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New Insight into Research in In Vitro Plants Propagation
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New Insight into Research in In Vitro Plants Propagation

A special issue of Plants (ISSN 2223-7747). This special issue belongs to the section "Plant Development and Morphogenesis".

Deadline for manuscript submissions: closed (20 October 2022) | Viewed by 31566

Special Issue Editors

Dr. Monika Tuleja

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Guest Editor
Department of Plant Cytology and Embryology, Institute of Botany, Jagiellonian University, Gronostajowa 9, 30-387 Kraków, Poland
Interests: in vitro plant cultures; aspects of in vitro morphogenesis of selected species; including apomictic species; stress in vitro; the role of ethylene and other regulators of plant growth and development
Special Issues, Collections and Topics in MDPI journals
Dr. Hasan Mehraj

E-Mail Website
Guest Editor
Graduate School of Agricultural Science, Kobe University, Rokkodai, Nada-ku, Kobe 657-8501, Japan
Interests: plant breeding; genotypes to phenotype mechanism; heterosis; epigenetics; plant tissue culture
Special Issues, Collections and Topics in MDPI journals
Dr. Saroj Kumar Sah

E-Mail Website
Guest Editor
Department of Biology, Brookhaven National Laboratory, Upton, NY, USA
Interests: plant stress responses (abiotic stress); ABA signaling; plant physiology; plant tissue culture; genetic transformation

Special Issue Information

Dear Colleagues,

Plants possess the ability to callus tissue and cause tumor induction and proliferation. In vivo tumor formation is induced by plant wounding and is intend to heal the injuries. This often results in the accumulation of pathogen-related compounds and is thus thought to prevent infection and water loss. Tumors induced by pathogens called "crown gall” are a unique source of totipotent cells. In vitro callus, an unorganized tissue consisting of an amorphous mass of loosely arranged thin-walled parenchymatous cells, can be obtained from almost any part of the plant through the dedifferentiation of differentiated plant tissue in response to various biotic and abiotic stimuli. Callus culture provides information about the plant morphogenic potency and serves as a good source material for protoplast isolation, as well as the inoculum for the initiation of suspension cultures, and can be maintained indefinitely in in vitro conditions. Presently, callus is widely and succesful used in biotechnology; in the production of different kinds of primary and secondary metabolities; and to scale up bioactive compounds production for pharmaceutical, cosmetic, and food applications. The goals of callus culture are also embryogenesis and organogenesis production, which is possible becouse many callus cells are totipotent, thus being able to regenerate the whole plant body. Based on this ability, callus culture facilitates the amplification of limiting plant material and offers tools for genetic cell transformation, which are not only much more rapid than conventional breeding, but also give rise to novel genes and genotypes. What is more, such genetically modified callus cultures can be a source of novel bioactive secondary metabolites and can lead to the generation of plants with improved resistance against salt, draft, diseases, and pests. Callus cultures also find applications in stress response studies at a cellular level. Despite the extensive use of callus, knowledge about its molecular and physiological regulation along with developmental aspects is still unsufficient. That is why the understanding of genetic and physiological mechanisms that underline the callus induction and proliferation should be the ultimate goal of modern botany, which will allow for the more effective use of callus in scientific research and in numerous industries.

Dr. Monika Tuleja
Dr. Hasan Mehraj
Dr. Saroj Kumar Sah
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. 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

  • biotechnology
  • in vitro callus culture
  • morphogenetic response
  • secondary metabolites
  • totiopotent
  • stress reaction
  • transformation

Published Papers (7 papers)

