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Plant Tissue Culture and Plant Somatic Embryogenesis

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A special issue of Agronomy (ISSN 2073-4395). This special issue belongs to the section "Plant-Crop Biology and Biochemistry".

Deadline for manuscript submissions: closed (10 February 2024) | Viewed by 11298

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Dr. Justyna Lema-Rumińska

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Department of Environmental Biology, Faculty of Biological Science, Kazimierz Wielki University,12 Ossoliński Av., PL-85-093 Bydgoszcz, Poland
Interests: somatic embryogenesis; secondary metabolites; molecular markers
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Dr. Danuta Kulpa

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Department of Plant Genetics, Breeding and Biotechnology, West Pomeranian University of Technology, Szczecin, 17 Słowackiego Str., PL-71-434 Szczecin, Poland
Interests: micropropagation; secondary metabolites; suspension cultures
Special Issues, Collections and Topics in MDPI journals

Dr. Alina Trejgell

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Department of Plant Physiology and Biotechnology, Nicolaus Copernicus University, Toruń, 1 Lwowska Str., 87-100 Torun, Poland
Interests: Asteraceae; plant in vitro culture; cold storage; slow growth; elicitation

Special Issue Information

Dear Colleagues,

In vitro plant cultures and somatic embryogenesis are the focus of this Special Issue. Scientists around the world are developing and improving methods of plant propagation and regeneration in vitro cultures. Micropropagation is used on a large scale for the production of high-quality cuttings. In addition, this technique is used in gene banks or for the production of important secondary metabolites. The most efficient plant regeneration methods include somatic embryogenesis. During the micropropagation and regeneration of plants, the genetic stability of the plant can be disturbed. To ensure that the plants obtained through this method are true-to-type, their genetic stability must be confirmed. Molecular markers that can be used at any stage of plant development work best here. The genetic variability created in this way, in addition to mutagenesis or genetic transformation in in vitro cultures, facilitate breeding new cultivars of crops.

With this in mind, scientists are encouraged to publish original research and review articles that present methods of plant propagation and regeneration in vitro cultures, especially somatic embryogenesis. Manuscripts on the determination of the genetic stability of plants after regeneration, micropropagation, or the use of in vitro culture methods for the production of secondary metabolites or plant breeding are also welcome.

Dr. Justyna Lema-Rumińska
Dr. Danuta Kulpa
Dr. Alina Trejgell
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. Agronomy is an international peer-reviewed open access monthly journal published by MDPI.

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Keywords

micropropagation
regeneration
slow growth
somatic embryos
molecular markers
genetic stability
breeding
elicitation

Related Special Issue

Plant Tissue Culture and Plant Somatic Embryogenesis–2nd Edition in Agronomy

Published Papers (9 papers)

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Research

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19 pages, 2986 KiB

Open AccessArticle

Chemical Profile of Cell Cultures of Kalanchoë gastonis-bonnieri Transformed by Agrobacterium rhizogenes

by María Guadalupe Barrera Núñez, Mónica Bueno, Miguel Ángel Molina-Montiel, Lorena Reyes-Vaquero, Elena Ibáñez and Alma Angélica Del Villar-Martínez

Agronomy 2024, 14(1), 189; https://doi.org/10.3390/agronomy14010189 - 15 Jan 2024

Viewed by 918

Abstract

Kalanchoë gastonis-bonnieri Raym.-Hamet & Perrier is a plant used for medicinal purposes in the treatment of several ailments. The aim of this study was to analyze the chemical profile of extracts from K. gastonis-bonnieri embryogenic calli, generated from genetically transformed roots by Agrobacterium rhizogenes. Putative transformants were verified by PCR. Hydroalcoholic extracts were obtained and the chemical profile was analyzed by LC-ESI-MS/MS. Root formation was obtained from 80% of infected seedlings. Fifteen root lines were isolated, and two lines showed prominent longitudinal growth and profuse branching in the B5 semi-solid medium. In all lines, the formation of nodules and later embryogenic callus was observed. Putative transgenic root lines were cultivated in free-plant growth regulators B5 medium. In the two selected lines, the PCR amplification of rolA, rolB, rolC, rolD, and aux1 genes was detected. The extract of embryogenic calli showed 60 chemical compounds tentatively identified, such as ferulic acid, quinic acid, neobaisoflavone, and malic acid, among others, and the chemical profile was different in comparison to wild-type extracts. This is the first study reporting the analysis of the chemical profile of hairy root extracts derived from Kalanchoë gastonis-bonnieri. This work displays the great potential for obtaining chemical compounds of pharmacological importance from hairy roots and facilitates the identification of new useful drugs against human chronic-degenerative diseases. Full article

