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Plant Tissue Culture and Plant Regeneration
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Plant Tissue Culture and Plant Regeneration

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

Deadline for manuscript submissions: 30 June 2025 | Viewed by 18179

Special Issue Editors

Dr. Douglas A. Steinmacher

E-Mail Website
Guest Editor
Vivetech Agrociências, Marechal Cândido Rondon 85960-000, Brazil
Interests: plant tissue culture
Dr. Taras Pasternak

E-Mail Website
Guest Editor
Instituto de Bioingeniería, Universidad Miguel Hernández, 03202 Elche, Spain
Interests: physiology and cell biology; plant tissue culture; molecular biology; cytochemistry; microscopy
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

Plant tissue culture and plant regeneration constitute crucial facets of plant biotechnology, spanning across several scientific and industrial domains. In recent years, we have witnessed significant advancements in the comprehension of the underlying mechanisms controlling in vitro plant regeneration, accompanied by the rapid evolution of specialized equipment and strategies for enhancing this process. This development has garnered the attention of both plant scientists and industries. Nonetheless, there exists an ongoing demand to further develop our strategies in order to select and regenerate superior genotypes, thereby refining protocols for enhanced reproducibility. In this context, a meticulous fine-tuning process is imperative. In light of these developments, we are excited to announce a Special Issue welcoming contributions covering all aspects of plant tissue culture and biotechnological processes, including the use of algorithms for improving protocols, aspects of somatic embryogenesis and organogenesis, cryopreservation, the use of temporary immersion bioreactors and new gene-editing technology enhancing shoot proliferation.

This Special Issue will accept original research articles, short-communications as well as comprehensive reviews.

We look forward to receiving your contributions.

Dr. Douglas A. Steinmacher
Dr. Taras Pasternak
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. 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

  • plant biotechnology
  • micropropagation
  • organogenesis
  • somatic embryogenesis
  • temporary immersion system
  • data-driven model
  • shoot proliferation
  • gene edition

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Published Papers (6 papers)

