Morphoanatomical aspects of the starting material for the improvement of pineapple cryopreservation by the droplet-vitrification technique

Guerra, Patrícia Araújo; Souza, Everton Hilo de; Max, Daniela A.S.; Rossi, Mônica Lanzoni; Villalobos-Olivera, Ariel; Lédo, C.A.S.; Martínez-Montero, Marcos Edel; Souza, Fernanda Vidigal Duarte

doi:10.1590/0001-3765202120190555

Cited by 5 publications

(6 citation statements)

References 13 publications

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“…In comparison, smaller kiwi shoot tips (0.5-1 mm) from 2-week-old young shoots produced higher LN regeneration over larger shoot tips (1-2 mm) from 6-week-old donor plants acclimated at 4 • C [36]. Guerra et al [7] noticed that pineapple shoot tips from 30 days subcultures had good morphoanatomical appearances and thus showed a higher survival rate compared to 45 days or 60 days subcultures. Subculture conditions, i.e., aeration of culture vessels, sparse planting density, and higher light intensity, helped produce healthy donor plants and thus significantly affected on recovery of LNC and LN shoot-tips in potatoes [8].…”

Section: Subculture Conditions and Post-cryopreservation Regenerationmentioning

confidence: 98%

“…The triangle of cryopreservation would be plant material, protocol, and manipulation (operational skill). In terms of plant material, numerous studies investigated genotypedependent various responses [7], and a few kinds of literature examined the effect of donor plants and the selection of appropriate explants [4,7,8]. However, the importance of healthy donor plants and the choice of proper explants for cryopreservation have been pointed out [9,10].…”

Section: Introductionmentioning

confidence: 99%

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Vigorous Growing of Donor Plantlets by Liquid Overlay in Subcultures Is the Key to Cryopreservation of Endangered Species Pogostemon yatabeanus

Lee

Kim

2022

Plants

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Cryopreservation is a unique option for the long-term conservation of threatened plant species with non-orthodox or limitedly available seeds. However, the wide application of cryopreservation for the protection of wild flora is hampered by some reasons: limits of source material available, difficulties in in vitro propagation, needs to re-optimize protocol steps for new species, etc. In this study, using an endemic and endangered Korean species, Pogostemon yatabeanus, we investigated subculture medium and supplements on in vitro growth of donor plants: medium strength, gelling agents, liquid overlay, plant hormones, and activated charcoal. Subculture conditions of each cycle tested significantly impacted on height and dry weight of subcultured donor plantlets. Among the treatments tested, the overlay of the liquid medium on top of gellan gum-gelled medium significantly increased the growth of shoots and roots. In the droplet-vitrification procedure, the survival and regeneration of cryopreserved shoot tips were critically impacted by the dry weight of donor plantlets (CORELL = 0.85~0.95) which was affected by the following subculture conditions. Moreover, every subsequent subculture cycle before cryopreservation positively or negatively impacted post-cryopreservation regeneration. This study highlights the vigor of donor plantlets for post-cryopreservation regeneration and provides practices for the revitalization of donor plants during subcultures.

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Section: Subculture Conditions and Post-cryopreservation Regenerationmentioning

confidence: 98%

Section: Introductionmentioning

confidence: 99%

Vigorous Growing of Donor Plantlets by Liquid Overlay in Subcultures Is the Key to Cryopreservation of Endangered Species Pogostemon yatabeanus

Lee

Kim

2022

Plants

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“…The importance of healthy donor plants is common sense in the plant cryobiologist community. There is a lot of literature on the vigor of donor plants (subculture conditions and duration, growth), preconditioning of donor plants (cold acclimation, pre-growth, preculture), and explants (type and location, size) [81][82][83][84][85][86]. Engelmann [80] and Reed [87] also pointed out that choosing starting material and cultural conditions is as necessary as the cryopreservation protocol.…”

Section: Liquid Overlay-induced Donor Plant Vigor (Dpv)mentioning

confidence: 99%

“…Cytotoxicity has been considered a bottleneck in the vitrification-based cryopreservation of complex spatial explants [17]. It has been recognized that small, actively dividing juvenile tissue with dense cytoplasm and less vacuolized cells is prone to tolerate freezing injury [85,88]. Even with the liquid overlay subculture, the shoot tips of P. chinense plantlets we sensitive to the cytotoxicity of the PVS.…”

Section: Liquid Overlay-induced Donor Plant Vigor (Dpv)mentioning

confidence: 99%

Optimizing the Droplet-Vitrification Procedure by Balancing the Cryoprotection and Cytotoxicity of Alternative Plant Vitrification Solutions Based on the Nature of Donor Plant Vigor

