A simple and efficient protocol for cryopreservation of embryogenic calli of the medicinal plant Anemarrhena asphodeloides Bunge by vitrification

Senrong, Hong; Ming-hua, Yin

doi:10.1007/s11240-011-0094-5

Cited by 15 publications

(5 citation statements)

References 48 publications

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“…Additionally, in most studies, the loading fluid was different from the cryoprotective fluid used for cryopreservation. The loading fluid needs to be replaced before freezing; this increases the number of test steps and operational difficulties (Ashmore et al 2007;Cheong 2012;Sen-Rong and Ming-Hua 2012). Interestingly, in this study, the PVS1 loading fluid was not toxic to Eriobotrya plants and had a good effect.…”

Section: Discussionmentioning

confidence: 75%

Programmed Cooling and Vitrification Method Applied to the Germplasm Preservation of Eriobotrya Plants

Liu

Huang

Lin

et al. 2023

horts

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Long-term cryopreservation of germplasm is of great importance to maintaining genetic resources. A cryopreservation technology system has not yet been established for Eriobotrya, an important fruit tree in the Rosaceae. In this study, Eriobotrya plants were used as test materials for the following purposes: to study different types of vitrification solution, compare different methods involving programmed cooling vitrification and rapid freezing vitrification; and evaluate the effects of vitrification solution loading time, low-temperature acclimation culture, dark culture, and cryoprotectant use in preculture medium for ultra-low-temperature preservation of Eriobotrya germplasm resources. Plant vitrification solution 1 had the best comprehensive effect on different vitrification solution tests. The programmed cooling and vitrification method yielded a certain survival rate with different vitrification solutions and materials, but the survival rate under the rapid freezing method was 0%. The longer the vitrification solution was loaded, the higher the survival rate. The most suitable time for dark culture of Eriobotrya plants after cryopreservation was 20 days. The preculture medium supplemented with 5% dimethyl sulfoxide resulted in significantly higher preservation than the control medium, and the optimal sucrose concentration was 0.3 mol⋅L−1. A stable, high-survival, and novel cryopreservation procedure for loquat plants was established. The operating procedures are described in a procedural and standardized manner. These findings provide a theoretical basis for research of the cryopreservation of germplasm resources of Eriobotrya plants.

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Section: Discussionmentioning

confidence: 75%

Programmed Cooling and Vitrification Method Applied to the Germplasm Preservation of Eriobotrya Plants

Liu

Huang

Lin

et al. 2023

horts

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“…In contrast, the authors [42] observed that the regrowth crocus calli improved after loading treatment for 30 minutes which was comparable to this study. Embryogenic calli of Anemarrhena asphodeloides Bunge that was treated with a mixture of 2 M glycerol+ 0.4 M sucrose for 30 minutes displayed high regrowth rate (67.8%) with normal plantlets produced after cryopreservation [43]. Even though the accurate mechanism on how the loading treatment induces osmotolerance to PVS2 is uncertain, loading treatment does reduce the osmotic stress and protects the cell membranes from severe dehydration and freezing [44].…”

Section: Discussionmentioning

confidence: 99%

Cryopreservation ofRosa hybridacv. Helmut Schmidt by PVS2 vitrification method usingin vitrofragmented explants (IFEs)

