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Somatic Embryogenesis and Organogenesis on Tree Species

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A special issue of Forests (ISSN 1999-4907). This special issue belongs to the section "Genetics and Molecular Biology".

Deadline for manuscript submissions: 25 June 2024 | Viewed by 12888

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Special Issue Editors

Dr. Marcos Edel Martinez-Montero

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Guest Editor

Department of Plant Breeding and Plant Conservation, Bioplantas Center, University of Ciego de Avila, Ciego de Ávila 65200, Cuba
Interests: plant tissue culture; in vitro culture; plant cryopreservation; cryobionomics
Special Issues, Collections and Topics in MDPI journals

Dr. Judit Dobránszki

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Guest Editor

Centre for Agricultural Genomics and Biotechnology, Faculty of the Agricultural and Food Science and Environmental Management, University of Debrecen, 4400 Nyíregyháza, Hungary
Interests: plant tissue culture; in vitro culture; organogenesis; cytokinins; somatic embryogenesis; transcriptomics; epigenetics
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

Tree species are of invaluable importance to our environment and to human well-being. Their benefits can help cities and countries meet 15 internationally supported United Nations Sustainable Development Goals.

According to Bioversity International, forest and tree genetic diversity matters because trees are the foundation species of the forest, diverse trees provide diverse goods and services, forests and trees provide ecosystem services, tree genetic diversity is vital in landscape restoration efforts, and trees provide nutrition all year round.

In recent years, ever-rising human population growth has put a tremendous pressure on trees and tree products. The combination of biotechnology and conventional methods such as plant propagation and breeding has become a novel approach to improving and multiplying a large number of tree species.

The rapid progress of somatic embryogenesis on tree species and its prospects for potential applications is of importance to this Special Issue. Therefore, our issue aims to cover various aspects of somatic embryogenesis as a valuable tool in basic cell biology and tree genetic research and in the long-term ex situ conservation of genetic resources via cryopreservation techniques, as well as in mass propagation.

Potential topics include but are not limited to:

Somatic embryogenesis
Tree biotechnology
Tree products
Cryopreservation
Mass propagation
Genetic diversity
Climate change
Plant breeding

Dr. Marcos Edel Martinez-Montero
Dr. Judit Dobránszki
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. Forests 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 2600 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

somatic embryogenesis
tree biotechnology
tree products
cryopreservation
mass propagation
forest
genetic diversity
ecosystem
climate change
plant breeding

Published Papers (7 papers)

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Research

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12 pages, 1168 KiB

Open AccessArticle

Effects of a New Plant Growth Regulator on Callus Induction from Immature Embryo Explants of Korean Pine (Pinus koraiensis)

by Shuai Nie, Yue Wang, Yong Yan, Shanshan Liu, Wenhui Guo, Ling Yang and Hailong Shen

Forests 2023, 14(12), 2413; https://doi.org/10.3390/f14122413 - 11 Dec 2023

Viewed by 819

Abstract

Somatic embryogenesis is currently the most promising technique for the large-scale production and breeding of conifer species. Nonetheless, the low explant induction rate in this process hampers the development of improved materials. In this study, three immature Korean pine (Pinus koraiensis) embryos capable of induction were used as experimental materials. Various concentrations of brassinolide (BL), melatonin (Mel), salicylic acid (SA), glutathione (GSH), and ascorbic acid (AsA) were added to induce embryogenic callus tissue. The results show that BL had the most significant positive effect on promoting induction and increasing explant survival. Mel was also found to slightly increase the induction and survival rates of explants. When 1.00 mg·L⁻¹ BL was added to the explants derived from stock tree I, which had strong self-induction ability, for 30 days, the callus induction rate rose to 400% compared to the control group. Following the addition of 25 mg·L⁻¹ Mel to stock tree I for 30 days, the callus induction rate further increased to 450% of the control group. The effect of GSH on callus induction was insignificant. The addition of 0.10 g·L⁻¹ to stock tree I for 30 days resulted in only a 150% higher induction rate compared to the control group. When 1.00 mg·L⁻¹ BL was applied to explants with weak self-induction ability (stock tree III) for 15 days, the callus induction rate increased to 600% of that of the control group. The callus induction rate further increased to 800% of the control group after 50 mg·L⁻¹ BL were added to stock tree III for 15 days. This study presents a method to improve the induction of embryogenic callus tissue in Korean pine. Full article