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Research

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10 pages, 1657 KiB  
Article
Mulberroside F from In Vitro Culture of Mulberry and the Potential Use of the Root Extracts in Cosmeceutical Applications
by Jiralapat Thamrongwatwongsa, Nattaya Pattarapipatkul, Titiradsadakorn Jaithon, Ananya Jindaruk, Atchara Paemanee, Nattanan Panjaworayan T-Thienprasert and Wannarat Pornsiriwong Phonphoem
Plants 2023, 12(1), 146; https://doi.org/10.3390/plants12010146 - 28 Dec 2022
Cited by 1 | Viewed by 2126
Abstract
Mulberry (Morus spp.) is primarily used in sericulture, and its uses also extend to the food, pharmaceutical, and cosmetic industries. Mulberry extracts are rich in many bioactive compounds that exhibit a wide range of biological properties. Mulberroside F (Moracin M-6, 3′-di-O-β-D-glucopyranoside), one [...] Read more.
Mulberry (Morus spp.) is primarily used in sericulture, and its uses also extend to the food, pharmaceutical, and cosmetic industries. Mulberry extracts are rich in many bioactive compounds that exhibit a wide range of biological properties. Mulberroside F (Moracin M-6, 3′-di-O-β-D-glucopyranoside), one of the bioactive compounds found in mulberry, has previously been reported as a whitening agent by inhibiting melanin synthesis and exhibiting antioxidant effects. However, there is still limited information on the presence of this compound in plants cultured in vitro. In this study, the mulberroside F content, biochemical, and cytotoxic properties of the extracts from mulberry cultured in vitro were determined. The results revealed that both root and callus were found to be a potential source of mulberroside F. Furthermore, the mulberroside F content was positively correlated with the inhibitory effects on tyrosinase activity. Cell viability assay also revealed that crude extract of the mulberry root has no cytotoxicity in both human keratinocyte cell line (HaCaT) and Vero cells. Taken together, mulberry tissue culture represents a possible alternative and continuous production of mulberroside F, which could be further utilized in cosmeceutical applications. Full article
(This article belongs to the Special Issue New Insight into Research in In Vitro Plants Propagation)
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17 pages, 3039 KiB  
Article
Genetic Evaluation of In Vitro Micropropagated and Regenerated Plants of Cannabis sativa L. Using SSR Molecular Markers
by Kostas Ioannidis, Ioanna Tomprou, Vangelis Mitsis and Polyxeni Koropouli
Plants 2022, 11(19), 2569; https://doi.org/10.3390/plants11192569 - 29 Sep 2022
Cited by 7 | Viewed by 4227
Abstract
Simple sequence repeat (SSR) markers were used to evaluate the genetic stability of the acclimatized micropropagated and regenerated plants of a high cannabidiol (H-CBD) and a high cannabigerol (H-CBG) variety of Cannabis sativa L. Shoot regeneration and proliferation were achieved by culturing calli [...] Read more.
Simple sequence repeat (SSR) markers were used to evaluate the genetic stability of the acclimatized micropropagated and regenerated plants of a high cannabidiol (H-CBD) and a high cannabigerol (H-CBG) variety of Cannabis sativa L. Shoot regeneration and proliferation were achieved by culturing calli in Murashige and Skoog basal medium (MS) supplemented with several concentrations of 6-benzyladenine (BA) or thidiazuron (TDZ). Calli derived mostly from stem explants, rather than leaves, cultured on MS supplemented with 2,4-Dichlorophenoxyacetic acid (2,4-D) or combination of kinetin (KIN) with 1-Naphthaleneacetic acid (NAA) or 2,4-D. Rooting of the regenerated plantlets accomplished on half-strength MS medium supplemented with indole-3-butyric acid (IBA). Previous studies performed have developed an efficient in vitro micropropagation protocol for mass production. Both in vitro methodologies can be employed in genetic breeding via molecular techniques. The genetic stability of micropropagated and regenerated plants was accomplished using twelve SSR primer pairs that produced reproducible and clear bands, ranging from 90 to 330 bp in size, and resulted in amplification of one or two alleles, corresponding to homozygous or heterozygous individuals. The SSR amplification products were monomorphic across all the micropropagated and regenerated plants and comparable to mother plants. The monomorphic banding pattern confirmed the genetic homogeneity of the in vitro cultured acclimatized and mother plants as no somaclonal variation was detected in clones for these specific SSRs. Our results evidently suggest that the developed culture protocols for in vitro multiplication is appropriate and applicable for clonal mass propagation of the C. sativa varieties and demonstrate the reliability of this in vitro propagation system. Full article
