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17 pages, 4877 KiB

Open AccessArticle

In Vitro Morphogenesis of Tobacco: Modulation of Endogenous Growth Regulators by Tulsi (Holy Basil)

by Vanessa Vongnhay, Mukund R. Shukla, Murali-Mohan Ayyanath, Karthika Sriskantharajah and Praveen K. Saxena

Plants 2024, 13(14), 2002; https://doi.org/10.3390/plants13142002 - 22 Jul 2024

Viewed by 731

Abstract

Plant growth regulators (PGRs) play a vital role in the induction of morphogenesis in vitro. Synthetic PGRs are commonly used to induce organogenesis and somatic embryogenesis from various explants, while natural substances are rarely utilized. This study aimed to enhance the regenerative response [...] Read more.

Plant growth regulators (PGRs) play a vital role in the induction of morphogenesis in vitro. Synthetic PGRs are commonly used to induce organogenesis and somatic embryogenesis from various explants, while natural substances are rarely utilized. This study aimed to enhance the regenerative response in Nicotiana tabacum leaf explants using Tulsi (Ocimum sanctum) leaf extract and to elucidate the biochemical interactions during modulation of endogenous plant growth regulators, including indole-3-acetic acid (IAA), abscisic acid (ABA), zeatin, and 6-(γ, γ-dimethylallylamino) purine (2iP). Tulsi leaf extract significantly improved shoot production through interactions between endogenous hormones and those present in the extract, which enhanced stress mitigation. The 20% Tulsi leaf extract treatment produced significantly more shoots than the control, coinciding with increased endogenous IAA and zeatin levels starting on day 10 in culture. Furthermore, ABA and zeatin concentrations increased on days 15 and 25, respectively, in the 20% Tulsi extract treatment, suggesting their role in the induction of somatic embryo-like structures. ABA likely acts as an activator of stress responses, encouraging the development of these structures. Additionally, 2iP was involved in the induction of both forms of regeneration in the 10% and 20% extract treatments, especially in combination with ABA. These results suggest that Tulsi leaf extract holds promising potential as a natural supplement for increasing plant regeneration in vitro and advancing our understanding of how natural extracts of plant origin can be harnessed to optimize plant regeneration processes in vitro. Full article

(This article belongs to the Special Issue In Vitro Morphogenesis of Plants)

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17 pages, 2115 KiB

Open AccessArticle

Enhanced In Vitro Plant Morphogenesis of Tobacco: Unveiling Indoleamine-Modulated Adaptogenic Properties of Tulsi (Ocimum sanctum L.)

by Vanessa Vongnhay, Mukund R. Shukla, Murali-Mohan Ayyanath, Karthika Sriskantharajah and Praveen K. Saxena

Plants 2024, 13(10), 1370; https://doi.org/10.3390/plants13101370 - 15 May 2024

Cited by 1 | Viewed by 1058

Abstract

The medicinal plant tulsi (Ocimum sanctum L.) is acknowledged for its invigorating and healing properties that enhance resilience to stress in various human and animal models by modulating antioxidant compounds. While extensive research has documented these effects in humans, the adaptogenic potential [...] Read more.

The medicinal plant tulsi (Ocimum sanctum L.) is acknowledged for its invigorating and healing properties that enhance resilience to stress in various human and animal models by modulating antioxidant compounds. While extensive research has documented these effects in humans, the adaptogenic potential of tulsi in stressful in vitro plant systems has not been explored. This study aimed to elucidate the adaptogenic properties of tulsi leaf extract on the in vitro regeneration of tobacco leaf explants through an investigation of the indoleamines at different developmental stages. Shoot regeneration from leaf explants on the medium supplemented with tulsi extract (20%) was compared to the control, and the differences in indoleamine compounds were analyzed using ultra-performance liquid chromatography. Treatment of the explants with the extract resulted in an almost two-fold increase in the number of regenerants after four weeks of culture, and 9% of the regenerants resembled somatic embryo-like structures. The occurrence of browning in the extract-treated explants stopped on day 10, shoots began to develop, and a significant concentration of tryptamine and N-acetyl-serotonin accumulated. A comparative analysis of indoleamine compounds in intact and cut tobacco leaves also revealed the pivotal role of melatonin and 2-hydroxymelatonin functioning as antioxidants during stress adaptation. This study demonstrates that tulsi is a potent adaptogen that is capable of modulating plant morphogenesis in vitro, paving the way for further investigations into the role of adaptogens in plant stress biology. Full article

