V. SIMPULAN DAN SARAN
5.2. Saran
Perlunya dilakukan penelitian lanjutan mengenai tebu transforman untuk pengujian stabilitas dan ekspresi gen fitase.
Daftar Pustaka
Ananda, RR Woro Umayi. 2004. Studi transformasi pada tebu dengan perantara
Agrobacterium tumafaciens GV 2260 (pMA) serta regenerasi kalus transgenik. Tesis. Sekolah Pascasarjana. IPB. 63 hlm.
Beyl AC. 2005. Getting started with tissue culture, media preparation, sterile technique, and laboratory equipment. Di dalam Trigiano, N.Robert and Gray, J. Dennis (ed). Plant Development and Biotechnology. CRC Press. P : 19 – 38.
Blanco AM, Nieves N, Sanchez M, Borroto GC, Castillo R, Gonzalez LJ, Escalona M, Baez E dan Hernandez Z. 1997. Protein changes associated with plant regeration in embryogenic calli of sugarcane (Saccharum sp). Plant Cell, Tissue and Organ Culture 51. P : 153 – 158.
Brown TA. 1996. Gene cloning an introduction. Champan & Hall.
Caponetti DJ, Gray JD and Trigiano NR. 2005. History of plant tissue and cell culture. Di dalam Trigiano, NR dan Gray JD (ed) Plant Development and Biotechnology. CRC Press. P : 9 – 15.
Cheng M, Fry JE, Peng S, Zou H, Hironaka R, Duncan DR, Connor TW dan Wan Y. 1997. Genetic transformation of wheat mediated by Agrobacterium tumefaciens. Plant Physiol 115. P : 971 – 980.
Chengalrayan K dan Gallo-Meagher M. 2001. Effect of various growth regulators on shoot regeneration of sugarcane. In Vitro Cell. Dev. Biol. Plant 37. P : 434 – 439.
Cosgrove DJ. 1980. Inositol phosphates, their chemistry, biochemistry and physiology. New York, Elsevier Scientific Publishing Company.
Davies JP. 1995. The plant hormones : their nature, occurrence, and functions. Di dalam Davies JP (ed). Plant hormones : physiology, biochemistry and molecular biology. Kluwer Academic Publishers. p. 1 – 12.
Dewan Gula Nasional. 2003. Statistik perkembangan gula Indonesia 2002-2003. Direktorat Jenderal Bina Produksi Perkebunan. Deptan. Jakarta.
Gaba PV. 2005. Plant growth regulators in plant tissue culture and development. Di dalam Trigiano, NR dan Gray JD (ed). Plant development and biotechnology. CRC Press. P : 241 – 250.
Gelvin BS. 2003. Agrobacterium-mediated plant transformation : the Biology behind the “Gene-Jockeying” tool. Microbiology and Molecular Biology Reviews. Vol. 67. No. 1. p. 16-37.
Gunawan LW. 1992. Teknik kultur jaringan tumbuhan. Direktorat Jenderal Pendidikan Tinggi. Pusat Antar Universitas Bioteknologi. IPB. Bogor.
Gustavo AR, Gonzales-Cabrera J, Vazquez-Padron R dan Ayra-Pardo C. 1998.
Agrobacterium tumefaciens : A natural tool for plant transformation. molecular biology and genetic. Plant Biotechnology Electric Jurnal Biotechnology. Vol. 1. No. 3.
Greiner R. 2005. Procedure phytase activity from callus. [Tidak dipublikasikan]. Greiner R, Konietzny U, Jany KD. 1993. Purification and characterization of two
phytases from Escherichia coli. Arch Biochem Biophys 303. P : 107 – 113.
Haris N dan. Mathius NT. 1995. Teknologi in vitro untuk pengadaan bibit tanaman perkebunan. Warta Pusat Penelitian Bioteknologi Perkebunan. 1(1):hal 2-9.
Hartatik S. 2000. Studi genetik plasma nutfah tebu (Saccharum spp) berdasarkan penanda morfologi, agronomi dan isozim. Disertasi. Program Pascasarjana IPB. Bogor.
Hegeman EC and Elizabeth AG. 2001. A novel phytase with sequence similarity to purple acid phosphatases is expressed in cotyledons of germinating soybean seedlings. Plant Physiology 126. P : 1598 – 1608.