(This article belongs to the Special Issue New Insight into Research in In Vitro Plants Propagation)
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15 pages, 3724 KiB  
Article
Induction, Proliferation, Regeneration and Kinsenoside and Flavonoid Content Analysis of the Anoectochilus roxburghii (Wall.) Lindl Protocorm-like Body
by Hongzhen Wang, Xinying Chen, Xiaoyun Yan, Zhixia Xu, Qingsong Shao, Xueqian Wu, Lingjuan Tou, Li Fang, Mingming Wei and Huasen Wang
Plants 2022, 11(19), 2465; https://doi.org/10.3390/plants11192465 - 21 Sep 2022
Cited by 2 | Viewed by 1688
Abstract
Anoectochilus roxburghii (Wall.) Lindl has been used in Chinese herbal medicine for treating various ailments. However, its wild resources are endangered, and artificial cultivation of the plant is limited by the low regeneration rate of conventional propagation methods. The lack of A. roxburghii [...] Read more.
Anoectochilus roxburghii (Wall.) Lindl has been used in Chinese herbal medicine for treating various ailments. However, its wild resources are endangered, and artificial cultivation of the plant is limited by the low regeneration rate of conventional propagation methods. The lack of A. roxburghii resources is detrimental to the commercial production of the plant and kinsenoside, which is unique to Anoectochilus species. To develop highly efficient methods for A. roxburghii micropropagation and find alternative resources for kinsenoside production, we created an induction, proliferation, and regeneration of PLBs (IPR-PLB) protocol for A. roxburghii. We also analyzed the kinsenoside and flavonoid contents during the induction and proliferation of PLBs. The best media of IPR-PLB for PLB induction and proliferation (secondary PLB induction and proliferation), shoot formation, and rooting medium were Murashige and Skoog (MS) + 3 mg/L 6-benzylaminopurine (6-BA) + 0.5 mg/L naphthaleneacetic acid (NAA) + 0.8 mg/L zeatin (ZT) + 0.2 mg/L 2,4-dichlorophenoxyacetic acid (2, 4-D), MS + 3 mg/L 6-BA + 0.5 mg/L NAA, and MS + 0.5 mg/L NAA, respectively. On these optimized media, the PLB induction rate was 89 ± 2.08%, secondary PLB induction rate was 120 ± 5%, secondary PLB proliferation rate was 400 ± 10% and 350 ± 10 % in terms of the quantity and biomass at approximately 1 month, shoot induction rate was 10.5 shoots/PLB mass, and root induction rate was 98%. All plantlets survived after acclimation. Darkness or weak light were essential for PLB proliferation, and light was crucial for PLB differentiation on these optimized media. The kinsenoside contents of PLBs and secondary PLBs were 10.38 ± 0.08 and 12.30 ± 0.08 mg/g fresh weight (FW), respectively. Moreover, the peak kinsenoside content during the proliferation of secondary PLBs was 34.27 ± 0.79 mg/g FW, which was slightly lower than that of the whole plant (38.68 ± 3.12 mg/g FW). Two flavonoids exhibited tissue- or temporal-specific accumulation patterns, and astragalin accumulated exclusively during the first 2 weeks of cultivation. The IPR-PLB protocol for A. roxburghii may facilitate the efficient micropropagation of A. roxburghii plants. Furthermore, the PLBs are a good alternative resource for kinsenoside production. Full article
(This article belongs to the Special Issue New Insight into Research in In Vitro Plants Propagation)
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17 pages, 3183 KiB  
Article
An Alternative In Vitro Propagation Protocol of Cannabis sativa L. (Cannabaceae) Presenting Efficient Rooting, for Commercial Production
by Kostas Ioannidis, Ioanna Tomprou and Vangelis Mitsis
Plants 2022, 11(10), 1333; https://doi.org/10.3390/plants11101333 - 18 May 2022
Cited by 4 | Viewed by 5097
Abstract
