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Applications of Plant Tissue Culture: New Insight into Plant Improvement Research Using Plant Tissue Culture and Plant Modification Techniques

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

Deadline for manuscript submissions: closed (10 September 2023) | Viewed by 19018

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Special Issue Editors

Dr. David W. M. Leung

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Guest Editor

School of Biological Sciences, University of Canterbury, Private Bag 4800, Christchurch 8140, New Zealand
Interests: plant biotechnology; in vitro plant breeding; micropropagation; phytoremediation; genetic modifications of plants; bio-printing
Special Issues, Collections and Topics in MDPI journals

Dr. Rambod Abiri

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Guest Editor

Department of Forestry Science and Biodiversity, Faculty of Forestry and Environment, Universiti Putra Malaysia, UPM Serdang 43400, Selangor, Malaysia
Interests: Plant genetics and biotechnology; plant-environment interactions; secondary metabolites; medicinal and aromatic plants; biotic and abiotic resistance
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

The global population increase puts a huge pressure on the availability of food and water resources. Traditionally, plant breeders have played a key role in the genetic improvement of the quality and quantity of crops to feed the world’s growing population. Traditional plant breeding is, however, an ineffective method for sustained genetic improvement and has many limitations. The classical plant breeding procedure is time-consuming, relying on limited natural genetic resources, and labour-intensive. Plant biotechnology has emerged as a reliable set of tools that empowers plant breeders to use genes of beneficial traits from diverse sources and insert them into plant genomes for genetic modifications. In addition, plant tissue culture (PTC) is a core biotechnology for maintaining or growing plant cells, tissues or organs under aseptic conditions. There have been remarkable advances in PTC research on multiple fronts including the regulation of plant developmental processes in vitro, discoveries of biologically active compounds, gene transfer and production of secondary metabolites of medicinal significance. Most recently, the integration of PTC with novel techniques such as genome editing has opened a new window to improve the desirable traits in plants. Therefore, this Special Issue titled “New Insight into Plant Improvement Research using Plant Tissue Culture and Plant Modification Techniques” aims to attract contributions for new advances in PTC focusing on plant growth regulators, gene transformation, genome editing, molecular farming and other applications of plant culture research in vitro. We welcome the submission of original research papers, reviews and novel in vitro protocols including, but not limited to, the following topics:

Micropropagation;
Cell totipotency;
Phytochemical production by in vitro cultures;
Somaclonal variation;
Somatic embryogenesis;
De novo organogenesis;
Synthetic seeds;
Virus-free plants;
Cell suspension culture;
Hairy root culture;
Cellular reprogramming;
Callus formation;
Plant transformation;
Molecular plant breeding;
Genetics;
Genome editing;
DNA (transgene)-free editing;
Base/prime editing;
HDR-based editing;
The CRISPR/dCas9 system;
Off-target effects;
Transcription factors;
Rewiring of plant defence genes in vitro;
Bioreactors;
Secondary metabolites in plant tissue cultures;
Elicitors;
Plant defence mechanisms;
Biotic and abiotic stress in vitro culture;
Plant growth regulators;
Quality improvement;
Phytoremediation.

Dr. David W. M. Leung
Dr. Rambod Abiri
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

micropropagation
molecular plant breeding
genome editing
biotic and abiotic stress in vitro culture
secondary metabolites in plant tissue cultures

Related Special Issue

Applications of Plant Tissue Culture: New Insights into Plant Improvement Research Using Plant Tissue Culture and Plant Modification Techniques―2nd Edition in Plants

Published Papers (8 papers)

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Research

9 pages, 3553 KiB

Open AccessCommunication

In Vitro-Based Production of Virus-Tested Babaco (Vasconcellea x heilbornii, syn. Carica pentagona) in Ecuador: An Integrated Approach to an Endangered Crop

by Valeria Muñoz, Diana Curillo, Sebastián Gómez, Lissette Moreno-Peña, Denisse Peña, Eduardo Chica, Viviana Yánez, Eduardo Sánchez-Timm and Diego F. Quito-Avila

