Path: Top S2-Theses Biology 1998

STUDI FUSI PROTOPLAS Vigna radiata (L.) Wilczek var. radiata DENGAN Vigna radiata (L.) Wilczek var. sublobata (Roxb.) Verdcourt

STUDY OF PROTOPLAST FUSION BETWEEN Vigna radiata (L.) WILCZEK var. radiata AND ITS WILD RELATIVE Vigna radiata (L.) WILCZEK var. sublobata (ROXB.) VERDCOURT

Master Theses from JBPTITBPP / 2001-08-25 10:03:00
Oleh : Wiwid Angraini, S2 - Biology (v_wied@lycos.com)
Dibuat : 20001-04-3, dengan 1 file

Keyword : Vigna radiata, fusi protoplas, PEG

Beberapa jenis larutan PEG telah digunakan untuk menginduksi fusi antara protoplas hipokotil Vigna radiata (L.) Wilczek var. radiata dengan protoplas daun Vigna radiata (L.) Wilczek var. sublobata (Roxb.) Verdcourt dengan tujuan mendapatkan hasil fusi protoplas tertinggi dan kemudian mengamati perkembangan tahap awal protoplas hasil fusi serta protoplas kedua induknya. Protoplas dari kedua macam eksplan diisolasi secara enzimatis dengan menggunakan larutan enzim yang terdiri dari cellulase YC 2%, macerozyme R-10 1%, mannitol 0,4 M dan CaCl2.2H2O 2,5 mM. Protoplas dari hipokotil dan daun difusikan dengan menggunakan tiga macam larutan (P1, P2, P3) yang mengandung polyethylene glycol (PEG) BM 4000 dan senyawa-senyawa lain. Persentase fusi tertinggi (15,27%) diperoleh dari larutan P2, yang mengandung PEG 33%, sukrosa 1,8%, KH2PO4 1mM dan CaCl2.2H2O 10 mM. Pengamatan jumlah inti protoplas hasil fusi yang diwarnai dengan pewarnaan 4-6 diamino 2-phenilindole (DAPI) di bawah mikroskop fluoresensi menunjukkan 58,82% uninukleat, 18,23% binukleat, dan 22,95% multinukleat. Viabilitas protoplas hasil fusi relatif tinggi pada awal kultur sampai terjadinya pembelahan pertama. Permukaan protoplas hipokotil dan daun telah tertutup sempurna dengan materi dinding sel setelah 8 jam kultur sedangkan pada protoplas hasil fusi setelah 24 jam kultur. Pembelahan sel pertama terjadi setelah usia kultur 2 hari pada protoplas hipokotil, serta 3 hari usia kultur pada protoplas daun dan protoplas hasil fusi. Kemampuan sel untuk membelah dan membentuk koloni pada protoplas hasil fusi sangat kecil (0,32%) dibanding protoplas hipokotil (3,11%) dan protoplas daun (4,28%). Berdasarkan hasil yang diperoleh dapat disimpulkan bahwa persentase hasil fusi tertinggi diinduksi dengan larutan P2, dan hasil fusi mampu beregenerasi hingga terbentuk mikrokoloni.

Deskripsi Alternatif :

Various PEG solutions were used to induce protoplasts fusion between hypocotyl protoplast Vigna radiata var. radiata and leaf protoplast Vigna radiata var. sublobata, with purposes to find the highest fusion product, and the early stage of development of fusion product and its parents protoplasts were then observed. Protoplasts from hypocotyl of V. radiata var. radiata and leaf V. radiata var. sublobata were isolated enzymatically with enzyme solutions consisted of 2% cellulase YC, 1% macerozyme R-10, 0,4 M mannitol and 2,5 mM CaCl2.2H2O. The protoplasts of hypocotyl and leaf were fused by using various solutions of polyethylene glycol (PEG) MW 4000 and other compounds namely P1, P2, P3. The highest yield of fusion was 15,27% obtained from P2 solution, which was consisted of 33% PEG, 1,8% sucrose, 1mM KH2PO4 and 10 mM CaCl2.2H2O. Observation of nuclear of fusion product which was stained by 4-6 diamino 2-phenilindole (DAPI) under fluorescence microscope indicated that 58,82% uninucleate, 18,23% binucleate, and 22,95% multinucleate. The viability of fusion products high at the beginning of culture until first division. The surface of hypocotyls and leaves protoplasts has been covered with cell wall material completely after 8 hours of culture, whilst in fusion product after 24 hours. The first cell division was observed after 2 days of culture in hypocotyl and after 3 days of culture in leaf protoplast as well as in fusion products. The capability of fusion product to divide and to form colonies was smaller (0,32%) than hypocotyls protoplast (3,11%) and leaf protoplast (4,28%). Based on the results, it is concluded that the highest protoplast fusion product was induced by P2 solution and the fusion product was able to regenerate cell walls and to form microcolonies.

Beri Komentar ?#(0) | Bookmark

PropertiNilai Properti
ID PublisherJBPTITBPP
OrganisasiS
Nama KontakUPT Perpustakaan ITB
AlamatJl. Ganesha 10
KotaBandung
DaerahJawa Barat
NegaraIndonesia
Telepon62-22-2509118, 2500089
Fax62-22-2500089
E-mail Administratordigilib@lib.itb.ac.id
E-mail CKOinfo@lib.itb.ac.id

Print ...

Kontributor...

  • Editor:

Download...