Sains Malaysiana 40(4)(2011): 339–343

An Effective Protocol for Callus Induction with Milk Clotting Activity from Solanum dubium Seeds

(Protokol Berkesan bagi Induksi Kalus dengan Aktiviti Pembekuan Susu daripada Biji Benih Solanum dubium)

Fatima Musbah Abbas¹ *, Elbushra Elsheikhelnur², Normah Mohd Noor¹, Eisa Algaali³, Zainon Mohd Ali⁴ & Roohaida Othman⁴

¹Institute of Systems Biology, Universiti Kebangsaan Malaysia

43600 Bangi, Selangor D.E., Malaysia

²Faculty of Science, University of Khartoum

P. O. Box 321, 11115 Khartoum, Sudan

³Commissions of Biotechnology and Genetic Engineers

Ministry of Science and Technology, Sudan

⁴School of Biosciences and Biotechnology, Faculty of Science and Technology

University Kebangsaan Malaysia, 43600 Bangi, Selangor D.E., Malaysia

Received: 9 December 2009 / Accepted: 24 June 2010

ABSTRACT

The aim of this study was to establish an effective protocol for callus induction from the seed explants of Solanum dubium and to investigate the callus extract ability in milk clotting activity. The effects of growth regulator, basal media strength and sucrose were studied using different concentrations (0.5, 1.0, 1.5, and 2.0 mg/L) of indole-3-acetic acid (IAA), indole-3-butyric acid (IBA), naphthalene acetic acid (NAA) and 2,4-dichlorophenoxy-acetic acid (2,4-D) alone or in combination with 0.5 mg/L of 6-benzylamino purine (BAP). For milk clotting activity, about 50 or 100 μL extracts of seed callus was mixed with 2 mL 50% milk held at 55°C for 5 and 10 min until milk clotting occurred. The results showed that NAA alone or in combination with BAP gave a higher callusing percentage (80 to100%) compared to the other plant growth regulators at the same concentrations. When an auxin was supplied in combination with BAP, a significant increase in callusing percentage or degree of callusing was observed. The time required for callus to be developed was shortened and the quality of the induced callus improved. An increase in callus growth in low sucrose (10 g/L) concentration was found be comparable (88%) to high sucrose concentration (30 g/L; 60%). Crude extracts obtained from S. dubium callus were shown to exhibit milk clotting activity.

Keywords: Auxin; callus; Gubbien; milk clotting activity; Solanum dubium

ABSTRAK

Kajian ini dijalankan untuk mendapatkan protokol yang berkesan bagi penginduksian kalus daripada eksplan biji benih Solanum dubium dan untuk mengkaji keupayaan ekstrak kalus dalam aktiviti pembekuan susu. Kesan pengawalatur pertumbuhan, medium asas, kekuatan medium dan kepekatan sukrosa telah dikaji menggunakan kepekatan pengawal atur pertumbuhan (0.5, 1.0, 1.5, dan 2.0 mg/L) asid indol-3-asetik (IAA), asid indol-3-butirik (IBA), asid naftalena asetik (NAA) dan asid 2,4-diklorofenoksi–asetik (2,4-D) sendiri atau berkombinasi dengan 0.5 mg/L 6-benzilamino purina (BAP) digunakan. Untuk pembekuan susu, 50 atau 100 μL ekstrak kalus biji benih dicampur dengan 2 mL 50% susu pada 55°C untuk 5 dan 10 min sehingga pembekuan susu berlaku. Hasil menunjukkan NAA sendiri atau bersama dengan BAP memberi peratus pembentukan kalus yang lebih tinggi berbanding pengawal atur pertumbuhan lain pada kepekatan sama. Apabila auksin diberi bersama BAP, pertambahan yang signifikan dilihat dalam peratus atau tahap pembentukan kalus. Masa untuk pembentukan kalus telah disingkatkan dan kualiti kalus didapati adalah lebih baik. Peningkatan pembentukan kalus (88%) yang dicerap pada medium yang mengandungi kepekatan sukrosa yang rendah (10 g/L) didapati setanding dengan medium yang mengandungi sukrosa lebih tinggi (30 g/L; 60%). Ekstrak kasar daripada kalus S. dubium didapati menunjukkan aktiviti pembekuan susu.

Kata kunci: Aktiviti pembekuan susu; auksin; Gubbien; kalus; Solanum dubium

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*Corresponding author; email: fatimamisbah@hotmail.com