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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
IZZATI1*,
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 Diserahkan: 14
November 2016/Diterima: 23 Mei 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 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 khusu spesies;
mikrosatelit RUJUKAN
Abd-Elsalam, K.A.,
Bahkali, A.H., Moslem, M.A. & Verreet, J.A. 2011. Development
of di- and tetranucleotide repeat primer for discrimination of
Fusarium species. Journal of Food Safety 31: 35-40.
Bahkali, A.H.,
Abd-Elsalam, K.A., Jian-Rong, G., Khiyami, M.A. & Verreet,
J.A. 2012. Characterization of di-, tri-, and tetranucleotide
microsatellite primers suitable for genotyping various plant pathogenic
fungi with special emphasis on Fusaria and Mycospherella
graminicola. International Journal of Molecular Sciences 13:
2951-2964. Cummings, M.P.
2004. PAUP* (Phylogenetic Analysis Using Parsimony and other
methods. Dictionary of Bioinformatics and Computational Biology.
New York: John Wiley & Sons, Ltd. Darnetty, D.M.M.N.,
Nur Ain Izzati, M.Z., Nor Azliza, I., Nik Mohd Izham, M.N. &
Baharuddin, S. 2008. Diversity of Fusarium species associated
with ear rot of corn in Indonesia and Malaysia. Proceeding
of 6th Regional IMT-GT UNINET Conference. pp. 422-425. Ellegren, H. 2000.
Microsatellite mutations in the germline: Implications for evolutionary
inference. Trends Genetics 16: 551-558. Gohari, A.M., Sedaghat,
N., Javan-Nikkah, M. & Riseh, R.S. 2007. Mycoflora of wheat
grains in the main production area in Kerman Province, Iran. International
Journal of Agriculture & Biology 9: 635-637. Jahan Quazi, S.A.,
Sariah, M., Hawa, J. & Zainal Abidin, B.M.A. 2013. Characterization
of Fusarium proliferatum through species-specific primers
and its virulence on rice seeds. International Journal of Agriculture
& Biology 15: 649-656. Jany, J.L., Bousquet,
J., Gagne, A. & Khasa, D.P. 2006. Simple sequence repeats
(SSR) markers in the ectomycorrhizal fungus Laccaria bicolor
for environmental monitoring of introduced strains and molecular
ecology applications. Mycological Research 110: 51-59.
Karaoglu, H., Lee,
C.M.Y. & Meyer, W. 2005. Survey of simple sequence repeats
in completed fungal genomes. Molecular and Biology Evolution
22: 639-649. Lu, G., Cannon,
P.F., Reid, A. & Simmons, C.M. 2004. Diversity and molecular
relationships of endophytic Colletotrichum isolates from
the Iwokrama Forest Reserve, Guyana. Mycological Research 108:
53-63. Nur Ain Izzati,
M.Z. & Salleh, B. 2010. Variability of Fusarium species
associated with Bakanae disease of rice based on their virulence,
vegetative and biological compatibilities. Sydowia 62:
89-104. Nur Ain Izzati,
M.Z., Azmi, A.R., Atta, M.A.A. & Norazlina, J. 2011. Contribution
to the knowledge of diversity of Fusarium associated with
maize in Malaysia. Plant Protection Science 47(1): 20-24.
Nur Ain Izzati,
M.Z., Siti Nordahliawate, M.S., Nor Azliza, I. & Salleh, B.
2009. Distribution and diversity of Fusarium species associated
with grasses in Peninsular Malaysia. Biotropika 16: 55-64.
Patino, B., Mirete,
S., Gonzalez-Jaen, T., Mule, G., Rodriguez, M.T. & Vazquez,
C. 2004. PCR detection assays of fumonisin-producing Fusarium
verticillioides strains. Journal of Food Protection 67:
1278-1283. Peakall, R. &
Smouse, P.E. 2012. GenAlEx 6.5: Genetic analysis in Excel. Population
and research-an update. Bioinformatics 28: 2537-2539. Rampersad, S.N.,
Perez-Brito, D., Torres-Calzada, C., Tapia- Tussell, R. &
Carrington, C.V.F. 2013. Genetic structure of Colletotrichum
gloeosporioides sensu lato isolates infecting papaya inferred
by multilocus ISSR markers. Phytopathology 103: 182-189.
Reid, L.M., Zhu,
X. & Ma, B.L. 2001. Crop rotation and nitrogen effects on
maize susceptibility to Gibberella (Fusarium graminearum)
ear rot. Plant and Soil 237: 1-14. Ren, Y., Zhao,
H., Kou, Q., Jiang, J., Guo, S., Zhang, H., Hou, W., Zou, X.,
Sun, H., Gong, G., Levi, A. & Xu, Y. 2012. A high resolution
genetic map anchoring scaffolds of the sequenced watermelon genome.
PLoS ONE 7(1): 29453. Toth, G., Gaspari,
Z. & Jurka, J. 2000. Microsatellites in different eukaryotic
genome: Survey and analysis. Genome Research 10(7): 1967-1981.
Yogeshwar, D.K.,
Noelle, S., Hile, S.E., Chiaromonte, F., Eckert, K.A. & Makova,
K.D. 2010. What is a microsatellite: A computational and experimental
definition based upon repeat mutation behaviour at A/T and GT/AC
repeats. Genome Biology and Evolution 2: 620-635. Zheng, Q. &
Ploetz, R. 2002. Genetic diversity in the mango malformation pathogen
and development of a PCR assay. Plant Pathology 51: 208-216.
*Pengarang untuk surat-menyurat; email: ainizzati@upm.edu.my
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