV-1800 Spectrophotometer,
Shimadzu, Japan). A calibration curve was established with glucose (POCH, Poland).
Final cellu-lase activity was expressed in filter paper units (FPU) defined as the
amount of enzyme which forms 1 μmol of glucose per minute under the assay
conditions. Three Independent experiments were run for each strain (includ-ing
blanks).
Data Write the data analysis used in the study.
Analysis
The data analysis used in the study was ANOVA and was determined using Tukey’s
test. The statistical analysis was performed using the program Statistica version 10.0
(StatSoft Inc.) and Microsoft Excel version 2010.
Attached https://www.researchgate.net/publication/
the link 284243058_Trichoderma_species_occurring_on_wood_with_decay_symptoms_in_m
here ountain_forests_in_Central_Europe_genetic_and_enzymatic_characterization?
fbclid=IwAR2PKuh4E_ZEtdGwjp5mlZKg3rPCg0tr9mDmNUSD64sdwQuvJOhZcgsXxPw
Source Follow correct referencing
Błaszczyk, L., Strakowska, J., Chełkowski, J., Gąbka-Buszek, A., &
Kaczmarek, J. (2015b). Trichoderma species occurring on wood with
decay symptoms in mountain forests in Central Europe: genetic and
enzymatic characterization. Journal of Applied Genetics, 57(3), 397–
407. https://doi.org/10.1007/s13353-015-0326-1