Plants 2023, 12(13), 2560; https://doi.org/10.3390/plants12132560 - 6 Jul 2023

Viewed by 1534

Abstract

Babaco (Vasconcellea x heilbornii), a fruit-bearing vegetatively propagated crop native to Ecuador, is appreciated for its distinctive flavor and nutritional properties. The aim of this research was to determine a functional protocol for tissue culture propagation of virus-free babaco plants including in vitro establishment, multiplication, rooting, and acclimation. First, symptomless babaco plants from a single commercial nursery were analyzed for virus detection and cared for using different disinfection treatments in the greenhouse to reduce contamination during the in vitro establishing step, and three cytokinins, 6-(γ,γ-Dimethylallylamino) purine (2IP), 6-Benzylaminopurine (BAP), and Thidiazuron (TDZ), were used to determine the best hormone for multiplication. The best treatment for plant disinfection was the weekly application of copper sulfate at the greenhouse and a laboratory disinfection using ethanol (EtOH) (70%), Clorox (2%), and a solution of povidone iodine (2.5%), with an 80% survival during in vitro plant establishment. TDZ showed a better multiplication rate when compared with other hormones, and 70% of the rooted plants were successfully acclimated at the greenhouse. Generated plants were virus-free when tested against babaco mosaic virus (BabMV) and papaya ringspot virus (PRSV), two of the most important viruses that can affect babaco. An efficient protocol to produce virus-free babaco plants was elaborated with an integrated use of viral diagnostic tools to ensure the production of healthy start material to farmers. Full article

(This article belongs to the Special Issue Applications of Plant Tissue Culture: New Insight into Plant Improvement Research Using Plant Tissue Culture and Plant Modification Techniques)

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17 pages, 3140 KiB

Open AccessArticle

Investigation of the Effect of the Auxin Antagonist PEO-IAA on Cannabinoid Gene Expression and Content in Cannabis sativa L. Plants under In Vitro Conditions

by Josef Baltazar Šenkyřík, Tereza Křivánková, Dominika Kaczorová and Nikola Štefelová

Plants 2023, 12(8), 1664; https://doi.org/10.3390/plants12081664 - 15 Apr 2023

Cited by 3 | Viewed by 2936

Abstract

The in vitro shoot propagation of Cannabis sativa L. is an emerging research area for large-scale plant material production. However, how in vitro conditions influence the genetic stability of maintained material, as well as whether changes in the concentration and composition of secondary metabolites can be expected are aspects that need to be better understood. These features are essential for the standardised production of medicinal cannabis. This work aimed to find out whether the presence of the auxin antagonist α-(2-oxo-2-phenylethyl)-1H-indole-3-acetic acid (PEO-IAA) in the culture media influenced the relative gene expression (RGE) of the genes of interest (OAC, CBCA, CBDA, THCA) and the concentrations of studied cannabinoids (CBCA, CBDA, CBC, ∆⁹-THCA, and ∆⁹-THC). Two C. sativa cultivars, ‘USO-31’ and ‘Tatanka Pure CBD’, were cultivated by in vitro conditions with PEO-IAA presence and then analysed. The RT-qPCR results indicated that even though some changes in the RGE profiles could be observed, no differences were statistically significant compared with the control variant. The results of the phytochemical analyses demonstrate that although there were some differences from the control variant, only the cultivar ‘Tatanka Pure CBD’ showed a statistically significant increase (at a statistical significance level α = 0.05) in the concentration of the cannabinoid CBDA. In conclusion, it would appear that using PEO-IAA in the culture medium is a suitable approach to improve in vitro cannabis multiplication. Full article

(This article belongs to the Special Issue Applications of Plant Tissue Culture: New Insight into Plant Improvement Research Using Plant Tissue Culture and Plant Modification Techniques)

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13 pages, 6114 KiB

Open AccessArticle

Promising Application of Automated Liquid Culture System and Arbuscular Mycorrhizal Fungi for Large-Scale Micropropagation of Red Dragon Fruit

by Yaser Hassan Dewir, Muhammad M. Habib, Ahmed Ali Alaizari, Jahangir A. Malik, Ali Mohsen Al-Ali, AbdulAziz A. Al-Qarawi and Mona S. Alwahibi

