THE INFLUENCE OF PLANT GROWTH REGULATORS AND LIGHT SUPPLY ON BITTER CASSAVA CALLUS INITIATION FOR STARCH PRODUCTION

Abstract

The application of cassava starch in the biopolymers industry has been growing significantly due to its low cost, good oxygen barrier in the dry state, and biodegradability. Plant culture technology is an alternative to traditional propagation as it does not require large areas for production, has a higher rate of multiplication, and produces disease-free plants. However, the application of cassava callus culture for starch production is limited. This study focuses on identifying the significant culture parameters for a maximum Rayong cultivar cassava callus growth and evaluation of its starch content. Cassava stems petiole and leaf explants were cultured on MS medium containing different combinations of 2,4-D (8, 12, and 15 mg/L) and BAP (1, 3, and 5 mg/L) under three light conditions (0, 16, and 24 h). The screening of the most influential parameter was done using the 2-level Factorial Design in Design Expert v13 by analyzing the frequency of callus formation. All leaf explant turned brown with no callus induction. The highest frequency of callus formation derived from stem petiole explant was achieved by the combination of 8 mg/L 2,4-D and 1 mg/L BAP under the light condition (75%) followed by 8 mg/L 2,4 D + 1 mg/L BAP under the dark condition (50%). Based on the ANOVA analysis, the individual supply of 2,4-D and BAP respectively, have a negative effect on callus formation while the combination of 2,4-D and BAP has a positive effect. Light supply did not significantly affect cassava callus formation. The amount of starch in the cassava callus was then investigated using an iodine test which yielded 0.21% of the total weight of the callus (0.0101g). The amount of starch is relatively low considering that the callus was not grown under the optimum condition for starch production. The findings of this study open prospects for future research in cassava cultures in favor of starch production. ABSTRAK: Penggunaan kanji ubi kayu dalam industri biopolimer telah meningkat secara mendadak disebabkan oleh faktor kosnya yang rendah, rintangan oksigen yang baik dan sifat keterbiodegradasi. Teknologi kultur tisu tumbuhan merupakan alternatif kepada pembiakan secara tradisional yang mana teknologi ini tidak memerlukan kawasan penanaman yang luas, penghasilan yang lebih singkat dan menghasilkan tumbuhan bebas penyakit. Walau bagaimanapun, kajian mengenai penggunaan kanji dari kultur ubi kayu masih belum meluas. Kajian ini bertujuan mengenal pasti parameter penting dalam pertumbuhan maksimum kalus kultivar ubi kayu Rayong dan kandungan kanjinya. Eksplan dari tangkai petiol dan daun ubi kayu dikulturkan dalam medium MS yang mengandungi pelbagai kombinasi hormon tumbuhan 2,4-D (8, 12 dan 15 mg/L) dan BAP (1, 3 dan 5 mg/L) di bawah tiga tempoh masa pencahayaan (0, 16 dan 24 jam). Saringan parameter paling berpengaruh dilakukan menerusi reka bentuk faktorial 2-peringkat perisian Design Expert v13 melalui analisa frekuensi pembentukan kalus. Semua eksplan daun telah bertukar dari hijau ke perang dan tidak menunjukkan induksi kalus. Bagi eksplan batang daun, frekuensi pembentukan kalus tertinggi diperoleh dari kombinasi 8 mg/L 2,4-D dan 1 mg/L BAP di bawah pencahayaan 24 jam (75%) diikuti dengan 8 mg/L 2,4 D + 1 mg/L BAP di bawah pencahayaan malap (50%). Berdasarkan analisis ANOVA, medium 2,4-D dan BAP masing-masing menunjukkan kesan negatif kepada pembentukan kalus, sementara kombinasi 2,4-D dan BAP memberikan kesan positif. Dapatan ini menunjukkan bahawa pencahayaan tidak signifikan terhadap pembentukan kalus kultur ubi kayu. Kandungan kanji di dalam kalus ubi kayu kemudiannya diuji dengan larutan iodin, dan menunjukkan dapatan sebanyak 0.21% dari berat kalus (0.0101 g). Secara relatif, nilai ini adalah rendah memandangkan eksperimen ini bukan dihasilkan di bawah keadaan optimum bagi pembentukan kanji. Hasil kajian ini membuka peluang kepada kajian-kajian lain di masa hadapan dalam penghasilan kanji dari kultur tisu ubi kayu.

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