Introduction to the in Vitro Preservation of Plant Cells, Tissues and Organs

Grout, B. W. W.

doi:10.1007/978-3-642-78661-7_1

Cited by 34 publications

(25 citation statements)

References 40 publications

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“…Menurut Grout (1995), teknik kriopreservasi merupakan teknik penyimpanan bahan tanaman dalam suhu yang sangat rendah, yaitu -160 hingga -180 o C (nitrogen fase uap) bahkan sampai -196 o C (nitrogen fase cair). Penyimpanan dengan pembekuan dalam nitrogen cair merupakan metode yang potensial untuk penyimpanan jangka panjang plasma nutfah tumbuhan (Towill dan Jarret 1992).…”

Section: Pendahuluanunclassified

“…Formasi es intraseluler mutlak bersifat lethal sedangkan for- masi es ekstraseluler juga dapat merusak sel karena daya mekanis dari kristal es yang tumbuh, gaya adesi kristal es terhadap membran, interaksi elektris yang disebabkan oleh perbedaan solubilitas ion pada fase es dan cair, formasi gelembung udara intraseluler, luka khemis yang berhubungan dengan peroksidase lipid dan perubahan pH pada lokasi tertentu (Grout 1995).…”

Section: Perlakuan Dehidrasi Dan Pembekuan Jaringanunclassified

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Kriopreservasi Tanaman Obat Langka Purwoceng dengan Teknik Enkapsulasi-Vitrifikasi

Roostika¹,

Rahayu²,

Sunarlim³

2016

BPN

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Pruatjan (Pimpinella pruatjan Molk.) is an Indonesian endangered medicinal plant, so that it is highly protected. Cryopreservation can be applied to this plant for long-term preservation. The aim of this research was to obtain a method of encapsulation-vitrification by optimizing each step in cryopreservation protocol i.e. preculture, loading, dehydration with and without freezing in liquid nitrogen. The best treatment of each step would be applied in the following step. On preculture experiment, in vitro shoots were planted on the Driver and Kuniyaki (DKW) basal media containing 0.3 M sucrose and incubated for 1, 2, 3, 4, and 5 days. After those incubation period, shoot tips were encapsulated with 2.5% Na-alginate and soaking for 15 minutes in 100 ppm CaCl 2 solution before planting. On loading experiment, precultured explants were loaded in DKW basal solution containing 2 M glycerol and 0.4 M sucrose for 0, 30, 60, and 90 minutes. On dehydration experiment, preculturead and loaded explants were dehydrated with PVS2 solution PVS2 (DKW + 30% glycerol + 15% DMSO + 15% ethyleneglicol + 0.4 M sucrose) for 0, 30, 60, 90, and 120 minutes. The parts of them were freezed in liquid nitrogen (-196 o C). The result showed that cryopreservation through encapsulation-vitrification technique could be applied on pruatjan. The best preculture treatment was 5 days incubation period. The best loading treatment was 30 minutes. The best dehydration treatment was 90 minutes. The successful level of this research was still low (10%) so that it needs optimization method.Key words: Pimpinella pruatjan Molk., cryopreservation, encapsulation-vitrification. ABSTRAKPurwoceng (Pimpinella pruatjan Molk.) adalah tanaman obat langka asli Indonesia yang hampir punah sehingga harus dilindungi. Kriopreservasi dapat diterapkan pada tanaman ini untuk penyimpanan jangka panjang. Tujuan penelitian adalah untuk memperoleh teknik enkapsulasi-vitrifikasi dengan melakukan optimasi dari tiap-tiap tahapan kriopreservasi yang meliputi perlakuan prakultur, loading, dehidrasi sebelum dan setelah pembekuan dalam nitrogen cair. Perlakuan yang terbaik kemudian diterapkan pada tahapan percobaan berikutnya. Pada perlakuan prakultur, tunas in vitro ditanam pada media Driver dan Kuniyaki (DKW) dengan penambahan sukrosa 0,3 M dengan masa inkubasi 1, 2, 3, 4, dan 5 hari. Setelah itu, pucuk yang berukuran 0,5 cm dienkapsulasi dengan Na-alginat 2,5% (yang mengandung media regenerasi) dalam larutan CaCl 2 100 ppm selama 15 menit sebelum penanaman kembali. (-196 o C) dan sebagian lainnya tidak dibekukan. Hasil penelitian menunjukkan bahwa kriopreservasi secara enkapsulasi-vitrifikasi berpeluang diterapkan pada tanaman purwoceng. Perlakuan prakultur terbaik adalah 5 hari. Perlakuan loading terbaik adalah 30 menit dan perlakuan dehidrasi terbaik 90 menit. Tingkat keberhasilan ini masih rendah (10%) sehingga diperlukan optimasi metode.Kata kunci: Pimpinella pruatjan Molk., kriopreservasi, enkapsulasi-vitrifikasi.

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Section: Pendahuluanunclassified

Section: Perlakuan Dehidrasi Dan Pembekuan Jaringanunclassified

Kriopreservasi Tanaman Obat Langka Purwoceng dengan Teknik Enkapsulasi-Vitrifikasi

Roostika¹,

Rahayu²,

Sunarlim³

2016

BPN

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“…Pembelahan sel dan proses metabolisme di dalam sel, jaringan, atau organ tanaman dapat dihentikan dalam waktu yang tidak terbatas melalui kriopreservasi (Bhojwani dan Razdan, 1983;Ashmore, 1997). Kondisi suhu penyimpanan secara kriopreservasi dalam nitrogen sangat rendah, yaitu -160 hingga - (Grout, 1995;Towill dan Jarret, 1992). Teknik kriopreservasi sangat potensial dikembangkan untuk penyimpanan plasma nutfah tanaman dalam jangka panjang hingga puluhan tahun (Bajaj, 1979;Withers, 1985;Towill dan Jarret, 1992).…”

Section: Pendahuluanunclassified

Perkembangan Aplikasi Teknik Kriopreservasi untuk Konservasi dan Mendukung Program Pemuliaan Tanaman

Roostika

2016

J. AgroBiogen

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Development of the Application of CryopreservationTechniques for Conservation and Supporting Plant Breeding Program. Ika Roostika. In the beginning, cryopreservation technique was primarily used for germplasms long-term storage as passive collection because cell division and metabolism process can be stopped at super low temperature, commonly in liquid nitrogen. The technique is suitable for vegetative propagated plants and recalcitrant seeds. Recently, its application is extending for storing many species and orthodox seeds. In this paper, the development of cryopreservation application is discussed. In Indonesia, cryopreservation is being studied but the application of cryopreservation has been significantly developed abroad. The application of cryopreservation technique is not only for preserving passive collections but also for storing active collections, including to provide plant materials for hybridization, cellular engineering, genetic transformation, as well as pathogens eradication or cryotherapy. It is concluded that cryopreservation plays an important role in conventional and modern plant breeding program.

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“…Below -120°C the rate of chemical or biophysical reactions is too slow to cause biological deterioration (Kartha 1985). Only in the long term might there be a small risk of ionising radiation causing genetic changes in materials stored at cryogenic temperatures (Grout 1995). …”

Section: Cryo Preservationmentioning

confidence: 99%