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Plants
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In Vitro Plant Regeneration and Biotechnological Approaches for Crop Improvement
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In Vitro Plant Regeneration and Biotechnological Approaches for Crop Improvement

A special issue of Plants (ISSN 2223-7747). This special issue belongs to the section "Plant Genetics, Genomics and Biotechnology".

Deadline for manuscript submissions: 28 September 2026 | Viewed by 3641

Special Issue Editor

Dr. Marga Pérez-Jiménez

E-Mail Website
Guest Editor
Department of Plant Biotechnology, Genomics and Breeding, Instituto Murciano de Investigación y Desarrollo Agrario y Alimentario (IMIDA), 30150 Murcia, Spain
Interests: genetic transformation and genome editing in woody and stone fruit species; in vitro regeneration systems in fruit crops; plant tissue culture and micropropagation; production of secondary metabolites through in vitro systems; elicitation strategies to enhance metabolite accumulation
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

I am pleased to announce a forthcoming Special Issue in Plants, focusing on "In Vitro Plant Regeneration and Biotechnological Approaches for Crop Improvement". This issue will showcase cutting-edge research and comprehensive reviews, highlighting recent advances and innovative methodologies in plant tissue culture, genetic engineering, and relevant biotechnological strategies to enhance crop productivity, resilience, and quality.

Given the importance of in vitro technologies and biotechnological tools in modern plant science and agriculture, this Special Issue will provide a valuable platform for researchers to present novel findings, protocols, and perspectives on improving crop performance under diverse conditions.

Contributions, including original research articles, review papers, and short communications on (but not limited to) the following topics, are welcome for submission:

  • Somatic embryogenesis and organogenesis;
  • Genetic transformation and genome editing;
  • Micropropagation and synthetic seed technology;
  • Tissue culture-based stress tolerance studies;
  • Metabolite production through in vitro systems;
  • Advances in culture media and growth regulators.

If you or your collaborators are working on additional, relevant topics, we would welcome discussions to consider your work for submission. Feel free to share this opportunity within your networks as well.

For further details on submission guidelines and deadlines, please refer to the journal’s official website.

I look forward to receiving your valuable contributions.

Dr. Marga Pérez-Jiménez
Guest Editor

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 250 words) can be sent to the Editorial Office for assessment.

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

  • agrobacterium
  • plant tissue culture
  • plant editing
  • organogenesis
  • embryogenesis

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Published Papers (4 papers)

