Optimasi Metode Cryotherapy untuk Mengeliminasi Virus pada Tunas Kentang In Vitro (Optimation of Cryotherapy Method to Eliminate Virus on In Vitro Potato Shoot Tips)

Ida Ayu Astarini, Angel Chappell, Douglas Scheuring, Sean Michael Thompson, Julian Creighton Miller Jr.

Abstract


Penggunaan benih kentang generasi awal dan bebas virus merupakan kunci keberhasilan produksi kentang berkualitas. Cryotherapy (perendaman dalam nitrogen cair) merupakan teknik terbaru untuk mengeliminasi virus pada benih kentang. Salah satu kendala dalam penerapan teknologi cryotherapy ialah tingkat daya hidup eksplan yang masih rendah. Penelitian ini bertujuan untuk mengetahui efektifitas teknik enkapsulasi-dehidrasi untuk mendapatkan tunas yang sehat setelah perendaman dalam nitrogen cair. Ujung tunas in vitro ukuran 2–3 mm dari empat genotipe kentang di prakultur selama 3 hari secara bertahap pada media MS dengan penambahan gula 0,25 M, 0,5 M, dan 0, 75 M. Kemudian tunas dienkapsulasi, didehidrasi selama 5 jam, lalu direndam dalam nitrogen cair selama 60 menit lalu dihangatkan kembali dalam waterbath selama 3 menit. Tunas dalam kapsul kemudian dikulturkan pada media MS +30 g/l sukrosa + 8 g/l agar + 0,4 mg/l BAP + 1 mg/l GA3 untuk pemulihan, lalu dipelihara di ruang kultur dengan suhu 24oC. Daya hidup ujung tunas diamati pada minggu ke-8 dengan menggunakan kriteria skoring sebagai berikut: (1) pemutihan jaringan dan tidak ada respons pertumbuhan, (2) kalus mencokelat, (3) kalus hijau, (4) tumbuh tunas, dan (5) planlet sehat. Hasil penelitian menunjukkan daya hidup ujung tunas bervariasi antargenotipe. Skor daya hidup berkisar 1–2 (frekuensi 2–10) pada perlakuan nitrogen cair, yang menunjukkan tidak ada respons pertumbuhan tunas, beberapa memperlihatkan pertumbuhan kalus. Tunas pada perlakuan kontrol (tanpa perendaman dalam nitrogen cair) menunjukkan skor daya hidup 5 (frekuensi 1–7), di mana ujung tunas mampu beregenerasi menjadi planlet.

Keywords

Cryopreservation; Solanum tuberosum; Kultur jaringan tanaman; Eliminasi virus

Abstract

Virus-free, early generation seed is a key in the production of high quality potatoes. Cryotherapy (exposure to liquid nitrogen) is a new and promising method of virus elimination. One bottleneck in cryotheraphy method is survival of the explants after treatment with liquid nitrogen. This study investigated the effectiveness of enkapsulasi-dehidrasi method to obtain survival explants. Shoot tips were precultured for 3 days in MS media with sucrose addition of 0.25 M, 0.5 M and 0.75 M. Shoot tips were then encapsulate, dehydrate for 5 hours, expose to liquid nitrogen for 60 minutes and rewarm in waterbath for 3 minutes. Beads with shoot tips were then cultured in MS media + 30 g/l sucrose + 8 g/l agar + 0.4 mg/l BAP + 1 mg/l GA3 for recovery, and placed in 24oC culture room. Shoot tip survival was assessed at 8 weeks using the following scoring criteria: (1) tissue bleaching and no growth response, (2) brown callus, (3) green callus, (4) shoot growth, and (5) plantlet establishment. Survival was varied among genotypes. Survival scored between 1–2 (frequency 2–10) on liquid nitrogen treatment, showing shoot tips are mostly has no growth response, only some callus growth. Shoot tips on control treatment (without exposure in liquid nitrogen) shows survival scored 5 (frequency 1–7), i.e. shoot tips able to regenerate into plantlets.


Keywords


Cryopreservation; Solanum tuberosum; Plant tissue culture; Virus elimination

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DOI: http://dx.doi.org/10.21082/jhort.v26n1.2016.p97-102

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