Aplikasi Teknik Embriogenesis Somatik dan Krioterapi untuk Eliminasi Virus SCSMV pada Tanaman Tebu

Abstract

The use of virus-free seedlings is an effective option to control viral disease. It was reported that several tissue culture methods could eliminate virus infection from plant tissues. The objective of the study was to know the in vitro responses of three sugarcane varieties to the organogenesis and cryotherapy techniques and to know the potential rate of both techniques in eliminating Sugarcane streak mosaic virus (SCSMV). The plant materials were virus-infected sugarcane of three varieties of sugarcane (PS862 from Cirebon, PS881 from Jember, and PSJK922 from Malang). Callus induction was conducted on MS medium with addition of 3 mg/l 2,4-D and 3 g/l casein hydrolysate. There were three treatments of subculture (SK0, SK1, and SK2). Regeneration of shoots was conducted on MS medium with addition of 0.3 mg/l BA, 0.5 mg/l IBA, and 100 mg/l PVP. Cryotheraphy was done by using vitrification technique, starting with preculture on 0.3 M sucrose-containing medium for 3 days, loading with LS solution for 10 minutes, dehydration with PVS2 solution for 40 minutes, plunging in liquid nitrogen for 1 hour, deloading with RS solution for 30 minutes, recovery, and regeneration. Virus indexing was conducted by RT-PCR assay using specific primer of SCSMV. The result showed that all varieties were able to form for calli and shoots, both with and without subculture. Within the three varieties, only PS862 from Cirebon could survive post cryotherapy treatment. Organogenesis regeneration up to twice subculture was not able to eliminate SCSMV. Cryotherapy could eliminate SCSMV virus with the proportion of one third (33.3%).Keywords: Saccharum officinarum L., embryogenic callus, vitrification, liquid nitrogen, virus elimination. ABSTRAKPenggunaan bibit bebas virus menjadi salah satu cara pengendalian penyakit virus yang efektif. Beberapa macam teknik kultur jaringan dilaporkan mampu mengeliminasi virus di dalam jaringan tanaman. Tujuan penelitian adalah mengetahui respons tiga varietas tebu terhadap teknik organogenesis dan krioterapi serta untuk mengetahui kemampuan kedua teknik tersebut dalam mengeliminasi Sugarcane streak mosaic virus (SCSMV). Bahan tanaman yang digunakan adalah varietas tebu PS862 asal Cirebon, PS881 asal Jember, dan PSJK922 asal Malang yang terinfeksi SCSMV. Induksi kalus dilakukan dengan menggunakan media MS yang ditambah dengan 2,4-D 3 mg/l dan kasein hidrolisat 3 g/l. Terdapat tiga macam perlakuan subkultur (SK0, SK1, dan SK2). Regenerasi tunas dilakukan pada media MS dengan penambahan BA 0,3 mg/l, IBA 0,5 mg/l, dan PVP 100 mg/l. Teknik krioterapi yang diterapkan adalah vitrifikasi dengan tahapan prakultur menggunakan sukrosa 0,3 M selama 3 hari, loading dengan larutan LS selama 10 menit, dehidrasi dengan larutan PVS2 selama 40 menit, krioterapi dengan nitrogen cair selama 1 jam, deloading dengan larutan RS selama 30 menit, pemulihan, dan regenerasi. Indeksing virus dilakukan secara RT-PCR menggunakan primer spesifik SCSMV. Hasil penelitian menunjukkan bahwa ketiga varietas mampu membentuk kalus dan t...