(This article belongs to the Special Issue In Vitro Morphogenesis of Plants)

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16 pages, 3128 KiB

Open AccessArticle

The WOX Genes from the Intermediate Clade: Influence on the Somatic Embryogenesis in Medicago truncatula

by Daria V. Yakovleva, Elena P. Efremova, Kirill V. Smirnov, Veronika Y. Simonova, Zakhar S. Konstantinov, Varvara E. Tvorogova and Ludmila A. Lutova

Plants 2024, 13(2), 223; https://doi.org/10.3390/plants13020223 - 13 Jan 2024

Cited by 1 | Viewed by 1456

Abstract

Transcription factors from the WOX family are well-known regulators of cell proliferation and differentiation in plants. Herein, we focused on several WOX genes from the intermediate clade and checked their impact on somatic embryogenesis using the model legume object Medicago truncatula. As [...] Read more.

Transcription factors from the WOX family are well-known regulators of cell proliferation and differentiation in plants. Herein, we focused on several WOX genes from the intermediate clade and checked their impact on somatic embryogenesis using the model legume object Medicago truncatula. As a result, we show that MtWOX9-1 overexpression not only stimulates somatic embryogenesis in the embryogenic M. truncatula line, as it was shown previously, but can also induce somatic embryogenesis in the non-embryogenic line. Other intermediate clade WOX, including the close paralog of MtWOX9-1, as well as WOX11 homologs, did not have any significant impact on somatic embryogenesis in our in vitro cultivation system. Together, our results give new information about the diversity of the WOX family proteins and their specific functions. These data can be used for the search of new regeneration stimulators. Full article

(This article belongs to the Special Issue In Vitro Morphogenesis of Plants)

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14 pages, 6682 KiB

Open AccessArticle

Physiological and Physical Strategies to Minimize Damage at the Branch–Stem Junction of Trees: Using the Finite Element Method to Analyze Stress in Four Branch–Stem Features

by Tung-Chi Liu, Yi-Sen Peng and Bai-You Cheng

Plants 2023, 12(23), 4060; https://doi.org/10.3390/plants12234060 - 3 Dec 2023

Viewed by 1344

Abstract

This study analyzed the mechanical and physiological strategies associated with four features in the branch–stem junction of a tree, namely the U-shaped branch attachment, the branch collar, the branch bark ridge, and the roughened lower stem. Models were established for each stage of [...] Read more.

This study analyzed the mechanical and physiological strategies associated with four features in the branch–stem junction of a tree, namely the U-shaped branch attachment, the branch collar, the branch bark ridge, and the roughened lower stem. Models were established for each stage of tree growth by adding these four features sequentially to a base model, and the finite element method (FEM) was employed to create three-dimensional models of an Acer tree’s branch–stem structure for static force analysis. According to the results, the development of the branch collar shifted the point of breakage to the outer part of the collar and, thus, constituted a physiological strategy that prevented decay in the stem. Additionally, the concentration of stress in the branch bark ridge limited the area of tear in the bark following breakage. Finally, the U-shaped branch attachment reduced stress and shifted the point of peak stress toward the branch, while the thickening of the lower stem reduced the overall stress. The development of these features, including the spatial positioning of the branch bark ridge and branch collar, resulted in two breakage points constituting a physical and a physiological strategy that limited damage to the tree and protected the xylem structure. This is the part that has been challenging to decipher in previous discussions of tree-related self-protection mechanisms. Full article

(This article belongs to the Special Issue In Vitro Morphogenesis of Plants)

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13 pages, 4162 KiB

Open AccessArticle

Biochemical and Molecular Characterization of Musa sp. Cultured in Temporary Immersion Bioreactor

by Christopher A. Sambolín Pérez, Rosalinda Aybar Batista, Sullymar Morales Marrero, Dinorah Andino Santiago, Axel Reyes Colón, Juan A. Negrón Berríos, Ángel Núñez Marrero and Alok Arun

Plants 2023, 12(21), 3770; https://doi.org/10.3390/plants12213770 - 4 Nov 2023

Cited by 1 | Viewed by 1679

Abstract

The genus Musa sp. contains commercially important fleshy fruit-producing plants, including plantains and bananas, with a strong potential for providing food security and sources of revenue to farmers. Concerns with the quality of vegetative tissues along with the possibility of the transmission of [...] Read more.