(This article belongs to the Special Issue Plant Tissue Culture and Plant Somatic Embryogenesis)

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10 pages, 1802 KiB

Open AccessArticle

Propagation of Clematis ‘Warszawska Nike’ in In Vitro Cultures

by Danuta Kulpa and Marcelina Krupa-Małkiewicz

Agronomy 2023, 13(12), 3056; https://doi.org/10.3390/agronomy13123056 - 14 Dec 2023

Viewed by 940

Abstract

A micropropagation protocol for growing Clematis ‘Warszawska Nike’ was developed. The MS medium supplemented with 1 mg∙dm⁻³ BAP showed good results in the case of microshoot initiation (80%). The addition of BAP to the medium at higher concentrations resulted in the formation of a large amount of callus tissue at the base of the explant. Of the explants growing on the medium with the lowest cytokinin concentration, 8% flowered. Very quickly, after just 14 days, the explants began to die: some of the leaves that developed in in vitro cultures began to turn yellow and wither. The propagation of shoots was performed in two steps. In the first step, cytokinin BAP and Kin in various concentrations (0.5–2 mg∙dm⁻³) were added to the MS medium. In the second step, MS medium with the combinations of BAP (0.5 and 1 mg∙dm⁻³) with IAA or GA₃ (1 and 2 mg∙dm⁻³) was used. The MS medium with 0.5 mg∙dm⁻³ BAP and 2 mg∙dm⁻³ GA₃ was the best medium for the multiplication stage of clematis. Plants growing on this medium had the largest number of leaves, shoots, and internodes, and were also heavier compared to plants propagated on other media. The proliferated clematis explants were rooted on MS medium with the addition of IAA or IBA in different concentrations (0.5 to 4 mg∙dm⁻³). Of the concentrations tested, 0.5 mg∙dm⁻³ IAA was the most effective one for in vitro root induction. The highest percentage of acclimatized plants (75%) was observed when the shoots were rooted on MS medium with 0.5 mg∙dm⁻³ IAA. Full article

(This article belongs to the Special Issue Plant Tissue Culture and Plant Somatic Embryogenesis)

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19 pages, 3024 KiB

Open AccessArticle

Effect of X-rays on Seedling Pigment, Biochemical Profile, and Molecular Variability in Astrophytum spp.

by Piotr Licznerski, Justyna Lema-Rumińska, Emilia Michałowska, Alicja Tymoszuk and Janusz Winiecki

Agronomy 2023, 13(11), 2732; https://doi.org/10.3390/agronomy13112732 - 30 Oct 2023

Viewed by 1219

Abstract

Cacti are important in agricultural economies and one of the most popular horticultural plant groups. The genus Astrophytum is one of the most valuable and desirable cacti for growers and collectors around the world. By selecting the appropriate breeding methods to induce variations in combination with modern biotechnology tools for rapid change detection, it is possible to meet the challenges of the modern world in creating new variability in plants. However, there exists a lack of research concerning the impact of ionizing radiation on cacti. The aim of the study was to assess the effects of X-rays at different doses (0 Gy—control, 15, 20, 25, and 50 Gy) on the dynamics of seed germination in vitro, changes in the color of seedlings, biochemical changes in the content of metabolites and changes at the molecular level in Astrophytum spp. ‘Purple’. A significant effect of X-rays on the induction of genetic variation was observed. Remarkably high polymorphism rates were observed, ranging from 59.09% for primer S12 to a full 100.0% for S3 and S8, as determined by the SCoT (Start-Codon-Targeted) marker. In addition, a large variation in the content of plant pigments (anthocyanins, carotenoids, chlorophyll a, and chlorophyll b) was noted. Additionally, discernible alterations in the color of the tested cactus seedlings, assessed by the RHSCC catalog, were attributed to the impact of ionizing radiation. These findings hold promise for the application of radiomutation breeding in acquiring new cactus cultivars. Full article

