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Application of Plant Biotechnology in Forestry

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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: closed (1 March 2023) | Viewed by 20497

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Special Issue Editors

Dr. Ricardo Javier Ordás

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Guest Editor

Institute of Biotechnology of Asturias, University of Oviedo, 33009 Oviedo, Spain
Interests: plant development; plant biotechnology

Dr. José Manuel Álvarez Díaz

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Guest Editor

Institute of Biotechnology of Asturias, University of Oviedo, 33009 Oviedo, Spain
Interests: plant development; plant biotechnology

Special Issue Information

Dear Colleagues,

Forest trees constitute the majority of continental biomass and terrestrial biodiversity. They provide various ecological services and are a source of raw material for many essential purposes. In recent years, the need for wood and its derivatives has increased, and will continue to do so. Biotechnological tools such as genomic selection, micropropagation and genetic engineering will play an essential role in meeting this demand with the smallest possible environmental impact.

Forest regeneration after harvest is often left to natural processes, although prompt artiﬁcial regeneration with selected genotypes provides the most effective means to increase forest yield. Domestication of forest species through traditional plant breeding is technically more difﬁcult and time-consuming than other crops due to their large size, long generation times, and prolonged juvenile stage. Genomic selection has the potential to accelerate breeding cycles, increase selection intensity, and improve the accuracy of breeding values. Currently, micropropagation techniques, together with the rooting of cuttings, are considered the most effective tools for the propagation of elite forest varieties at a large scale. Micropropagation exploits the characteristic developmental plasticity of plants to adapt to variable environmental conditions; in particular, their high regeneration capacity. Genetic engineering provides the potential to allow gene function analysis or to transfer specific traits into selected genotypes without affecting their desirable genetic background. Thus, genetic engineering provides a powerful tool to rapidly increase yield and wood quality in forest species, and allows the breeding process to be shortened substantially.

The objective of this Special Issue is to deepen our knowledge of the latest biotechnological approaches to forestry, such as rejuvenation through clonal propagation, including cuttings, micrografting and tissue culture; haploid generation; somatic embryogenesis; protoplast fusion; cryopreservation of germplasm; genetic engineering; and functional mapping of quantitative trait loci.

Dr. Ricardo Javier Ordás
Dr. José Manuel Álvarez Díaz
Guest Editors

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Keywords

micropropagation
genetic engineering
genomic selection
somatic embryogenesis
protoplasts
cryopreservation

Published Papers (11 papers)

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Editorial

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2 pages, 656 KiB

Open AccessEditorial

Application of Plant Biotechnology in Forestry

by José Manuel Alvarez and Ricardo Javier Ordás

Forests 2023, 14(11), 2148; https://0-doi-org.brum.beds.ac.uk/10.3390/f14112148 - 28 Oct 2023

Viewed by 1022

Abstract

Forests, often referred to as the lungs of our planet, stand as a testament to the incredible diversity and significance of our terrestrial ecosystems [...] Full article

(This article belongs to the Special Issue Application of Plant Biotechnology in Forestry)

Research

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12 pages, 1634 KiB

Open AccessEditor’s ChoiceArticle

Identification of Candidate Genes Involved in Bud Growth in Pinus pinaster through Knowledge Transfer from Arabidopsis thaliana Models

by José Manuel Alvarez, Sonia María Rodríguez, Francisco Fuente-Maqueda, Isabel Feito, Ricardo Javier Ordás and Candela Cuesta

Forests 2023, 14(9), 1765; https://0-doi-org.brum.beds.ac.uk/10.3390/f14091765 - 31 Aug 2023