Udain

et al. 2019

Preprint

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1415 This is the first report on cryopreservation via PVS2 vitrification method on roses using in vitro 16 fragmented explants (IFEs) as the starting material. The aim of this study is to optimize the efficient 17 plant recovery and regeneration system for cryopreservation of Rosa hybrida cv. Helmut Schmidt 18 using IFEs. Some important parameters have been optimized in this study are the effect of ascorbic 19 acid (0.3 mM) examined separately and in combination at all steps in cryopreservation procedure 20 (preculture, loading, unloading and growth recovery), loading type, loading duration, and PVS2 21 duration. The highest growth recovery of 43.33% was obtained when 3-4 mm size IFEs precultured 22 on 0.25 M sucrose media supplemented with full-strength MS for one (1) day, followed by loading 23 treatment supplemented with 1.5 M glycerol + 0.4 M sucrose + 5% DMSO + 0.3 mM ascorbic acid 24 for 20 minutes, dehydration with PVS2 solution for 30 minutes and then treated with unloading 25 solution supplemented with 1.2 M sucrose + 0.3 mM ascorbic acid for 20 minutes. This finding 26 implies that long-term storage of Rosa Hybrida cv. Helmut Schmidt by PVS2 vitrification method 27 was successful with essential biomolecules. 28 29 30 Keywords: In vitro fragmented explants (IFEs), PVS2 vitrification, ascorbic acid, Biomolecules, 31 Rose 32 34 Roses are one of the most important ornamental plants in many parts of the world. This 35 is because they bloom beautiful flowers and as such, they are widely used in the horticultural 36 industry. Cultivation of rose plants via tissue culture micropropagation has disadvantages in 37 terms of contamination, somaclonal variations and also due to biotic stresses [1]. Hence, 38 cryogenic storage offers a stable and safe method for long term storage of plant materials 39 while safeguarding genetic stability [2]. Cryopreservation at temperatures of -196°C has been 40 considered as an ideal means for long term storage of plant germplasm. Through 41 cryopreservation, hybrids that are in danger of being extinct can be conserved with low 42 maintenance apart from being easy for international exchange. Cryopreservation through 43 vitrification reduces injury caused by intracellular crystallization and provides high 44 regeneration rates prior to immersion into liquid nitrogen. Basically, the most appropriate 45 plant part utilized is the meristem culture since it generates virus free lines [3]. For PVS2 46 cryopreservation study in roses, in vitro fragmented explants (IFE) represent a new source of 47 explant as starting material. IFEs were first introduced by authors [4] who conducted research 48 on Vanilla planifolia "Andrews". The study discovered that IFEs allows the production of a 49 larger number of shoots than micro cuttings after 4 months in culture. However, every 50 species of plants has different genetic makeup and respond differently to various treatments. 51 Therefore, improvement of the cryopreservation method is crucial in each step for growth 52 after recovery. Vitrification ...

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“…All the conditions of vitrification cryopreservation were designed by a single factor experiment. The basic culture conditions were the same as the cryopreservation of EC of Anemarrhena asphodeloides Bunge by vitrification [17]. The specific methods were as follows:…”

Section: Cryopreservation Of Ec By Vitrificationmentioning

confidence: 99%

“…For the fresh weight measurement and 2,3,5-Triphenyltetrazolium chloride (TTC) staining method of callus refer to Yu et al [16]. The relative survival percentage of cells after cryopreservation is expressed as the ratio of absorbance of treatment and control [17]. The following formula was used: (2015).…”

Section: Determination Of the Cell Relative Survival Percentage And O...mentioning

confidence: 99%

Improved Method for Cryopreservation of Embryogenic Callus of Fraxinus mandshurica Pupr. by Vitrification

Liu

et al. 2022

Forests

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In order to simplify the experimental procedure and treatment procedure, we preserved the embryonic callus (EC) of Fraxinus mandshurica more efficiently. In this paper, we established a method for cryopreservation of EC of F. mandshurica by vitrification. EC was subcultured for 7–10 days (d). Vigorous EC with good growth conditions were selected, and cryopreservation was performed by vitrification. The best pre-culture method was to pre-culture EC on 0.5 mol·L−1 sucrose medium for 3 d, load and culture in the liquid woody plant medium (WPM) supplemented with 2 mol·L−1 glycerol and 0.4 mol·L−1 sucrose for 60 min, then dehydrate in 2 mL of plant vitrification solution 2 (PVS2) (30% glycerol + 15% dimethyl sulfoxide (DMSO) + 15% ethylene glycol + 0.4 mol·L−1 sucrose + liquid WPM). EC was rewarmed in a 40 °C water bath for 2 min after cooling in liquid nitrogen. The procedure for cryopreservation of F. mandshurica EC by the vitrification method established in this experiment is relatively reliable. The results from the present study provide a technical reference for improving the cryopreservation of F. mandshurica EC.

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A simple and efficient protocol for cryopreservation of embryogenic calli of the medicinal plant Anemarrhena asphodeloides Bunge by vitrification

Cited by 15 publications

References 48 publications

Programmed Cooling and Vitrification Method Applied to the Germplasm Preservation of Eriobotrya Plants

Programmed Cooling and Vitrification Method Applied to the Germplasm Preservation of Eriobotrya Plants

Cryopreservation ofRosa hybridacv. Helmut Schmidt by PVS2 vitrification method usingin vitrofragmented explants (IFEs)

Improved Method for Cryopreservation of Embryogenic Callus of Fraxinus mandshurica Pupr. by Vitrification

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