(This article belongs to the Special Issue Somatic Embryogenesis and Organogenesis on Tree Species)

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18 pages, 3736 KiB

Open AccessArticle

Effects of Long-Term Subculture on Maturation Ability and Plant Conversion in Pinus radiata: Using FT-IR Spectroscopy to Determine Biomarkers of Embryogenic Tissue Aging

by Yenny Lineros, Macarena Rojas-Rioseco, Martha Hernández, Darcy Ríos, Ximena Muñoz and Rodrigo Hasbún

Forests 2023, 14(7), 1446; https://doi.org/10.3390/f14071446 - 14 Jul 2023

Cited by 1 | Viewed by 1052

Abstract

The forestry industry has integrated somatic embryogenesis into its clonal programs due to the generation of a high number of plants from selected genotypes at low cost. Somatic embryos are generated in a stressful environment after multiplication of the proembryogenic masses; thus, it is critical to determine the degree of stability of the embryogenic cultures and their potential for mass propagation. Maturation ability in cultures of different ages was evaluated in conjunction with the integrity of the proembryogenic masses, germination rate, hypocotyl and root length, plant conversion, and ex vitro survival. To identify differences in embryogenic tissue from different subcultures, their DNA was analyzed using FT-IR spectroscopy. A significant decrease in the production of somatic embryos was detected from week 15, and some lines even stopped producing embryos. Germination rate, hypocotyl length, and plant conversion were negatively affected by long-term cultivation, while root length and ex vitro survival were not significantly affected. The results obtained from the FT-IR spectroscopy analysis indicate that it is feasible to use mid-infrared spectroscopy to differentiate between embryogenic tissues with different cumulative subculture times based on the spectra obtained from their DNA, which is directly related to maturation ability. Full article

(This article belongs to the Special Issue Somatic Embryogenesis and Organogenesis on Tree Species)

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17 pages, 5043 KiB

Open AccessFeature PaperArticle

Somatic Embryogenesis Induction and Genetic Stability Assessment of Plants Regenerated from Immature Seeds of Akebia trifoliate (Thunb.) Koidz

by Yiming Zhang, Yunmei Cao, Yida Wang and Xiaodong Cai

Forests 2023, 14(3), 473; https://doi.org/10.3390/f14030473 - 26 Feb 2023

Cited by 3 | Viewed by 1671

Abstract

Akebia trifoliata is a perennial woody plant with considerable potential in nutrition, food, and health, and the production of seedlings with high quality is critical for its economic utilization. Plant regeneration through somatic embryogenesis is a powerful alternative for propagating many plant species. In this study, a simple and practicable protocol was developed for plant regeneration from immature seeds of A. trifoliata via somatic embryogenesis, and the genetic stability of regenerated plants was also assessed. In the somatic embryo (SE) induction stage, the highest frequency of somatic embryogenesis (35.2%) was observed on the WPM medium containing 1.0 mg L⁻¹ of thidiazuron (TDZ) and 1.0 mg L⁻¹ of 6-benzyladenine (6-BA). The concentration of 6-BA was optimized at 1.0 mg L⁻¹ for the proliferation and maturation of the induced SEs, and the combination of 2.0 mg L⁻¹ of indole-3-butyric acid (IBA) and 0.5 mg L⁻¹ of TDZ was the most responsive for root development and plant growth. The leaf morphological characteristics greatly varied among the established plants, and they could be grouped into three plant types, namely the normal type, Type Ι, and Type ΙΙ. Remarkable differences in the number, size, shape, and color of the leaflets were observed among the three types, while their ploidy level was the same via flow cytometry analysis. The Type ΙΙ and the Type Ι plants had the highest and the lowest net photosynthesis rate, transpiration rate, and stomatal conductance among the three groups, respectively, and both had a smaller size of stomatal guard cells than the normal type. Simple sequence repeat (SSR) analysis detected that 41 bands (43.62%) were different from those observed in the wild, indicating a high degree of polymorphism between the regenerants and their donor parent. The obtained plants might hold potential for future genetic improvement and breeding in A. trifoliata, and the established regeneration protocol might serve as a foundation for in vitro propagation and germplasm preservation of this crop. Full article

