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In Vitro Propagation and Cryopreservation of Plants

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A special issue of Plants (ISSN 2223-7747). This special issue belongs to the section "Plant Physiology and Metabolism".

Deadline for manuscript submissions: 31 July 2024 | Viewed by 2092

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Special Issue Editor

Prof. Haeng-hoon Kim

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Guest Editor

Department of Agricultural Life Science, Sunchon National University, Suncheon 57922, Republic of Korea
Interests: biodiversity conservation; cryopreservation; plant biotechnology
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

Call upon the importance of plant biodiversity for humankind and their threatening level. Cryopreservation, the storing of biological samples in liquid nitrogen (LN), can offer valuable options for non-orthodox seeds, vegetatively propagated species, and cell cultures. In vitro propagation is also helpful for preparing plant materials for cryopreservation, especially threatened wild species. Moreover, as is common sense among cryobiologists, the success of cryopreservation depends on the vigor of plant materials provided by the in vitro culture and the regrowth protocol. In the era of cryobanking germplasm collections of food and agriculture, we still need to develop cryo-biotechnology through principle studies, systematic approaches, and practical applications. Since cryopreservation is a multidisciplinary process, approaches for tuning the whole process or focusing on specific stages, i.e., plant material preparation, pre-LN, cooling/rewarming and unloading, post-LN regrowth, etc., are welcome. This Special Issue of Plants will highlight all aspects of in vitro propagation and cryopreservation technologies to solve plant conservation problems.

Prof. Dr. Haeng-hoon Kim
Guest Editor

Manuscript Submission Information

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. All submissions that pass pre-check are peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short communications are invited. For planned papers, a title and short abstract (about 100 words) can be sent to the Editorial Office for announcement on this website.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. Plants is an international peer-reviewed open access semimonthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 2700 CHF (Swiss Francs). Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

Keywords: Breeding; cryoconservation; Humulus lupulus; pollination; Solanum tuberosum; variety

Published Papers (2 papers)

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Research

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21 pages, 5353 KiB

Open AccessArticle

Cryopreservation of Indigenous Plums and Monitoring of Multiplication and Rooting Capacity of Shoots Obtained from Cryopreserved Specimens

by Tatjana Vujović, Tatjana Anđelić, Bojana Vasilijević, Darko Jevremović and Florent Engelmann

Plants 2023, 12(17), 3108; https://doi.org/10.3390/plants12173108 - 30 Aug 2023

Cited by 1 | Viewed by 870

Abstract

The objective of this study is to assess the suitability of vitrification cryo-plate (V cryo-plate) and dehydration cryo-plate (D cryo-plate) methods for the long-term conservation of eight autochthonous Prunus domestica L. genotypes originating from the Balkan Peninsula region. In vitro shoot tips were briefly pre-cultured for 1 day at 23 °C in the dark on a medium containing 0.3 M sucrose and then embedded in calcium alginate gel within the wells of the aluminum cryo-plates. In the V cryo-plate protocol, dehydration was carried out at room temperature using the following vitrification solutions: original plant vitrification solution 2 (PVS2) and 90% PVS2 solution (for 20 and 40 min) and plant vitrification solution 3 (PVS3) (for 60 and 80 min). In the D cryo-plate protocol, desiccation was performed for 2, 2.5, or 3 h over silica gel at 23 °C. The effect of different treatments was evaluated by monitoring the regrowth of both non-frozen and cryo-preserved explants. After cryo-preservation, five genotypes achieved regrowth rates over 40% in at least one of the applied protocols, while two genotypes showed regrowth rates of around 10%. A significant improvement in regrowth success for all genotypes using both cryo-plate methods was achieved by pre-culturing shoot tips for 7 days on a medium containing 0.5 M sucrose in complete darkness at 4 °C. Shoots regenerated from cryo-preserved explants were further monitored in vitro. By the third subculture, they had not only regained but had even exceeded the multiplication capacity (index of multiplication, length of axial, and lateral shoots) of shoots regenerated from dissection controls. Following multiplication, the cryo-preserved shoots were successfully rooted and rooting ability was assessed by monitoring the percentage of rooting, number and length of roots, and height of rooted plantlets. Full article

(This article belongs to the Special Issue In Vitro Propagation and Cryopreservation of Plants)

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Review

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22 pages, 760 KiB

Open AccessReview

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

by Haenghoon Kim

Plants 2023, 12(23), 4040; https://doi.org/10.3390/plants12234040 - 30 Nov 2023

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Abstract

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. Full article

(This article belongs to the Special Issue In Vitro Propagation and Cryopreservation of Plants)

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Planned Papers

The below list represents only planned manuscripts. Some of these manuscripts have not been received by the Editorial Office yet. Papers submitted to MDPI journals are subject to peer-review.

Analysis of thermal characteristics of potato and hop pollen for their cryopreservation and cross-breeding

Miloš Faltus 1*, Jaroslava Domkářová 2, Petr Svoboda 3, Vendulka Horáčková 2, Vladimír Nesvadba 3, Vladislav Klička 4, Alois Bilavčík 2 and Jiří Zámečník 2

Abstract: This study investigated the thermal properties of potato and hop pollen for cryopreservation and following crossbreeding purposes. Using a differential scanning calorimeter, the phase transitions and water content of fresh and dehydrated pollen samples were measured. Potato pollen showed high variability in water content (0.1-0.6 g/g) and crystalline fraction (0-14%), and a wide range of glass transition temperatures (-80 to +40 °C). Hop pollen had lower water content (0.1-0.3 g/g) and no detectable freezing or melting events, with glass transition temperatures between -53 and +15 °C. Both types of pollen maintained high viability before and after cryopreservation, except when exposed to unfavorable conditions. The cryopreserved pollen was successfully used to produce hybrids of potato and hop. The results suggest that cryopreservation is a feasible technique for preserving and utilizing pollen of these crops.