In vitro regeneration of bulblet using two and four bulb-scales explants of summer snowflake (Leucojum aestivum L.)

Abstract Leucojum aestivum is a valuable and endangered plant species with bulb scales best suited as explants in micropropagation. In the current study, its micropropagation was investigated by using two different explants and various concentrations and combinations of plant growth regulators (PGRs). Bulbs were first disinfected with benomyl® for 5 hours. After meeting the chilling requirements, two-scale and four-scale explants were provided for direct and indirect organogenesis. Explants were exposed to hot water, 70% ethanol and 2.5% sodium hypochlorite for further disinfestation. Four-scale explants were treated with different concentrations and combinations of naphthaleneacetic acid (NAA), 6-benzyladenine (BA), and kinetin (Kin) for bulblet regeneration. For callogenesis, 0.5 mg L-1 of BA combined with 1, 2, 3, 4, 5 or 6 mg L-1 of 2,4-dichlorophenoxyacetic acid (2,4-D) were applied. Regarding two-scale explants, different combinations and concentrations of BA, Indole-3-butyric acid (IBA) and NAA were used for bulblet induction, and various combinations of Indoleacetic acid (IAA), NAA, 2, 4-D and BA were used for callus induction. None of the two-scale explants responded to the bulblet regeneration and callus induction media. Unlike, four-scale explants regenerated bulblets and roots in the control medium and MS media enriched with different PGRs. Callus was generated on MS medium supplemented with 2,4-D and BA, and indirect regeneration was observed in some cases. On the control medium, the regenerated roots had a natural form, but in PGRs-rich media, they were deformed. Regarding the regeneration percentage, bulblet number and length and root length, no significant differences were found between the control and the best PGR-treatment in each case. Therefore, it seems logical suggesting not to use PGRs, which will considerably reduce the costs at large-scale production.

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Callus induction and betacyanin quantification by HPLC/MS-MS in Alternanthera brasiliana (L.) Kuntze

Hoehnea ◽

10.1590/2236-8906-75/2016 ◽

2017 ◽

Vol 44 (1) ◽

pp. 90-95 ◽

Cited By ~ 1

Author(s):

Andressa Reis ◽

Alítcia Moraes Kleinowski ◽

Fátima Rosane Schuquel Klein ◽

Renata Trevizan Telles de Souza ◽

Luciano do Amarante ◽

...

Keyword(s):

High Performance Liquid Chromatography ◽

Callus Induction ◽

High Performance ◽

Large Scale ◽

Induction Medium ◽

Naphthaleneacetic Acid ◽

Scale Production ◽

Future Studies ◽

Large Scale Production ◽

Equilibrium Concentrations

ABSTRACT The objective of this study was to establish a protocol for callus induction and betacyanin production in plants of Alternanthera brasiliana. Explants of A. brasiliana and five combinations of cytokinin and auxin were used for callus induction. Calli were transferred to a Betacyanin Induction Medium (MIB), composed of MS, with 0.5 mg L-1 of thidiazuron (TDZ) and 1 mg L-1 of naphthaleneacetic acid (NAA), and kept in the light for 45 days. The aspect and intensity of pigments were assessed and total betacyanins were quantified in high performance liquid chromatography (HPLC). The combination of internodal segments and a medium containing equilibrium concentrations of auxins and cytokinins was the most efficient metod to induce calli and increased production of betacyanins. The presence of amaranthine in calli of A. brasiliana justifies its medical use and the consequent need for future studies for the large-scale production of this molecule.

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Phytosynthesis of Silver and Gold Nanoparticles Using the Hot Water Extract of Mixed Woodchip Powder and Their Antibacterial Efficacy

Journal of Nanomaterials ◽

10.1155/2017/8734758 ◽

2017 ◽

Vol 2017 ◽

pp. 1-19 ◽

Cited By ~ 1

Author(s):

Min-Ho Joe ◽

Hyun-Tak Jeong ◽

Hyung-Min Lee ◽

Hae-Jun Park ◽

Dong-Ho Kim ◽

...

Keyword(s):

Gold Nanoparticles ◽

Large Scale ◽

Water Extract ◽

Hot Water ◽

Scale Production ◽

Antibacterial Efficacy ◽

External Energy ◽

X Ray ◽

Silver And Gold Nanoparticles ◽

Hot Water Extract

This study investigates the phytosynthesis, characterization, and antibacterial efficacy of silver and gold nanoparticles (NPs) produced using the hot water extract of mixed woodchip powder. The woodchip extract (WCE) was successfully used as both a reducing and stabilizing agent for the phytosynthesis of both crystalline metal NPs. The effects of different physicochemical factors affecting the formation of the metal NPs including reaction pH, concentration of the precursor metal salts, amount of WCE, and external energy input were evaluated. The characterization of the metal NPs was performed by transmission electron microscopy, selected area electron diffraction (SAED), energy dispersive X-ray (EDX) spectroscopy, and X-ray diffraction (XRD) pattern analysis. In addition, the antibacterial efficacy of the phytosynthesized NPs was measured. The AgNPs showed clear antibacterial activity against four representative bacterial strains. However, the AuNPs did not exhibit bactericidal activity, probably due to their surface modifications and relatively large size. These results suggest that the phytosynthesis of the metal NPs using WCE is highly efficient, and its convenience makes it suitable for use in large-scale production.

