131. Development of a cryopreservation protocol for Loxocarya cinerea – A recalcitrant plant species used in post-mining restoration

Kaczmarczyk, Anja; Turner, Shane R.; Funnekotter, Bryn; Bunn, Eric

doi:10.1016/j.cryobiol.2010.10.135

Cited by 6 publications

(13 citation statements)

References 2 publications

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“…In contrast with this experiment data, shoot tips of Carica papaya cultivars "70" and "B2" exposed to 20% PVS2 solution for 60 min, followed by 100% PVS2 for 20 min and stored at LN during 2, 6, 12 and 18 months showed no effects on leaf and shoot development from shoot tips [32]. It was reported that the amount of ice formation is much lower using 60 min of PVS2 exposure time [33]. However, in the present experiment, it was recorded a better embryo viability using lower concentration of PVS2 and short exposure time.…”

Section: Discussioncontrasting

confidence: 91%

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Cryopreservation by Vitrification of Vitis vinifera cv. “Red Globe” Zygotic Embryos and Effect on the Expression of DNA Methyltransferase Genes

García-Coronado¹,

Báez-Flores²,

Troncoso‐Rojas³

et al. 2016

JAST-B

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Mexico is a large producer of table grape (Vitis vinifera L.) and therefore it is important to develop protocols to store the grape varieties germplasm. The objective of the present work was to design a protocol for the cryopreservation by vitrification of zygotic embryos of V. vinifera cv. "Red Globe" and evaluate possible epigenetics changes. The plant vitrification solution 2 (PVS2) was utilized before the utilization of liquid nitrogen (LN). The effect of this protocol on embryo viability was tested by the triphenyl-tetrazolium chloride solution, as well as by the in vitro development of grape embryos into plantlet. A cDNA expression library of grape zygotic embryos was created to isolate expressed sequence tags of several DNA methyltrasferases. Gene expression of domains rearranged methyltransferase type 1 (DMR1) and DNA (cytosine-5)-methyltransferase 1 (MET1-2) isozymes was analyzed by quantitative reverse transcriptase PCR. The optimal conditions for vitrification were 10 min in 50% PVS2, followed by 10 min in 100% PVS2. Under these conditions, about 30% of plantlet was obtained from embryos after cryopreservation. It was recorded a reduction in the MET1-2 gene expression, which plays a role in the maintenance of DNA methylation. It is possible to cryopreserve viable grape zygotic embryos, although the treatment seems to induce alterations in the normal DNA methylation pattern of the zygotic embryo genome.

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Section: Discussioncontrasting

confidence: 91%

“…Besides, it appears that 15% of dimethyl sulfoxide in the PVS2 solution can damage the plant tissues [35]. Indeed, the reduction in the amount of dimethyl sulfoxide from 15% to 10% and 5% in the PVS2 solution, improved the regeneration of Loxocarya cinerea shoot tips genotype SXH404 from 20% to almost 60%, respectively [33].…”

Section: Discussionmentioning

confidence: 99%

Cryopreservation by Vitrification of Vitis vinifera cv. “Red Globe” Zygotic Embryos and Effect on the Expression of DNA Methyltransferase Genes

García-Coronado¹,

Báez-Flores²,

Troncoso‐Rojas³

et al. 2016

JAST-B

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“…Shoot tips, containing the apical meristem, from where growth emerges, are organized and genetically stable structures. Cells within the apical meristem are slight differentiated, small and unvacuolated, and can resist desiccation and freezing better than highly vacuolated and differentiated cells [77,78]. Therefore, after cryostorage and rewarming, these characteristics enable direct shoot formation from cryopreserved shoot tips and maintenance of the genetic integrity [14,77,78].…”

Section: Explant Typementioning

confidence: 99%

“…Cells within the apical meristem are slight differentiated, small and unvacuolated, and can resist desiccation and freezing better than highly vacuolated and differentiated cells [77,78]. Therefore, after cryostorage and rewarming, these characteristics enable direct shoot formation from cryopreserved shoot tips and maintenance of the genetic integrity [14,77,78]. Several endemic plant species, such as Paraisometrum mileense W. T. Wang (endemic to Yunan, China) [79], Thymus moroderi Pau ex Martínez (endemic to South-eastern Spain) [80,81], and Tuberaria major (Willk.)…”

Section: Explant Typementioning

confidence: 99%

“…Despite being an unorganized tissue and more prone to genetic abnormalities, when compared to shoot tips, calli are easier and faster to handle, and the final amount of regenerated material can be higher [77,78]. Cryopreservation of shoot tips of Loxocarya cinerea R.Br., an endemic species to the southwest Western Australia, allowed a very low regeneration rate post-cryopreservation (below 5%) [78]. On the other hand, applying the same protocol to callus tissues, authors obtained survival percentages above 90%.…”

Section: Explant Typementioning

confidence: 99%

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Endemic Plant Species Conservation: Biotechnological Approaches

Coelho

Gonçalves

Romano

2020

Plants

120

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Endemic plant species are usually more vulnerable to anthropogenic threats and natural changes and, therefore, hold a higher extinction risk. The preservation of these species is a major concern on a worldwide context and in situ protection alone will not guarantee their conservation. Ex situ conservation measures must be undertaken to support the conservation of these species, and seed banking is the more efficient and cost-effective method. However, when seed banking is not an option, alternative approaches should be considered. Biotechnological tools provide new and complementary options for plant conservation including short-, medium-, and long-term strategies, and their application for plant species conservation has increased considerably in the last years. This review provides information about the status of the use biotechnology-based techniques for the conservation of endemic plant species. Particular attention is given to cryopreservation, since is the only long-term ex situ conservation strategy that can complement and support the other conservation measures. The cryopreservation of plant genetic resources is, however, more focused on crop or economically important species and few studies are available for endemic plant species. The plant material used, the cryopreservation methods employed, and the assessment of cryogenic effects are reviewed. The reasons to explain the difficulties in cryopreserving these species are discussed and new strategies are proposed to facilitate and increase the interest on this matter. We expect that further studies on the conservation of endemic plant species will increase in a near future, thus contributing to maintain these valuable genetic resources.

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The need for novel cryoprotectants and cryopreservation protocols: Insights into the importance of biophysical investigation and cell permeability

Raju

Bryant

Wilkinson

et al. 2021

Biochimica et Biophysica Acta (BBA) - General Subjects

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131. Development of a cryopreservation protocol for Loxocarya cinerea – A recalcitrant plant species used in post-mining restoration

Cited by 6 publications

References 2 publications

Cryopreservation by Vitrification of Vitis vinifera cv. “Red Globe” Zygotic Embryos and Effect on the Expression of DNA Methyltransferase Genes

Cryopreservation by Vitrification of Vitis vinifera cv. “Red Globe” Zygotic Embryos and Effect on the Expression of DNA Methyltransferase Genes

Endemic Plant Species Conservation: Biotechnological Approaches

The need for novel cryoprotectants and cryopreservation protocols: Insights into the importance of biophysical investigation and cell permeability

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