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Horticulturae
Special Issues
Micropropagation and In Vitro Techniques: Theory, Methods and Applications
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Micropropagation and In Vitro Techniques: Theory, Methods and Applications

A special issue of Horticulturae (ISSN 2311-7524). This special issue belongs to the section "Propagation and Seeds".

Deadline for manuscript submissions: closed (28 February 2023) | Viewed by 13288

Special Issue Editors

Dr. Orsolya Borsai

E-Mail Website
Guest Editor
Faculty of Horticulture, University of Agricultural Sciences and Veterinary Medicine Cluj-Napoca, Mănăștur St. 3-5, 400372 Cluj-Napoca, Romania
Interests: micropropagation; abiotic stress; salinity; drought; fruit trees; fruit production; orchard management
Dr. Clapa Doina

E-Mail Website
Guest Editor
Institute of Life Sciences, University of Agricultural Sciences and Veterinary Medicine Cluj-Napoca, Mănăștur St. 3-5, 400372 Cluj-Napoca, Romania
Interests: micropropagation; tissue culture; nanoparticles; plant growth regulators
Prof. Dr. Mirela Irina Cordea

E-Mail Website
Guest Editor
Faculty of Horticulture, University of Agricultural Sciences and Veterinary Medicine Cluj-Napoca, Mănăștur St. 3-5, 400372 Cluj-Napoca, Romania
Interests: plant genetics; plant breeding; abiotic stress; genetically modified organisms; phenotyping; genetic engineering
Dr. Monica Harta

E-Mail Website
Guest Editor
Faculty of Horticulture, University of Agricultural Sciences and Veterinary Medicine Cluj-Napoca, Mănăștur St. 3-5, 400372 Cluj-Napoca, Romania
Interests: molecular markers; plant biotechnology; in vitro plant cultures; marker assisted selection; plant genetics; genetic diversity
Dr. Songling Bai

E-Mail Website
Guest Editor
Zhejiang Provincial Key Laboratory of Integrative Biology of Horticultural Plants, Hangzhou 310058, China
Interests: breeding of horticultural plant; physiology and molecular biology of horticultural plants

Special Issue Information

Dear Colleagues,

Micropropagation was first developed in the 1960s and has since found application in various scientific and economic fields with high impact. Due to the mesmerizing ability of most plants to regenerate from a single cell, micropropagation and in vitro techniques became one of the most efficient and ‘secure’ plant propagation methods. Growing plant organs or tissues in aseptic conditions requires special attention and full control of the whole process of plant regeneration to obtain high-quality and virus-free stocks of crop plants. Micropropagation can sucessfully be applied for mass propagation, new cultivar development, preservation of wild, old, or rare plant species, and also for the propagation of certain crops which are difficult or slow to be propagated using traditional methods. The resulting plants must undergo extensive hardening treatments, preferably in greenhouse conditions to prevent any rate of plant death after their transfer to ex vitro conditions. Currently, micropropagation and in vitro techniques are practiced on a large scale as an accelerated version of clonal propagation to satisfy the increasing demand for modern crops or to regenerate genetically modified plants. However, before application, some disadvantages must be considered, such as production and labor costs and the potential risk of plant contamination or plant adaptation failure to normal growing environment.

Dr. Orsolya Borsai
Dr. Clapa Doina
Prof. Dr. Mirela Irina Cordea
Dr. Monica Harta
Dr. Songling Bai
Guest Editors

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. Horticulturae is an international peer-reviewed open access monthly 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 2200 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

  • in vitro
  • propagation
  • plant growth regulators
  • contamination
  • multiplication rate
  • hardening
  • root formation
  • callus
  • ex vitro

Published Papers (5 papers)