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Research

17 pages, 9520 KB  
Article
Two Optimized Methods for Efficient, Stable and Transient Transformation of Broccoli (Brassica oleracea Var. Italica)
by Alberto Coronado-Martín, Alejandro Atarés, Rosa Porcel, Lynne Yenush and José M. Mulet
Plants 2026, 15(6), 978; https://doi.org/10.3390/plants15060978 - 22 Mar 2026
Viewed by 1193
Abstract
Broccoli (Brassica oleracea var. italica) is an important crop valued for its nutritional and health-promoting properties, yet its biotechnological improvement is limited by low effectivity and genotype-dependent transformation protocols. The absence of reliable transient expression systems further constrains functional genomics and genome-editing [...] Read more.
Broccoli (Brassica oleracea var. italica) is an important crop valued for its nutritional and health-promoting properties, yet its biotechnological improvement is limited by low effectivity and genotype-dependent transformation protocols. The absence of reliable transient expression systems further constrains functional genomics and genome-editing applications. Here, we optimized regeneration and transformation protocols for different broccoli genotypes. Endoreduplication patterns in young tissues were analyzed by flow cytometry to identify suitable explants, and combinations of plant growth regulators were tested to develop an efficient organogenic medium. Stable transformation was achieved via Agrobacterium tumefaciens using nptII and eGFP markers. Cotyledons and hypocotyls up to day 7 showed similar endoreduplication patterns, with abundant 2n cells, but hypocotyls exhibited higher regeneration capacity. The optimized medium supported efficient organogenesis while maintaining diploidy. Transformation efficiency reached 10.4% in ‘S1’ and 2.8% in ‘Naxos’, highlighting genotype dependence. In parallel, a transient expression system was established using cotyledon-derived protoplasts and electroporation-mediated DNA delivery. GFP expression was confirmed through fluorescence microscopy, confocal imaging, and Western blotting. These protocols provide a robust toolkit for broccoli genetic manipulation, facilitating molecular biology studies in the native plant, functional genomics and genome-editing strategies, including CRISPR-based approaches. Full article
(This article belongs to the Special Issue In Vitro Plant Regeneration and Biotechnological Approaches for Crop Improvement)
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19 pages, 7259 KB  
Article
Red and Far-Red LED Lighting Enhances Protoplast-to-Plant Regeneration in Broccoli (Brassica oleracea var. italica)
by Miriam Romero-Muñoz, José Manuel Gambín-Sánchez, Francisco José Vidal-Sánchez, José E. Cos-Terrer and Margarita Pérez-Jiménez
Plants 2026, 15(6), 905; https://doi.org/10.3390/plants15060905 - 14 Mar 2026
Viewed by 585
Abstract
Plants have a remarkable ability to regenerate tissues and organs from single cells, a property that underpins in vitro protoplast regeneration. Efficient protoplast-to-plant regeneration remains a major bottleneck for genome engineering in many crop species, including broccoli (Brassica oleracea var. italica). [...] Read more.
Plants have a remarkable ability to regenerate tissues and organs from single cells, a property that underpins in vitro protoplast regeneration. Efficient protoplast-to-plant regeneration remains a major bottleneck for genome engineering in many crop species, including broccoli (Brassica oleracea var. italica). In this study, we established and optimized a regeneration system for broccoli cv. Claremont by evaluating enzyme composition, light quality, and culture media at successive stages of development. Among the tested enzyme mixtures, 1.5% Cellulase R-10 combined with 0.4% Macerozyme R-10 yielded the highest protoplast viability and recovery. Alginate-embedded protoplasts were cultured under control (dark), blue, and red + far-red LED illumination. Red + far-red treatment significantly enhanced microcolony formation, plating efficiency, and shoot regeneration compared with blue light, whereas blue illumination consistently reduced regenerative performance. The inclusion of activated charcoal in the regeneration medium further increased shoot production. The generalized linear model analyses identified light quality as a significant predictor of both shoot number and regeneration. To our knowledge, this study provides one of the first demonstrations of LED-assisted enhancement of protoplast regeneration in broccoli. The optimized protocol enables whole-plant recovery within approximately 5 months and offers a practical platform for CRISPR-based genome editing and advanced breeding applications in B. oleracea. Full article
(This article belongs to the Special Issue In Vitro Plant Regeneration and Biotechnological Approaches for Crop Improvement)
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20 pages, 2008 KB  
Article
Development of an Efficient Somatic Embryogenesis Protocol for Carica papaya L. Var. TNAU Papaya CO 8 on Different Basal Media
by Shalini Chandrasekar, Kavitha Chinnasamy, Ganga Mathian, Krish K Kumar, Babu Rajendra Prasad, Manoranjitham S. Karuppannan, Selvaraju Kanagarajan and Saraladevi Muthusamy
Plants 2026, 15(6), 893; https://doi.org/10.3390/plants15060893 - 13 Mar 2026
Viewed by 571
Abstract
