Pollen Cryopreservation: Advances and Prospects

Rajasekharan, P. E.; Rohini, M. R.

doi:10.1007/978-1-0716-2843-0_1

Cited by 4 publications

(3 citation statements)

References 34 publications

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“…The importance of pollen cryopreservation for the long-term conservation of genetic resources has been highlighted in many plant species, including ornamental, fruit, and forest [ 76 , 77 , 78 , 79 ]. Pollen grains are suitable genetic materials for preservation for their small size and desiccation tolerance, and their conservation is beneficial in breeding programs overcoming seasonal and geographical restrictions [ 25 ].…”

Section: Resultsmentioning

confidence: 99%

Long-Term Conservation for the Safeguard of Abies nebrodensis: An Endemic and Endangered Species of Sicily

Benelli,

Tarraf,

İzgü

et al. 2024

Plants

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The combined approaches between ex situ and in situ conservation are of great importance for threatened species in urgent need of protection. This study aims to develop concrete actions to preserve the relic of 30 adult trees of the Sicilian fir (Abies nebrodensis) from extinction using long-term germplasm conservation in liquid nitrogen (LN, −196 °C). Pollen grains were collected, and their moisture content (MC) was measured. Then, viability (2,3,5-tryphenyl tetrazolium chloride, TTC), in vitro germinability, and enzymatic antioxidant activity (ascorbate peroxidase, APX; catalase, CAT) were evaluated before and after cryopreservation. Seeds collected from mature cones underwent X-ray analysis, and only full seeds were used to excise the zygotic embryos (ZEs) for cryopreservation. The MC percentage of ZEs was determined, and then they were plunged in LN with (+PVS2) or without (−PVS2) Plant Vitrification Solution 2; untreated ZEs were used as a control. Viability (TTC test) and in vitro germination were assessed for all ZEs (+PVS2, −PVS2, and control). Embryogenic callus (EC) lines obtained from mature ZEs were cryopreserved applying the ‘encapsulation-dehydration’ technique. This study has allowed, after optimizing cryopreservation protocols for pollen, ZEs, and EC of A. nebrodensis, to establish the first cryobank of this endangered species in Polizzi Generosa (Palermo, Italy), inside the ‘Madonie Regional Park’. The strategy developed for Sicilian fir conservation will pave the way for similar initiatives for other critically endangered conifer species.

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Section: Resultsmentioning

confidence: 99%

Long-Term Conservation for the Safeguard of Abies nebrodensis: An Endemic and Endangered Species of Sicily

Benelli,

Tarraf,

İzgü

et al. 2024

Plants

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“…The success of long-term conservation of pollen samples relies on many factors, such as the stage of pollen collection, the pretreatment of the samples, and the methodology applied [62]. Furthermore, post-thawing is yet another key factor that greatly influences the retrieval of stored pollen because it is related to pollen metabolism and the reactivation of post-cryopreserved metabolic processes [121]. Gladiolus pollen conservation was reported by Rajasekharan et al [116]: in this study, five selected cultivars were tested, and the mean post-germination rate recorded was 52.8% compared to fresh pollen (without storage) at 58.1%.…”

Section: Iridaceae Familymentioning

confidence: 99%

The Cryopreservation of Medicinal and Ornamental Geophytes: Application and Challenges

Merzougui

Benelli̇

Boullani

et al. 2023

Plants

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Nowadays, plant genetic resources are often at risk of loss and destruction. Geophytes are herbaceous or perennial species that are annually renewed by bulbs, rhizomes, tuberous roots, or tubers. They are often subject to overexploitation, which, combined with other biotic and abiotic stresses, can make these plants more vulnerable to a decline in their diffusion. As a result, multiple endeavors have been undertaken to establish better conservation strategies. Plant cryopreservation at ultra-low temperatures in liquid nitrogen (−196 °C) has proven to be an effective, long-term, low-cost, and suitable conservation method for many plant species. Over the last two decades, major advances in cryobiology studies have enabled successful explants of multiple genera and types, including pollen, shoot tips, dormant buds, and zygotic and somatic embryos. This review provides an update on recent advances and developments in cryopreservation and its application to medicinal and ornamental geophytes. In addition, the review includes a brief summary of factors limiting the success of bulbous germplasm conservation. The critical analysis underpinning this review will benefit biologists and cryobiologists in their further studies on the optimization of geophyte cryopreservation protocols and will support a more complete and wider application of knowledge in this area.

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“…However, pollen can only be stored for short- to medium-term periods (hours to days to weeks to months) at these temperatures ( Fayos et al., 2022 ; Wu et al., 2022 ; Shankar et al., 2023 ). Cryopreservation is the storage of pollen at ultra-low temperature in liquid nitrogen (LN) either in the liquid phase (−196°C) or in the vapor phase (−150 to −180°C) retaining its original viability and offers long-term storage ( Rajasekharan and Rohini, 2023 ). Thus, in the present study, four species of Luffa genus, namely, L. acutangula , L. cylindrica , L. echinata , and L. graveolens , were utilized for pollen cryopreservation studies with the objectives of (a) standardizing the optimum in vitro pollen germination medium; (b) identifying the reliable pollen viability assessment method; (c) studying the effect of different storage temperatures and duration on pollen viability; (d) identifying the optimum moisture content, desiccation period, and thawing temperature for cryopreservation; and (e) standardizing the species-wide pollen cryopreservation protocol.…”

Section: Introductionmentioning

confidence: 99%

Simple cryopreservation protocol for Luffa pollen: enhancing breeding efficiency

Nagar,

Gowthami,

Sureja

et al. 2023

Front. Plant Sci.

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This study aimed to develop a long-term pollen storage protocol for Luffa species (L. acutangula, L. cylindrica, L. echinata, and L. graveolens) and assess its potential for crop improvement. The optimal medium for in vitro pollen germination varied by species, with Brewbaker and Kwack (BK) medium with 10% sucrose suitable for L. acutangula, L. cylindrica, and L. echinata, and BK medium with 3% sucrose ideal for L. graveolens. Overestimation in staining tests compared to in vitro pollen germination was observed. The best results for cryopreservation were achieved with desiccation periods of 20, 30, and 40 min, maintaining moisture content between 14.04% and 18.55%. Pollen viability was negatively correlated with storage temperature (25, 4, and −20°C) and duration. Cryopreserved pollen at −196°C exhibited the highest viability over a prolonged period (2 months) and was comparable to fresh pollen in terms of germination, ovule fertilization, and fruit and seed set. This study presents a simple and reproducible pollen cryopreservation protocol applicable across Luffa species, facilitating long-term storage and its use in crop improvement efforts.

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Pollen Cryopreservation: Advances and Prospects

Cited by 4 publications

References 34 publications

Long-Term Conservation for the Safeguard of Abies nebrodensis: An Endemic and Endangered Species of Sicily

Long-Term Conservation for the Safeguard of Abies nebrodensis: An Endemic and Endangered Species of Sicily

The Cryopreservation of Medicinal and Ornamental Geophytes: Application and Challenges

Simple cryopreservation protocol for Luffa pollen: enhancing breeding efficiency

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