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http://dx.doi.org/10.17576/jsm-2018-4712-12
Isolation and Identification of Endophytic Fungi
from UiTM Reserve Forest, Negeri Sembilan (Pemencilan dan Pengenalpastian Kulat Endofit
dari Hutan Simpan UiTM Negeri Sembilan)
SITI NURSYAZWANI
MAADON1, SARINI AHMAD WAKID1, IWANA IZNI ZAINUDIN1,
LILI SYAHANI RUSLI1, MOHD SYAHRIL MOHD ZAN1,
NOR'AISHAH HASAN1, NOR'AISHAH ABU SHAH1 &
EMELDA ROSSELEENA ROHANI2*
1Faculty of Applied Sciences, Universiti
Teknologi MARA, Cawangan Negeri Sembilan,72000 Kuala Pilah, Negeri Sembilan
Darul Khusus, Malaysia
2Institute of Systems Biology, Universiti Kebangsaan Malaysia,
43600 UKM Bangi, Selangor Darul Ehsan, Malaysia
Received: 30 May 2018 /Accepted: 18 September
2018
ABSTRACT
Endophytic fungi are those living inside
the host plant without causing any apparent negative effect on the
host plant. Two isolates endophytic fungi from leaves and two isolates
from root at Universiti Teknologi MARA (UiTM) Reserve Forest, Negeri
Sembilan were successfully isolated and identified by morphology
and molecular characteristic. Samples were surface sterilized and
sub-cultured to obtain a pure culture. Characteristics of the isolates
such as colony appearance, mycelial texture, conidia/spores and
pigmentation were studied to explore their morphology. Isolates
were also subjected to a PCR-based
genotyping test. There were noticeable differences in morphological
characteristics among the four isolates. Microscopic analysis showed
four isolates consist of septa and conidia/spores. The pigmentation
result showed that colony in A1leaf samples demonstrated an orange color on potato dextrose
agar (PDA) media, colony in A1root
demonstrate a black texture in PDA media
while hairy colonies in the others two isolates showed a white color
on
PDA media. Based on molecular analyses the fungal
genera showed 99-100% similarity with the related fungi recorded
in the GenBank. Both morphology and molecular sequencing of internal
transcribed spacer (ITS)
regions of endophytic fungi showed that three isolates (A1root,
C2leaf, and C3root) were grouped in Basidiomycota
while one isolate (A1leaf) belonged to Ascomycota.
The endophyte funguses were identified as Daldinia sp. (A1leaf),
Polyporales sp. (A1root,) Lentinus sp. (C2leaf,)
and Rigidoporus sp. (C3root). Overall, the new discoveries
of isolated endophyte fungal have dyeing potential of fungal pigments
which offer a viable alternative to natural vegetable and harmful
synthetic dyes.
Keyword: Endophyte fungi;
identification; morphological characteristic
ABSTRAK
Kulat endofit yang hidup di dalam tumbuhan
perumah tanpa menyebabkan sebarang kesan negatif yang jelas terhadap
tumbuhan perumah. Dua pencilan kulat endofit daripada daun dan dua
pencilan daripada akar dari Hutan Simpan Universiti Teknologi MARA (UiTM),
Negeri Sembilan telah berjaya dipencilkan dan dikenal pasti daripada
segi morfologi dan ciri molekul. Sampel disterilkan pada permukaan
dan di sub-kultur untuk memperoleh kultur tulen. Ciri-ciri pencilan
seperti rupa koloni, tekstur miselium, conidia/spora dan pigmentasi
dikaji untuk meneroka morfologi mereka. Pencilan juga dijalankan
untuk ujian genotip berasaskan PCR.
Terdapat perbezaan yang ketara di dalam ciri-ciri morfologi dalam
kalangan empat pencilan. Analisis mikroskopik menunjukkan empat
pencilan terdiri daripada septum dan conidia/spora. Keputusan pigmentasi
menunjukkan bahawa koloni di dalam sampel A1daun menunjukkan warna jingga pada media
dekstrose agar (PDA), koloni di dalam A1akar
menunjukkan tekstur hitam di dalam media PDA manakala
koloni berbulu di dua lagi pencilan menunjukkan media PDA berwarna
putih. Berdasarkan analisis molekul, genus kulat menunjukkan persamaan
99-100% dengan kulat yang berkaitan yang direkodkan di dalam bank
Gen. Kedua-dua morfologi dan jujukan molekul di kawasan mentranskripsi
penjarak dalaman (ITS)
kulat endofit mendedahkan bahawa tiga pencilan (A1akar,
C2daun, dan C3akar) dikelompokkan di dalam Basidiomycota
manakala satu pencilan (A1daun) tergolong di dalam Ascomycota.
Kulat endofit dikenal pasti sebagai Daldinia sp. (A1daun),
Polyporales sp. (A1akar,) Lentinus sp. (C2daun,)
dan Rigidoporus sp. (C3akar). Secara keseluruhannya,
penemuan baru pencilan kulat endofit mempunyai potensi pewarnaan
pigmen daripada pigmen kulat yang menawarkan alternatif terkedepan
kepada sayur-sayuran semula jadi dan pewarna sintetik yang merbahaya.
Kata
kunci: Ciri morfologi; kulat endofit; pengenalpastian
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*Corresponding author; email: emelda@ukm.edu.my
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