Papaya (Carica papaya L.) is a highly cross-pollinated crop that exhibits considerable genetic variability when propagated through seeds, resulting in non-true-to-type progeny. Therefore, the development of an efficient in vitro regeneration system is essential for large-scale clonal propagation of elite cultivars. In [...] Read more.
Papaya (Carica papaya L.) is a highly cross-pollinated crop that exhibits considerable genetic variability when propagated through seeds, resulting in non-true-to-type progeny. Therefore, the development of an efficient in vitro regeneration system is essential for large-scale clonal propagation of elite cultivars. In the present study, a highly efficient and reproducible somatic embryogenesis protocol was developed for C. papaya var. TNAU Papaya CO 8 using immature zygotic embryos as explants. This study provides the first comprehensive comparative evaluation of three basal media, viz., Murashige and Skoog Medium, N6 Medium, and Woody Plant Medium, for somatic embryogenesis and plant regeneration in this variety, along with the optimization of polyamine-enriched media for enhanced plantlet recovery. The embryogenic potential of explants was assessed across different stages, including callus induction, somatic embryo development, plant regeneration, shoot elongation, rooting, and acclimatization. Maximum callus induction (81.96%) was observed on half-strength MS medium supplemented with 2,4-Dichlorophenoxyacetic acid under dark conditions, followed by ½ N6 (63.00%) and ½ WPM (58.02%). Somatic embryo initiation was highest on ½ MS medium containing 2.0 mgL−1 2,4-D (77.82%). Somatic embryos developed through distinct globular, heart, torpedo, and cotyledonary stages. Embryo maturation was significantly enhanced on MS medium supplemented with abscisic acid, polyethylene glycol, benzylaminopurine, and proline. The highest plantlet regeneration (85.02%) was achieved on MS medium enriched with putrescine, whereas comparatively lower regeneration was recorded on N6 (75.99%) and WPM (57.97%). Shoot elongation was significantly improved by supplementation with gibberellic acid (1.0 mgL−1). Root induction was optimal on half-strength MS medium containing Indole-3-butyric acid, 1-Naphthaleneacetic acid, phloroglucinol, and activated charcoal, resulting in well-developed roots. Regenerated plantlets were successfully acclimatized in a cocopeat–vermicompost substrate with a survival rate of 74.01%. The optimized protocol provides a reliable and efficient system for large-scale clonal propagation and offers promising applications in genetic transformation and commercial production of papaya var. TNAU papaya CO 8. Full article
(This article belongs to the Special Issue In Vitro Plant Regeneration and Biotechnological Approaches for Crop Improvement)
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18 pages, 23218 KB  
Article
Induction of Embryogenic Callus, Protoplast Isolation, and PEG-Mediated Transformation Protocols in Eucommia ulmoides
by Hongrun Zhou, Zibo Zhou, Jiangyuan Zhang, Haoran Kan, Mengqi Yin, Han Zhang, Luyao Wang, Jie Zhao and Jing Ye
Plants 2026, 15(2), 194; https://doi.org/10.3390/plants15020194 - 8 Jan 2026
Viewed by 899
Abstract
Eucommia ulmoides, a tree species native to China, holds considerable medicinal, ecological, and industrial importance. However, the absence of an efficient and stable genetic transformation system poses significant challenges to gene function studies and molecular breeding in E. ulmoides. Protoplasts, which [...] Read more.
Eucommia ulmoides, a tree species native to China, holds considerable medicinal, ecological, and industrial importance. However, the absence of an efficient and stable genetic transformation system poses significant challenges to gene function studies and molecular breeding in E. ulmoides. Protoplasts, which lack cell walls, serve as effective receptors for transient transformation and are thus ideal for genetic engineering research. In this study, the optimal conditions for callus induction were identified, and formation of the embryogenic callus was confirmed by histological analysis. Furthermore, we developed an efficient protoplast isolation and PEG-mediated transient transformation system using suitable embryogenic callus as the starting material. Our findings revealed that the optimal medium for inducing embryogenic callus was B5 + 1.5 mg/L 6-BA + 0.5 mg/L NAA + 30 g/L sucrose + 7 g/L agar (pH = 5.8). In this medium, the induction rate of callus achieved 97.50%, and the rate of embryogenic callus formation was 86.30%. For protoplast isolation, the best conditions involved enzymatic digestion with 1.5% cellulase R-10 and 1.0% macerozyme R-10 at an osmotic pressure of 0.6 M for 4 h, resulting in 1.82 × 106 protoplasts/g FW with 91.13% viability. The highest transfection efficiency (53.23%) was attained when protoplasts were cultured with 10 µg of plasmid and 40% PEG4000 for 20 min. This study successfully established a stable and efficient system for protoplast isolation and transient transformation in E. ulmoides, offering technical support for exploring somatic hybridisation and transient gene expression in this species. Full article
(This article belongs to the Special Issue In Vitro Plant Regeneration and Biotechnological Approaches for Crop Improvement)
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