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Plant Tissue and Organ Cultures for Crop Improvement in Omics Era

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A special issue of Horticulturae (ISSN 2311-7524). This special issue belongs to the section "Genetics, Genomics, Breeding, and Biotechnology (G2B2)".

Deadline for manuscript submissions: closed (10 November 2024) | Viewed by 45436

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Dr. Anca Butiuc-Keul

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1. Department of Molecular Biology and Biotechnology, Faculty of Biology and Geology, Babeş-Bolyai University, M. Kogalniceanu St. 1, 400084 Cluj-Napoca, Romania
2. Center of Systems Biology, Biodiversity and Bioresources, Babeş-Bolyai University, Clinicilor St. 5-7, 400006 Cluj-Napoca, Romania
Interests: plant tissue culture; conservation of plant biodiversity; gene transfer

Special Issue Information

Dear Colleagues,

Plant tissue and organ cultures have been employed over the years for plant regeneration, multiplication, germplasm conservation, production of secondary metabolites and improvement of genetic characteristics by somaclonal and gametoclonal variations. In the last few decades, plant tissue cultures have also been used as experimental systems for genetic transformation and genome editing of several plant species for the development of smart crops with improved quality and yield. Thus, the plant tissue and organ culture constitute one of the most promising tools in research and biotechnology. Therefore, this Special Issue of ‘Plant Tissue and Organ Cultures for Crop Improvement in Omics Era’ will be focused on the most recent studies on the following topics (but not exclusively): gene expression during organogenesis and somatic embryogenesis, transcription and translation studies in gene regulation, genetic and epigenetic changes associated with in vitro culture conditions, gene transfer, genome editing, metabolic engineering, molecular breeding.

Dr. Anca Butiuc-Keul
Guest Editor

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Keywords

bioactive compounds
gene regulation
gene transfer
genome editing
horticultural crops
micropropagation
organogenesis
plant tissue culture
plant transformation

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Published Papers (10 papers)

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Research

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18 pages, 5747 KiB

Open AccessArticle

Comparative Transcriptome Analysis of Non-Organogenic and Organogenic Tissues of Gaillardia pulchella Revealing Genes Regulating De Novo Shoot Organogenesis

by Yashika Bansal, A. Mujib, Mahima Bansal, Mohammad Mohsin, Afeefa Nafees and Yaser Hassan Dewir

Horticulturae 2024, 10(11), 1138; https://doi.org/10.3390/horticulturae10111138 - 25 Oct 2024

Viewed by 619

Abstract

Gaillardia pulchella is an important plant species with pharmacological and ornamental applications. It contains a wide array of phytocompounds which play roles against diseases. In vitro propagation requires callogenesis and differentiation of plant organs, which offers a sustainable, alternative synthesis of compounds. The morphogenetic processes and the underlying mechanisms are, however, known to be under genetic regulation and are little understood. The present study investigated these events by generating transcriptome data, with de novo assembly of sequences to describe shoot morphogenesis molecularly in G. pulchella. The RNA was extracted from the callus of pre- and post-shoot organogenesis time. The callus induction was optimal using leaf segments cultured onto MS medium containing α-naphthalene acetic acid (NAA; 2.0 mg/L) and 6-benzylaminopurine (BAP; 0.5 mg/L) and further exhibited a high shoot regeneration/caulogenesis ability. A total of 68,366 coding sequences were obtained using Illumina150bpPE sequencing and transcriptome assembly. Differences in gene expression patterns were noted in the studied samples, showing opposite morphogenetic responses. Out of 10,108 genes, 5374 (53%) were downregulated, and there were 4734 upregulated genes, representing 47% of the total genes. Through the heatmap, the top 100 up- and downregulating genes’ names were identified and presented. The up- and downregulated genes were identified using the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway. Important pathways, operative during G. pulchella shoot organogenesis, were signal transduction (13.55%), carbohydrate metabolism (8.68%), amino acid metabolism (5.11%), lipid metabolism (3.75%), and energy metabolism (3.39%). The synthesized proteins displayed phosphorylation, defense response, translation, regulation of DNA-templated transcription, carbohydrate metabolic processes, and methylation activities. The genes’ product also exhibited ATP binding, DNA binding, metal ion binding, protein serine/threonine kinase -, ATP hydrolysis activity, RNA binding, protein kinase, heme and GTP binding, and DNA binding transcription factor activity. The most abundant proteins were located in the membrane, nucleus, cytoplasm, ribosome, ribonucleoprotein complex, chloroplast, endoplasmic reticulum membrane, mitochondrion, nucleosome, Golgi membrane, and other organellar membranes. These findings provide information for the concept of molecular triggers, regulating programming, differentiation and reprogramming of cells, and their uses. Full article