Hoy WJ, Bischoff PK, Miligan BS dan Gravois AK. 2003. Effect of tissue culture explant source on sugarcane yield components. Euphytica 129. P : 237 – 240.
Indarwati S. 2000. Isolasi dan modifikasi media produksi bakteri penghasil fitase. Tesis. Program Pascasarjana IPB. 56 hlm.
Kane EM. 2005. Shoot culture procedures. Di dalam Trigiano, NR dan Gray JD (ed). Plant development and biotechnology. CRC Press. P : 241 – 250. Khudori. 2005. Posisi Indonesia di pertemuan WTO. Koran Tempo. 13
Desember 2005.
Kilmer OL,. Serb PA dan. Hoseney RC. 1994. Effects of mineral and apparent phytase activity in the development of hard to cook state of beans. Cereal Chem 71 (5). P : 476 – 482.
Lenhninger AL. 1990. Dasar-dasar mikrobiologi. Rajawali Press. Jakarta. Li TZ dan Gray JD. 2005. Genetic engineering technologies. Di dalam Trigiano,
NR dan Gray JD (ed). Plant development and biotechnology. CRC Press. P : 241 – 250.
Liu MC. 1981. In vitro methods applied to sugarcane improvement. Di dalam Thorpe TA (eds.) Plant tissue culture methods and application in agriculture. Academic Press. New York. P.23.
Mamun MA, Sikdar MBH, Paul DK, Rahman MM, dan Islam MR. 2004. In vitro micropropagation of some important sugarcane varieties of Bangladesh. Asian Journal of Plant Sciences 3 (6): 666-669.
Martin JR, Golves G, de Armas R, Espinosa R, Vigoa R, Leon A. 1982. La Cana de Azucar en Cuba. Di dalam Enriquez-Obregon GA, Vazquez-Padron RI, Prieto-Samsonov DL (ed). Genetic transformation of sugarcane by
Agrobacterium tumefaciens using antioxidant compound. Biotechnologia Aplicada. 14. P : 169 – 174.
Minarsih H. 2003. Rekayasa genetik tebu (Saccharum officinarum L.) untuk toleransi kekeringan. Laporan Riset. Riset Unggulan Terpadu VIII Bidang Bioteknologi. Kementerian Riset dan Teknologi RI Lembaga Ilmu Pengetahuan Indonesia.
Naik GR. 2001. Sugarcane biotechnology. Science Publisher, Inc. Enfield (NH), USA.
Nester WE, Kemner J, Deng W, Lee YW, Fullner K, Liang X, Pan S dan Heath DJ. 1996. Agrobacterium : A natural genetic engineer exploited for plant biotechnology. Di dalam Stacey G, Mullin B dan Gresshoff MP (ed). Bioloy of plant-microbe interactions. International Society for Molecular Plant-Microbe Interaction. St. Paul, Minnesota, USA.
Santosa DA. 2002. IPB memiliki tebu transgenik. Kompas 11 (Kolom 1 – 5). 24 September 2002.
Santosa DA, Hendroko R, Farouk A dan Greiner R. 2004. A rapid and higly efficient method for transformation of sugarcane callus. Molecular Biotechnology. Vol.24. P. 113 – 119.
Sastrowijono S. 1991. Aspek fisiologis dalam mikropropagasi tebu. Di dalam : Diktat Pelatihan Penangan Mikropropagasi Tanaman Tebu. Pasuruan 4 – 9 Februari 1991. Pusat Penelitian Perkebunan Gula Indonesia. Pasuruan. Shieh TR, dan Ware JH. 1968. Survey of microorganism for the production of
extracelluler phytase. Appl. Microbiol 16 P : 1348 – 1351.
Sheng J dan Citovsky V. 1996. Agrobacterium – Plant cell DNA transport : Have virulence proteins will travel. The Plant Cell 8. P : 1699 – 1740.
Singh N, Somai BM dan Pillay D. 2005. In vitro screening of sugarcane to evaluate smut susceptibility. Plant Cell, Tissue and Organ Culture 80. P : 259-266.
Sreevinasan TV, Ahloowalia BS dan. Heinz DJ. 1987. Cytogenetics. Di dalam Heinz DJ (ed). Sugarcane improvement through breeding. Elsevier Amsterdam. Pp: 211-252.
