Sains Malaysiana 47(12)(2018): 3025–3030

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

 

Diserahkan: 30 Mei 2018 /Diterima: 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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*Pengarang untuk surat-menyurat; email: emelda@ukm.edu.my