(This article belongs to the Special Issue Plant Tissue and Organ Cultures for Crop Improvement in Omics Era)

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18 pages, 5275 KiB

Open AccessArticle

Genetic Transformation of Potato without Antibiotic-Assisted Selection

by Dmitry Miroshnichenko, Anna Klementyeva, Tatiana Sidorova, Alexander S. Pushin and Sergey Dolgov

Horticulturae 2024, 10(3), 222; https://doi.org/10.3390/horticulturae10030222 - 26 Feb 2024

Viewed by 1671

Abstract

The genetic engineering of plants often relies on the use of antibiotic or herbicide resistance genes for the initial selection of primary transgenic events. Nevertheless, the commercial release of genetically modified crops containing any marker gene encounters several challenges stemming from the lack of consumer acceptance. The development of strategies enabling the generation of marker-free transgenic plants presents an alternative to address public concerns regarding the safety of biotech crops. This study examined the capabilities of highly regenerative potato cultivars to develop transgenic plants without the presence of selective substances in their media. Internodal segments of in vitro potato plants were inoculated with the Agrobacterium strain AGL0 carrying plasmids, which contained the GFP or RFP gene driven by the CaMV 35S promoter to monitor the transformation process by observing in vivo green or red fluorescence. Despite the absence of selective pressure, inoculated explants demonstrated comparable or even higher transient expression compared to experiments based on antibiotic assistant selection. Consequently, under non-selective conditions, non-transgenic, chimeric, and fully fluorescent potato plantlets were concurrently developed. Among the five tested cultivars, the regeneration efficiency of non-chimeric transgenic plants varied from 0.9 (‘Chicago’) to 2.7 (#12-36-42) plants per 100 detached plantlets. Depending on the regenerative characteristics of potato varieties (early, intermediate, or late), a specific time interval can be determined when a blind collection of transgenic plantlets is more successful, streamlining the transformation procedure. The results indicate that the outlined procedure is simple and reproducible, consistently achieving the transformation efficiency of 7.3–12.0% (per 100 inoculated explants) in potato cultivars without selective pressure. The described transformation procedure holds the potential for obtaining cisgenic or intragenic potato plants with new valuable traits that do not carry marker genes. Full article

(This article belongs to the Special Issue Plant Tissue and Organ Cultures for Crop Improvement in Omics Era)

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11 pages, 2144 KiB

Open AccessArticle

Plant Regeneration via Somatic Embryogenesis and Indirect Organogenesis in Blue Honeysuckle (Lonicera caerulea L.)

by Yu Liu, Ying Zhan, Qiang Fu, Songlin Li, Xinyu Sun, Yaru Wang, Min Yu, Dong Qin, Junwei Huo and Chenqiao Zhu

Horticulturae 2023, 9(9), 996; https://doi.org/10.3390/horticulturae9090996 - 4 Sep 2023

Cited by 3 | Viewed by 2303

Abstract

Blue honeysuckle (Lonicera caerulea L.), which belongs to the Caprifoliaceae family, is an emerging fruit crop worldwide. For the development of a transgenic system and multipurpose tissue culture, this study for the first time established an in vitro regeneration system via somatic embryogenesis, as well as improving the previously established indirect organogenesis-based regeneration system. For embryogenesis, Murashige and Skoog (MS) medium supplemented with 1.0 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) showed the highest induction rate of the embryogenic callus (97.6%), and MS supplemented with 0.1 mg/L 6-benzyladenine (6-BA), 0.1 mg/L α-naphthaleneacetic acid (NAA), and 0.5 g/L activated carbon (AC) achieved the highest somatic embryo rate (28.3%). For indirect organogenesis, MS medium supplemented with 1.0 mg/L 6-BA and 0.1 mg/L NAA resulted in the highest non-embryogenic callus induction rate (98.9%) and adventitious shoot induction rate (51.6%). For adventitious root induction, MS supplemented with 1.0 mg/L indole-3-butyric acid (IBA) achieved the highest root induction rate (96.0%) and average root length (4.6 cm), whereas MS supplemented with 0.5 mg/L indole-3-acetic acid (IAA) resulted in the highest average regenerated root number (8.8). The total time for the regeneration from explants to soil-planted seedlings (10 euphylla) was 105 and 150 days with an efficiency of 44.1% and 23.9% through organogenesis and somatic embryogenesis, respectively. This study provides a powerful tool for rapid propagation, proliferation, and transformation, as well as laying a technological foundation for gene function research and genetic improvement of blue honeysuckle. Full article