Plants 2023, 12(5), 1037; https://doi.org/10.3390/plants12051037 - 24 Feb 2023

Cited by 2 | Viewed by 1953

Abstract

Red dragon fruit (Hylocereus polyrhizus) is an economic and promising fruit crop in arid and semi-arid regions with water shortage. An automated liquid culture system using bioreactors is a potential tool for micropropagation and large-scale production. In this study, axillary cladode multiplication of H. polyrhizus was assessed using cladode tips and cladode segments in gelled culture versus continuous immersion air-lift bioreactors (with or without a net). Axillary multiplication using cladode segments (6.4 cladodes per explant) was more effective than cladode tip explants (4.5 cladodes per explant) in gelled culture. Compared with gelled culture, continuous immersion bioreactors provided high axillary cladode multiplication (45.9 cladodes per explant) with a higher biomass and length of axillary cladodes. Inoculation of H. polyrhizus micropropagated plantlets with arbuscular mycorrhizal fungi (Gigaspora margarita and Gigaspora albida) significantly increased the vegetative growth during acclimatization. These findings will improve the large-scale propagation of dragon fruit. Full article

(This article belongs to the Special Issue Applications of Plant Tissue Culture: New Insight into Plant Improvement Research Using Plant Tissue Culture and Plant Modification Techniques)

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17 pages, 11447 KiB

Open AccessArticle

In Vitro Propagation of Three Date Palm (Phoenix dactylifera L.) Varieties Using Immature Female Inflorescences

by Ahmed M. Abdelghaffar, Said. S. Soliman, Tarek A. Ismail, Ahmed M. Alzohairy, Arafat Abdel Hamed Abdel Latef, Khadiga Alharbi, Jameel M. Al-Khayri, Nada Ibrahim M. Aljuwayzi, Diaa Abd El-Moneim and Abdallah. A. Hassanin

Plants 2023, 12(3), 644; https://doi.org/10.3390/plants12030644 - 1 Feb 2023

Cited by 3 | Viewed by 2897

Abstract

Immature female inflorescences are promising materials for use as explants for the tissue culture of date palm. Four types of MS media were used in this study during the four micropropagation stages—starting media (SM), maturation media (MM), multiplication media (PM) and rooting media (RM)—to micropropagate three elite date palm varieties, Amri, Magdoul and Barhy using the immature female inflorescences as explant. The highest percentage of callus induction in all the varieties studied was obtained on the SM1 (9 µM 2,4-D + 5.7 µM IAA + 10 µM NAA). Culturing on the MM1 (4.5 µM 2,4-D + 9.8 µM 2-iP + 1.5 AC) allowed us to obtain the best value in terms of callus weight. After culturing on the PM1 (4.4 µM BA + 9.8 µM 2-iP) produced the highest numbers of somatic embryos and shoots. The explants on RM2 (0.5 µM NAA + 1.25 µM IBA + 3 g AC) showed the highest root numbers and root lengths, while the highest shoot length was achieved on RM3 (0.5 µM NAA + 0.5 µM IBA + 3 g AC). The Amri variety presented the best response among the three varieties in all parameters, followed by the Magdoul and Barhy varieties. In all the stages of micropropagation, the analysis of variance revealed highly significant variations among varieties and culture media, and a significant difference in the number of roots during the rooting stage. The results also showed non-significant differences in the interaction between varieties and culture media, except for shoot length in the rooting stage. The results also reveal the broad sense heritability ranging from low to high for the measured parameters. It can be concluded that the immature female inflorescences can be used as a productive explant source for successful date palm micropropagation using the SM1, MM1, PM1 and RM2 culture media. It can also be concluded that the success of date palm micropropagation not only depends on the concentrations of growth regulators, but also on their types. Full article

(This article belongs to the Special Issue Applications of Plant Tissue Culture: New Insight into Plant Improvement Research Using Plant Tissue Culture and Plant Modification Techniques)

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15 pages, 2905 KiB

Open AccessArticle

Stress-Induced Intensification of Deoxyshikonin Production in Rindera graeca Hairy Root Cultures with Ester-Based Scaffolds

by Kamil Wierzchowski, Mateusz Kawka, Michał Wrzecionek, Julia Urbanek, Agnieszka Pietrosiuk, Katarzyna Sykłowska-Baranek, Agnieszka Gadomska-Gajadhur and Maciej Pilarek