(This article belongs to the Special Issue Plant Tissue Culture and Plant Somatic Embryogenesis)

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16 pages, 12929 KiB

Open AccessArticle

Production of Black Cumin via Somatic Embryogenesis, Chemical Profile of Active Compounds in Callus Cultures and Somatic Embryos at Different Auxin Supplementations

by Ahmed E. Higazy, Mohammed E. El-Mahrouk, Antar N. El-Banna, Mosaad K. Maamoun, Hassan El-Ramady, Neama Abdalla and Judit Dobránszki

Agronomy 2023, 13(10), 2633; https://doi.org/10.3390/agronomy13102633 - 17 Oct 2023

Viewed by 1095

Abstract

Black cumin or Nigella sativa L. is a medicinal plant of the Ranunculaceae family that has enormous importance. It has traditionally been used to cure a lot of diseases since ancient times. In the current study, the effects of different auxins on callus induction and subsequent somatic embryo formation of N. sativa L. cv. Black Diamond were examined. The best result of callus induction was observed when cotyledon explants were incubated in a Murashige and Skoog (MS) medium supplemented with 1.0 mg L⁻¹ α-naphthaleneacetic acid (NAA). The formation of somatic embryos was achieved efficiently from cotyledon-derived calli cultured on a 2 mg L⁻¹ Indole-3-butyric acid (IBA)-containing medium. Furthermore, histological analysis of embryogenic calli was used to detect the presence of different developmental stages of somatic embryos. In contrast to the calli and embryos of N. sativa ‘Black Diamond’, which initiated in the dark, light was necessary for the complete differentiation of callus and embryo cultures into shoots/developed plants. Hypocotyl-derived calli and embryos were successfully differentiated on IBA at 2.0, 1.0 mg L⁻¹, and NAA at 2.0 mg L⁻¹. To the best of our knowledge, this work can be considered the first report on the differentiation of N. sativa ‘Black Diamond’ somatic embryos into developed plants. Moreover, the metabolic profiles of secondary products of N. sativa ‘Black Diamond’ callus and embryo cultures originated from the best auxin treatments identified and were compared with that of intact seeds. Callus cultures of N. sativa ‘Black Diamond’ contained thymoquinone (TQ) in a significant percentage of the peak area (2.76%). Therefore, callus cultures could be used as a perfect alternative source of TQ for pharmaceutical and therapeutic purposes. In addition, fatty acids and/or their esters were recorded as the major components in callus and embryo cultures. These vital compounds could be isolated and used for numerous industrial applications. Full article

(This article belongs to the Special Issue Plant Tissue Culture and Plant Somatic Embryogenesis)

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18 pages, 4848 KiB

Open AccessArticle

Antioxidant Response in the Salt-Acclimated Red Beet (Beta vulgaris) Callus

by Jarosław Tyburski and Natalia Mucha

Agronomy 2023, 13(9), 2284; https://doi.org/10.3390/agronomy13092284 - 30 Aug 2023

Viewed by 812

Abstract

Callus cultures initiated from red beet tubers were acclimated to 75 or 100 mM NaCl salinity by exposing them to gradually increasing NaCl concentrations. The acclimated callus lines displayed growth rates comparable to the control culture cultivated on the NaCl-free medium. Several antioxidant system components were analyzed to assess the role of the antioxidant defense in the acclimated callus’s ability to proliferate on salt-supplemented media. It was found that proline and ascorbate concentrations were increased in salt-acclimated callus lines with respect to the control line. On the other hand, glutathione concentration was unchanged in all tested callus lines. Total activities of the antioxidant enzymes, namely superoxide dismutase (SOD, EC 1.15.1.1), catalase (CAT, EC 1.11.1.6), ascorbate peroxidase (APX, EC 1.11.1.11), and class III peroxidase (POX, EC 1.11.1.7) were increased in salt-acclimated cultures. The enzymatic components of the antioxidant systems were upregulated in a coordinated manner during the initial phases of the culture cycle when the increase in callus fresh mass occurs. Full article