Cited by 1 | Viewed by 1608

Abstract

Pinus pinaster is a plant species of great ecological and economic importance. Understanding the underlying molecular mechanisms that govern the growth and branching of P. pinaster is crucial for enhancing wood production and improving product quality. In this study, we describe a simple methodology that enables the discovery of candidate genes in Pinus pinaster by transferring existing knowledge from model species like Arabidopsis thaliana and focusing on factors involved in plant growth, including hormonal and non-hormonal pathways. Through comparative analysis, we investigated the main genes associated with these growth-related factors in A. thaliana. Subsequently, we identified putative homologous sequences in P. pinaster and assessed the conservation of their functional domains. In this manner, we can exclude sequences that, despite displaying high homology, lack functional domains. Finally, we took an initial approach to their validation by examining the expression levels of these genes in P. pinaster trees exhibiting contrasting growth patterns. This methodology allowed the identification of 26 candidate genes in P. pinaster. Our findings revealed differential expression patterns of key genes, such as NCED3, NRT1.2, PIN1, PP2A, ARF7, MAX1, MAX2, GID1, AHK4, AHP1, and STP1, in relation to the different growth patterns analyzed. This study provides a methodological foundation for further exploration of these genes involved in the growth and branching processes of P. pinaster. This will contribute to the understanding of this important tree species and open new avenues for enhancing its utilization in sustainable forestry practices. Full article

(This article belongs to the Special Issue Application of Plant Biotechnology in Forestry)

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16 pages, 2509 KiB

Open AccessArticle

How to Capture Thousands of Genotypes—Initiation of Somatic Embryogenesis in Norway Spruce

by Saila Varis, Mikko Tikkinen, Jaanika Edesi and Tuija Aronen

Forests 2023, 14(4), 810; https://0-doi-org.brum.beds.ac.uk/10.3390/f14040810 - 14 Apr 2023

Cited by 2 | Viewed by 1142

Abstract

Somatic embryogenesis (SE) is considered the most effective method for vegetative propagation of Norway spruce (Picea abies L. Karst). When the aim is commercial production, the process needs scaling up. This includes many initiations to increase the number of available genotypes in the cryo-bank. Numerous genotypes are needed to maintain genetic diversity in reforestation and, at the same time, are a prerequisite for the efficient improvement of breeding traits. Norway spruce is also highly susceptible to Heterobasidion root rot. We analysed the data from the SE initiations of Norway spruce from six different years, including a total of 126 families and almost 13,000 initiations, and used several genetic (including allele PaLAR3B improving Heterobasidion resistance), environmental, and operational variables to explain the initiation success and the number of cryopreserved embryogenic tissue (ET). Overall, the cone collection date was the best and most comprehensive single variable for predicting the initiation success and the number of cryopreserved ET in the logistic regression models. PaLAR3B allele did not interfere with SE initiation or the cryopreservation. In the optimal scenario, according to the current data, Norway spruce cones would be collected in southern Finland during the first two weeks of July (in approximately 800 d.d. accumulation) from the seed orchard or greenhouse and delivered quickly to the laboratory, and the cones would be cold-stored for five days or less before initiations on mLM media. Lower initiation frequencies in some families can be compensated by increasing the number of explants—however, taking operational limitations into account. Full article

(This article belongs to the Special Issue Application of Plant Biotechnology in Forestry)

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12 pages, 6584 KiB

Open AccessArticle

Castanea crenata Ginkbilobin-2-like Recombinant Protein Reveals Potential as an Antimicrobial against Phytophthora cinnamomi, the Causal Agent of Ink Disease in European Chestnut

by Maria Belén Colavolpe, Fernando Vaz Dias, Susana Serrazina, Rui Malhó and Rita Lourenço Costa

Forests 2023, 14(4), 785; https://0-doi-org.brum.beds.ac.uk/10.3390/f14040785 - 11 Apr 2023

Cited by 2 | Viewed by 1605

Abstract

The European chestnut tree (Castanea sativa Mill.) is widely cultivated throughout the world’s temperate regions. In the Mediterranean region, it has a significant economic role mainly because of the high quality of its edible nuts. The Oomycete Phytophthora cinnamomi is one of the most severe pathogens affecting European chestnuts, causing ink disease and significant losses in production. Ginkgobilobin-2 (Gnk2) in Ginkgo biloba is a secreted protein with a plant-specific cysteine-rich motif that functions as a lectin, and its carbohydrate-binding properties are closely related to its antifungal activity. The binding of lectins to mannose residues of the cell wall of Phytophthora species may disturb and disrup the cell wall structure. This work determined that the amino acid sequence has a signal peptide that directs the final protein peptide to the apoplast. The Cast_Gnk2-like expression was performed and optimized, and different in vitro antagonism tests were done against P. cinnamomi using different purified protein concentrations. As a result of one of these assays, Cast_Gnk2-like significantly reduced the mycelia growth of P. cinnamomi in liquid medium as shown by the mycelia weight (g) in control treatments was 377% higher than in the treatments. These insights reveal the potential of Cast_Gnk2-like for agricultural uses and biotechnological developments for the pathosystem chestnut/P. cinnamomi. Full article