(This article belongs to the Special Issue Plant Tissue and Organ Cultures for Crop Improvement in Omics Era)

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15 pages, 2774 KiB

Open AccessFeature PaperEditor’s ChoiceArticle

Screening of Cork Oak for Resistance to Phytophthora cinnamomi and Micropropagation of Tolerant Seedlings

by María Teresa Martínez, Beatriz Cuenca, Fátima Mosteiro, Pablo Piñeiro, Felipe Pérez, Alejandro Solla and Elena Corredoira

Horticulturae 2023, 9(6), 692; https://doi.org/10.3390/horticulturae9060692 - 12 Jun 2023

Cited by 2 | Viewed by 1730

Abstract

Massive propagation of cork oak (Quercus suber) individuals tolerant to Phytophthora cinnamomi (Pc) is probably the most important challenge for cork production. Screening for resistance to Pc of ca. 200 seedlings obtained from a single cork oak tree that has survived the epidemic was performed by soil infestation. Twenty months after Pc inoculation, 33 seedlings survived from Pc infection and the four most vigorous seedlings were selected. The plants were forced to produce new shoots under controlled climatic conditions, and the new shoots were used to establish the plants in vitro by axillary budding. High axillary shoot proliferation rates were achieved by culturing the new shoots on Lloyd and McCown (WPM) medium, followed by subculturing for 2 weeks on 0.22 µM benzyladenine (BA) and for 2 weeks further on 0.04 µM BA. Addition of 20 µM silver thiosulphate (STS) increased the proliferation rates and improved the appearance and development of shoots. Rooting rates of 80–100% were obtained by culturing the shoots for 24 or 48 h on Gresshoff and Doy medium with ⅓ macronutrients plus 122.5 µM indole-3-butyric acid and subsequent transfer to root expression medium containing 20 µM STS. The results of this study optimize the micropropagation of a relevant and recalcitrant tree species in forestry. Full article

(This article belongs to the Special Issue Plant Tissue and Organ Cultures for Crop Improvement in Omics Era)

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16 pages, 4588 KiB

Open AccessEditor’s ChoiceArticle

Influence of the 135 bp Intron on Stilbene Synthase VaSTS11 Transgene Expression in Cell Cultures of Grapevine and Different Plant Generations of Arabidopsis thaliana

by Konstantin V. Kiselev, Zlata V. Ogneva, Olga A. Aleynova, Andrey R. Suprun, Alexey A. Ananev, Nikolay N. Nityagovsky and Alexandra S. Dubrovina

Horticulturae 2023, 9(4), 513; https://doi.org/10.3390/horticulturae9040513 - 20 Apr 2023

Cited by 1 | Viewed by 1348

Abstract

Modern plant biotechnology often faces the problem of obtaining a stable and powerful vector for gene overexpression. It is known that introns carry different regulatory elements whose effects on transgene expression have been poorly studied. To study the effect of an intron on transgene expression, the stilbene synthase 11 (VaSTS11) gene of grapevine Vitis amurensis Rupr. was selected and overexpressed in grapevine callus cell cultures and several plant generations of Arabidopsis thaliana as two forms, intronless VaSTS11c and intron-containing VaSTS11d. The STS genes play an important role in the biosynthesis of stilbenes, valuable plant secondary metabolites. VaSTS11d contained two exons and one intron, while VaSTS11c contained only two exons, which corresponded to the mature transcript. It has been shown that the intron-containing VaSTS11d was better expressed in several generations of transgenic A. thaliana than VaSTS11c and also exhibited a lower level of cytosine methylation. As a result, the content of stilbenes in the VaSTS11d-transgenic plants was much higher than in the VaSTS11c-transgenic plants. Similarly, the best efficiency in increasing the content of stilbenes was also observed in grapevine cell cultures overexpressing the intron-containing VaSTS11d transcript. Thus, the results indicate that an intron sequence with regulatory elements can have a strong positive effect on both transgene expression level and its biological functions in plants and plant cell cultures. Full article

(This article belongs to the Special Issue Plant Tissue and Organ Cultures for Crop Improvement in Omics Era)

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9 pages, 1035 KiB

Open AccessArticle

Optimized Regeneration of Petunia Protoplast and Its Association with Tissue Identity Regulators

by Luhua Tu, Saminathan Subburaj, Kayoun Lee, Yongsam Jeon, Fanzhuang Yan, Jian Yao, Young-Sun Kim, Ok-Jae Koo and Geung-Joo Lee