(This article belongs to the Special Issue Plant Tissue Culture and Plant Somatic Embryogenesis)

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24 pages, 8400 KiB

Open AccessArticle

Effect of Different Culture Conditions on Anthocyanins and Related Genes in Red Pear Callus

by Wantian Yao, Diya Lei, Xuan Zhou, Haiyan Wang, Jiayu Lu, Yuanxiu Lin, Yunting Zhang, Yan Wang, Wen He, Mengyao Li, Qing Chen, Ya Luo, Xiaorong Wang, Haoru Tang and Yong Zhang

Agronomy 2023, 13(8), 2032; https://doi.org/10.3390/agronomy13082032 - 31 Jul 2023

Viewed by 1084

Abstract

Red pears are appreciated for their abundant nutritional benefits and visually striking red hue, rendering them a favored option among consumers and stimulating substantial market demand. The present study employs the flesh of a red pear as the explant, subjecting the flesh callus to varying sugar sources, MS concentrations, light qualities, and temperatures to investigate the alterations in secondary metabolites, including anthocyanins, within the callus. It was found that sucrose can induce more anthocyanins, and its related metabolites and genes also increase as the sucrose and MS concentrations increase. Under the conditions of red-blue light and a temperature of 15 °C, it can further induce the production of more anthocyanins and secondary metabolites and can also upregulate the synthesis of anthocyanin-related genes. As such, this investigation serves to elucidate the factors that contribute to anthocyanin accumulation in red pears, thereby providing a theoretical foundation for understanding the mechanisms underlying color change. Full article

(This article belongs to the Special Issue Plant Tissue Culture and Plant Somatic Embryogenesis)

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13 pages, 2618 KiB

Open AccessArticle

Magnesium Oxide Nanoparticles: An Influential Element in Cowpea (Vigna unguiculata L. Walp.) Tissue Culture

by Rabia Koçak, Melih Okcu, Kamil Haliloğlu, Aras Türkoğlu, Alireza Pour-Aboughadareh, Bita Jamshidi, Tibor Janda, Azize Alaylı and Hayrunnisa Nadaroğlu

Agronomy 2023, 13(6), 1646; https://doi.org/10.3390/agronomy13061646 - 19 Jun 2023

Cited by 4 | Viewed by 1735

Abstract

Nanotechnology is a rapidly growing field of science and technology that deals with the development of new solutions by understanding and controlling matter at the nanoscale. Since the last decade, magnesium oxide nanoparticles (MgO-NPs) have gained tremendous attention because of their unique characteristics and diverse applications in materials sciences and because they are non-toxic and relatively cheaply available materials. MgO-NPs can improve plant growth and contribute to plant tolerance of heavy metal toxicity. The effects of MgO-NPs on cowpea (Vigna unguiculata L. Walp.) plants were surveyed under in vitro conditions to find the optimum combination for cowpea tissue culture. The MgO-NPs used in the study were synthesized using walnut shell extract by the green synthesis method. MgO nanoparticles with 35–40 nm size was used in this research. When the size distribution of the MgO-NPs’ structure was examined, two peaks with 37.8 nm and 78.8 nm dimensions were obtained. The zeta potential of MgO-NPs dispersed in water was measured around −13.3 mV on average. The results showed that different doses of MgO-NPs applied to cowpea plant on all in vitro parameters significantly affected all measured parameters of cowpea plantlets under in vitro condition in a positive way. The best results in morphogenesis were MS medium supplemented with high MgO-NP applications (555 mg/L), resulting in a 25% increase in callus formation. The addition of Mg-NPs in the induction medium at concentrations at 370 mg/L increased shoot multiplication. The highest root length with 1.575 cm was obtained in MS medium containing 370 mg/L MgO. This study found that MgO-NPs greatly influenced the plantlets’ growth parameters and other measured traits; in addition, our results indicate that the efficiency of tissue culture of cowpea could be improved by increased application of MgO in the form of nanoparticles. In conclusion, the present work highlights the possibility of using MgO-NPs in cowpea tissue culture. Full article