(This article belongs to the Special Issue Application of Plant Biotechnology in Forestry)

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16 pages, 2647 KiB

Open AccessArticle

Adult Trees Cryptomeria japonica (Thunb. ex L.f.) D. Don Micropropagation: Factors Involved in the Success of the Process

by Alejandra Rojas-Vargas, Itziar A. Montalbán and Paloma Moncaleán

Forests 2023, 14(4), 743; https://0-doi-org.brum.beds.ac.uk/10.3390/f14040743 - 05 Apr 2023

Cited by 1 | Viewed by 1120

Abstract

Cryptomeria japonica (Thunb. ex L.f.) D. Don is a commercial tree native to Japan and is one of the most important forest species in that country and the Azores (Portugal). Because of the quality of C. japonica timber, several genetic improvement programs have been performed. Recently, some studies focusing on C. japonica somatic embryogenesis have been carried out. However, in this species, this process uses immature seeds as initial explants, and for this reason, it is not possible to achieve the maximum genetic gain (100% genetic of the donor plant). Although some studies have been made applying organogenesis to this species, the success of the process in adult trees is low. For this reason, our main goal was to optimize the micropropagation method by using trees older than 30 years as a source of plant material. In this sense, in a first experiment, we studied the effect of different types of initial explants and three basal culture media on shoot induction; then, two sucrose concentrations and two light treatments (LEDs versus fluorescent lights) were tested for the improvement of rooting. In a second experiment, the effects of different plant growth regulators (6-benzylaminopurine, meta-topolin, and thidiazuron) on shoot induction and the subsequent phases of the organogenesis process were analyzed. The cultures produced the highest number of shoots when QL medium (Quoirin and Lepoivre, 1977) and long basal explants (>1.5 cm) were used; the shoots obtained produced a higher number of roots when they were grown under red LED lights. Moreover, root induction was significantly higher in shoots previously induced with meta-topolin. Full article

(This article belongs to the Special Issue Application of Plant Biotechnology in Forestry)

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14 pages, 1336 KiB

Open AccessArticle

Efficient Procedure for Induction Somatic Embryogenesis in Holm Oak: Roles of Explant Type, Auxin Type, and Exposure Duration to Auxin

by María Teresa Martínez and Elena Corredoira

Forests 2023, 14(2), 430; https://0-doi-org.brum.beds.ac.uk/10.3390/f14020430 - 19 Feb 2023

Cited by 1 | Viewed by 1377

Abstract

Holm oak is the dominant tree species in the Mediterranean climate. Currently, worrisome degradation of its ecosystems has been observed, produced, among other factors, by changes in land use, extreme weather events, forest fires, climate change, and especially the increasingly frequent episodes of high tree mortality caused by “oak decline”, which has brought with it a social concern that transcends the productive interest. Breeding and conservation programs for this species are necessary to ensure the prevalence of these ecosystems for future generations. Biotechnological tools such as somatic embryogenesis (SE) have great potential value for tree improvement and have been shown to be highly efficient in the propagation and conservation of woody species. One challenge to this approach is that SE induction in holm oak has not yet been optimized. Here, we present a new reproducible procedure to induce SE in holm oak; we evaluated the responsiveness of different initial explants exposed to different types, concentrations, and durations of auxin. SE rates were significantly improved (37%) by culturing nodal segments for two weeks in induction medium. In addition, a significant auxin–genotype interaction was observed. Full article

(This article belongs to the Special Issue Application of Plant Biotechnology in Forestry)

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13 pages, 3085 KiB

Open AccessArticle

Potential and Constraints on In Vitro Micropropagation of Juniperus drupacea Labill.