Horticulturae 2023, 9(2), 216; https://doi.org/10.3390/horticulturae9020216 - 6 Feb 2023

Cited by 4 | Viewed by 2469

Abstract

The popular ornamental plant Petunia is also a valuable model plant in tissue culture. Cellular conversions during differentiation and regeneration have been investigated using various combinations of phytohormones; however, studies on genes for reprogramming toward desired tissue identities have been limited. In this study, we isolated Petunia protoplasts and cultured them in the callus, rooting, or shooting stages, which were used to establish the optimal protoplast culture conditions and to identify genes that epigenetically function as tissue identifiers. The optimal conditions for plasmolysis and enzyme digestion to obtain healthy protoplasts were compared, in which combinations of Viscozyme, Celluclast, and Pectinex (VCP) enzymes were more efficient in isolating protoplasts when followed by 21 to 25% sucrose purification and washing processes. The filtered and washed protoplasts started to divide at 1 day and developed into colonies after 3 weeks of culture, which showed higher efficiency in the Murashige and Skoog (MS) salt culture media compared to that in the Kao and Michayluk (KM) salt media. The pluripotent colonies formed calli on the solid medium supplemented with 3% sucrose after 4 weeks, and were destined to the same cell mass, rooting, or shooting on the regeneration medium. Three epigenetic controllers, ATXR2, ATX4A, and ATX4B, were highly expressed in calli, shoots, and organs of shoots and roots, respectively, confirming that dedifferentiation and regeneration of tissue identity is plastic. Full article

(This article belongs to the Special Issue Plant Tissue and Organ Cultures for Crop Improvement in Omics Era)

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15 pages, 1280 KiB

Open AccessArticle

In Vitro Propagation and Phytochemistry of Thymol-Producing Plants from a Horticultural Form of Thymus × josephi-angeli Mansanet & Aguil. (Lamiaceae)

by Esther Asensio, Roberto de Medinacelli Juan-Méndez and Jorge Juan-Vicedo

Horticulturae 2022, 8(12), 1188; https://doi.org/10.3390/horticulturae8121188 - 13 Dec 2022

Cited by 5 | Viewed by 1941

Abstract

Thymus L. is of great interest in horticulture as ornamentals, spices, and medicinal plants, as well as in the extracts industry due to the richness in bioactive specialized metabolites. The natural hybrid T. × josephi-angeli Mansanet & Aguil. is produced in Spain, as its horticultural forms are very popular for domestic uses and gardening. However, its micropropagation and chemical composition have not been studied yet. Therefore, the main objective of this work was to develop a micropropagation procedure for T. × josephi-angeli, and to check whether the in vitro culture had an impact on the chemical profile of the plants. The results showed a high initiation rate (>91%) after two sterilization treatments were applied. Moreover, a micropropagation rate of around 21 new rooted explants per culture cycle was obtained in treatment M7 (Murashige and Skoog with 0.064 µM 6-(γ,γ-Dimethylallylamino)purine) when compared to the other 10 treatments performed. Acclimatization was successful in all three approaches tested (>75%), and all plants kept growing after 4 months of outdoor cultivation. Finally, 36 volatiles were identified, and the content of major compounds remained not statistically different in acclimatized plants when compared to the wild-type plants according to the analyses made by HS-SPME-GC/MS and SPME-GC/MS. This chemical stability points out the uniformity of the microplants and the suitability of the procedure applied in this study for T.×josephi-angeli horticultural production using in vitro techniques. Full article

(This article belongs to the Special Issue Plant Tissue and Organ Cultures for Crop Improvement in Omics Era)

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18 pages, 3993 KiB

Open AccessArticle

Micropropagation of Vaccinium corymbosum L.: An Alternative Procedure for the Production of Secondary Metabolites

by Doina Clapa, Silvia-Amalia Nemeș, Floricuța Ranga, Monica Hârța, Dan-Cristian Vodnar and Lavinia-Florina Călinoiu

Horticulturae 2022, 8(6), 480; https://doi.org/10.3390/horticulturae8060480 - 28 May 2022