(This article belongs to the Special Issue Plant Tissue Culture and Plant Somatic Embryogenesis)

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16 pages, 2415 KiB

Open AccessArticle

Elicitation and Enhancement of Phenolics Synthesis with Zinc Oxide Nanoparticles and LED Light in Lilium candidum L. Cultures In Vitro

by Piotr Pałka, Bożena Muszyńska, Agnieszka Szewczyk and Bożena Pawłowska

Agronomy 2023, 13(6), 1437; https://doi.org/10.3390/agronomy13061437 - 23 May 2023

Cited by 2 | Viewed by 1291

Abstract

In this study, we identified and determined the content of phenolic compounds in Lilium candidum adventitious bulbs formed in vitro. HPLC analysis revealed the presence of four phenolic acids: chlorogenic, caffeic, p-coumaric, and ferulic acid. Phenolic acid content was assessed in adventitious bulbs formed in vitro on media supplemented with zinc oxide nanoparticles (ZnO NPs at 25, 50, and 75 mg/L) under fluorescent light (FL) or in darkness (D). The second experiment analyzed the effects of light-emitting diodes (LEDs) of variable light spectra on the formation of adventitious bulbs and their contents of phenolic acids. Spectral compositions of red (R; 100%), blue (B; 100%), red and blue (RB; 70% and 30%, respectively), a mix of RB and green (RBG) in equal proportions (50%), and white light (WLED, 33.3% warm, neutral, and cool light, proportionately) were used in the study. FL and D conditions were used as controls for light spectra. Bulbs grown in soil served as control samples. The most abundant phenolic acid was p-coumaric acid. Treatment with LED light spectra, i.e., RB, RBG, WLED, and B, translated into the highest p-coumaric acid concentration as compared with other treatments. Moreover, all the bulbs formed in light, including those grown on the media supplemented with ZnO NPs and under FL light, contained more p-coumaric acid than the bulbscales of the control bulbs grown in soil. On the other hand, control bulbs grown in soil accumulated about two to three times higher amounts of chlorogenic acid than those formed in vitro. We also found that the levels of all examined phenolics decreased under FL, R, and D conditions, while the bulblets formed in vitro under RB light showed the highest phenolic content. The use of ZnO NPs increased the content of p-coumaric, chlorogenic, and caffeic acid in the bulblets formed under FL as compared with those grown in darkness. Full article

(This article belongs to the Special Issue Plant Tissue Culture and Plant Somatic Embryogenesis)

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Review

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24 pages, 399 KiB

Open AccessReview

Oat (Avena sativa L.) In Vitro Cultures: Prospects and Challenges for Breeding

by Marzena Warchoł, Edyta Skrzypek, Katarzyna Juzoń-Sikora and Dragana Jakovljević

Agronomy 2023, 13(10), 2604; https://doi.org/10.3390/agronomy13102604 - 12 Oct 2023

Viewed by 1374

Abstract

Plant in vitro cultures have been a crucial component of efforts to enhance crops and advance plant biotechnology. Traditional plant breeding is a time-consuming process that, depending on the crop, might take up to 25 years before an improved cultivar is available to farmers. This is a problematic technique since both beneficial qualities (such as pest resistance) and negative ones (such as decreased yield) can be passed down from generation to generation. In vitro cultures provide various advantages over traditional methods, including the capacity to add desirable characteristics and speed up the development of new cultivars. When it comes to oat (Avena sativa L.), the efficient method of plant regeneration is still missing compared to the most common cereals, possibly because this cereal is known to be recalcitrant to in vitro culture. In this review, an effort has been made to provide a succinct overview of the various in vitro techniques utilized or potentially involved in the breeding of oat. The present work aims to summarize the crucial methods of A. sativa L. cultivation under tissue culture conditions with a focus on the progress that has been made in biotechnological techniques that are used in the breeding of this species. Full article

(This article belongs to the Special Issue Plant Tissue Culture and Plant Somatic Embryogenesis)

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