KONDISI OPTIMUM FUSI PROTOPLAS ANTARA JAMUR TIRAM PUTIH {Peurotus floridae) DAN JAMUR TIRAM COKLAT {Pleurotus cystidiosusy

Ira N Djajanegara, Korri El-khobar
| Abstract views: 911 | PDF views: 2281

Abstract

Genetic crossing of white oyster mushroom to introduce longer storage life trait can only be done within individuals in this particular species. However, longer storage life trait is possessed by brown oyster mushroom (Pleurotus cystidiosus) which is other species within this genus. Thefeore, protoplast fusion between white oyster mushroom (Peurotus floridae) and brown oyster mushroom (Pleurotus cystidiosus) was conducted to hopefully obtain an oyster mushroom strain that has higher production and longer storage life. Protoplast fusion was done by isolating protoplast from 5-days old monokaryotic mycelia grown in PDB. As much as 3.15 x 10 protoplasts/ml were harvested using mixture of cellulase Onozuka R-10 (1%) and macerozyme R-10 (1%) from brown oyster mushroom with 80.61% viability. Similarly, 3.71 x 10 protoplasts/ml were harvested using lysing enzyme (2%)from brown oyster mushroom with 83.68% viability. Protoplast fusion were conducted using 0% (control), 30%, 40% and 50% of PEG6000. Fusion periods were done at 10, 20 and 30 minutes. The candidate fusants were then screened using MRM (minimum regeneration media) media. Based on this experiment, the optimum conditions for protoplast fusion is 10 minutes incubation using 40% PEG6000 that yielded 121 colonies grown on MRM media as candidate fusants.

Keywords

Fusi , MRM, PEG, Pleurotus cystidiosus, Pleurotus florida, protoplas, regenerasi.

Full Text:

PDF

References

Bhandal MS dan KB Mehta. 1994. Geentic improvement of Pleurotus species. Dalam: Nair MC (ed.). 1994. Advances in mushroom biotechnology. Scientific Publisher, Jodhpur: 127-133.

Budiwati TA dan Pudjiraharti. 2001. Improvement of tempe inoculum quality through protoplast fusion of Rhizopus oligosporus L36 spores and Brevibacterium epidermis cell to produce B-glucosidase. The 2nd Indonesian Biotechnology Conference: 711-719.

Cahyana YA, Muchrodji dan M Bakrun. 2002. Jamur tiram: Pembibitan, pembudidayaan, dan analisi usaha. Penebar Swadaya, Jakarta. 23-24.

Constabel F dan Aj Cutler. 1985. Protoplast fusion. Dalam: Fowke, LC dan F Constabel. 1985. Plant protoplast. SRC Press Inc., Florida: 53-62.

Davis B. 1985. Factors influencing protoplast isolation. Dalam: Peberdy, JF & L Ferenczy (eds.) 1985. Fungal protoplasts: Application in biochemistry and genetics. Marcell Dekker, Inc., New York: 45-71.

Endress R. 1994. Palnt Cell biotechnology. Springer-Verlag, Nerlin. 211-215.

Eriksson TR. 1985. Protoplast isolation and culture. Dalam: Fowke LC dan F Constabel. 1985. Palnt protoplast. CRC Press Inc., Florida: 1-15.

Fadlia I. 1993. Kajian awal penerapan fusi protoplas Volvariella volvacea (BULL. EX. FR) SING. dengan Auricularia polytricha (MONT.) SACC. Tesis Sarjana Biologi ITB, Bandung. 22-24.

Gomez KA dan AA Gomez. 1995. Prosedur statistik untuk penelitian pertanian. Ed. ke-2. Terj. dari Statistical procedures for agricultural research, oleh E. Syamsuddin & JS Baharsyah. Penerbit Universitas Indonesia, Jakarta. 455 - 457.

Hadini H. 1997. Hibridisasi somatik pisang ambon dankavendis secara in vitro. Tesis Pascasarjana UGM, Yogyakarta. 65 -67.

Irawan D. 2004. Optimasi isolasi protoplas jamur tiram putih (P. ostreatus floridae) dan jamur tiram coklat (P. ostreatus cystidious). Laporan Kerja Praktek Departemen Biologi FMIPA UI, Depok. 18 -19.

Kao KN. 1991a. Fusi protoplas tanaman dan isolasi heterokariosit. Dalam: Wetter, LR & F Constabel (eds.). 1991. Metode kultur jaringan tanaman. Ed. ke-2. Penerbit ITB, Bandung. 65-74.