Cited by 8 | Viewed by 4023

Abstract

In vitro culture has become a dependable approach for the mass production of plant material as the market for innovative plant-derived medicinal approaches has grown significantly. Furthermore, because it permits manipulation of biosynthetic routes to boost the production and accumulation of certain compounds, this technology has enormous potential for the manufacture of natural bioactive chemicals. As a result, the goal of this study was to develop an efficient micropropagation system for biomass production and to investigate the accumulation of bioactive compounds from Vaccinium corymbosum L., Duke and Hortblue Petite cultivars. Two in vitro plant tissue culture systems were used for shoots production: a solid medium (5 g/L Plant agar) and liquid medium (Plantform bioreactor). The culture medium used was Woddy Plant Medium (WPM) supplemented with two growth regulators: 0.5 mg/L and 1 mg/L zeatina (Z) and 5 mg/L N6-(2-Isopentenyl) adenine (2iP). The content of phenolic compounds, carotenoids, and chlorophylls of the in vitro shoot extracts were examined via the HPLC-DAD-MS/MS technique. The results showed that cv. Hortblue Petite produced a higher amount of biomass compared with cv. Duke, on all variants of culture media in both systems (solid and liquid), while the shoots extract of the Duke variety in the liquid culture system (under all concentrations of growth regulators) had the highest content of total phenolic compounds (16,665.61 ± 424.93 μg/g). In the case of the lipophilic compounds analysed (chlorophylls and carotenoids), the solid medium reported the highest values, whereas media supplemented with 0.5 mg/L Z was proved to have the richest total content for both cultivars. Full article

(This article belongs to the Special Issue Plant Tissue and Organ Cultures for Crop Improvement in Omics Era)

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27 pages, 855 KiB

Open AccessEditor’s ChoiceReview

Biotechnologies and Strategies for Grapevine Improvement

by Anca Butiuc-Keul and Ana Coste

Horticulturae 2023, 9(1), 62; https://doi.org/10.3390/horticulturae9010062 - 4 Jan 2023

Cited by 13 | Viewed by 5290

Abstract

Grapevine (Vitis vinifera subsp. vinifera) is one of the most widespread and economically important perennial fruit crops in the world. Viticulture has changed over the years in response to changing environmental conditions and market demands, triggering the development of new and improved varieties to ensure the crop’s sustainability. The aim of this review is to provide a perspective on the recent developments in biotechnology and molecular biology and to establish the potential of these technologies for the genetic improvement of grapevine. The following aspects are discussed: (i) the importance of molecular marker-based methods for proper cultivar identification and how NGS-based high-throughput technologies have greatly benefited the development of genotyping techniques, trait mapping, and genomic selection; (ii) the recent advances in grapevine regeneration, genetic transformation, and genome editing, such as new breeding technology approaches for enhanced grapevine yield, quality improvement, and the selection of valuable varieties and cultivars. The specific problems and challenges linked to grapevine biotechnology, along with the importance of integrating classical and new technologies, are highlighted. Full article

(This article belongs to the Special Issue Plant Tissue and Organ Cultures for Crop Improvement in Omics Era)

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28 pages, 5354 KiB

Open AccessEditor’s ChoiceReview

An Academic and Technical Overview on Plant Micropropagation Challenges

by Neama Abdalla, Hassan El-Ramady, Mayada K. Seliem, Mohammed E. El-Mahrouk, Naglaa Taha, Yousry Bayoumi, Tarek A. Shalaby and Judit Dobránszki

Horticulturae 2022, 8(8), 677; https://doi.org/10.3390/horticulturae8080677 - 25 Jul 2022

Cited by 46 | Viewed by 22166

Abstract

The production of micropropagated plants in plant-tissue-culture laboratories and nurseries is the most important method for propagation of many economic plants. Micropropagation based on tissue-culture technology involves large-scale propagation, as it allows multiplication of a huge number of true-to-type propagules in a very short time and in a very limited space, as well as all year round, regardless of the climate. However, applying plant-tissue-culture techniques for the commercial propagation of plants may face a lot of obstacles or troubles that could result from technical, biological, physiological, and/or genetical reasons, or due to overproduction or the lack of facilities and professional technicians, as shown in the current study. Moreover, several disorders and abnormalities are discussed in the present review. This study aims to show the most serious problems and obstacles of plant micropropagation, and their solutions from both scientific and technical sides. This review, as a first report, includes different challenges in plant micropropagation (i.e., contamination, delay of subculture, burned plantlets, browning, in vitro rooting difficulty, somaclonal variations, hyperhydricity, shoot tip necrosis, albino plantlets, recalcitrance, shoot abnormalities, in vitro habituation) in one paper. Most of these problems are related to scientific and/or technical reasons, and they could be avoided by following the micropropagation protocol suitable for each plant species. The others are dominant in plant-tissue-culture laboratories, in which facilities are often incomplete, or due to poor infrastructure and scarce funds. Full article

(This article belongs to the Special Issue Plant Tissue and Organ Cultures for Crop Improvement in Omics Era)

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