Plant Genetic Resources In Vitro Culture: Present Situation and Prospects for Propagation, Conservation and Sustainable Use

A special issue of Plants (ISSN 2223-7747). This special issue belongs to the section "Plant Genetic Resources".

Deadline for manuscript submissions: closed (20 September 2023) | Viewed by 19223

Special Issue Editors


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Guest Editor
Institute of BioEconomy (IBE)– Biology, Agriculture and Food Sciences Department– National Research Council (CNR), Via Madonna del Piano, 10 – 50019 Sesto Fiorentino, FI, Italy
Interests: conservation and enhancement of plant biodiversity; in vitro culture, traditional and innovative culture systems; in vitro conservation, through cryopreservation and conservation in slow growth storage; genetic stability of in vitro propagated and conserved material; ethylene production in the plant in response to stress conditions; identification and quantification of signal molecules and volatile organic compounds (VOC); analysis of plant biochemical and physiological mechanisms in response to environmental stress

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Guest Editor
Institute of BioEconomy (IBE)– Biology, Agriculture and Food Sciences Department– National Research Council (CNR), Research Campus of Florence– Via Madonna del Piano, 10 – 50019 Sesto Fiorentino, FI, Italy
Interests: sustainable agriculture; organic agriculture; biofertilizers; arbuscular mycorrhizal fungi; medicinal and aromatic plants; essential oils; biocidal activities; plant tissue culture; cryopreservation; in vitro storage

Special Issue Information

Dear Colleagues,

Plant genetic resources comprise a wide range of plant species such as herbs, shrubs and trees that are of a great importance to ensuring food, nutrition and health security. Biotic or abiotic stresses, human activities and climate change have long threatened the biodiversity in their natural habitat, causing an exponential loss of plant germplasm throughout the world. Thus, there is an urgent need to slow down genetic erosion and instigate their conservation and sustainable utilization. Advances in biotechnology can generate new opportunities for the rational use and regeneration of valuable plant material by adopting techniques such as in vitro culture and conservation. This Special Issue will focus on novel approaches of in vitro culture and ex vitro acclimatization that can positively influence the use of plant genetic resources for biomass or secondary metabolites production, as well as efficient in vitro storage methods and cryopreservation technologies applied to ensure a sustainable supply of high-quality plants as raw materials for different uses.

Potential topics include but are not limited to:

  • Role of in vitro techniques for biodiversity conservation;
  • Protocols for micropropagation;
  • Optimization of ex vitro acclimatization;
  • Use of liquid culture;
  • Methods for medium- and long-term conservation;
  • Production of bioactive compounds in vitro;
  • Future prospects for the sustainable use of genetic resources.

Dr. Anna De Carlo
Dr. Waed Tarraf
Guest Editors

Manuscript Submission Information

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Keywords

  • biodiversity
  • biotechnology
  • plant tissue culture
  • micropropagation
  • temporary immersion system
  • bioreactor
  • bioactive compounds
  • synthetic seeds
  • ex situ conservation
  • slow growth storage
  • cryopreservation
  • cryobanks

Published Papers (9 papers)