(This article belongs to the Special Issue Somatic Embryogenesis and Organogenesis on Tree Species)

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19 pages, 2123 KiB

Open AccessArticle

Optimizing Somatic Embryogenesis Initiation, Maturation and Preculturing for Cryopreservation in Picea pungens

by Xi Cao, Fang Gao, Caiyun Qin, Shigang Chen, Jufeng Cai, Changbin Sun, Yuhui Weng and Jing Tao

Forests 2022, 13(12), 2097; https://doi.org/10.3390/f13122097 - 8 Dec 2022

Cited by 7 | Viewed by 1346

Abstract

Picea pungens (Engelm.), known for its blue-green needles, has become a likable ornamental species in northeast China since 2000. Nonetheless, a lack of propagation methods that can maintain genetic fidelity and develop seedlings at a large scale prevents the further expansion of the species. Somatic embryogenesis (SE), paired with cryopreservation technologies, may provide a valid alternative. Picea pungens SE is not new, but its practical application has been limited due to low efficiencies in SE initiation and maturation as well as a lack of effective cryopreservation technology. In this study, experiments were carried out to overcome the limitations by modifying culture media. For initiation, the efficiency was enhanced by adjusting concentrations of 2.4-dichlorophenoxy acetic acid (2,4-D), 6-benzyl amino–purine (6-BA) or sucrose supplemented to the induction medium. The concentrations of 4.0 mg/L 2,4-D, 2 mg/L 6-BA, and 5 to 10 g/L sucrose were found optimal in maximizing initiation efficiency. For maturation, the efficiency, expressed as the number of mature somatic embryos per gram of fresh mass cultured (E/gFM), varied greatly with the choices of the basal medium and concentration of abscisic acid (ABA) of the maturation medium. Based on our results, the judicial choices were using the DCR medium as the basal medium and 10 mg/L ABA. The maturation efficiency could also be improved by adjusting the maturation medium’s osmotic pressure by manipulating the concentrations of carbohydrate and Gelrite and culture density. While the maturation medium, using sucrose as carbohydrate source or supplemented with a low (<8 g/L) Gelrite concentration, facilitated maturation, optimal selections were truly genotype-dependent. Our results also suggest that, while the optimal culture density varied with genotype, in general it is needless to culture more than 100 mg embryogenesis tissues per dish (size: 10 × 1.5 cm). Based on this study, the optimum pretreatment for embryogenesis tissue cryopreservation was culturing the tissues on the proliferation medium with 0.4 mol/L sorbitol for 24 h, followed by treatment with 5% Dimethyl sulfoxide. This study significantly improved the initiation (achieved a frequency of 0.56) and embryo maturation efficiencies (achieved 1030 E/gFM) and established an effective preculturing protocol for cryopreservation (recovered 1354 E/gFM) for the species. The protocols developed here, paired with the available ones for other SE steps in the literature, form a well-refined SE technology intended for commercial application to Picea pungens. Full article

(This article belongs to the Special Issue Somatic Embryogenesis and Organogenesis on Tree Species)

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12 pages, 2679 KiB

Open AccessArticle

Interaction of Subculture Cycle, Hormone Ratio, and Carbon Source Regulates Embryonic Differentiation of Somatic Cells in Pinus koraiensis

by Yuhui Ren, Xiaoqian Yu, Honglin Xing, Iraida Nikolaevna Tretyakova, Alexander Mikhaylovich Nosov, Ling Yang and Hailong Shen

Forests 2022, 13(10), 1557; https://doi.org/10.3390/f13101557 - 23 Sep 2022

Cited by 6 | Viewed by 1295

Abstract

During somatic embryogenesis of Pinus koraiensis, the ability of the embryogenic callus to produce embryos gradually decreases with long-term proliferative culture, which seriously affects large-scale application of this technology. In this study, embryogenic calli of Korean pine 1–100 cell line were used as materials. It was found that in the 7-day subculture cycle of embryogenic calli the somatic embryogenic potential of Korean pine remained for the longer time. In comparison with a subculture cycle of 14 days, indoleacetic acid (IAA), soluble sugar, and starch contents in embryogenic callus were higher with a subculture cycle of 7 days, while hydrogen peroxide (H₂O₂) content showed the opposite trend. Further, low levels of endogenous abscisic acid (ABA) and high levels of IAA could inhibit somatic embryogenesis of P. koraiensis calli. Among different carbon sources, maltose produced more storage materials and higher superoxide dismutase (SOD) and catalase (CAT) activities than sucrose, which was conducive to cell differentiation and somatic embryogenesis. The results revealed the physiological characteristics of the loss of embryogenic capacity of Korean pine embryogenic callus and provided a basis for the long-term maintenance of its somatic embryogenic capacity. Full article