Kao KN. 1991b. Metode pewarnaan untuk protoplas dan sel. Dalam: Wetter, LR & F Constabel (eds.). 1991. Metode kultur jaringan tanaman. Ed. ke-2. Penerbit ITB, Bandung. 87-93.

Kiyohara H, T Watanabe, J Imai, N Takizawa, T Hatta, K Nagao dan A Yamamoto. 1990. Intergeneric hybridization between Monascus anka and Aspergillus oryzae by protoplast fusion. Applied Microbiology Biotechnology 33: 671-676.

Larraya LM, G Perez, MM Penas, JJP Baars, TP Mikosch, AG Pisabarro dan L Ramirez. 1999. Molecular karyotype of the white rot fungus Pleurotus ostreatus. Applied and Environmental Microbiology 65(8): 3413-3417.

Larraya LM, G Perez, E Ritter, AG Pisabarro dan L Ramirez. 2000. Genetic linkage map of the edible basidiomycete Pleurotus ostreatus. Applied and Environmental Microbiology 66(12): 5290-5300.

Necas O dan A Svoboda. 1985. Cell wall regeneration and protoplast reversion. Dalam: Peberdy, JF & L Ferenczy (eds.) 1985. Fungal protoplasts: Applications in biochemistry and genetics. Marcell Dekker, Inc., New York. 115-133.

Purwito A. 1999. Fusi protoplas intra dan interspesies pada tanaman kentang. Disertasi Pascasarjana IPB, Bogor. 211 -214.

Ramirez L, L. Larraya dan AG Pisabarro. 2000. Molecular tools for breeding basidiomycetes. International Microbiology 3: 147-152.

Santiago Jr, CM. 19S2. Production of Volvariella protoplasts by use of Trichoderma enzyme. Mushroom Newsletter for the Tropics 3(2): 3-6.

Santoso U dan F Nursandi. 2003. Kultur jaringan tanaman. UMM Press, Malang. 190 - 193.

Sarkar S dan G Saunders. 1997. The role of genetics in the effective utilisation of filamentous fungi. International Conference on Biodiversity and Bioresources: Conservation and Utilization, 23-27 November 1997, Phuket: 1-9.

Setiari N. 2001. Studi tentang fusi protoplas pada tanaman kacang hijau(vigna radiata (L.) Wilczek) dan kacang hijau liar (Vigna sublobata L): Struktur anatomi protoplas induk dan protoplas hasil fusi. Tesis Magister Sains ITB, Bandung. 4 7 - 5 1 .

Setiawati MR dan Juhiya. 2005. Jamur tiram. 4 him. http:/ Ayarintek.progressio.or.id/pertanian/jtiram.htm. 15 Maret 2005, pk. 11.15.

Smith JE. 1993. Prinsip bioteknotogi. Terj. dari Biotechnology principle, oleh Sumo, U.F., B. Sumantri & A. Sumantri. PT. Gramedia Pustaka Utama, Jakarta. 202 - 211.

Sudjana. 1991. Desain dan analisis eksperimen. Ed. ke-3. Penerbit Tarsito, Bandung. 415 - 423.

Sudjana. 1992. Metoda statistika. Ed. ke-5. Penerbit Tarsito, Bandung. 386 - 387.

Sufiani N. 1993. Fusi protoplas intergenera antara Agrocybe cyllindracea (D.C. ex Fr.) Gillet dengan Lentinus edodes (Berk.) Sing. Tesis Sarjana Biologi ITB, Bandung. 42 - 43.

Suryowinoto M. 1996. Pemuliaan tanaman secara in vitro. Penerbit Kanisius, Yogyakarta. 2 1 2 - 2 1 3 .

Suriawira U. 1993. Pengantar untuk mengenal dan menanam jamur. Penerbit Angkasa, Bandung. 10 -13.

Varavallo MA, MV de Quieroz, JF Pereira dan EF de Araujo. 2004. Isolation and regeneration of Penicillium brevicompactum protoplasts. Acta Scientarum 26(4): 475-479.

Ye GN dan ED Earle. 1991. Effect of cellulases on spontaneous fusion of maize protoplasts. Plant Cell Reports 10: 213-216.

Zhao KD, Zhou W P dan J Ge. 2004. Study on the preparation and regeneration of protoplast from taxolproducing fungus Nodulisporium sylviforme. Nature and Science 2(2): 52-59.


Refbacks

  • There are currently no refbacks.