by Kostas Ioannidis, Ioanna Tomprou, Danae Panayiotopoulou, Stefanos Boutsios and Evangelia N. Daskalakou

Forests 2023, 14(1), 142; https://0-doi-org.brum.beds.ac.uk/10.3390/f14010142 - 12 Jan 2023

Cited by 7 | Viewed by 1722

Abstract

Juniperus drupacea Labill. (Cupressaceae) is a species with ecological and medicinal value. In Europe, it is native only in southern Greece, and is listed as endangered. Due to its uniqueness, this study attempted, for the first time, an in vitro propagation effort of Syrian juniper. Explants of the lateral shoot tips were surface-sterilized and cultured on Murashige and Skoog (MS) medium. The cultures were subcultured on MS, woody plant medium (WPM), and Driver and Kuniyaki Walnut (DKW) supplemented with different concentrations of 6-benzylaminopurine (BA), thidiazuron (TDZ), or meta-topolin [6-(3-hy-droxybenzylamino)purine] for shoot induction. Explants derived from female trees exhibited 54.17% bud proliferation on DKW medium with 4 μM meta-topolin or 4 μM TDZ and on WPM with 4 μM meta-topolin or 4 μM BA. A total of 62.50% of the male tree derived explants produced multiple shoots on DKW with 4 μM BA. The maximum average number of shoots per explant were 1.17 per explant in both cases. The length of the shoot derived from explants of female origin was 2.94 mm compared to 2.69 mm of the in vitro shoots from the explants of male trees. Overall, the best medium and plant growth regulator combination for the explants derived from both female and male trees, for the traits under study, was proven to be DKW + 4 µM TDZ. Our experiments show that Juniperus drupacea, under in vitro conditions, shows recalcitrance in rooting, as the applications of IBA, NAA, and IAA concentrations were proven to be ineffective treatments. Although the results show low values, this avant-garde study provides a foundation for further research on the in vitro regeneration of Juniperus drupacea. Full article

(This article belongs to the Special Issue Application of Plant Biotechnology in Forestry)

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13 pages, 2047 KiB

Open AccessArticle

Improved Method for Cryopreservation of Embryogenic Callus of Fraxinus mandshurica Pupr. by Vitrification

by Xueqing Liu, Yingying Liu, Xiaoqian Yu, Iraida Nikolaevna Tretyakova, Alexander Mikhaylovich Nosov, Hailong Shen and Ling Yang

Forests 2023, 14(1), 28; https://0-doi-org.brum.beds.ac.uk/10.3390/f14010028 - 23 Dec 2022

Cited by 2 | Viewed by 1489

Abstract

In order to simplify the experimental procedure and treatment procedure, we preserved the embryonic callus (EC) of Fraxinus mandshurica more efficiently. In this paper, we established a method for cryopreservation of EC of F. mandshurica by vitrification. EC was subcultured for 7–10 days (d). Vigorous EC with good growth conditions were selected, and cryopreservation was performed by vitrification. The best pre-culture method was to pre-culture EC on 0.5 mol·L⁻¹ sucrose medium for 3 d, load and culture in the liquid woody plant medium (WPM) supplemented with 2 mol·L⁻¹ glycerol and 0.4 mol·L⁻¹ sucrose for 60 min, then dehydrate in 2 mL of plant vitrification solution 2 (PVS2) (30% glycerol + 15% dimethyl sulfoxide (DMSO) + 15% ethylene glycol + 0.4 mol·L⁻¹ sucrose + liquid WPM). EC was rewarmed in a 40 °C water bath for 2 min after cooling in liquid nitrogen. The procedure for cryopreservation of F. mandshurica EC by the vitrification method established in this experiment is relatively reliable. The results from the present study provide a technical reference for improving the cryopreservation of F. mandshurica EC. Full article

(This article belongs to the Special Issue Application of Plant Biotechnology in Forestry)

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16 pages, 3770 KiB

Open AccessArticle

Improvement of Rooting Performance in Stem Cuttings of Savin Juniper (Juniperus sabina L.) as a Function of IBA Pretreatment, Substrate, and Season

by Maliheh Abshahi, Francisco Antonio García-Morote, Hossein Zarei, Bahman Zahedi and Abdolhossein Rezaei Nejad

