ISOLASI DAN KULTUR PROTOPLAS MESOFIL DAUN DARI BEBERAPA GENOTIP UBI KAYU (Manihot esculenta Crantz) - Isolation and Protoplas Culture of Leaf Mesophyll from Some Genotypes of Cassava (Manihot Esculenta Crantz)

Hani Fitriani; Nurhamidar Rahman; Siti Kurniawati; Pramesti Dwi Aryaningrum; N Sri Hartati

doi:10.36974/jbi.v10i1.4145

ISOLASI DAN KULTUR PROTOPLAS MESOFIL DAUN DARI BEBERAPA GENOTIP UBI KAYU (Manihot esculenta Crantz) - Isolation and Protoplas Culture of Leaf Mesophyll from Some Genotypes of Cassava (Manihot Esculenta Crantz)

Hani Fitriani, Nurhamidar Rahman, Siti Kurniawati, Pramesti Dwi Aryaningrum, N Sri Hartati

Abstract

Superior cassava seedlings can be obtained through protoplast fusion techniques. This study aimed to obtain protoplasts from mesophyll leaves of three types of cassava namely Mentega 2, Ubi Kuning and Gajah. Protoplast was successfully obtained from three cassava genotypes after incubation of pre-plasmolysis cells in a mixture of LE3 treatment enzymes, i.e 0.4 M Mannitol, 2% cellulase, 1% maserozyme and 0.1% pectoliase for 18 hours. Protoplast succeeded in forming microcaluses in Murashige & Skoog (MS) semi-solid media with the addition of vitamin Kao & Michayluk (KM) and KM organic matter and enriched with 2 µM CuSO₄, 2 mg/L BAP, 1 mg/L IAA, 2% sucrose with Phytagel agar variations were 0.5% (M1) and 1% (M2). Callus from microcalus occurred in all treatment media. The best treatment was shown on P4 media, namely MS media containing 1.0 mg/L 2,4-D and 2 mg/L BAP, where the callus obtained was larger than other media.

Keywords: cassava, cellulase, microcalus, protoplasts

ABSTRAK

Bibit ubi kayu unggul sangat dibutuhkan oleh petani atau penggiat ubi kayu dan dapat diperoleh melalui teknik fusi protoplas. Penelitian ini bertujuan untuk mendapatkan protoplas dari mesofil daun tiga jenis ubi kayu yaitu Gajah, Mentega 2 dan Ubi Kuning koleksi in vitro dengan perlakuan komposisi enzim selulase, maserozim dan pektoliase serta media kultur protoplas. Isolasi protoplas dilakukan dengan menginkubasi daun ubi kayu in vitro dari genotip Mentega 2, Ubi Kuning dan Gajah pada media cair yang terdiri dari beberapa campuran enzim. Protoplas berhasil diperoleh dari ketiga genotip ubi kayu setelah inkubasi pra-plasmolisis sel pada larutan campuran enzim perlakuan LE3 yaitu 0,4 M manitol, 2% selulase, 1% maserozim dan 0,1% pektoliase selama 18 jam. Protoplas berhasil membentuk mikrokalus pada media semi padat Murashige & Skoog (MS) dengan penambahan vitamin Kao & Michayluk (KM) dan bahan organik KM serta diperkaya dengan 2 µM CuSO₄, 2 mg/L BAP, 1 mg/L IAA, 2% sukrosa dengan variasi agar Phytagel 0,5% (M₁) dan 1% (M₂). Pembentukan kalus dari mikrokalus terjadi pada semua media perlakuan. Secara keseluruhan, perlakuan terbaik ditunjukkan pada media P4, yaitu media MS yang mengandung1,0 mg/L 2,4-D dan 2mg/L BAP, dimana kalus yang diperoleh berukuran lebih besar dibanding media lainnya.

Kata kunci: mikrokalus, protoplas, selulase, ubi kayu

Keywords

cassava; cellulase; microcalus; protoplasts

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DOI: http://dx.doi.org/10.36974/jbi.v10i1.4145