Cryopreservation of an endangered Hladnikia pastinacifolia Rchb. by shoot tip encapsulation-dehydration and encapsulation-vitrification

Ciringer, Terezija; Martı́n, Carmen; Šajna, Nina; Kaligarič, Mitja; Ambrožič-Dolinšek, Jana

doi:10.1007/s11627-018-9917-y

Cited by 15 publications

(12 citation statements)

References 27 publications

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“…Gord (73% regrowth) (Kalita et al 2012), Gentiana scabra Bunge var. buergeri Maxim (73-78% regrowth) (Suzuki et al 2008) and Hladnikia pastinacifolia Rchb (60% regrowth) (Ciringer et al 2018). In our study, VSL as a cryoprotectant was very effective for both cultivars, conferring high survival and regrowth levels, especially − LN (Fig.…”

Section: Discussionsupporting

confidence: 53%

First report on cryopreservation of mature shoot tips of two avocado (Persea americana Mill.) rootstocks

O’Brien

Hiti-Bandaralage

Folgado

et al. 2020

Plant Cell Tiss Organ Cult

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Cryopreservation combined with in vitro culture offers a safe and cost-effective method to conserve germplasm. Conservation of Persea spp. has been limited to heterozygous somatic embryos that are not true-to-type. A method for shoot-tip cryopreservation is vital to preserve the exact gene pool of interest. For the first time cryopreservation protocols for mature shoot tips of two avocado cultivars (cvs) 'Velvick' and 'Reed', were established. In vitro shoots were subjected to two different optimised pre-treatments; (1) cv 'Velvick'-high sucrose (0.3 M) or (2) cv 'Reed'-low temperature (10 °C) incubation, over a 2-week period prior shoot tip dissection. Two different plant vitrification solutions, plant vitrification solution 2 (PVS2) and vitrification solution L (VSL) were tested at 0 °C for 0, 10, 20, 30 and 40 min. Vitrified shoots were evaluated for survival and regrowth at 2 and 8 weeks after vitrification treatment and either with or without liquid nitrogen exposure. The study revealed that the optimal exposure time for each cultivar varied with the cryoprotectant used. After liquid nitrogen cv 'Velvick' highest regrowth levels were observed with 20 min exposure to either PVS2 or VSL, however, vigorous plants were produced only from VSL treated shoots. In the case of cv 'Reed' highest regrowth levels were observed with 10 min exposure to PVS2 however only morphologically normal plants were recovered from VSL treated shoots. Key messageCryopreservation of avocado shoot tips was successful using PVS2 and VSL with both recording similar recovery rates for 'Velvick' and 'Reed'; although only vigorous and morphologically normal plants were developed from VSL treatments.

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Section: Discussionsupporting

confidence: 53%

First report on cryopreservation of mature shoot tips of two avocado (Persea americana Mill.) rootstocks

O’Brien

Hiti-Bandaralage

Folgado

et al. 2020

Plant Cell Tiss Organ Cult

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“…Protocols must be optimized individually according to the species/clone and explant typeto be cryopreserved to obtain the best recovery rates after cryostorage [20,61]. There are many cryopreservation studies concerning crops or economically relevant plant species, due to their value and the large knowledge of their growing conditions [14,62]. In the case of most endemic and wild species, they are highly variable and the information about their biology is scarce, which makes the process of developing a cryopreservation protocol more complex.…”

Section: Cryopreservationmentioning

confidence: 99%

Endemic Plant Species Conservation: Biotechnological Approaches

Coelho

Gonçalves

Romano

2020

Plants

143

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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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“…A similarly low genomic variation-at the level of 5%-was reported in LN-stored Hladnikia pastinacifolia Rchb. [18] and Chrysanthemum × grandiflorum [36]. The results obtained here suggest that, with no altered genotypes detected, the encapsulation-vitrification technique not only secures the viability of the highest number of explants, but is also effective in preserving the genetic stability of plants.…”

Section: Stability Of Plant Materials After Various (Cryo)treatmentsmentioning

confidence: 62%

“…Stability and uniformity of the recovered plant after cryo-exposure is a major concern in the cryopreservation of plant genetic resources. No or minor variation is often detected after tissue storage in LN [18]. However, Martín et al [19] reported significant genetic variation after applying the encapsulation-dehydration cryopreservation protocol (based on osmotic dehydration of tissues in sucrose gradient) in Chrysanthemum × morifolium Ramat.…”

Section: Introductionmentioning

confidence: 99%

Shoot Tip Cryopreservation of Lamprocapnos spectabilis (L.) Fukuhara Using Different Approaches and Evaluation of Stability on the Molecular, Biochemical, and Plant Architecture Levels

Kulus

2020

IJMS

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The aim of this study is to optimize and evaluate the effectiveness of vitrification, droplet-vitrification, and encapsulation-vitrification techniques in the cryopreservation of Lamprocapnos spectabilis (L.) Fukuhara ‘Gold Heart’, a popular medicinal and ornamental plant species. In vitro-derived shoot tips were used in the experiments. All three techniques were based on explant dehydration with plant vitrification solution 3 (PVS3; 50% glycerol and 50% sucrose) for 0, 30, 60, 90, 120, 150, or 180 min. The recovered microshoots were subjected to morphometric, biochemical, and molecular analyses (RAPD, ISSR, SCoT). The highest recovery level was reported with the encapsulation-vitrification protocol based on 150 min dehydration (73.1%), while the vitrification technique was the least effective (maximum 25.8% recovery). Explants cryopreserved with the encapsulation-vitrification technique produced the highest mean number of shoots (4.9); moreover, this technique was optimal in terms of rooting efficiency. The highest fresh weight of shoots, on the other hand, was found with the vitrification protocol based on a 30-min PVS3 treatment. The concentrations of chlorophyll a and b were lower in all cryopreservation-derived plants, compared to the untreated control. On the other hand, short dehydration and cryopreservation of non-encapsulated explants stimulated the synthesis of anthocyanins. A small genetic variation in 5% of all samples analyzed was detected by RAPD and ISSR marker systems. Only plants recovered from the encapsulation-vitrification protocol had no DNA sequence alternations.

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Cryopreservation of an endangered Hladnikia pastinacifolia Rchb. by shoot tip encapsulation-dehydration and encapsulation-vitrification

Cited by 15 publications

References 27 publications

First report on cryopreservation of mature shoot tips of two avocado (Persea americana Mill.) rootstocks

First report on cryopreservation of mature shoot tips of two avocado (Persea americana Mill.) rootstocks

Endemic Plant Species Conservation: Biotechnological Approaches

Shoot Tip Cryopreservation of Lamprocapnos spectabilis (L.) Fukuhara Using Different Approaches and Evaluation of Stability on the Molecular, Biochemical, and Plant Architecture Levels

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