In Vitro Propagation of Selected Sugarcane ( Saccharum Officinarum L . ) Varieties ( C 86-165 and C 86-12 ) Through Shoot Apical Meristem
Sugarcane (Saccharum officinarum L.) is monocotyledonous crop plant that belongs to the Poaceae family that mostly propagates through conventional methods. However, conventional propagation lacks rapid multiplication procedures to commercialize newly released varieties within a short period of time. Hence, the objective of this work was to optimize in vitro micro propagation protocol for two sugarcane varieties (C 86-165 and C 86-12) using shoot apical meristem explants. The two varieties were cultured on MS medium supplemented with different concentrations of growth regulators on shoot initiation, multiplication and root induction stages. Analysis of variance (ANOVA) revealed that the two varieties showed statistically significant difference in their response to the various hormonal treatments with regard to the parameters measured. For initiation stage vars. C 86-165 and C 86-12 performed best on 1.0 mg/l and 0.5 mg/l of BAP, respectively. On the other hand, multiplication stage was best in MS media enriched with 2.0 mg/l BAP + 0.5 mg/l NAA and 1.5 mg/l BAP + 0.5 NAA as manifested in terms of number of shoots and shoot length per explants after four weeks of culture for vars. C 86-165 and C 86-12, respectively. With regard to root induction, best rooting response in terms of mean root number, mean root length, mean fresh weight and mean dry weight was achieved in 1/2 MS media enriched with 2.0.mg/l NAA and 1.0 mg/l NAA after four weeks of culture for vars. C 86-165 and C 86-12, respectively.
Induksi Kalus Morfogenik dan Regenerasi Tanaman Tebu (Saccharum officinarum L.) secara In Vitro Induction of Morphogenic Callus and In Vitro Regeneration of Sugarcane
ABSTRAK Propagasi in vitro merupakan cara memperoleh tanaman baru yang sehat secara cepat dan massal, Percobaan in vitro dilaksanakan untuk menginduksi kalus morfogenik dan regenerasi dari varietas tebu domestik. Bagian terdalam gulungan daun tebu digunakan sebagai eksplan dan diinkubasi pada medium MS yang dimodifikasi dan dilengkapi dengan 2,4-D pada konsentrasi 1 mg/L, 2.5 mg/L, dan 5 mg/L untuk menginduksi pembentukan kalus. Kalus yang terbentuk di medium induksi selanjutnya dipindahkan ke medium MS yang diperkaya dengan zat pengatur tumbuh untuk inisiasi dan perbanyakan tunas. Persentase pembentukan kalus tertinggi dicapai pada penggunaan medium yang mengandung 2.5 mg/L 2,4-D dan maksimal proliferasi kalus morfogenik diperoleh setelah subkultur ketiga. Dua tipe kalus yang terbentuk pada medium induksi yaitu berstruktur kering, remah dan struktur halus kompak. Jenis kalus yang morfogenik tersebut berwarna putih dan putih kekuningan serta mudah terpisah-pisah. Hasil proliferasi tunas dari kalus tertinggi diperoleh pada medium MS dengan kombinasi hormon tumbuh 2 mg/L Kinetin + 1 mg/L IAA dan tidak ada pertumbuhan tunas pada medium yang ditambahkan zat pengatur tumbuh tunggal, yaitu Kinetin. Dengan demikian, penelitian ini menyimpulkan bahwa hormon tumbuh jenis sitokinin yang dikombinasi dengan auksin sangat dibutuhkan untuk regenerasi kalus tebu secara in vitro. Kata Kunci:
Thidiazuron stimulates shoot regeneration of sugarcane embryogenic callus
SummaryEfficient shoot regeneration of sugarcane (Saccharum spp. hybrid cv. CP84-1198) from embryogenic callus cultures has been obtained using thidiazuron (TDZ). Callus was placed on modified Murashige and Skoog (MS) medium containing 2.3 μM 2,4-dichlorophenoxyacetic acid (2,4-D), or 9.3 μM kinetin and 22.3 μM naphthaleneacetic acid (NAA) and compared with the same MS medium supplemented with 0.5, 1.0, 2.5, 5.0 or 10.0 μMTDZ, A11 TDZ treatments resulted in faster shoot regeneration than the kinetin/NAA treatment, and more shoot production than either the 2,4-D or kinetin/NAA treatments. Maximum response, as determined by total number of shoots (26 per explant) and number of shoots greater than 1 cm (4 per explant) 4 wk after initiation, was obtained with 1.0 μM TDZ. The shoots rooted efficiently on MS medium supplemented with 19.7 μM indole-3-butyric acid (IBA). These results indicate that TDZ effectively stimulates sugarcane plant regeneration from embryogenic callus, and may be suitable to use in genetic transformation studies to enhance regeneration of transgenic plants.