Plants 2022, 11(24), 3462; https://doi.org/10.3390/plants11243462 - 10 Dec 2022

Cited by 3 | Viewed by 1243

Abstract

In vitro plant cell and tissue culture systems allow for controlling a wide range of culture environmental factors selectively influencing biomass growth and the yield of secondary metabolites. Among the most efficient methods, complex supplementation of the culture medium with elicitors, precursors, and other functional substances may significantly enhance valuable metabolite productivity through a stress induction mechanism. In the search for novel techniques in plant experimental biotechnology, the goal of the study was to evaluate stress-inducing properties of novel biodegradable ester-based scaffolds made of poly(glycerol sebacate) (PGS) and poly(lactic acid) (PLA) influencing on the growth and deoxyshikonin productivity of Rindera graeca hairy roots immobilized on the experimental constructs. Rindera graeca hairy roots were maintained under the dark condition for 28 days in three independent systems, i.e., (i) non-immobilized biomass (a reference system), (ii) biomass immobilized on PGS scaffolds, and (iii) biomass immobilized on PLA scaffolds. The stress-inducing properties of the applied polymerized esters selectively impacted R. graeca hairy roots. The PGS scaffolds caused the production of deoxyshikonin, which does not occur in other culture systems, and PLA promoted biomass proliferation by doubling its increase compared to the reference system. Full article

(This article belongs to the Special Issue Applications of Plant Tissue Culture: New Insight into Plant Improvement Research Using Plant Tissue Culture and Plant Modification Techniques)

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15 pages, 2027 KiB

Open AccessArticle

A New Approach for Controlling Agrobacterium tumefaciens Post Transformation Using Lytic Bacteriophage

by Fiqih Ramadhan, Yuzer Alfiko, Sigit Purwantomo, Andhika Faisal Mubarok, Widyah Budinarta, Antonius Suwanto and Sri Budiarti

Plants 2022, 11(22), 3124; https://doi.org/10.3390/plants11223124 - 16 Nov 2022

Cited by 3 | Viewed by 2389

Abstract

Overgrowth of Agrobacterium tumefaciens has frequently been found in Agrobacterium-mediated plant transformation. This overgrowth can reduce transformation efficiency and even lead to explant death. Therefore, this research investigates an alternative way to mitigate or eliminate Agrobacterium after transformation using a bacteriophage. To develop this alternative method, we conducted effectiveness studies of two lytic bacteriophages (ΦK2 and ΦK4) and performed an application test to control Agrobacterium growth after transformation. According to plaque morphological characterization and molecular analysis, the two bacteriophages used in this experiment were distinct. Moreover, some stability physicochemical and growth kinetics, such as adsorption time and susceptibility test, also showed that both bacteriophages differed. On the other hand, the optimum temperature and pH of both phages were the same at 28–30 °C and pH 7. Further investigation showed that both ΦK2 and ΦK4 were able to reduce the overgrowth of A. tumefaciens post transformation. Moreover, applying the cocktail (mixture of ΦK2 and ΦK4) with antibiotic application eradicated A. tumefaciens (0% overgrowth percentage). This result indicates that the application of bacteriophage could be used as an alternative way to eradicate the overgrowth of A. tumefaciens subsequent to transformation. Full article

(This article belongs to the Special Issue Applications of Plant Tissue Culture: New Insight into Plant Improvement Research Using Plant Tissue Culture and Plant Modification Techniques)

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16 pages, 3829 KiB

Open AccessArticle

Exogenous Glutathione Promotes the Proliferation of Pinus koraiensis Embryonic Cells and the Synthesis of Glutathione and Ascorbic Acid

by Fang Gao, Yujie Shi, Ruirui Wang, Iraida Nikolaevna Tretyakova, Alexander Mikhaylovich Nosov, Hailong Shen and Ling Yang

