INDUKSI MUTASI DENGAN KOLKISIN DAN SELEKSI IN VITRO TEBU TOLERAN KEKERINGAN MENGGUNAKAN POLYETHYLENE GLYCOL / Induced Mutation using Colchicine and In vitro Selection using Polyethylene glycol for Drought-Tolerant Sugarcane

RR. Sri Hartati, Sri Suhesti, Rossa Yunita, Syafaruddin Syafaruddin

Abstract


Creating of varieties can be done through mutation breeding at the cellular level, combined with in vitro selection. This research was conducted at the UPBUP from January until December 2017 to find out colchicine concentration and treatment duration which effectively produced tolerant mutant through in vitro drought selection using polyethylene glycol (PEG). The study consists of two stages. The first was mutation induction on sugarcane calli using colchicine, which was arranged factorially with a completely randomized environment design. The first factor was varieties (BL, PS 862, and PSJT 941), the second was colchicine concentration (0,01,0,0 and 0,05%), and the third was colchicine duration treatment (1 and 3 days). Observations were made on the percentage of callus survival. The second stage was in vitro selection of droughts using a PEG 6000, which was arranged factorially with a complete randomized design. The first factor was the concentration of colchicine (0, 0.01, 0.03, and 0.05%), the second was the colchicine duration treatment (1 and 3 days), and the third was PEG concentration (0, 10 and 20%). Selection was done for 4 weeks. Percentage of live callus, regenerated callus, number and height of shoots were observed as a selected criteria. Colchicine treatment in the 0.01 - 0.05% for 3 days on PS 862 and 0.01 - 0.03% for 3 days on PSJT 941 callus resulted mutant passing in vitro drought selection at 10% PEG concentration level. Mutant selection will be continued through in vivo. The optimum mutation treatment for BL has not been obtained.

Keywords: chemical mutagen, colchicine, mutation, selection agent, PEG 6000

 

Abstrak

Perakitan varietas tebu toleran kekeringan dapat dilakukan melalui pemuliaan mutasi pada tingkat sel, dikombinasikan dengan seleksi in vitro. Penelitian dilakukan di Laboratorium Unit Pengelola Benih Unggul Pertanian (UPBUP), Bogor, mulai Januari sampai Desember 2017 dengan tujuan mengetahui konsentrasi dan lama perlakuan mutagen kimia kolkisin, yang dapat menghasilkan mutan tebu yang lolos seleksi kekeringan secara in vitro menggunakan agen penyeleksi polyethylen glycol (PEG). Penelitian terdiri dari 2 tahap. Tahap pertama adalah induksi mutasi pada kalus tebu menggunakan mutagen kimia kolkisin. Penelitian disusun secara faktorial dengan rancangan lingkungan Acak Lengkap. Faktor pertama varietas tebu (BL, PS 862, dan PSJT 941), faktor kedua konsentrasi kolkisin (0, 0,01, 0,03, dan 0,05%), dan faktor ketiga lama perlakuan kolkisin (1 dan 3 hari). Pengamatan dilakukan terhadap persentase kalus hidup. Tahap kedua adalah seleksi kekeringan secara in vitro menggunakan PEG 6000. Penelitian disusun secara faktorial dengan rancangan Acak Lengkap. Faktor pertama konsentrasi kolkisin (0; 0,01; 0,03; dan 0,05%), faktor kedua lama perlakuan kolkisin (1 dan 3 hari), dan faktor ketiga konsentrasi PEG (0; 10; dan 20%). Seleksi dilakukan selama 4 minggu. Persentase kalus hidup, kalus yang berregerenerasi, jumlah dan tinggi tunas, diamati sebagai kriteria kalus mutan lolos seleksi. Perlakuan kolkisin pada kisaran konsentrasi 0,01 – 0,05% selama 3 hari pada kalus PS 862 dan 0,01 – 0,03% selama 3 hari pada PSJT 941 dapat menginduksi kalus mutan yang lolos seleksi kekeringan in vitro pada tingkat konsentrasi PEG 10%. Seleksi mutan akan dilanjutkan secara in vivo. Perlakuan mutasi yang optimum untuk BL belum diperoleh.

Kata kunci: mutagen kimia, kolkisin, mutasi, agen penyeleksi, PEG 6000


Keywords


chemical mutagen, colchicine, mutation, selection agent, PEG 6000

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DOI: http://dx.doi.org/10.21082/littri.v24n2.2018.93-104

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