Sains Malaysiana 45(3)(2016): 329–337
Gamma Irradiation Effect on Embryogenic
Callus Growth of Citrus reticulata cv. Limau Madu
(Kesan Sinaran Gamma ke atas Pertumbuhan
Kalus Embriogenik Citrus reticulata
cv. Limau Madu)
DITA AGISIMANTO1,2,
NORMAH
MOHD
NOOR3*,
RUSLI
IBRAHIM4
& AZHAR MOHAMAD4
1School of Biosciences and Biotechnology,
Faculty of Science and Technology, Universiti Kebangsaan Malaysia,
43600 Bangi, Selangor Darul Ehsan, Malaysia
2Indonesian Citrus and Subtropical
Crops Research Institute, Jl. Raya Tlekung 1 Junrejo, Batu
East Java 65301 Indonesia
3Institute of Systems Biology (INBIOSIS), Universiti Kebangsaan Malaysia, 43600 Bangi,
Selangor Darul Ehsan, Malaysia
4Agrotechnology and Biosciences Division,
Malaysian Nuclear Agency, 43000 Bangi,
Selangor Darul Ehsan, Malaysia
Diserahkan: 8 Disember 2014/Diterima:
28 Ogos 2015
ABSTRACT
Induced mutagenesis using gamma
ray has been proven applicable to improve varieties of many genotypes
of crop species. The effects of 60oC gamma ray dosage
on growth and callus induction of nucellus segments of Citrus
reticulata cv. limau madu were investigated. The nucelli were
exposed to gamma rays at doses of 10, 20, 40, 60, 80, 100 and 120
Gy, followed by embryogenic callus (EC) induction on Murashige and Skoog
medium supplemented with 500 mg L-1 malt extract (ME),
146 mM sucrose, 0.8% (w/v) agar and 13.3 μM benzyl amino purine
(BAP).
Survival, callus type and colour, degree of callus formation, time
of callus formation and total fresh weight of callus varied among
the treatments. All untreated explants (controls) survived and produced
friable EC in
the 2nd or 3rd week of culture, whereas the irradiated nucelli showed
delayed response. EC derived from the nucelli irradiated at 10, 20 and 40
Gy appeared in the 3rd week of culture, whereas EC from
the 60 and 80 Gy doses appeared in the 4th week. Exposure to higher
doses (100 and 120 Gy) completely suppressed callus formation. After
35 days of culture, an average of 697 and 660 mg of EC were harvested from the nucelli
irradiated at 10 and 20 Gy, respectively, which was higher than
those at 40 Gy (441 mg), 60 Gy (436 mg) and 80 Gy (380 mg). EC were
initiated and proliferated and subsequently regenerated into plantlets.
DNA
of plantlets from the 20, 40 and 60 Gy exposure were
individually amplified and compared to the control for early detection
of mutagenesis using retrotransposon, inter simple sequence repeat
and markers related to seedlessness. No variants were observed from
the plantlets produced.
Keywords: Embryogenic callus;
gamma-rays; LD50; limau madu; nucellus
ABSTRAK
Mutagenesis aruhan menggunakan
sinaran gamma dibuktikan berupaya untuk menambah baik pelbagai genotip
tanaman. Kesan dos sinar 60oC gamma pada pertumbuhan
dan induksi kalus pada segmen nuselus Citrus reticulata cv. limau madu dikaji. Nuselus
didedahkan kepada sinar gamma pada dos 10, 20, 40, 60, 80, 100 dan
120 Gy , diikuti dengan aruhan kalus embriogenik (EC)
di atas medium Murashige dan Skoog dengan penambahan 500 mg L-1 ekstrak
malt (ME), 146 mM sukrosa, 0.8% (w/v) agar dan 13.3 μM benzil
amino purin (BAP). Kemandirian, jenis dan warna kalus,
tahap pembentukan kalus; masa pembentukan kalus dan berat segar
kalus berbeza antara rawatan. Semua eksplan tidak dirawat (kawalan)
hidup dan menghasilkan EC rapuh dalam masa 2 hingga 3 minggu kultur, manakala nuselus
radiasi menunjukkan tindak balas yang lambat. EC daripada
nuselus diradiasi pada 10, 20 dan 40 Gy muncul pada minggu ke-3
kultur, manakala EC daripada dos Gy 60 dan 80 muncul pada minggu ke-4. Pendedahan
kepada dos lebih tinggi (100 dan 120 Gy) telah menghalang pembentukan
kalus sepenuhnya. Selepas 35 hari dikultur, purata 697 dan 660 mg
EC
dicatatkan daripada nuselus yang diradiasi masing-masing
pada 10 dan 20 Gy, iaitu lebih tinggi daripada 40 Gy (441 mg), 60
Gy (436 mg) dan 80 Gy (380 mg). EC
dimulakan, berkembang biak dan seterusnya dijana semula
menjadi anak pokok. DNA anak
pokok daripada pendedahan 20, 40 dan 60 Gy telah diamplifikasi secara
individu dan dibandingkan dengan kawalan untuk pengesanan awal mutagenesis
menggunakan retrotransposon, antara ulangan jujukan ringkas dan
penanda berkaitan ketidakhadiran biji benih. Tiada variasi diperhatikan
daripada anak pokok yang dihasilkan.
Kata kunci: Kalus embriogenik; LD50;
limau madu; nuselus; sinar gamma
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*Pengarang untuk surat-menyurat;
email: normah@ukm.edu.my
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