Induksi Tanaman Haploid Dianthus sp. melalui Pseudofertilisasi Menggunakan Polen yang Diiradiasi dengan Sinar Gamma
Abstract
ABSTRACT
Haploid plants of Dianthus sp. were obtained by pseudofertilization using irradiated pollen. Gamma ray of 100 gray was used to inactivate pollen. Crossing were made using 131 female of Dianthus chinensis Dchi-11 as pod parents. After pollination 59% fruits were harvested, 41% were dropped and only 51 ovaries were cultured on the medium. Ovaries were explanted 1 – 2 weeks after pollination and cultured on solid MS medium containing 400 mg L-1glutamine + 103. 77 mg L- 1 proline and growth regulator including 1-Naphthalene acetic acid (NAA) and 6-Benzylaminopurine (BAP). The first ploidy seedling observation was made by number of chloroplast in each side of guard cells of stomata. Four haploid plants were obtained from this screening, but based on chromosome counting and DNA analysis using flow cytometry only three haploid plants and two haploid plants revealed respectively.
Key words: BAP, medium, pollination, proline, stomata
ABSTRAK
Tanaman haploid Dianthus sp diperoleh melalui pseudofertilisasi menggunakan serbuk sari yang diiradiasi. Sinar gamma dari senyawa cobalt 60Co pada dosis 100 Gy digunakan untuk menonaktifkan serbuk sari. Persilangan semu dilakuan menggunakan 131 bunga betina dari Dianthus chinensis Dchi- 11 sebagai donor ovul. Setelah penyerbukan 41% buah gugur dan hanya 51 ovari dapat dikulturkan pada medium. Ovari ditanam pada umur 1 – 2 minggu setelah penyerbukan dan dikulturkan pada dua media MS yang mengandung 400 mg L-1glutamin + 103. 77 mg L-1 prolin dan dengan penambahan 1-Naphthaleneacetic acid (NAA) dan 6-Benzylaminopurine (BAP). Observasi ploidi awal dilakukan dengan melihat jumlah kloroplas pada sel penjagastomata pada daun. Berdasarkan pengamatan ini empat tanaman haploid diperoleh, namun berdasarkan penghitungan jumlah kromosom hanya tiga tanaman yang haploid, dan berdasarkan an alisis DNA dengan flowcytometer diperoleh dua tanaman haploidKata
Kunci: BAP, media, penyerbukan, prolin, stomata