Sains Malaysiana 46(12)(2017): 2425–2432

http://dx.doi.org/10.17576/jsm-2017-4612-18

 

Characterization of Fusarium proliferatum and Fusarium verticillioides based on Species-Specific Gene and Microsatellites Analysis

(Perincian Fusarium proliferatum dan Fusarium verticillioides Berdasarkan Gen Khusus Spesies dan Analisis Mikrosatelit)

 

A.      NAJIHAH¹, M.Z. NUR AIN IZZATI^1*, S.Y.C. YONG¹ & M.N. NIK MOHD IZHAM²

 

¹Department of Biology, Faculty of Science, Universiti Putra Malaysia, 43400 Serdang, Selangor Darul Ehsan, Malaysia

 

²School of Biological Sciences, Universiti Sains Malaysia, 11800 Penang, Pulau Pinang

Malaysia

 

Received: 14 November 2016/Accepted: 23 May 2017

 

ABSTRACT

Fusarium species are known to cause various diseases on plantations including fruits and vegetables. The most common Fusarium that can cause plant diseases are Fusarium proliferatum and Fusarium verticillioides. Ear rot disease on maize, wilt disease on cucurbits and fruit rot disease on tomato as well as banana are example of diseases caused by these two species. The objectives of this study were to identify F. proliferatum and F. verticillioides based on species-specific primers and polymerase chain reaction (PCR) amplification and to evaluate the genetic diversity of both species based on microsatellite markers. Fifty isolates of Fusarium species that were previously collected throughout Malaysia from different hosts were identified by using species-specific PCR amplification. Twenty-nine isolates were identified as F. proliferatum and 21 isolates were identified as F. verticillioides based on species-specific primer. The genetic diversity of all the fungal isolates was evaluated by using microsatellite analysis with six established primers. Five out of six primers amplified polymorphic bands with the most effective primer showing high polymorphism were (AG)7C and (TCC)5 meanwhile one primer (TTTC)4 gave negative result with no band amplified. The phylogenetic tree that was constructed showing two different clades distinguished between F. proliferatum and F. verticillioides.

 

Keywords: Fusarium proliferatum; Fusarium verticillioides; microsatellite; species-specific gene

 

ABSTRAK

Spesies Fusarium dikenali sebagai penyebab pelbagai penyakit terhadap tumbuhan termasuk buah-buahan dan sayur-sayuran. Antara spesies Fusarium yang paling kerap menyebabkan penyakit pokok adalah Fusarium proliferatum dan Fusarium verticillioides. Penyakit reput tongkol pada jagung, penyakit layu pada mentimun dan penyakit buah reput pada tomato dan pisang adalah contoh penyakit disebabkan oleh dua spesis ini. Objektif kajian ini adalah untuk mengenal pasti F. proliferatum dan F. verticillioides berdasarkan khusus spesies reaksi rantai polimerase dan untuk menilai kepelbagaian genetik kedua-dua spesies berdasarkan mikrosatelit. Lima puluh isolat spesies Fusarium yang terlebih dahulu dipencilkan daripada pelbagai perumah yang diperoleh dari serata Malaysia telah diuji dan dikenal pasti dengan menggunakan gen khusus spesies reaksi rantai polimerase. Dua puluh sembilan isolat telah dikenal pasti sebagai F. proliferatum dan dua puluh satu isolat telah dikenal pasti sebagai F. verticillioides berdasarkan primer gen khusus spesies. Setelah proses pengenalpastian dilakukan, kepelbagaian genetik telah dinilai dengan menggunakan analisis mikrosatelit menggunakan enam primer. Lima daripada enam primer menghasilkan band polimorfik dan primer yang paling tinggi kadar polimorfisme adalah (AG)7C dan (TCC)5 manakala satu primer (TTTC)4 memberikan hasil negatif dan tiada amplifikasi. Pohon filogenetik yang telah dihasilkan menunjukkan dua klad yang membezakan antara F. proliferatum dan F. verticillioides.

 

Kata kunci: Fusarium proliferatum; Fusarium verticillioides; gen khusus spesies; mikrosatelit

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*Corresponding author; email: ainizzati@upm.edu.my