The genus Musa sp. contains commercially important fleshy fruit-producing plants, including plantains and bananas, with a strong potential for providing food security and sources of revenue to farmers. Concerns with the quality of vegetative tissues along with the possibility of the transmission of phytopathogens makes the availability of healthy plantlets limited for farmers. Micropropagation of plantains offers an alternative to producing large numbers of plantlets. However, conventional methods of micropropagation have high production costs and are labor-intensive. Recently, the temporary immersion bioreactor (TIB) has emerged as an alternative to conventional micropropagation (CM) methods. Our work utilized SEM microscopy (scanning electron microscope) and molecular and biochemical tools (qRT-PCR and ICP-OES) to characterize and compare the morphological properties, elemental composition, and photosynthetic gene expression of plantains cultured on TIB. Additionally, morphological features of growth and propagation rates were analyzed to compare outputs obtained from TIB and CM. Results showed higher growth and multiplication rates for plantlets cultivated in TIB. Gene expression analysis of selected photosynthetic genes demonstrated high transcript abundance of phosphoenolpyruvate carboxylase (PEPC) in plantain tissues obtained by TIB. Elemental composition analysis showed higher content of iron in plantains grown in TIB, suggesting a potential correlation with PEPC expression. These results demonstrate that micropropagation of Musa sp. via the liquid medium in TIB is an efficient and low-cost approach in comparison with solid media in CM. Full article

(This article belongs to the Special Issue In Vitro Morphogenesis of Plants)

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15 pages, 8945 KiB

Open AccessEditor’s ChoiceArticle

Optimization of Callus Induction and Shoot Regeneration from Tomato Cotyledon Explants

by Olha Yaroshko, Taras Pasternak, Eduardo Larriba and José Manuel Pérez-Pérez

Plants 2023, 12(16), 2942; https://doi.org/10.3390/plants12162942 - 14 Aug 2023

Cited by 5 | Viewed by 6726

Abstract

Cultivated tomato (Solanum lycopersicum L.) is one of the most important horticultural crops in the world. The optimization of culture media for callus formation and tissue regeneration of different tomato genotypes presents numerous biotechnological applications. In this work, we have analyzed the [...] Read more.

Cultivated tomato (Solanum lycopersicum L.) is one of the most important horticultural crops in the world. The optimization of culture media for callus formation and tissue regeneration of different tomato genotypes presents numerous biotechnological applications. In this work, we have analyzed the effect of different concentrations of zeatin and indole-3-acetic acid on the regeneration of cotyledon explants in tomato cultivars M82 and Micro-Tom. We evaluated regeneration parameters such as the percentage of callus formation and the area of callus formed, as well as the initiation percentage and the number of adventitious shoots. The best hormone combination produced shoot-like structures after 2–3 weeks. We observed the formation of leaf primordia from these structures after about 3–4 weeks. Upon transferring the regenerating micro-stems to a defined growth medium, it was possible to obtain whole plantlets between 4 and 6 weeks. This hormone combination was applied to other genotypes of S. lycopersicum, including commercial varieties and ancestral tomato varieties. Our method is suitable for obtaining many plantlets of different tomato genotypes from cotyledon explants in a very short time, with direct applications for plant transformation, use of gene editing techniques, and vegetative propagation of elite cultivars. Full article

(This article belongs to the Special Issue In Vitro Morphogenesis of Plants)

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18 pages, 1775 KiB

Open AccessArticle

Morphological and Physiological Responses of Hybrid Aspen (Populus tremuloides Michx. × Populus tremula L.) Clones to Light In Vitro

by Toms Kondratovičs, Mārtiņš Zeps, Diāna Rupeika, Pauls Zeltiņš, Arnis Gailis and Roberts Matisons

Plants 2022, 11(20), 2692; https://doi.org/10.3390/plants11202692 - 12 Oct 2022

Viewed by 1672

Abstract

Micropropagation of fast-growing tree genotypes such as the hybrid aspen (Populus tremuloides Michx. × Populus tremula L.) is increasing. The efficiency of micropropagation depends on the luminaires, hence luminescent electric diodes (LED), which emit light of a narrow spectrum, are gaining popularity. [...] Read more.