An alternative in vitro propagation protocol for medical Cannabis sativa L. cultivars for pharmaceutical industrial use was established. The aim of the protocol was to reduce the culture time, offering healthy and aseptic propagating material, while making the whole process more economic for [...] Read more.
An alternative in vitro propagation protocol for medical Cannabis sativa L. cultivars for pharmaceutical industrial use was established. The aim of the protocol was to reduce the culture time, offering healthy and aseptic propagating material, while making the whole process more economic for industrial use. The propagation procedure was performed using plastic autoclavable vented and non-vented vessels, containing porous rooting fine-milled sphagnum peat moss-based sponges, impregnated in ½ Murashige and Skoog liquid growth medium, supplemented with indole-3-butyric acid (IBA) at various concentrations (0, 2.46, 4.92, and 9.84 µM) or by dipping nodal cuttings into 15 mM IBA aqueous solution. The highest average root numbers per cutting, 9.47 and 7.79 for high cannabidiol (H_CBD) and high cannabigerol (H_CBG) varieties, respectively, were achieved by dipping the cuttings into IBA aqueous solution for 4 min and then placing them in non-vented vessels. The maximum average root length in H_CBD (1.54 cm) and H_CBG (0.88 cm) was ascertained using 2.46 μM filter sterilized IBA in non-vented vessels. Filter-sterilized IBA at concentrations of 2.46 μM in vented and 4.92 μM in non-vented vessels displayed the maximum average rooting percentages in H_CBD (100%) and H_CBG (95.83%), respectively. In both varieties, maximum growth was obtained in non-vented vessels, when the medium was supplemented with 4.92 μM filter-sterilized IBA. Significant interactions between variety and vessel type and variety and IBA treatments were observed in relation to rooting traits. Approximately 95% of plantlets were successfully established and acclimatized in field. This culture system can be used not only for propagating plant material at an industrial scale but also to enhance the preservation and conservation of Cannabis genetic material. Full article
(This article belongs to the Special Issue New Insight into Research in In Vitro Plants Propagation)
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14 pages, 2678 KiB  
Article
Low Concentration of Anti-Auxin and Anti-Fungal Agent Accelerates the PLB Regeneration of Dendrobium okinawense under Green LED
by Hasan Mehbub, Kazuhiko Shimasaki and Hasan Mehraj
Plants 2022, 11(8), 1082; https://doi.org/10.3390/plants11081082 - 15 Apr 2022
Cited by 1 | Viewed by 2177
Abstract
Dendrobium okinawense is an endangered epiphytic orchid, and there has been no scientific report so far on its propagation. Protocorm is a mass of cells, and protocorm-like bodies (PLBs) are lookalike protocorms produced by vegetative explants in vitro. Regeneration of PLBs is a [...] Read more.
Dendrobium okinawense is an endangered epiphytic orchid, and there has been no scientific report so far on its propagation. Protocorm is a mass of cells, and protocorm-like bodies (PLBs) are lookalike protocorms produced by vegetative explants in vitro. Regeneration of PLBs is a widely used technique for orchid micropropagation. We used different light-emitting diodes (LEDs) for the PLB regeneration of D. okinawense. The number of PLBs and fresh weight were increased by 81.1% and 80.8%, respectively, under green LED over the white fluorescent (WF) light. We added different concentrations of PCIB (p-Chlorophenoxyisobutyric acid, an anti-auxin) and HMI (3-Hydroxy-5-methyl isoxazole, an anti-fungal agent) in culture media. The number of PLBs was increased in media having 0.01 mg/L of PCIB (35.9%) compared to control (no PCIB), whereas 19.3% increased in media having 0.01 mL/L of HMI compared to control (no HMI). Green LED in combination with 0.01 mg/L of PCIB significantly increased the number of PLBs (69.0%) compared to the WF–without PCIB combination. In LEDs-PCIB and LED-HMI combinations, HMI did not show better PLBs regeneration compared with PCIB. The results suggested that a combination of low concentrations of PCIB and green LED have the potential to accelerate PLB regeneration of D. okinawense. Full article
(This article belongs to the Special Issue New Insight into Research in In Vitro Plants Propagation)
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Review

