Sains Malaysiana 48(8)(2019): 1593–1600

http://dx.doi.org/10.17576/jsm-2019-4808-04

Phylogenetic Analysis of Fusarium oxysporum f. sp. cubense Associated with Fusarium Wilt of Bananas from Peninsular Malaysia

(Analisis Filogenetik Fusarium oxysporumf. sp. cubense Berkait dengan Penyakit Layu Fusarium Pisang dari Semenanjung Malaysia)

CLEMENT KIING FOOK WONG¹, DZARIFAH ZULPERI¹*, GANESAN VADAMALAI¹, NOOR BAITY SAIDI² & CHUI YAO TEH²

¹Department of Plant Protection, Faculty of Agriculture, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor Darul Ehsan, Malaysia

²Department of Cell and Molecular Biology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400, UPM Serdang, Selangor Darul Ehsan, Malaysia

Diserahkan: 3 November 2018/Diterima: 12 Mei 2019

ABSTRACT

Fusarium wilt of bananas caused by Fusarium oxysporum f. sp. cubense (Foc) has been known to hamper banana production. In Malaysia, most of the edible banana cultivars are susceptible to the fungal strain Foc tropical race 4 (Foc TR4). To date, molecular characterisation of Foc TR4 in this country remains elusive. Thus, this study aimed to characterise the diversity of Foc TR4 in Peninsular Malaysia based on the phylogenetic analysis of transcription elongation factor (TEF-1α) sequences. From March 2016 to December 2016, 17 isolates of Foc were isolated from nine outbreak states in Peninsular Malaysia consisting of Penang, Perak, Selangor, Negeri Sembilan, Melaka, Johor, Pahang, Kelantan and Terengganu, with disease incidence exceeding 70% in several plantations. Morphological, molecular identification using TR4 specific primers and pathogenicity assay confirmed the identity of the fungal pathogen. Pathogenicity assays indicated that the aggressiveness level varies among the isolates. Phylogenetic analyses of TEF1-α sequences showed that there was no genetic variation among the Foc isolates by clustering them into four vegetative compatibility groups (VCGs) of 01213 (TR4), 01216 (TR4), 01213/16 (TR4) and 0121 (R4). PCR amplification using VCG specific primers have further categorised 16 Foc isolates as VCG01213 and a single isolate as VCG01213/16. In addition, no correlation between TEF-1α sequences and aggressiveness of isolates could be established.

Keywords: Aggressiveness; banana; fusarium wilt; phylogenetic analysis; TEF-1α

ABSTRAK

Penyakit layu Fusarium pisang disebabkan oleh kulatFusarium oxysporum f. sp. cubense (Foc) telah menjejaskan pengeluaran pisang. Kebanyakan kultivar pisang yang boleh dimakan di Malaysia mudah dijangkiti oleh strain kulat Foc tropical race 4 (Foc TR4). Sehingga kini, pencirian Foc TR4 di peringkat molekul dalam negara ini masih tidak dikaji. Justeru, kajian ini bertujuan untuk mencirikan kepelbagaian Foc TR4 di Semenanjung Malaysia berdasarkan analisis filogenetik jujukan faktor transkripsi pemanjangan (TEF-1α). Dari Mac 2016 hingga Disember 2016, sebanyak 17 pencilan Foc telah dipencilkan dari sembilan kawasan berpenyakit di Semenanjung Malaysia terdiri daripada Pulau Pinang, Perak, Selangor, Negeri Sembilan, Melaka, Johor, Pahang, Kelantan dan Terengganu dengan kejadian penyakit melebihi 70% di beberapa ladang pisang. Pengenalpastian morfologi dan molekul dengan menggunakan primer khusus TR4 dan ujian kepatogenan telah mengesahkan identiti patogen kulat. Ujian kepatogenan menunjukkan bahawa tahap agresif antara pencilan Foc adalah berbeza. Analisis filogenetik jujukan TEF-1α menunjukkan bahawa tiada variasi genetik antara pencilan Foc dengan dibahagikan kepada empatvegetative compatibility groups (VCGs) iaitu 01213 (TR4), 01216 (TR4), 01213/16 (TR4) dan 0121 (R4). Amplifikasi PCR dengan menggunakan primer khusus VCG telah selanjutnya mengkategorikan 16 pencilan Foc sebagai VCG01213 dan satu pencilan sebagai VCG01213/16. Tambahan pula, tiada korelasi antara jujukan TEF-1α dan tahap agresif pencilan Foc dapat ditunjukkan.

Kata kunci: Analisis filogenetik; penyakit layu fusarium; pisang; tahap agresif; TEF-1α

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*Pengarang untuk surat-menyurat; email: dzarifah@upm.edu.my