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Research

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14 pages, 2452 KiB  
Article
In Vitro Conservation of Mexican Garlic Varieties by Minimal Growth
by Bibiana Tirado, Víctor Manuel Gómez-Rodríguez, Carlos Iván Cruz-Cárdenas, Lily Xochilt Zelaya-Molina, Humberto Ramírez-Vega and Gabriela Sandoval-Cancino
Plants 2023, 12(23), 3929; https://doi.org/10.3390/plants12233929 - 22 Nov 2023
Viewed by 1078
Abstract
Garlic (Allium sativum L.) is one of the 30 crops that are essential for world food; therefore, its conservation should be considered a priority. There are two main plant conservation strategies: in situ and ex situ conservation. Both strategies are important; nevertheless, [...] Read more.
Garlic (Allium sativum L.) is one of the 30 crops that are essential for world food; therefore, its conservation should be considered a priority. There are two main plant conservation strategies: in situ and ex situ conservation. Both strategies are important; nevertheless, ex situ field conservation is affected by biotic and abiotic factors. A complementary strategy to preserve garlic germplasm in the medium term is through in vitro culture by minimal growth. The aim of this study was to evaluate the in vitro conservation of three Mexican garlic varieties by minimal growth. Garlic plants obtained from in vitro garlic bulbs were preserved in six culture media at 25, 18, and 5 °C. A randomized design was used and an analysis of the variance of the survival, contamination, and shoot height of the explants was performed at 30, 60, 90, 180, 270, and 365 days of culture. The results showed that the in vitro conservation of Pebeco, Tacátzcuaro Especial, and Huerteño garlic varieties was optimally obtained for one year at 5 °C in a basal Murashige and Skoog (MS) culture medium with 68.46 g L−1 sucrose and 36.43 g L−1 sorbitol. Thus, the achieved protocol can be adapted to other varieties of garlic for medium-term storage in germplasm banks. Full article
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17 pages, 12207 KiB  
Article
In Vitro Induction of Interspecific Hybrid and Polyploidy Derived from Oryza officinalis Wall
by Meimei Tan, Ruoxin Chen, Xingran Chen, Muhammad Qasim Shahid, Xiangdong Liu and Jinwen Wu
Plants 2023, 12(16), 3001; https://doi.org/10.3390/plants12163001 - 20 Aug 2023
Viewed by 944
Abstract
Oryza officinalis Wall is a potential genetic resource for rice breeding; however, its distant genome limits its crossing ability with cultivated rice. The interspecific hybridization of O. officinalis and cultivated rice, establishment of its tissue culture, and induction of polyploidy are ways to [...] Read more.
Oryza officinalis Wall is a potential genetic resource for rice breeding; however, its distant genome limits its crossing ability with cultivated rice. The interspecific hybridization of O. officinalis and cultivated rice, establishment of its tissue culture, and induction of polyploidy are ways to improve O. officinalis’s poor crossability. We developed an interspecific hybrid and studied its reproductive pollen development process in this work, and the results showed that abortive pollens (81.94%) and embryo sac abnormalities (91.04%) were the key causes of its high sterility. In order to induce callus formation in interspecific hybrid explants, two different culture media, namely Chu’s N-6 medium (N6) and 1/2 Murashig and Skoog medium (1/2 MS), were employed. Additionally, two plant growth regulators (PGRs), namely 2,4-dichlorophenoxyacetic acid (2,4-D) and 6-benzylaminopurine (BA), along with L-proline (Pro) and acid hydrolyzed casein, were utilized in the experiment. The optimal N6 medium, supplemented with 2.0 mg·L−1 2,4-D, produced the highest induction rate (80.56 ± 5.44)%. For callus differentiation and proliferation, the MS medium supplemented with 2.0 mg·L−1 BA + 0.2 mg·L−1 NAA produced the highest differentiation rate (75.00 ± 4.97)% and seedling emergence ratio (28.97 ± 4.67)%. The optimal combination for seedling rooting was the 1/2 MS medium supplemented with 2.0 mg L−1 NAA and 0.2 mg L−1 BA, which produced an average of 13.95 roots per plant. For polyploidy induction in the interspecific hybrid, the concentration of colchicine treatment at 400 mg·L−1 for three days is an ideal protocol. We devised tissue culture and interspecific hybrid polyploidy induction to overcome O. officinalis’ poor crossability and introduce its favorable features into cultivated rice. Full article