Forests 2022, 13(10), 1705; https://0-doi-org.brum.beds.ac.uk/10.3390/f13101705 - 16 Oct 2022

Cited by 5 | Viewed by 2172

Abstract

Juniperus sabina is an interesting species for forest restoration and ornamental purposes. The seeds of this plant have several types of dormancies; therefore, seed propagation is difficult and time consuming. The production of cuttings can be an alternative way to produce plants more quickly. The main objective of this experiment was to determine the best propagation conditions (indole butyric acid dose, substrate, and season) for this species using stem cuttings. Rooting performance of the cuttings was evaluated based on the rooting percentage (%), root biomass, and specific root length (SRL). In addition, we examined the internal composition (auxin and peroxidase content) in treated stem cuttings. Cuttings were pretreated with five doses of indole butyric acid (IBA; 0 (control), 1000, 2000, 4000, and 8000 ppm) and were rooted in four substrates (perlite, perlite-cocopeat, pumice, and mixed substrate) during the four seasons (winter, spring, summer, and autumn). The best treatments, with more than 60% rooting, were applied in spring, and IBA at 1000 ppm in perlite–cocopeat substrate obtained 62% rooting. The highest rooting percentage correlated with the highest root biomass production and the lowest SRL. IBA pretreatment decreased the concentration of peroxidase in spring (coinciding with maximum rooting), representing an indicator of rooting performance. Based on these results, we recommend a new protocol for Juniperus sabina production: (i) prepare cuttings in spring, (ii) treat cutting bases with 1000 ppm IBA, and (iii) plant cuttings in a substrate of perlite–cocopeat (1:1). Full article

(This article belongs to the Special Issue Application of Plant Biotechnology in Forestry)

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Review

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21 pages, 881 KiB

Open AccessReview

Conifer Biotechnology: An Overview

by Sonia María Rodríguez, Ricardo Javier Ordás and José Manuel Alvarez

Forests 2022, 13(7), 1061; https://0-doi-org.brum.beds.ac.uk/10.3390/f13071061 - 05 Jul 2022

Cited by 3 | Viewed by 2600

Abstract

The peculiar characteristics of conifers determine the difficulty of their study and their great importance from various points of view. However, their study faces numerous important scientific, methodological, cultural, economic, social, and legal challenges. This paper presents an approach to several of those challenges and proposes a multidisciplinary scientific perspective that leads to a holistic understanding of conifers from the perspective of the latest technical, computer, and scientific advances. This review highlights the deep connection that all scientific contributions to conifers can have in each other as fully interrelated communicating vessels. Full article

(This article belongs to the Special Issue Application of Plant Biotechnology in Forestry)

17 pages, 1280 KiB

Open AccessReview

Somatic Embryogenesis of Norway Spruce and Scots Pine: Possibility of Application in Modern Forestry

by Teresa Hazubska-Przybył, Mikołaj Krzysztof Wawrzyniak, Joanna Kijowska-Oberc, Aleksandra Maria Staszak and Ewelina Ratajczak

Forests 2022, 13(2), 155; https://0-doi-org.brum.beds.ac.uk/10.3390/f13020155 - 20 Jan 2022

Cited by 10 | Viewed by 3235

Abstract

Somatic embryogenesis (SE) is an important method for the vegetative propagation of trees. SE is the developmental in vitro process in which embryos are produced from somatic cells. This method can be integrated with other biotechnological techniques, genomic breeding and cryopreservation, which enables commercial-scale sapling production of selected high-yielding genotypes in wood production combined with fast breeding cycles. The SE is potential tool to improve plant stock in comparison with seed orchards. It can be useful for ecologically and economically important species, such as Norway spruce (Picea abies L. Karst.) and Scots pine (Pinus sylvestris L.), ensuring stable production in the era of climate change and biodiversity crisis. In this review, we summarize the current state of research on problems associated with somatic embryogenesis in P. abies and P. sylvestris. Full article

(This article belongs to the Special Issue Application of Plant Biotechnology in Forestry)

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