Cryopreservation and post-thaw genetic integrity of Viola stagnina Kit., an endangered species of wet habitats – A useful tool in ex situ conservation

Żabicki, Piotr; Mikuła, Anna; Śliwińska, Elwira; Migdałek, Grzegorz; Nobis, Agnieszka; Żabicka, Justyna; Kuta, Elżbieta

doi:10.1016/j.scienta.2021.110056

Cited by 6 publications

(6 citation statements)

References 50 publications

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“…During dehydration of vitrification cryopreservation, the treatment time of plant vitrification solution is the key to cell vitrification cryopreservation [27]. In this study, long dehydration time led to a decrease in the cell survival percentage of F. mandshurica, which is consistent with the results of the shoot tip of Gentiana kurroo [28] and the shoot tip of Viola stagnina [29]. When the dehydration time is too short, the cell dehydration is not less, and it is difficult to reach the vitrification state quickly in the process of cooling treatment.…”

Section: Effect Of Dehydration On Vitrification Cryopreservationsupporting

confidence: 88%

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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Section: Effect Of Dehydration On Vitrification Cryopreservationsupporting

confidence: 88%

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

Liu

et al. 2022

Forests

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“…The next step in conservation practice is developing an effective procedure for cryopreservation of shoot tips, shoots or somatic embryos of P. patens and introducing them for long-term storage in liquid nitrogen, including individuals from different European populations. Although several rare, endemic and endangered species have already been introduced to cryogenic conditions 32,[52][53][54] , thousands still need to be cryopreserved, including all Pulsatilla species. It should be emphasized that in situ protection alone of endangered species will not guarantee their conservation, and thus must be supported by ex situ management with the use of biotechnological techniques and optionally seed banking as complementary for plant conservation including short-, medium-and long-term strategies 54 .…”

Section: Discussionmentioning

confidence: 99%

“…In vitro culture serves as an excellent biotechnological tool for ex situ conservation of different endangered species 31,32 , including Pulsatilla vulgaris 33 , and also species growing at metalliferous sites [34][35][36] . However, the reintrodution of in vitro propagated plants to the field sites, will be successful provided the areas are properly managed before the transplantation 22,25,37 .…”

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confidence: 99%

Re-introduction of an extinct population of Pulsatilla patens using different propagation techniques

Żabicka

Żabicki

Słomka

et al. 2022

Sci Rep

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The study focuses on the propagation of a rare and endangered plant species (Pulsatilla patens) to re-introduce an extinct population from calamine area in Southern Poland. The plants were propagated from seeds, rhizome cuttings, or regenerated in vitro from shoot tips, hypocotyls with roots or cotyledons of seedlings on Murashige & Skoog (MS) medium supplemented with 0.25 or 0.50 mg L−1 BAP (Benzylaminopurine) via direct and indirect organogenesis or somatic embryogenesis (SE). The most efficient micropropagation method was with shoot tips as an explant on MS + 0.25 mg L−1 BAP where 97% of the explants produced multiple shoots, mass SE was observed after transfer on ½ MS with 2% saccharose; 267 (35%) shoots rooted on ½ MS + 2% saccharose were acclimatized to ex vitro conditions. Flow cytometry revealed genome size stability of propagated plantlets. Low genetic differentiation between micropropagated plantlets and initial material was indicated by ISSR (Inter Simple Sequence Repeat) markers. Totally, 132 vigorous plantlets obtained on various pathways were introduced to the field plots in 2020; 30.33% survived the winter, and several reached the generative stage and flowered in the spring 2021. In next season (March/April 2022) the number of introduced plants decreased to 25% while the number of flowering and fruiting shoots in different clumps increased in some plots. This is the first report of successful re-introduction of the endangered P. patens based on micropropagation, rhizome cuttings, and seed germination.

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“…According to results, by using PVS3 solution for durations 30 to 120 min, high recovery as (65%) was obtained. Whereas, by using PVS2 for 60 min, 40 % recovery was observed [32,33].…”

Section: Survival Percentage After Thawing and Regenerationmentioning

confidence: 92%

In vitro PROPAGATION OF BANANA SHOOT TIPS AND CRYOPRESERVATION BY VITRIFICATION

VATS¹,

AGRAWAL²

2022

PCBMB

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Micropropagation of banana is limited as explants are difficult to maintain in culture. The present study was conducted to investigate different factors for banana cryopreservation. This study has two parts. In the first part of study in vitro propagation of banana was done to propagate large number of explants for cryopreservation. Whereas, in the second part, banana shoot tips were cryopreserved by vitrification technique. During this study, different parameters such as exposure to plant vitrification solution PVS2 and PVS3, rewarming time were optimized. For cryopreservation by vitrification method, shoot tips were precultured on solidified MS medium supplemented with 0.3 M sucrose for 16 h at 25C. Then they were loaded in loading solution containing 2M glycerol and 0.4 M sucrose for 30 minutes at room temperature. After loading step, shoot tips were dehydrated with plant vitrification solution (PVS2) and (PVS3) with different exposure times (10,30,40,60,90,120 min) respectively. The best result was obtained with shoot tips treated with PVS2 for 60 min and 120 minutes for PVS3 with survival rates of cryopreserved shoot tips as 85% and 33% respectively. Furthermore, effect of different rewarming times was also investigated on shoot tip viability. Among tested different rewarming times (0,2,4 and 6min), 2 min of rewarming time in PVS2 and 4 min of rewarming time in PVS3 were found suitable for cryopreservation as they retained survival rate as 97% and 75% respectively.

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Cryopreservation and post-thaw genetic integrity of Viola stagnina Kit., an endangered species of wet habitats – A useful tool in ex situ conservation

Cited by 6 publications

References 50 publications

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

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

Re-introduction of an extinct population of Pulsatilla patens using different propagation techniques

In vitro PROPAGATION OF BANANA SHOOT TIPS AND CRYOPRESERVATION BY VITRIFICATION

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