A simple and efficient method for onion pollen preservation: Germination, dehydration, storage conditions, and seed production

Fayos, Oreto; Echávarri, B.; Vallés, M. P.; Mallor, C.; Garcés-Claver, Ana; Castillo, A. M.

doi:10.1016/j.scienta.2022.111358

Cited by 5 publications

(6 citation statements)

References 23 publications

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“…The normal development of pollen as the male gametophyte is crucial for successful fertilization during reproduction [24]. Previous studies showed that the pollen source and vitality could affect the seed setting rate [25] and the nutritional content of the seeds in cross-pollination [1,3]. In production, the flowering period of P. ostii is inconsistent, and understanding the pollen viability of P. ostii in vitro is important for improving the breeding efficiency through hybridization.…”

Section: Discussionmentioning

confidence: 99%

“…The medium was placed at different temperatures (15,20,25,30,35 • C) for germination, and after 30, 60, 90, 120, and 180 min, the germination percentage and pollen tube length were measured (Tables 3 and 4). The germination percentage of pollen grains was found to be temperature-dependent, with variations in temperatures significantly affecting the germination of pollen grains.…”

Section: Pollen Germination and Pollen Tube Growth In Vitro At Differ...mentioning

confidence: 99%

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Optimization of In Vitro Germination, Viability Tests and Storage of Paeonia ostii Pollen

Jiang

et al. 2023

Plants

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Paeonia ostii is an important woody oil crop mainly cross-pollinated. However, the low yield has become an important factor restricting the industrial development of P. ostii. Cross-pollination has become one of the important measures to increase the seed yield. Therefore, conservation of pollen with high vitality is crucial to ensure successful pollination of P. ostii. In this study, we found an effective methodological system to assess the viability, ability to germinate, and optimal storage conditions of P. ostii pollen grains. The optimal medium in vitro was 50 g/L sucrose, 100 mg/L boric acid, 50 g/L PEG6000, 100 mg/L potassium nitrate, 300 mg/L calcium nitrate, and 200 mg/L magnesium sulfate at pH 5.4. Optimal germination condition in vitro was achieved at 25 °C for 120 min, allowing easy observation of the germination percentage and length of the pollen tubes. In addition, the viability of pollen grains was assessed by comparing nine staining methods. Among them, MTT, TTC, benzidine-H2O2, and FDA were effective to distinguish between viable and non-viable pollen, and the results of the FDA staining method were similar to the pollen germination percentage in vitro. After evaluation of pollen storage, thawing and rehydration experiments showed that thawing at 4 °C for 30 min and rehydration at 25 °C for 30 min increased the germination percentage of pollen grains stored at low temperatures. The low-temperature storage experiments showed that 4 °C was suitable for short-term storage of P. ostii pollen grains, while −80 °C was suitable for long-term storage. This is the first report on the in vitro germination, viability tests, and storage of P. ostii pollen grains, which will provide useful information for P. ostii germplasm conservation and artificial pollination.

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Section: Discussionmentioning

confidence: 99%

Section: Pollen Germination and Pollen Tube Growth In Vitro At Differ...mentioning

confidence: 99%

Optimization of In Vitro Germination, Viability Tests and Storage of Paeonia ostii Pollen

Jiang

et al. 2023

Plants

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“…Storing in an ultra-low temperature freezer at −80 °C using a routine pre-treatment method in semi-solid media gave surprisingly low regeneration rates compared to storing in LN. Storage at −80 °C of ET has only been reported for P. radiata [ 27 ], although this storage temperature has been used for other types of germplasm, such as the pollen of different angiosperms [ 37 , 38 ], or macroalgal tissues [ 39 ]. The complexity of the ET and its high-water content may hinder the application of preservation at this temperature for Norway spruce ET.…”

Section: Discussionmentioning

confidence: 99%

Reducing Pre- and Post-Treatments in Cryopreservation Protocol and Testing Storage at −80 °C for Norway Spruce Embryogenic Cultures

Varis

Virta

Montalbán

et al. 2022

IJMS

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Somatic embryogenesis (SE) is considered the most effective method for vegetative propagation of Norway spruce (Picea abies L. Karst). For mass propagation, a storage method that is able to handle large quantities of embryogenic tissues (ETs) reliably and at a low cost is required. The aim of the present study was to compare freezing at −80 °C in a freezer to cryopreservation using liquid nitrogen (LN) as a method for storing Norway spruce ETs. The possibility of simplifying both the pre-treatment and thawing processes in cryopreservation was also studied. The addition of abscisic acid (ABA) to the pre-treatment media and using polyethylene glycol PEG4000 instead of PEG6000 in a cryoprotectant solution were tested. Both the pre-and post-treatments on semi-solid media could be simplified by reducing the number of media, without any loss of genotype or embryo production capacity of ETs. On the contrary, the storage of ETs in a freezer at −80 °C instead of using LN was not possible, and the addition of ABA to the pre-treatment media did not provide benefits but increased costs. The lower regeneration rate after using PEG4000 instead of PEG6000 in a cryoprotectant solution in cryovials was unexpected and unwanted. The simplified pre-and post-treatment protocol will remarkably reduce the workload and costs in the mass-cryopreservation of future forest regeneration materials and in thawing the samples for mass propagations, respectively.

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“…Pollen storage methods range from room temperature storage (25°C) to low-temperature storage in household refrigerators (4°C), freezers (−20°C), and deep freezers (−80°C), with options for storing fresh, desiccated, vacuum- or freeze-dried pollen. 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 ).…”

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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A simple and efficient method for onion pollen preservation: Germination, dehydration, storage conditions, and seed production

Cited by 5 publications

References 23 publications

Optimization of In Vitro Germination, Viability Tests and Storage of Paeonia ostii Pollen

Optimization of In Vitro Germination, Viability Tests and Storage of Paeonia ostii Pollen

Reducing Pre- and Post-Treatments in Cryopreservation Protocol and Testing Storage at −80 °C for Norway Spruce Embryogenic Cultures

Simple cryopreservation protocol for Luffa pollen: enhancing breeding efficiency

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