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Research

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9 pages, 1597 KiB  
Communication
Rapid and Efficient In Vitro Propagation Protocol of Endangered Wild Prickly Pear Growing in Eastern Morocco
by Ahmed Marhri, Aziz Tikent, Laurine Garros, Othmane Merah, Ahmed Elamrani, Christophe Hano, Malika Abid and Mohamed Addi
Horticulturae 2023, 9(4), 491; https://doi.org/10.3390/horticulturae9040491 - 14 Apr 2023
Cited by 1 | Viewed by 1822
Abstract
The Opuntia genus is widely recognized as a significant member of the Cactaceae family. The eastern Moroccan prickly pear’s wild ecotype is renowned for its production of fruits of superior quality, which are in high demand. Nonetheless, the white cochineal (Dactylopius opuntiae [...] Read more.
The Opuntia genus is widely recognized as a significant member of the Cactaceae family. The eastern Moroccan prickly pear’s wild ecotype is renowned for its production of fruits of superior quality, which are in high demand. Nonetheless, the white cochineal (Dactylopius opuntiae) has emerged as a significant hazard to the persistence of the indigenous wild prickly pear population in the majority of the country’s territories. Conventional plant propagation techniques may facilitate the transmission of pathogens to successive generations and thus fail to satisfy market requirements. Therefore, the primary goal of this study was to develop a rapid and efficient protocol for large-scale production of the eastern Moroccan wild ecotype using axillary buds as the starting material. Sterilization of the starting material is a crucial but challenging step in this species, as the meristem is located just beneath the spine. The protocol developed for this study produced moderately satisfactory results, with 20 to 30% contamination in each experiment. The obtained shoots were incubated on Murashige and Skoog medium supplemented with varying concentrations of BAP (0, 1.5, 3, 5, and 7.5 mg/L). The treatment with BAP at 5 mg/L exhibited a statistically significant increase in the average number of regenerated shoots per explant (19.42). The effect of kinetin on the rooting of prickly pear was evaluated by transferring the shoots to a MS medium supplemented with varying concentrations of kinetin (0, 0.5, 1, 1.5, 2, and 2.5 mg/L). The use of kinetin increased the number and length of roots while also shortening the root development period from 21 days to 10 days. The best results were obtained at a concentration of 1.5 mg/L of kinetin. Furthermore, satisfactory acclimatization of plants was achieved by using plastic containers with a gradually increasing opening of the lids. The outcomes of this experimentation have significant potential to facilitate the preservation of this botanical variety, reduce the risk of white cochineal infestation, and address the need for superior quality fruit supply in the market. Full article
(This article belongs to the Special Issue Micropropagation and In Vitro Techniques: Theory, Methods and Applications)
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20 pages, 3116 KiB  
Article
In Vitro Cultivation of Purple-Fleshed Potato Varieties: Insights into Their Growth and Development
by Alexandra Mihaela Nagy, Paula Oros, Corina Cătană, Maria Mihaela Antofie and Camelia Sava Sand
Horticulturae 2023, 9(4), 425; https://doi.org/10.3390/horticulturae9040425 - 25 Mar 2023
Viewed by 1623
Abstract
Purple-fleshed potatoes (PFP) are varieties of Solanum tuberosum L., which recently have been recorded to be more and more cultivated and consumed in all European countries, including Romania, as they are promoted for their content in bioactive compounds and benefits to human health. [...] Read more.
Purple-fleshed potatoes (PFP) are varieties of Solanum tuberosum L., which recently have been recorded to be more and more cultivated and consumed in all European countries, including Romania, as they are promoted for their content in bioactive compounds and benefits to human health. This paper presents a micropropagation protocol study for PFP varieties already traded into the Romanian market, namely Blue Danube (BD), Salad Blue (SB), Violet Negretin (VN), and Violet Queen (VQ). These varieties were tested for in vitro micropropagation also considering asepsis, initiation, callus formation, and microtuberization. To establish the optimum asepsis treatment, a preliminary experiment was performed and, the best results were obtained by using 70% EtOH (1 min) followed by 20% (v/v) Domestos® (20 min). The MS formula (Murashige and Skoog 1962) was tested as the basic culture medium without growth regulators for all tested stages except for callus initiation and its further multiplication stages. The effect of glycine on direct organogenesis and shoot multiplication was evaluated for propagated micro-cuttings. We emphasize that the addition of glycine at a concentration of 15 mg/L to the culture medium induced a better plantlet vigor for all four varieties. Regarding the indirect organogenesis, culture medium supplemented with NAA (5.00 mg/L), GA3 (1.00 mg/L), TDZ (1.00 mg/L) and glycine (15.00 mg/L) induced the best results for shoot cluster regeneration as well as turning of white callus from control to purple callus. Further, the microtuberization was successfully produced when sucrose was supplemented at 8% (w/v) into the culture medium. Among all four tested PFP varieties, SB has proven to give the best results regarding the adaptability for in vitro cultivation. Full article
(This article belongs to the Special Issue Micropropagation and In Vitro Techniques: Theory, Methods and Applications)
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14 pages, 2485 KiB  
Article
High Frequency Direct Organogenesis in Five Romanian Tomato (Lycopersicon esculentum Mill.) Cultivars
by Adela Halmagyi, Ana Coste, Constantin Deliu and Ioan Băcilă
Horticulturae 2023, 9(3), 411; https://doi.org/10.3390/horticulturae9030411 - 22 Mar 2023
Cited by 2 | Viewed by 1412
Abstract
Tomato (Lycopersicon esculentum Mill.) as the most economically important vegetable crop worldwide has been investigated intensively for the development of new and improved varieties. Most of these technologies require efficient protocols for in vitro regeneration and propagation of plant material. In the [...] Read more.
Tomato (Lycopersicon esculentum Mill.) as the most economically important vegetable crop worldwide has been investigated intensively for the development of new and improved varieties. Most of these technologies require efficient protocols for in vitro regeneration and propagation of plant material. In the present study, an efficient and reproducible in vitro regeneration system for five Romanian tomato genotypes (cvs. ‘Capriciu’, ‘Darsirius’, ‘Kristin’, ‘Pontica’ and ‘Siriana’) has been established. The tomato genotypes were selected based on their horticultural and economically valuable traits. To study the in vitro morphogenic response, various explants, such as cotyledons, cotyledonary nodes, hypocotyls, leaf explants, internodes, stem nodes and apical buds have been selected. The highest efficiency in terms of direct shoot organogenesis was obtained in cv. ‘Capriciu’ (98% for apical buds and 94% for stem nodes) on culture media with zeatin and indole-3-butyric acid. One advantage of this regeneration procedure is beside its feasibility in handling, the high percentage of regenerated shoots and their rooting. The present protocol contributes to the existing information regarding the response of tomato cultivars to in vitro culture conditions. Full article
(This article belongs to the Special Issue Micropropagation and In Vitro Techniques: Theory, Methods and Applications)
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15 pages, 1439 KiB  
Article
Establishment of Direct Organogenesis Protocol for Arachis hypogaea cv. Virginia in Liquid Medium by Temporary Immersion System (TIS)
by Elif Aylin Ozudogru, Elif Karlik, Doaa Elazab and Maurizio Lambardi
Horticulturae 2022, 8(12), 1129; https://doi.org/10.3390/horticulturae8121129 - 01 Dec 2022
Cited by 1 | Viewed by 1522
Abstract
Peanuts (Arachis hypogaea L.) are a rich source of herbal oil, proteins, minerals, vitamins, fibers, essential amino acids, as well as bioactive compounds, and are thus widely used for human nutrition and animal feed, and for prevention from certain diseases. However, the [...] Read more.
Peanuts (Arachis hypogaea L.) are a rich source of herbal oil, proteins, minerals, vitamins, fibers, essential amino acids, as well as bioactive compounds, and are thus widely used for human nutrition and animal feed, and for prevention from certain diseases. However, the in vitro regeneration response of the species is generally low, and it also displays a significant variability among its varieties. Thus, the development of advanced protocols and approaches for the in vitro propagation of peanut is still of immense importance. A recently developed in vitro propagation technique, TIS; Temporary Immersion Bioreactor System, provides a new approach for the mass propagation of plants. Accordingly, the present study provides an efficient de novo regeneration protocol for Arachis hypogaea L. cv. Virginia by using a TIS. Different concentrations of cytokinins, i.e., benzyladenine (BA) or thidiazuron (TDZ), were tested with several combinations of dry and medium immersion periods of TIS, corresponding to a total of 8, 12, 16, 24, 32, 36, 48, 64, 72, and 96 min daily immersions for the induction of direct organogenesis. The study exhibited that an MS medium added to 110 µM BA or 10 µM TDZ are the most appropriate medium formulations in TIS, when applied for 16 min every 16 h. The application of optimized procedures to cv. NC7 and two valuable Turkish autochthonous varieties, 7 × 77 and Com74, is also reported. To the best of our knowledge, the present study draws attention also for being the first study in which a TIS was used for peanuts. Full article
(This article belongs to the Special Issue Micropropagation and In Vitro Techniques: Theory, Methods and Applications)
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Review