Sugiyarta E. 1991. Persyaratan laboratorium dan penetapan media pada teknik kultur jaringan tebu. Di Dalam : Diktat Pelatihan Penangan Mikropropagasi Tanaman Tebu. Pasuruan 4 – 9 Februari 1991. Pusat Penelitian Perkebunan Gula Indonesia. Pasuruan.
Trigiano RN dan Gray JD. 2000. Plant tissue culture concepts and laboratory exercise. 2nd edition. CRC Press. New York.
Uchimiya H, Handa T dan Brar DS. 1989. Transgenic plant. biotech 12. P : 1 – 20.
Wattimena GA. 1988. Zat pengatur tumbuh tanaman. Pusat Antar Universitas Institut Pertanian Bogor Bekerjasama Dengan Lembaga Sumberdaya Informasi – IPB. Bogor.
Wattimena GA, Gunawan LW, Mattjik AN, Syamsudin E, Wiendi AMN dan Ernawati A. 1992. Bioteknologi tanaman. Depertemen Pendidikan Dan Kebudayaan Direktorat Jenderal Pendidikan Tinggi. Pusat Antar Universitas Bioteknologi. Institut Pertanian Bogor. Bogor.
Wulandari I. 2005. Studi beberapa metode transformasi genetik tanaman tebu (Saccharum offinarum L.) dengan gen fitase melalui perantara
Agrobacterium tumefaciens GV 2260. Tesis. Sekolah Pascasarjana IPB.54 hlm.
Lampiran 1. Komposisi media MS untuk induksi kalus dan regenerasi planlet yang telah dimodifikasi
Induksi Kalus Regenerasi Tunas Induksi Akar Persenyawaan MS-I mg/L MS-T mg/L MS-A mg/L A. NH4NO3 1650 1650 1650 B. KNO3 1900 1900 1900 170 170 170 6.2 6.2 6.2 0.83 0.83 0.83 0.250 0.250 0.250 C. KH2PO4 H3BO3 KI Na2Mo4.2H2O CoCl2.6H2O 0.025 0.025 0.025 D. CaCl.2H2O 440 440 440 370 370 370 22 22 22 8.6 8.6 8.6 E. MgSO4.7H2O MnSO4.4H2O ZnSO4.4H2O CuSO4.7H2O 0.025 0.025 0.025 37.3 37.3 37.3 F. Na2EDTA.2H2O FeSO4.7H2O 27.8 27.8 27.8 Myo-Inositol 1.0 1.0 1.0 0.1 0.1 0.1 0.5 0.5 0.5 0.5 0.5 0.5 Vit Thiamine Niacine Pyridoxine Glycine 2.0 2.0 2.0 Air kelapa* - 100 ml 100 ml Sukrosa 30000 30000 30000 Gelrite 2500 2500 - 2,4-D 3.0 - -
Kinetin 0.1 Sesuai dgn perlakuan -
BAP - Sesuai dgn perlakuan -
IBA - - Sesuai dgn perlakuan
Lampiran 2. Komposisi media LB Tripton 10 g/L Ekstrak Yeast 5 g/L NaCl 5 g/L Sukrosa 5 g/L MgSO4.7H2O 20 g/L MgCl2.6H2O 10 g/L
Lampiran 3. Komposisi media transformasi Metode Santosa, et al., (2004)
Media Komposisi MS MS salts; 1 mg/l asam nicotinat; 0.8
mg/l vitamin B1; 0.5 mg/l vitamin B6; 100 mg/l myo-inositol dan 20 g/l sukrosa
MS cd MS salts; 1 mg/l asam nicotinat; 0.8 mg/l vitamin B1; 0.5 mg/l vitamin B6; 100 mg/l myo-inositol dan 20 g/l sukrosa, 500 mg/l casein hidrolisat; 5 mg/l 2,4 D
MS cd-AO MS Induksi kalus ditambah dengan 15 mg/l asam askorbat; 40 mg/l sistein; 2 mg/l silver nitrat
MS Seleksi MS salts; 1 mg/l asam nicotinat; 0.8 mg/l vitamin B1; 0.5 mg/l vitamin B6; 100 mg/l myo-inositol dan 20 g/l sukrosa; 5 mg/l 2,4 D dan 150 mg/l kanamycin