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16 pages, 3483 KiB  
Article
Response of Rowan Berry (Sorbus redliana) Shoot Culture to Slow Growth Storage Conditions
by Nóra Mendler-Drienyovszki and Katalin Magyar-Tábori
Plants 2023, 12(6), 1287; https://doi.org/10.3390/plants12061287 - 12 Mar 2023
Viewed by 1445
Abstract
Slow growth storage can preserve the genetic resources of endangered species such as those of genus Sorbus. Our aim was to study the storability of rowan berry in vitro cultures, their morpho-physiological changes, and regeneration ability after different storage conditions (4 ± [...] Read more.
Slow growth storage can preserve the genetic resources of endangered species such as those of genus Sorbus. Our aim was to study the storability of rowan berry in vitro cultures, their morpho-physiological changes, and regeneration ability after different storage conditions (4 ± 0.5 °C, dark; and 22 ± 2 °C, 16/8 h light/dark). The cold storage lasted for 52 weeks, and observations were made every four weeks. Cultures showed 100% survival under cold storage, and those taken from the storage showed 100% regeneration capacity after the passages. A dormancy period lasting about 20 weeks was observed, followed by intensive shoot growth until the 48th week, which led to the exhaustion of the cultures. The changes could be traced to the reduction of the chlorophyll content and the Fv/Fm value, as well as in the discoloration of the lower leaves and the appearance of necrotic tissues. Long, etiolated shoots (89.3 mm) were obtained at the end of cold storage. Shoot cultures stored in a growth chamber as control (22 ± 2 °C, 16/8 h light/dark) senesced and died after 16 weeks. Explants from stored shoots were subcultured for four weeks. The number and length of newly developed shoots were significantly higher on explants from cold storage compared to those from control cultures if the storage was longer than one week. Full article
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16 pages, 3385 KiB  
Article
Cretan Dittany (Origanum dictamnus L.), a Valuable Local Endemic Plant: In Vitro Regeneration Potential of Different Type of Explants for Conservation and Sustainable Exploitation
by Virginia Sarropoulou, Eleni Maloupa and Katerina Grigoriadou
Plants 2023, 12(1), 182; https://doi.org/10.3390/plants12010182 - 1 Jan 2023
Cited by 5 | Viewed by 1882
Abstract
Origanum dictamnus L. is a medicinal local endemic to the Island of Crete, Greece. Its propagation through biotechnological tissue culture techniques is essential due to its augmented multi-industrial sector demand. For direct organogenesis, among different culture media variants (MS, Gamborg B5), and cytokinins [...] Read more.
Origanum dictamnus L. is a medicinal local endemic to the Island of Crete, Greece. Its propagation through biotechnological tissue culture techniques is essential due to its augmented multi-industrial sector demand. For direct organogenesis, among different culture media variants (MS, Gamborg B5), and cytokinins [6-benzyladenine (BA), kinetin (Kin), 2-isopentenyl adenine (2-iP)], the MS + added with BA (2.2 μM) was the most effective treatment for shoots and roots formation. For indirect organogenesis, all explant types (leaves, petioles, roots) showed a 100% callusing rate after 2 months in all media variants tested; ODK1: 20 μM thidiazuron (TDZ) + 5 μM indole-3-butyric acid (IBA) or ODK2: 0.5 μM kinetin + 5 μM 2,4-dichlorophenoxy acetic acid (2,4-D). The leaves and petiole explants assured a low rate of shoot regeneration (20%) in ODK1. Afterwards, leaf-, petiole-and root-callus derived from both media were transferred to four new media plant growth regulators—free or with BA + IBA + gibberellic acid (GA3). After 10 months from callus transferring, the petiole callus gave rise to roots (20–75%) while the leaf callus exhibited 10–30% shoot or 30% root regeneration. In this study, indirect organogenesis of O. dictamnus was carried out for the first time, thus various organs can be used for plant regeneration, and the developed protocol may be applicable in the horticulture industry. Full article
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17 pages, 2275 KiB  
Article
Photosynthetic Parameters and Oxidative Stress during Acclimation of Crepe-Myrtle (Lagerstroemia speciosa (L.) Pers.) in a meta-Topolin-Based Micropropagation System and Genetic Fidelity of Regenerated Plants
by Naseem Ahmad, Nigar Fatima, Mohammad Faisal, Abdulrahman A. Alatar and Ranjith Pathirana
Plants 2022, 11(9), 1163; https://doi.org/10.3390/plants11091163 - 26 Apr 2022