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16 pages, 1651 KiB  
Review
Managing Lethal Browning and Microbial Contamination in Musa spp. Tissue Culture: Synthesis and Perspectives
by Nandang Permadi, Mohamad Nurzaman, Arshad Naji Alhasnawi, Febri Doni and Euis Julaeha
Horticulturae 2023, 9(4), 453; https://doi.org/10.3390/horticulturae9040453 - 01 Apr 2023
Cited by 7 | Viewed by 5856
Abstract
Browning and contamination are regarded as the main constraints in the plant tissue culture of Musa spp. that can hinder the success of plant propagation in vitro. Browning is caused by enzymatic reactions due to explant injury, while microbial contamination is caused by [...] Read more.
Browning and contamination are regarded as the main constraints in the plant tissue culture of Musa spp. that can hinder the success of plant propagation in vitro. Browning is caused by enzymatic reactions due to explant injury, while microbial contamination is caused by phyllospheric, rhizospheric, and endophytic microorganisms that reside on, in, and inside the plants. When not properly addressed, they can cause decreased regenerative ability, decreased callus growth, inhibited adventitious shoot growth, and even tissue death. To overcome the browning problem, various attempts have been made in vitro, e.g., immersing the explants in an anti-browning solution, incorporating anti-browning compounds into the medium, and manipulating cultural practices. Correspondingly, to control the problem of contamination, efforts have been made, for example, using various methods, such as thermotherapy, chemotherapy, and cryotherapy, and chemical agents, such as disinfectants, antiseptics, and nanoparticles. This review aims to investigate and provide a comprehensive understanding of the causes of browning and contamination as well as the many approaches used to control browning and contamination problems in Musa spp. tissue cultures. Full article
(This article belongs to the Special Issue Micropropagation and In Vitro Techniques: Theory, Methods and Applications)
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