Cryopreservation of in vitro-grown shoot tips of taro (Colocasia esculenta (L.) Schott) by vitrification. 1. Investigation of basic conditions of the vitrification procedure

Takagi, H.; Thịnh, Nguyễn Tiến; Islam, O. M.; Senboku, Toshihiro; Sakai, A.

doi:10.1007/s002990050285

Cited by 21 publications

(25 citation statements)

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“…It was found that using loading solution was essential to allow shoot formation. A similar finding was reported by Takagi et al (1997), preserving tips of Colocasia esculenta. Increasing time of exposure to vitrification solutions leads to increase solute concentrations inside the plant tissue (Bachiri et al, 1995).…”

Section: Discussionsupporting

confidence: 73%

Clonal propagation and cryogenic storage of the medicinal plant Stevia rebaudiana

Shatnawi

Shibli

Abu-Romman

et al. 1970

Span. j. agric. res.

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Successful clonal propagation of Stevia rebaudiana was achieved using microshoots as a primary step for in vitro conservation. Maximum proliferation was obtained on Murashige and Skoog (MS) medium supplemented with 1.5 mg L -1 benzyl amino purine and 0.2 mg L -1 indole-3-butyric-acid (IBA). Auxin increased rooting percentage of shoots at concentration of 0.4 mg L -1 IBA, indole-3-acetic-acid or naphthalene acetic acid and no rooting occurred without plant growth regulator. A survival of 90% was achieved when rooted explants were acclimatized in vivo in 1 soil: 1 perlite: 1 peat. In vitro S. rebaudiana shoots were successfully stored for up to 32 weeks on MS medium supplemented with an appropriate concentration of sucrose, sorbitol or mannitol, at 24 ± 2°C. After 32 weeks, 93.6% of the shoots were able to survive. Moreover, 89.3% of them were able to regrow when stored under light conditions. Cryopreservation by vitrif ication was successfully achieved (65.6% regrowth) when shoot tips were precultured on a medium supplemented with 0.4 M sorbitol for 2 d, followed by loading shoot tips with 80% concentrated plant vitrification solution 2 (PVS2) for 20 min; then dehydrated with 100% PVS2 for 60 min at 0°C prior to storage in liquid nitrogen. This procedure is easy to handle and produced a high levels of shoot formation. This protocol could be useful for longterm storage of S. rebaudiana germplasm.Additional key words: cryopreservation; in vitro conservation; micropropagation; osmoticum; root formation. Resumen Propagación clonal y almacenamiento criogénico de la planta medicinal Stevia rebaudianaSe logró con éxito la propagación clonal de Stevia rebaudiana, utilizando microtallos como primer paso para su conservación in vitro. Se obtuvo la proliferación máxima en el medio Murashige y Skoog (MS) suplementado con 1,5 mg L -1 de bencil amino purina y 0,2 mg L -1 de ácido indol-3-butírico (IBA). Las auxinas aumentaron el porcentaje de enraizamiento de los brotes en la concentración de 0,4 mg L -1 de IBA, ácido indol-3-acético o el ácido naftalen acético, pero no se produjo enraizamiento sin regulador del crecimiento vegetal. Se logró una supervivencia del 90% cuando los explantes enraizados fueron aclimatizados in vivo en 1 suelo: 1 perlita: 1 turba. Se almacenaron con éxito brotes de S. rebaudiana in vitro hasta 32 semanas a 24 ± 2°C, en medio MS suplementado con una concentración apropiada de sacarosa, sorbitol o manitol. Después de 32 semanas, el 93,6% de los brotes fueron capaces de sobrevivir y el 89,3% fueron capaces de rebrotar cuando se almacenaron bajo condiciones de luz. Se logró con éxito (65,6% rebrotes) la criopreservación por vitrificación cuando los ápices se pre-cultivaron durante 2 días en un medio suplementado con sorbitol 0,4 M, y a continuación con una solución de vitrificación de la planta 2 (PVS2) al 80% durante 20 minutos, seguido de una deshidratación con PVS2 al 100% durante 60 min a 0°C antes de su almacenamiento en nitrógeno líquido. Este procedimiento es fácil de ejecutar y puede ser...

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Section: Discussionsupporting

confidence: 73%

Clonal propagation and cryogenic storage of the medicinal plant Stevia rebaudiana

Shatnawi

Shibli

Abu-Romman

et al. 1970

Span. j. agric. res.

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“…untuk meningkatkan fleksibilitas membran dan proses dehidrasi berjalan secara perlahan-lahan sehingga sel tidak akan pecah. Perendaman jaringan dalam larutan LS efektif meningkatkan osmotoleransi jaringan pada beberapa macam tanaman seperti talas (Takagi et al 1997), kentang (Hirai dan Sakai 1999a), mentha (Hirai dan Sakai 1999b), dan wasabi (Matsumoto et al 1994). Dari percobaan pemuatan, diketahui bahwa semakin lama durasi rendam dalam larutan LS maka semakin turun tingkat pertumbuhan tunas purwoceng yang terenkapsulasi (Gambar 4).…”

Section: Perlakuan Pemuatanunclassified

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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“…Panis et al (2005b) showed that regrowth of banana meristems is not influenced by loading solution exposure time (0 -5 hours). Although the precise mechanism of loading is not yet fully understood, it has been proven for different plant species that loading can dramatically enhance the tolerance of isolated meristems to dehydration by the vitrification solution (Matsumoto et al 1994, Takagi et al 1997, Panis 2009.…”

Section: IImentioning

confidence: 99%

Cryopreservation of banana’s cv Grand Naine in vitro rhizomes

Londe

Vendrame

Sanaei

et al. 2017

An. Acad. Bras. Ciênc.

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The preservation of banana genetic material is usually performed through seedlings. However, most banana cultivars do not produce seed and are propagated vegetatively. Therefore, cryopreservation is a feasible technique that allows the preservation of banana genotypes indefinitely. For the success of cryopreservation protocols, the selection of cryoprotectants and pre-freezing techniques are important factor. Therefore, the objective of this study was to verify the effects of different cryoprotectants with and without 1% phloroglucinol and pre-cooling periods on the development of a protocol for cryopreservation of in vitro rhizomes of Musa accuminata (AAA) cv Grand Naine banana. The addition of 1% phloroglucinol to the cryoprotective solutions, such as PVS2 enhanced recovery of cryopreserved banana rhizomes. In addition, pre-cooling of explants in ice for 3 hours in PVS2 + 1% of phloroglucinol allowed efficient cryopreservation of banana rhizomes, followed by successful recovery and regeneration of in vitro shoots of banana cv Grand Naine.

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Cryopreservation of in vitro-grown shoot tips of taro (Colocasia esculenta (L.) Schott) by vitrification. 1. Investigation of basic conditions of the vitrification procedure

Cited by 21 publications

References 0 publications

Clonal propagation and cryogenic storage of the medicinal plant Stevia rebaudiana

Clonal propagation and cryogenic storage of the medicinal plant Stevia rebaudiana

Kriopreservasi Tanaman Obat Langka Purwoceng dengan Teknik Enkapsulasi-Vitrifikasi

Cryopreservation of banana’s cv Grand Naine in vitro rhizomes

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