Cited by 4 | Viewed by 2425
Abstract
An improved and stable micropropagation system using the cytokinin, meta-Topolin (N6 (3-hydroxybenzylamino purine—mT), with nodal explants in Lagerstroemia speciosa L. was established. Among the different doses of mT, the maximum number of shoots with the highest shoot length was [...] Read more.
An improved and stable micropropagation system using the cytokinin, meta-Topolin (N6 (3-hydroxybenzylamino purine—mT), with nodal explants in Lagerstroemia speciosa L. was established. Among the different doses of mT, the maximum number of shoots with the highest shoot length was obtained using Murashige and Skoog’s (MS) medium supplemented with 5.0 µM mT. The results were consistent throughout the proliferation period, when recorded at week 4, 8, and 12 of being cultured, with an average of 16.4 shoots per nodal explant, and having a mean length of 4.10 cm at week 8. Shoot proliferation rates could be further improved by a combination of 5.0 µM mT with 0.5 µM α-naphthalene acetic acid in MS medium; nodal explants produced an average of 24.3 shoots with a mean length of 5.74 cm after 8 weeks of being cultured. Among the five different concentrations of three auxins tested for the rooting of microshoots in MS medium, a 1.0 µM indole-3-butyric acid treatment was the best, with an average of 10.3 roots per microshoot at an average length of 3.56 cm in 93% of microshoots within 4 weeks of being transferred to this medium. A significant reduction of both chlorophyll a and b in leaves during the first week of acclimation corresponded with a high accumulation of malondialdehyde (MDH), indicating that lipid peroxidation affected chlorophyll pigments. From the second week of acclimation, photosynthetic pigment content significantly increased and MDH content decreased. The net photosynthetic rate and leaf carotenoid content showed almost linear increases throughout the acclimation period. Activity of antioxidant enzymes, namely, superoxide dismutase, catalase, and peroxidases, consistently increased throughout the acclimation period, corresponding with the accumulation of photosynthetic pigments, thus demonstrating the role of the improved antioxidant enzymatic defense system during acclimation. A comparison of parent plant DNA with that of the greenhouse acclimated plants using random amplified polymorphic DNA and inter-simple sequence repeat markers showed a monomorphic pattern indicating genetic stability and the suitability of the method for micropropagation of L. speciosa. Full article
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21 pages, 6019 KiB  
Article
Treatment of Chrysanthemum Synthetic Seeds by Air SDBD Plasma
by Nikola Škoro, Suzana Živković, Slađana Jevremović and Nevena Puač
Plants 2022, 11(7), 907; https://doi.org/10.3390/plants11070907 - 29 Mar 2022
Cited by 6 | Viewed by 2490
Abstract
Herein, we present the effect of surface dielectric barrier discharge (SDBD) air cold plasma on regrowth of chrysanthemum synthetic seeds (synseeds) and subsequent plantlet development. The plasma system used in this study operates in air at the frequency of 50 Hz. The detailed [...] Read more.
Herein, we present the effect of surface dielectric barrier discharge (SDBD) air cold plasma on regrowth of chrysanthemum synthetic seeds (synseeds) and subsequent plantlet development. The plasma system used in this study operates in air at the frequency of 50 Hz. The detailed electrical characterization of SDBD was shown, as well as air plasma emission spectra obtained by optical emission spectroscopy. The chrysanthemum synseeds (encapsulated shoot tips) were treated in air plasma for different treatment times (0, 5 or 10 min). Plasma treatment significantly improved the regrowth and whole plantlet development of chrysanthemum synseeds under aseptic (in vitro) and non-aseptic (ex vitro) conditions. We evaluated the effect of SDBD plasma on synseed germination of four chrysanthemum cultivars after direct sowing in soil. Germination of synseeds directly sowed in soil was cultivar-dependent and 1.6−3.7 fold higher after plasma treatment in comparison with untreated synseeds. The study showed a highly effective novel strategy for direct conversion of simple monolayer alginate chrysanthemum synseeds into entire plantlets by plasma pre-conversion treatment. This treatment reduced contamination and displayed a considerable ex vitro ability to convert clonally identical chrysanthemum plants. Full article
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Review