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33 pages, 3966 KiB  
Review
Tissue Culture in Ornamentals: Cultivation Factors, Propagation Techniques, and Its Application
by Hasan Mehbub, Ayasha Akter, Mst. Arjina Akter, Mohammad Shamim Hasan Mandal, Md. Ashraful Hoque, Monika Tuleja and Hasan Mehraj
Plants 2022, 11(23), 3208; https://doi.org/10.3390/plants11233208 - 23 Nov 2022
Cited by 13 | Viewed by 11705
Abstract
Ornamentals come in a variety of shapes, sizes, and colors to suit a wide range of climates, landscapes, and gardening needs. Compared to demand, a shortage of plant materials and diversity force the search for solutions for their constant acquisition and improvement to [...] Read more.
Ornamentals come in a variety of shapes, sizes, and colors to suit a wide range of climates, landscapes, and gardening needs. Compared to demand, a shortage of plant materials and diversity force the search for solutions for their constant acquisition and improvement to increase their commercial value, respectively. In vitro cultures are a suitable solution to meet expectations using callus culture, somatic embryogenesis, protoplast culture, and the organogenesis of protocorm-like bodies; many of these techniques are commercially practiced. Factors such as culture media, explants, carbohydrates, plant growth regulators, and light are associated with the success of in vitro propagation. Techniques, especially embryo rescue and somatic hybridization, are widely used to improve ornamentals. The development of synthetic seed allows season-independent seed production and preservation in the long term. Despite the advantages of propagation and the improvement of ornamentals, many barriers still need to be resolved. In contrast to propagation and crop developmental studies, there is also a high scope for molecular studies, especially epigenetic changes caused by plant tissue culture of ornamentals. In this review, we have accumulated and discussed an overall update on cultivation factors, propagation techniques in ornamental plant tissue culture, in vitro plant improvement techniques, and future perspectives. Full article
(This article belongs to the Special Issue New Insight into Research in In Vitro Plants Propagation)
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19 pages, 794 KiB  
Review
Biotechnological Research Progress in Jatropha, a Biodiesel-Yielding Plant
by Jameel M. Al-Khayri, Wudali N. Sudheer, Thenmozhi R. Preetha, Praveen Nagella, Adel A. Rezk and Wael F. Shehata
Plants 2022, 11(10), 1292; https://doi.org/10.3390/plants11101292 - 12 May 2022
Cited by 6 | Viewed by 2785
Abstract
Environmental pollution is one of the most pressing challenges in today’s world. The main cause of this pollution is fuel emissions from automobiles and other sources. As industrialization progresses, we will be unable to compromise on the use of energy to power heavy [...] Read more.
Environmental pollution is one of the most pressing challenges in today’s world. The main cause of this pollution is fuel emissions from automobiles and other sources. As industrialization progresses, we will be unable to compromise on the use of energy to power heavy machines and will be forced to seek out the best options. As a consequence, utilizing green fuel, such as biodiesel derived from natural sources, is a realistic option. Jatropha curcas L. (Euphorbiaceae) is recognized as the greatest feedstock for biodiesel production throughout the world, and it has gained a huge market value in the recent years. Conventional cultivation alone will not be sufficient to meet the global need for the plant’s biomass for the production of biodiesel. Adoption of plant tissue culture techniques that improve the biomass availability is an immediate need. The present review provides detailed information regarding in-vitro plant propagation (direct and indirect organogenesis), somatic embryogenesis, and acclimatization protocols of plantlets for stabilized production of biomass. The review also focuses on biotechnological approaches such as gene transformation studies, production of haploids, and double haploids for developing elite germplasm for high biomass and improved traits for the production of biodiesel. Full article
(This article belongs to the Special Issue New Insight into Research in In Vitro Plants Propagation)
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