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Research

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22 pages, 6251 KiB  
Article
Importance of Media Composition and Explant Type in Cannabis sativa Tissue Culture
by Rekhamani Das, Tobias Kretzschmar and Jos C. Mieog
Plants 2024, 13(18), 2544; https://doi.org/10.3390/plants13182544 - 10 Sep 2024
Viewed by 1020
Abstract
Producing uniform Cannabis sativa (Cannabis) for medicinal/recreational flower production through sexual propagation has been problematic, leading to dominance of clonal propagation from “mother plants” in the cannabinoid industry, which also faces significant limitations. Cannabis tissue culture (TC) methods have been developed to overcome [...] Read more.
Producing uniform Cannabis sativa (Cannabis) for medicinal/recreational flower production through sexual propagation has been problematic, leading to dominance of clonal propagation from “mother plants” in the cannabinoid industry, which also faces significant limitations. Cannabis tissue culture (TC) methods have been developed to overcome these challenges, but the long-term health and maintenance of Cannabis explants in TC have been largely overlooked in previous studies. The current study focused on the development of an efficient and optimized micropropagation protocol covering the entire process, with a specific focus on the health and performance in the multiplication stage. Multiplication media were formulated hormone-free to avoid longer-term vitrification issues, resulting in single-main-shoot cultures rather than multiple-shoot cultures. This instigated the use of stage II explant types different from the standard shoot tips previously used for multiple shoot cultures. Multiplication media were further improved from the basal salt composition via nitrogen and calcium additives. The optimized protocol was used on eight diverse Cannabis cultivars to test its applicability across various genetic backgrounds. Results indicated that the protocol was effective for conservation purposes across all cultivars and achieved good long-term multiplication rates for some but not all. The outcomes of this study mark a significant stride towards an efficient Cannabis TC methodology ready for more comprehensive industrial applications. Full article
(This article belongs to the Special Issue Plant Tissue Culture and Plant Regeneration)
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19 pages, 6228 KiB  
Article
Induction and Suspension Culture of Panax japonicus Callus Tissue for the Production of Secondary Metabolic Active Substances
by Siqin Lv, Fan Ding, Shaopeng Zhang, Alexander M. Nosov, Andery V. Kitashov and Ling Yang
Plants 2024, 13(17), 2480; https://doi.org/10.3390/plants13172480 - 4 Sep 2024
Viewed by 753
Abstract
Using Panax japonicus as research material, callus induction and culture were carried out, and high-yielding cell lines were screened to establish a suspension culture system that promotes callus growth and the accumulation of the “total saponins” (total content of triterpenoid glycosides or ginsenosides). [...] Read more.
Using Panax japonicus as research material, callus induction and culture were carried out, and high-yielding cell lines were screened to establish a suspension culture system that promotes callus growth and the accumulation of the “total saponins” (total content of triterpenoid glycosides or ginsenosides). Using the root as an explant, the medium for callus induction and proliferation was optimized by adjusting culture conditions (initial inoculation amount, carbon source, shaking speed, hormone concentration, culture time) and a high-yielding cell line with efficient proliferation and high total saponins content was screened out. The conditions of suspension culture were refined to find out the most suitable conditions for the suspension culture of callus, and finally, the suspension culture system was established. We found that the lowest (5%) contamination rate was achieved by disinfecting the fresh roots with 75% alcohol for 60 s, followed by soaking in 10% NaClO for 15 min. The highest induction rate (88.17%) of callus was obtained using the medium MS + 16.11 μmol·L−1 NAA + 13.32 μmol·L−1 6-BA + 30.0 g·L−1 sucrose + 7.5 g·L−1 agar. The callus was loose when the callus subcultured on the proliferation medium (MS + 5.37 μmol·L−1 NAA + 13.32 μmol·L−1 6-BA + 30.0 g·L−1 sucrose + 3.8 g·L−1 gellan gum) for 21 days. The callus growth was cultured in a liquid growth medium (MS + 5.37 μmol·L−1 NAA + 13.32 μmol·L−1 6-BA + 30.0 g·L−1 sucrose) with an initial inoculation amount of 40 g·L−1, a shaking speed of 110 r/min and darkness. Cell growth was fastest with a culture period of 21 days. We replaced the growth medium with the production medium (MS + 5.37 μmol·L−1 NAA + 13.32 μmol·L−1 6-BA + 30.0 g·L−1 glucose) for maximum accumulation of total saponins. [Conclusion] A callus induction and suspension culture system for the root of P. japonicus was established. In this way, we can promote the accumulation of total saponins in callus cells and provide a basis for large-scale cell culture and industrial production of medicinal total saponins. Full article
(This article belongs to the Special Issue Plant Tissue Culture and Plant Regeneration)
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21 pages, 6232 KiB  
Article
Genome-Wide Association Analysis Identifies Candidate Loci for Callus Induction in Rice (Oryza sativa L.)
by Wintai Kamolsukyeunyong, Yeetoh Dabbhadatta, Aornpilin Jaiprasert, Burin Thunnom, Wasin Poncheewin, Samart Wanchana, Vinitchan Ruanjaichon, Theerayut Toojinda and Parichart Burns
Plants 2024, 13(15), 2112; https://doi.org/10.3390/plants13152112 - 30 Jul 2024
Viewed by 1141
Abstract
Callus induction (CI) is a critical trait for transforming desirable genes in plants. A genome-wide association study (GWAS) analysis was conducted on the rice germplasms of 110 Indica rice accessions, in which three tissue culture media, B5, MS, and N6, were used for [...] Read more.
Callus induction (CI) is a critical trait for transforming desirable genes in plants. A genome-wide association study (GWAS) analysis was conducted on the rice germplasms of 110 Indica rice accessions, in which three tissue culture media, B5, MS, and N6, were used for the CI of those rice panels’ mature seeds. Seven quantitative trait loci (QTLs) on rice chromosomes 2, 6, 7, and 11 affected the CI percentage in the three media. For the B5 medium, one QTL (qCI–B5–Chr6) was identified on rice chromosome 6; for the MS medium, two QTLs were identified on rice chromosomes 2 and 6 (qCI–MS–Chr2 and qCI–MS–Chr6, respectively); for the N6 medium, four QTLs were identified on rice chromosomes 6, 7, and 11 (qCI–N6–Chr6.1 and qCI–N6–Chr6.2, qCI–N6–Chr7, and qCI–N6–Chr11, respectively). Fifty-five genes were identified within the haplotype blocks corresponding to these QTLs, thirty-one of which showed haplotypes associated with different CI percentages in those media. qCI–B5–Chr6 was located in the same region as qCI–N6–Chr6.2, and the Caleosin-related family protein was also identified in this region. Analysis of the gene-based haplotype revealed the association of this gene with different CI percentages in both B5 and N6 media, suggesting that the gene may play a critical role in the CI mechanism. Moreover, several genes, including those that encode the beta-tubulin protein, zinc finger protein, RNP–1 domain-containing protein, and lysophosphatidic acid acyltransferase, were associated with different CI percentages in the N6 medium. The results of this study provide insights into the potential QTLs and candidate genes for callus induction in rice that contribute to our understanding of the physiological and biochemical processes involved in callus formation, which is an essential tool in the molecular breeding of rice. Full article
(This article belongs to the Special Issue Plant Tissue Culture and Plant Regeneration)
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11 pages, 13009 KiB  
Article
An Improved and Simplified Agrobacterium-Mediated Genetic Transformation Protocol for Solanum nigrum with a Shorter Growth Time
by Qianqian Li, Xiuyuan Wang, Chong Teng, Xuxia He, Xinyue Fu, Wentao Peng, Yinglun Fan and Shanhua Lyu
Plants 2024, 13(15), 2015; https://doi.org/10.3390/plants13152015 - 23 Jul 2024
Viewed by 760
Abstract
Solanum nigrum (Solanaceae family) is widely consumed as a fruit or local leafy vegetable after boiling; it also serves as a medicinal plant. Although Agrobacterium-mediated genetic transformation has been established in S. nigrum, the transformation period is long. Specifically, induction [...] Read more.
Solanum nigrum (Solanaceae family) is widely consumed as a fruit or local leafy vegetable after boiling; it also serves as a medicinal plant. Although Agrobacterium-mediated genetic transformation has been established in S. nigrum, the transformation period is long. Specifically, induction of roots takes approximately five weeks for tetraploid and hexaploid S. nigrum, and 7 weeks for diploid Solanum americanum. In this study, we developed an improved rooting-induced method that requires only about 1 week and avoids the use of tissue culture. After generating the transgenic shoots, they were directly transplanted into the soil to facilitate root formation. Remarkably, 100% of the transgenic shoots developed roots within 6 days. Our improved method is time-saving (saving more than 1 month) and simpler to operate. The improved rooting-induced step can be applied to induce roots in various plants using tissue culture, exemplified by the carnation (Dianthus caryophyllus L.). Furthermore, we applied the improved method to generate S. americanum plants expressing AcMYB110 from kiwifruit (Actinidia chinensis spp.). This method will contribute to speeding up gene functional analysis and trait improvement in S. nigrum and might have potential in fast plant molecular breeding processes in crops and rapid rooting induction in tissue culture. Full article
(This article belongs to the Special Issue Plant Tissue Culture and Plant Regeneration)
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9 pages, 1268 KiB  
Article
High-Efficiency In Vitro Root Induction in Pear Microshoots (Pyrus spp.)
by Jae-Young Song, Jinjoo Bae, Young-Yi Lee, Ji-Won Han, Ye-ji Lee, Sung Hee Nam, Ho-sun Lee, Seok Cheol Kim, Se Hee Kim and Byeong Hyeon Yun
Plants 2024, 13(14), 1904; https://doi.org/10.3390/plants13141904 - 10 Jul 2024
Viewed by 660
Abstract
Extensive research has been conducted on the in vitro mass propagation of pear (Pyrus spp.) trees through vegetative propagation, demonstrating high efficiency in shoot multiplication across various pear species. However, the low in vitro rooting rates remain a significant barrier to the [...] Read more.
Extensive research has been conducted on the in vitro mass propagation of pear (Pyrus spp.) trees through vegetative propagation, demonstrating high efficiency in shoot multiplication across various pear species. However, the low in vitro rooting rates remain a significant barrier to the practical application and commercialization of mass propagation. This study aims to determine the favorable conditions for inducing root formation in the in vitro microshoots of Pyrus genotypes. The base of the microshoots was exposed to a high concentration (2 mg L−1) of auxins (a combination of IBA and NAA) for initial root induction at the moment when callus formation begins. The microshoots were then transferred to an R1 medium (1/2 MS with 30 g L−1 sucrose without PGRs) to promote root development. This method successfully induced rooting in three European pear varieties, one Asian pear variety, and a European–Asian hybrid, resulting in rooting rates of 66.7%, 87.2%, and 100% for the European pear (P. communis), 60% for the Asian pear (P. pyrifolia), and 83.3% for the hybrid pear (P. pyrifolia × P. communis) with an average of 25 days. In contrast, the control group (MS medium) exhibited rooting rates of 0–13.3% after 60 days of culture. These findings will enhance in vitro root induction for various pear varieties and support the mass propagation and acclimatization of pear. The in vitro root induction method developed in this study has the potential for global commercial application in pear cultivation. Full article
(This article belongs to the Special Issue Plant Tissue Culture and Plant Regeneration)
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Review