Micropropagation of fast-growing tree genotypes such as the hybrid aspen (Populus tremuloides Michx. × Populus tremula L.) is increasing. The efficiency of micropropagation depends on the luminaires, hence luminescent electric diodes (LED), which emit light of a narrow spectrum, are gaining popularity. Mostly, different LEDs are combined to increase the photosynthetic efficiency. However, light also acts as an environmental signal, which triggers specific responses in plants, which are genotype specific, and regarding hybrid aspen, are likely affected by heterosis. In this study, morphological and physiological responses of clones of hybrid aspen with contrasting field performance to the spectral composition of illumination were studied in vitro. Among the 15 variables measured, area of leaves and concentration and ratio of chlorophyll a and b explained most of the variance (58.6%), thereby linking a specific combination of traits to productivity. These traits and their responses to light were affected by heterosis, as indicated by the clone-treatment interaction, particularly for the clone’s moderate productivity. The top-performing clones were little sensitive to illumination due to efficient photosystems. Nevertheless, illumination with wider spectral composition had generally positive effects on plantlet performance. Accordingly, clone-specific illumination protocols and luminaries capable of it are advantageous for the efficiency of micropropagation of hybrid aspen. Full article

(This article belongs to the Special Issue In Vitro Morphogenesis of Plants)

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Review

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40 pages, 569 KiB

Open AccessReview

Multifaceted Role of Nanomaterials in Modulating In Vitro Seed Germination, Plant Morphogenesis, Metabolism and Genetic Engineering

by Ashutosh Pathak, Shamshadul Haq, Neelam Meena, Pratibha Dwivedi, Shanker Lal Kothari and Sumita Kachhwaha

Plants 2023, 12(17), 3126; https://doi.org/10.3390/plants12173126 - 30 Aug 2023

Cited by 7 | Viewed by 2086

Abstract

The agricultural practices of breeding, farm management and cultivation have improved production, to a great extent, in order to meet the food demands of a growing population. However, the newer challenges of climate change, global warming, and nutritional quality improvement will have to [...] Read more.

The agricultural practices of breeding, farm management and cultivation have improved production, to a great extent, in order to meet the food demands of a growing population. However, the newer challenges of climate change, global warming, and nutritional quality improvement will have to be addressed under a new scenario. Plant biotechnology has emerged as a reliable tool for enhancing crop yields by protecting plants against insect pests and metabolic engineering through the addition of new genes and, to some extent, nutritional quality improvement. Plant tissue culture techniques have provided ways for the accelerated clonal multiplication of selected varieties with the enhanced production of value-added plant products to increase modern agriculture. The in vitro propagation method has appeared as a pre-eminent approach for the escalated production of healthy plants in relatively shorter durations, also circumventing seasonal effects. However, there are various kinds of factors that directly or indirectly affect the efficiency of in vitro regeneration like the concentration and combination of growth regulators, variety/genotype of the mother plant, explant type, age of seedlings and other nutritional factors, and elicitors. Nanotechnology as one of the latest and most advanced approaches in the material sciences, and can be considered to be very promising for the improvement of crop production. Nanomaterials have various kinds of properties because of their small size, such as an enhanced contact surface area, increased reactivity, stability, chemical composition, etc., which can be employed in plant sciences to alter the potential and performance of plants to improve tissue culture practices. Implementing nanomaterials with in vitro production procedures has been demonstrated to increase the shoot multiplication potential, stress adaptation and yield of plant-based products. However, nanotoxicity and biosafety issues are limitations, but there is evidence that implies the promotion and further exploration of nanoparticles in agriculture production. The incorporation of properly designed nanoparticles with tissue culture programs in a controlled manner can be assumed as a new pathway for sustainable agriculture development. The present review enlists different studies in which treatment with various nanoparticles influenced the growth and biochemical responses of seed germination, as well as the in vitro morphogenesis of many crop species. In addition, many studies suggest that nanoparticles can be useful as elicitors for elevating levels of important secondary metabolites in in vitro cultures. Recent advancements in this field also depict the suitability of nanoparticles as a promising carrier for gene transfer, which show better efficiency than traditional Agrobacterium-mediated delivery. This review comprehensively highlights different in vitro studies that will aid in identifying research gaps and provide future directions for unexplored areas of research in important crop species. Full article

(This article belongs to the Special Issue In Vitro Morphogenesis of Plants)

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