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31 pages, 940 KiB  
Review
The Cryopreservation of Medicinal and Ornamental Geophytes: Application and Challenges
by Soumaya El Merzougui, Carla Benelli, Rachida El Boullani and Mohammed Amine Serghini
Plants 2023, 12(11), 2143; https://doi.org/10.3390/plants12112143 - 29 May 2023
Cited by 1 | Viewed by 1617
Abstract
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, [...] Read more.
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. Full article
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15 pages, 2160 KiB  
Review
Development, Management and Utilization of a Kiwifruit (Actinidia spp.) In Vitro Collection: A New Zealand Perspective
by Jayanthi Nadarajan, Azadeh Esfandiari, Liya Mathew, Jasmine Divinagracia, Claudia Wiedow and Ed Morgan
Plants 2023, 12(10), 2009; https://doi.org/10.3390/plants12102009 - 17 May 2023
Cited by 1 | Viewed by 1548
Abstract
The New Zealand Institute for Plant and Food Research Limited (PFR) supports a large kiwifruit breeding program that includes more than twenty Actinidia species. Almost all the kiwifruit accessions are held as field collections across a range of locations, though not all plants [...] Read more.
The New Zealand Institute for Plant and Food Research Limited (PFR) supports a large kiwifruit breeding program that includes more than twenty Actinidia species. Almost all the kiwifruit accessions are held as field collections across a range of locations, though not all plants are at multiple locations. An in vitro collection of kiwifruit in New Zealand was established upon the arrival of Pseudomonas syringae pv. Actinadiae-biovar 3 in 2010. The value of an in vitro collection has been emphasized by restrictions on importation of new plants into New Zealand and increasing awareness of the array of biotic and abiotic threats to field collections. The PFR in vitro collection currently holds about 450 genotypes from various species, mostly A. chinensis var. chinensis and A. chinensis var. deliciosa. These collections and the in vitro facilities are used for germplasm conservation, identification of disease-free plants, reference collections and making plants available to users. Management of such a diverse collection requires appropriate protocols, excellent documentation, training, sample tracking and databasing and true-to-type testing, as well as specialized facilities and resources. This review also discusses the New Zealand biosecurity and compliance regime governing kiwifruit plant movement, and how protocols employed by the facility aid the movement of pathogen-free plants within and from New Zealand. Full article
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18 pages, 1110 KiB  
Review
In Vitro Conservation through Slow Growth Storage Technique of Fruit Species: An Overview of the Last 10 Years
by Carla Benelli, Waed Tarraf, Tolga Izgu and Anna De Carlo
Plants 2022, 11(23), 3188; https://doi.org/10.3390/plants11233188 - 22 Nov 2022
Cited by 7 | Viewed by 3627
Abstract
Plant genetic resources conservation may be a potential option for the improvement of agricultural crops through modern biotechnologies, and in vitro conservation is a tool available to safeguard plant biodiversity. Ex situ conservation of plant genetic resources using the in vitro procedures is [...] Read more.
Plant genetic resources conservation may be a potential option for the improvement of agricultural crops through modern biotechnologies, and in vitro conservation is a tool available to safeguard plant biodiversity. Ex situ conservation of plant genetic resources using the in vitro procedures is in progress in many countries. The slow growth storage (SGS) technique is a valid in vitro approach to preserve several vegetatively propagated species by controlling the growth and development of plantlets, economizing storage space and labor and reducing costs. Moreover, SGS prolongs the timing between subcultures, lowers the risk of losing germplasm through handling errors, such as contamination problems, and decreases the risk of genetic instability due to the reduction in the number of subcultures. SGS is applied by considering different factors: temperature, light or darkness conditions, medium composition, including mineral or sucrose concentrations, and the presence/absence of plant growth regulators, osmotic agents and growth inhibitors. SGS protocols for some fruit species have been well defined, others require additional research. The present review focuses on the effect of several factors that influence the SGS of in vitro shoots derived from temperate and tropical fruit species during the last ten years. Full article
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