Kim

2023

Plants

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Over 30 years of plant vitrification, droplet vitrification (DV) of in vitro propagules and slow freezing of dormant buds are typical methods of large-scale cryobanking worldwide. One-step sucrose preculture and Plant Vitrification Solution 2 (PVS2) cryoprotection in solution-based vitrification often face unacceptably low regeneration, and the results are on a case-by-case basis depending on the plant species, like a blind test. The absence of a universal protocol applicable across all plant diversity is considered one of the limiting factors. For wild flora, limits of source material available and difficulties in in vitro propagation make it worse to re-optimize the protocol steps for new species. Since cryoprotectant toxicity is the most crucial barrier to the vitrification of organized explants, selecting alternative plant vitrification solutions (PVS) based on the cytotoxicity of cryoprotectants is vital. This review proposes the concept of donor plant vigor (DPV), which refers to the donor plant properties that determine the potential to regenerate normal plantlets under various cryopreservation procedures. DV is a multi-stage procedure with many factors from stage (1) material preparation to (2) pre-liquid nitrogen (pre-LN) (preculture, osmoprotection, cryoprotection), (3) LN (cooling), (4) warming conditions (rewarming, unloading), and (5) regrowth. Since the cytotoxicity of PVS is a primary limiting factor in DV approaches, DPV is crucial for coping with the toxicity of PVS. The DPV is innate and can be maximized with appropriate material preparations, i.e., vigorously growing in subcultures aided by a liquid overlay on top of the gelled medium, selecting proper explants, optimizing the two-step preculture conditions, and media supplements. Developing the DV protocol starts with testing the material with a tentative standard protocol, which includes a two-step preculture (10% sucrose for 31 h and 17.5% sucrose for 16 h), osmoprotection with C4-35%, cryoprotection with A3-80% (60 min at 0 °C), cooling, and rewarming using aluminum foil strips. Using a three-step regrowth initially with ammonium-free regrowth medium, regrowth of shoot tips in one plate following the successive stages of the tentative standard protocol for shoot tips, i.e., fresh, PC, OP, CP (LNC), and LN, is a valuable tool to characterize the sensitivity of the material and to standardize the procedure by tuning the cryoprotection and cytotoxicity of cryoprotectants. A-series PVS (A3-90%, A3-80%, A3-70%) and B-series PVS (PVS3, B5-85%) can be tested based on the DPV. These alternative PVSs have been applied in over 30 pieces of literature with an 8.5~67.3% increase in LN regeneration compared to PVS2 and Plant Vitrification Solution 3 (PVS3) treatments. Using this approach as an alternative to blind condition screening would be influential in broadening the cryopreservation of diverse wild species and problem materials.

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“…At the Embrapa Cassava & Fruits, indexing for the wilt virus is routinely performed via RT-PCR (GUERRA et al, 2021;SILVA et al, 2021). As for the time to obtain the seedlings, micropropagation takes the most time but compensates for the number of plants produced.…”

Section: Micropropagationmentioning

confidence: 99%

Vegetative propagation strategies in commercial pineapple cultivars

Almeida,

Souza,

Costa

et al. 2023

Rev. Caatinga

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The production of healthy pineapple seedlings for commercial plantations has been one of the main limiting factors of the species, and the search for improvements to meet this demand has been constant. Therefore, the present study aimed to compare different propagation strategies (natural tillering, stem sectioning, and micropropagation) in three commercial cultivars 'BRS Imperial', 'Pérola', and Gold 'MD-2' of pineapple. Three methods of vegetative propagation were evaluated: conventional, stem sectioning, and micropropagation. The evaluations consisted of the number of seedlings produced by each method and the period for their development until planting in the field. According to the conventional propagation technique results, the young seedling type in the cultivar 'Pérola' was more abundant. In the propagation by sectioning the stem, the 'BRS Imperial' cultivar presented the highest number of seedlings. Regarding the micropropagation technique, the highest production of shoots was observed in the third subculture for all cultivars, especially for ‘BRS Imperial’, which presented the highest total number of shoots in all subcultures. It was concluded that the micropropagation technique proved efficient in producing uniform seedlings on a large scale, in addition to having an advantage in the greater number of seedlings produced compared to the other methods studied.

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Morphoanatomical aspects of the starting material for the improvement of pineapple cryopreservation by the droplet-vitrification technique

Cited by 5 publications

References 13 publications

Vigorous Growing of Donor Plantlets by Liquid Overlay in Subcultures Is the Key to Cryopreservation of Endangered Species Pogostemon yatabeanus

Vigorous Growing of Donor Plantlets by Liquid Overlay in Subcultures Is the Key to Cryopreservation of Endangered Species Pogostemon yatabeanus

Optimizing the Droplet-Vitrification Procedure by Balancing the Cryoprotection and Cytotoxicity of Alternative Plant Vitrification Solutions Based on the Nature of Donor Plant Vigor

Vegetative propagation strategies in commercial pineapple cultivars

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