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

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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 28076

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Dr. Anna De Carlo

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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

Dr. Waed Tarraf

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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

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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

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Published Papers (11 papers)

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Editorial

Jump to: Research, Review

5 pages, 198 KiB

Open AccessEditorial

In Vitro Biotechnology for Conservation and Sustainable Use of Plant Genetic Resources

by Waed Tarraf and Anna De Carlo

Plants 2024, 13(14), 1897; https://doi.org/10.3390/plants13141897 - 10 Jul 2024

Cited by 1 | Viewed by 1727

Abstract

This Special Issue contains novel contributions related to the role and use of in vitro culture in the conservation and sustainable use of plant genetic resources [...] Full article

(This article belongs to the Special Issue Plant Genetic Resources In Vitro Culture: Present Situation and Prospects for Propagation, Conservation and Sustainable Use)

Research

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25 pages, 8379 KiB

Open AccessArticle

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

by Carla Benelli, Waed Tarraf, Tolga İzgü, Monica Anichini, Cecilia Faraloni, Maria Cristina Salvatici, Nourhene Jouini, Maria Antonietta Germanà, Roberto Danti and Maurizio Lambardi

Plants 2024, 13(12), 1682; https://doi.org/10.3390/plants13121682 - 18 Jun 2024

Cited by 1 | Viewed by 930

Abstract

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. Full article

(This article belongs to the Special Issue Plant Genetic Resources In Vitro Culture: Present Situation and Prospects for Propagation, Conservation and Sustainable Use)

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14 pages, 2452 KiB

Open AccessArticle

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

Cited by 1 | Viewed by 1705

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, 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

(This article belongs to the Special Issue Plant Genetic Resources In Vitro Culture: Present Situation and Prospects for Propagation, Conservation and Sustainable Use)

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17 pages, 12207 KiB

Open AccessArticle

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

Cited by 1 | Viewed by 1415

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 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

(This article belongs to the Special Issue Plant Genetic Resources In Vitro Culture: Present Situation and Prospects for Propagation, Conservation and Sustainable Use)

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16 pages, 3483 KiB

Open AccessArticle

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

Cited by 2 | Viewed by 1841

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 ± 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 F_v/F_m 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

(This article belongs to the Special Issue Plant Genetic Resources In Vitro Culture: Present Situation and Prospects for Propagation, Conservation and Sustainable Use)

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16 pages, 3385 KiB

Open AccessArticle

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 7 | Viewed by 2414

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 [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

(This article belongs to the Special Issue Plant Genetic Resources In Vitro Culture: Present Situation and Prospects for Propagation, Conservation and Sustainable Use)

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17 pages, 2275 KiB

Open AccessArticle

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 8 | Viewed by 2899

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 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

(This article belongs to the Special Issue Plant Genetic Resources In Vitro Culture: Present Situation and Prospects for Propagation, Conservation and Sustainable Use)

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21 pages, 6019 KiB

Open AccessArticle

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 7 | Viewed by 2933

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 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

(This article belongs to the Special Issue Plant Genetic Resources In Vitro Culture: Present Situation and Prospects for Propagation, Conservation and Sustainable Use)

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Review

Jump to: Editorial, Research

31 pages, 940 KiB

Open AccessReview

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 14 | Viewed by 2397

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, 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

(This article belongs to the Special Issue Plant Genetic Resources In Vitro Culture: Present Situation and Prospects for Propagation, Conservation and Sustainable Use)

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15 pages, 2160 KiB

Open AccessReview

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 3 | Viewed by 2327

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 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

(This article belongs to the Special Issue Plant Genetic Resources In Vitro Culture: Present Situation and Prospects for Propagation, Conservation and Sustainable Use)

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18 pages, 1110 KiB

Open AccessEditor’s ChoiceReview

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 14 | Viewed by 4881

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 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

(This article belongs to the Special Issue Plant Genetic Resources In Vitro Culture: Present Situation and Prospects for Propagation, Conservation and Sustainable Use)

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