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24 pages, 1023 KiB  
Review
Plant Growth Regulation in Cell and Tissue Culture In Vitro
by Taras P. Pasternak and Douglas Steinmacher
Plants 2024, 13(2), 327; https://doi.org/10.3390/plants13020327 - 22 Jan 2024
Cited by 17 | Viewed by 12085
Abstract
Precise knowledge of all aspects controlling plant tissue culture and in vitro plant regeneration is crucial for plant biotechnologists and their correlated industry, as there is increasing demand for this scientific knowledge, resulting in more productive and resilient plants in the field. However, [...] Read more.
Precise knowledge of all aspects controlling plant tissue culture and in vitro plant regeneration is crucial for plant biotechnologists and their correlated industry, as there is increasing demand for this scientific knowledge, resulting in more productive and resilient plants in the field. However, the development and application of cell and tissue culture techniques are usually based on empirical studies, although some data-driven models are available. Overall, the success of plant tissue culture is dependent on several factors such as available nutrients, endogenous auxin synthesis, organic compounds, and environment conditions. In this review, the most important aspects are described one by one, with some practical recommendations based on basic research in plant physiology and sharing our practical experience from over 20 years of research in this field. The main aim is to help new plant biotechnologists and increase the impact of the plant tissue culture industry worldwide. Full article
(This article belongs to the Special Issue Plant Tissue Culture and Plant Regeneration)
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