Plants 2022, 11(19), 2586; https://doi.org/10.3390/plants11192586 - 30 Sep 2022

Cited by 5 | Viewed by 1409

Abstract

Somatic embryogenesis (SE), which leads to the formation of embryonic callus (EC) tissue, is the most promising method for large-scale production and selective breeding of woody plants. However, in many species, SE suffers from low proliferation rates, hindering the production of improved plant materials. One way of improving proliferation rates is achieved by improving the redox status of the culture medium. In this study, we investigated the effects of exogenous glutathione (GSH) and L-buthionine sulfoximine (BSO, the inhibitor of glutathione synthase) on the EC proliferation rate in Korean pine (Pinus koraiensis), using cell lines with both high (F: 001#-001) and low (S: 001#-010) proliferation potential. We found that exogenous GSH promoted cell proliferation in both cell lines, while exogenous BSO inhibited proliferation in both cell lines. At 35 d with exogenous GSH treatment, the fresh weight of F and S cell lines increased by 35.48% and 48.39%, respectively, compared with the control. The exogenous application of GSH increased the intracellular levels of GSH, total GSH (T-GSH), oxidized glutathione (GSSG), ascorbic acid (ASA), total ASA (T-ASA), and the ratios of GSH:T-GSH and ASA:T-ASA in both F and S cell lines. Furthermore, exogenous GSH increased the activity of both glutathione reductase (GR) and dehydroascorbate reductase (DHAR) while decreasing the activity of ascorbate peroxidase (APX) in both cell lines. It appears that the application of exogenous GSH promotes a reducing cultural environment, which is conducive to EC proliferation in Korean pine. By helping to reveal the mechanism whereby GSH regulates redox homeostasis in Korean pine EC cells, we have laid the foundation for a large-scale breeding of Korean pine somatic embryogenesis technology system. Full article

(This article belongs to the Special Issue Applications of Plant Tissue Culture: New Insight into Plant Improvement Research Using Plant Tissue Culture and Plant Modification Techniques)

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17 pages, 6547 KiB

Open AccessArticle

Morphological and Physiological Indicators for Screening Cell Lines with High Potential for Somatic Embryo Maturation at an Early Stage of Somatic Embryogenesis in Pinus Koraiensis

by Chunxue Peng, Fang Gao, Hao Wang, Iraida Nikolaevna Tretyakova, Alexander Mikhaylovich Nosov, Hailong Shen and Ling Yang

Plants 2022, 11(14), 1867; https://doi.org/10.3390/plants11141867 - 18 Jul 2022

Cited by 7 | Viewed by 1671

Abstract

Many cell lines in the embryogenic callus cannot produce somatic embryos (SEs) even if they meet the optimal SE maturation culture conditions during conifer somatic embryogenesis. This phenomenon hinders the progress of the industrial-scale reproduction of conifers. Therefore, there is an urgent need to obtain morphological and physiological markers to screen embryogenic calli in response to SE maturation conditions. To detect cell lines with high somatic embryogenesis potential during the proliferation process, we counted the number of pro-embryos and early SEs (ESEs) in different cell lines and storage substances, endogenous hormones, and polyamine contents. The results showed that the yield of P. koraiensis SEs was heavily dependent on genotype (p = 0.001). There were high levels of PE III (pro-embryo III) number, ESE number, and soluble protein content, in the response cell lines (R cell lines), which were 1.6-, 3-, and 1.1-fold those of the obstructive cell lines (B cell lines), respectively. The B cell line had high levels of starch, auxin (IAA), Put, Spd, and putrescine: spermine (Put: Spm) compared to the R cell line. In addition, the numbers of PE III, ESEs, and soluble protein content were significantly positively correlated with SE yield. In contrast, the contents of starch, abscisic acid (ABA), Put, Spm, and Spd were significantly negatively correlated with SE yield. To ensure the accuracy of the results, we used nine cell lines to test the results. The PE III and ESE numbers and the Spm and Spd contents were positively correlated with SE yield, while the levels of starch, ABA, IAA, Put: Spd, and Put: Spm were negatively correlated with SE yield. Thus, we recommend using high PE III and ESEs as morphological indicators and low levels of starch, IAA, ABA, and Put: Spm as physiological markers to screen cell lines with a high somatic embryogenesis potential. In addition, we also found that the relationship between Spd, Spm, and SE yield was opposite in the two experimental results. Therefore, we speculate that the differences in Spd and Spm content are mainly affected by genotype. In conclusion, this study obtained the morphological and physiological markers of some high-somatic embryogenic cell lines by comparing the differences between nine somatic embryogenic cell lines. Our results can guide the improvement of conifer somatic embryogenesis technology and can provide a theoretical basis for accelerating the application of biotechnology in large-scale artificial breeding. Full article

(This article belongs to the Special Issue Applications of Plant Tissue Culture: New Insight into Plant Improvement Research Using Plant Tissue Culture and Plant Modification Techniques)

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