Determination of the optimum initial callus weight for the efficient propagation of sugarcane in temporary immersion bioreactor
DOI:
https://doi.org/10.22302/iribb.jur.mp.v90i2.505Keywords:
in vitro culture, liquid culture, proliferationAbstract
Abstrak
Bioreaktor perendaman sesaat (BPS) telah digunakan secara luas untuk propagasi skala massal berbagai tanaman penting, termasuk tanaman tebu. BPS menyediakan sistem kultur semi-otomatis dan kondisi optimal bagi pertumbuhan tanaman. Beberapa faktor menentukan pertumbuhan tanaman pada BPS, salah satunya densitas dari eksplan. Oleh karena itu, penelitian dilakukan untuk menentukan bobot awal yang optimal untuk kalus tebu yang dikulturkan pada BPS, serta mengevaluasi pengaruh perbedaan bobot awal kalus tersebut terhadap proliferasi dan regenerasi kalus tebu. Kalus tebu diinduksi dari daun muda yang masih menggulung dari empat varietas tebu unggul Indonesia. Bobot awal kalus yang dikultur ke dalam bejana TIB yaitu 0,05 g; 0,1 g; 0,2 g; 0,5 g; dan 1,0 g untuk setiap bejana. Kalus kemudian melalui tahap proliferasi pada BPS sebanyak tiga siklus, kemudian kalus diregenerasi pada BPS dengan perlakuan auksin dan sitokinin. Hasil penelitian menunjukkan bahwa 0,2 g merupakan bobot awal kalus yang efisien untuk proliferasi kalus tebu pada TIB, dimana eksponensial multiplikasi kalus tercapai pada bobot awal tersebut, yaitu untuk masing-masing varietas 130,3 kali (PSKA 942), 136,8 kali (PS 094), 21,3 (PS 881), dan 12,9 kali (PS 091) setelah 12 minggu. Densitas kalus pada TIB berkorelasi negatif dengan karakteristik fisikokimia medium. Hal ini menggambarkan variasi intensitas pertumbuhan dan metabolisme kalus dengan adanya perbedaan densitas pada BPS. Penggunaan BAP 0,2 mg L-1 bersama kinetin 0,2 mg L-1 paling sesuai untuk memacu regenerasi kalus tebu dengan menghasilkan jumlah tunas terbanyak dalam waktu relatif lebih cepat (1 – 2 minggu lebih cepat) dibandingkan perlakuan lainnya dan dengan tingkat kejadian pencoklatan yang rendah.
[Kata kunci: kultur in vitro, kultur cair, proliferasi]
Abstract
Temporary immersion bioreactor (TIB) has been utilized for the mass-scale propagation of many important plants, including sugarcane. TIB facilitates a semiautomated culture system and provides optimal conditions for plant growth. Several factors determine plant growth in the TIB, such as explant density. Therefore, an experiment was carried out to determine the optimal initial weight of sugarcane calli and to evaluate its effect on the proliferation and regeneration in TIB. Sugarcane calli were induced from spindle leaves isolated from four Indonesian prime sugarcane varieties. The initial weights of the calli cultured in the TIB flasks were 0.05 g, 0.1 g, 0.2 g, 0.5 g and 1.0 g per flask. The calli were proliferated through three cycles in TIB, and subsequently regenerated in TIB with auxin and cytokinin treatments. The results of the experiments showed that 0.2 g was the most efficient initial weight for sugarcane callus proliferation in the TIB, resulting in an exponential multiplication rate of 130.3-fold (PSKA 942), 136.8-fold (PS 094), 21.3-fold (PS 881), and 12.9-fold (PS 091) within 12 weeks. In the TIB, callus density showed a negative correlation with the physicochemical properties of the medium, demonstrating various growth intensities or metabolic activities of calli at different densities in the TIB. The use of 0.2 mg L-1 BAP along with 0.2 mg L-1 kinetin was suitable for promoting the regeneration of sugarcane calli and producing the highest number of shoots in a relatively short amount of time (1 – 2 weeks faster) with low incidences of browning.
[Keywords: in vitro culture, liquid culture, proliferation]
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