(This article belongs to the Special Issue Somatic Embryogenesis and Organogenesis on Tree Species)

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15 pages, 5804 KiB

Open AccessArticle

Agrobacterium-Mediated Genetic Transformation of Larix kaempferi (Lamb.) Carr. Embryogenic Cell Suspension Cultures and Expression Analysis of Exogenous Genes

by Shaofei Dang, Lifeng Zhang, Suying Han and Liwang Qi

Forests 2022, 13(9), 1436; https://doi.org/10.3390/f13091436 - 8 Sep 2022

Cited by 1 | Viewed by 1542

Abstract

A simple and efficient protocol for the genetic transformation of Japanese larch (Larix kaempferi (Lamb.) Carr.) was developed by altering the infection method and duration and the bacterial removal process. More than 600 hygromycin-resistant embryonal masses with the vector pCAMBIA1301 were obtained, with an average of 20 transgenic lines per gram of fresh weight. Nine hygromycin-resistant transformation events (designated P1–P9) were analyzed using PCR, quantitative real-time PCR, and histochemical β-glucuronidase (GUS) assays. The GUS transcript abundance in each transformed cell line ranged from 10¹ to 10³ magnitudes, with a maximum abundance of 2.89 × 10³. In addition, the pLaTCTP::GUS vector, which contains GUS under the control of the L. kaempferi LaTCTP promoter, led to constitutive expression of GUS in embryonal-suspensor mass and somatic embryos. The transcript abundance of the exogenous genes HPT and GUS, driven by the CaMV 35S or LaTCTP promoter, ranged from 10¹ to 10⁴, which was equivalent to genes with moderate and low abundances in Japanese larch. The relatively low expression levels of exogenous genes in transformants might reflect the large genome of Japanese larch. Additional transgenic cell lines need to be screened to obtain transformants with higher expression levels of foreign genes for further functional research in Japanese larch. Full article

(This article belongs to the Special Issue Somatic Embryogenesis and Organogenesis on Tree Species)

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Review

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12 pages, 1516 KiB

Open AccessReview

Thin Cell Layer Tissue Culture Technology with Emphasis on Tree Species

by Vikas Sharma, Tanvi Magotra, Ananya Chourasia, Divye Mittal, Ujjwal Prathap Singh, Saksham Sharma, Shivika Sharma, Yudith García Ramírez, Judit Dobránszki and Marcos Edel Martinez-Montero

Forests 2023, 14(6), 1212; https://doi.org/10.3390/f14061212 - 12 Jun 2023

Cited by 1 | Viewed by 2639

Abstract

An increased dependency on plant-based resources for food, shelter, and medicinal usage has increased their sustainable and unsustainable exploitation. To use this resource sustainably, plant tissue culture (PTC) is one important technology. Among different PTC techniques, thin cell layer (TCL) technology is a relatively simple and easily adaptable technique for in vitro cultures of plants. This technique uses small explants about 0.5–2 mm in thickness excised from different plant organs. It has been successfully used in the large-scale propagation of vegetables, legumes, and plants with medicinal benefits. TCL technology has proven to be effective in stimulating various organogenic responses when combined with various new methods such as nanotechnology or microtome-based explantation, especially in tree species. It is considered an important tool in plant biotechnology. Although the morphogenetic response per explant is usually higher in conventional explants, the appropriate use of plant growth regulators and geometric factors in TCL has the potential to make it more efficient and beneficial. This article provides an overview of the concept of TCL as applied to different plant species, particularly trees, since there are few, if any, summaries of TCL technology, especially in trees. This review will certainly revitalize this important technology so that it can be used effectively for successful mass propagation in the field of plant tissue culture. Full article

(This article belongs to the Special Issue Somatic Embryogenesis and Organogenesis on Tree Species)

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