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Factors Affecting the Regeneration, via Organogenesis, and the Selection of Transgenic Calli in the Peach Rootstock Hansen 536 (Prunus persica × Prunus amygdalus) to Express an RNAi Construct against PPV Virus

Plants ◽

10.3390/plants8060178 ◽

2019 ◽

Vol 8 (6) ◽

pp. 178 ◽

Cited By ~ 3

Author(s):

Sabbadini ◽

Ricci ◽

Limera ◽

Baldoni ◽

Capriotti ◽

...

Keyword(s):

Shoot Regeneration ◽

Prunus Persica ◽

Fluorescent Protein ◽

In Vitro Regeneration ◽

Naphthaleneacetic Acid ◽

Prunus Amygdalus ◽

Indirect Organogenesis ◽

Egfp Gene ◽

Rnai Construct

Prunus spp. is one of the most recalcitrant fruit tree species in terms of in vitro regeneration and transformation, mostly when mature tissues are used as explants. The present study describes the in vitro regeneration via indirect organogenesis, and Agrobacterium tumefaciens-mediated transformation of the peach rootstock Hansen 536 (Prunus persica × Prunus amygdalus) through the use of meristematic bulks (MBs) as starting explants. Efficient adventitious shoot regeneration was obtained when Hansen 536 MBs were cultured on an optimized medium consisting of modified McCown Woody Plant medium (WPM) enriched with 4.4 M 6-Benzyladenine (BA), 0.1 M 1-Naphthaleneacetic acid (NAA) and 6.0 g L−1 plant agar S1000 (B&V). MB slices were used later as starting explants for Agrobacterium-mediated transformation to introduce an RNAi construct “ihp35S-PPV194” against PPV virus. Transgenic events were identified by both green fluorescent protein (GFP) screening and kanamycin selection at different concentrations (0, 17 or 42 M). GFP-fluorescent proliferating callus lines were selected and confirmed to stably express the ihp35S-PPV194::eGFP gene construct by molecular analysis. Although shoot regeneration from these transgenic calli has not been obtained yet, this represents one of the few examples of successful attempts in peach genetic transformation from somatic tissues, and also serves as a useful in vitro system for future gene functional analysis in peach.

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Cannabis sativa: From Therapeutic Uses to Micropropagation and Beyond

Plants ◽

10.3390/plants10102078 ◽

2021 ◽

Vol 10 (10) ◽

pp. 2078

Author(s):

Tristan K. Adams ◽

Nqobile A. Masondo ◽

Pholoso Malatsi ◽

Nokwanda P. Makunga

Keyword(s):

Tissue Culture ◽

Genetic Engineering ◽

Large Scale ◽

Gene Editing ◽

Cannabis Sativa ◽

Scale Production ◽

Indirect Organogenesis ◽

Large Scale Production ◽

Pharmaceutical Industries

The development of a protocol for the large-scale production of Cannabis and its variants with little to no somaclonal variation or disease for pharmaceutical and for other industrial use has been an emerging area of research. A limited number of protocols have been developed around the world, obtained through a detailed literature search using web-based database searches, e.g., Scopus, Web of Science (WoS) and Google Scholar. This article reviews the advances made in relation to Cannabis tissue culture and micropropagation, such as explant choice and decontamination of explants, direct and indirect organogenesis, rooting, acclimatisation and a few aspects of genetic engineering. Since Cannabis micropropagation systems are fairly new fields, combinations of plant growth regulator experiments are needed to gain insight into the development of direct and indirect organogenesis protocols that are able to undergo the acclimation stage and maintain healthy plants desirable to the Cannabis industry. A post-culture analysis of Cannabis phytochemistry after the acclimatisation stage is lacking in a majority of the reviewed studies, and for in vitro propagation protocols to be accepted by the pharmaceutical industries, phytochemical and possibly pharmacological research need to be undertaken in order to ascertain the integrity of the generated plant material. It is rather difficult to obtain industrially acceptable micropropagation regimes as recalcitrance to the regeneration of in vitro cultured plants remains a major concern and this impedes progress in the application of genetic modification technologies and gene editing tools to be used routinely for the improvement of Cannabis genotypes that are used in various industries globally. In the future, with more reliable plant tissue culture-based propagation that generates true-to-type plants that have known genetic and metabolomic integrity, the use of genetic engineering systems including “omics” technologies such as next-generation sequencing and fast-evolving gene editing tools could be implemented to speed up the identification of novel genes and mechanisms involved in the biosynthesis of Cannabis phytochemicals for large-scale production.

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Somatic Embryogenesis of the Cedars of Lebanon (Cedrus libani)

HortScience ◽

10.21273/hortsci.32.3.461g ◽

1997 ◽

Vol 32 (3) ◽

pp. 461G-462

Author(s):

S.N. Talhouk ◽

M. Shmoury ◽

R. Baalbaki ◽

S. Khuri

Keyword(s):

Somatic Embryogenesis ◽

Callus Induction ◽

Embryogenic Callus ◽

Large Scale ◽

Developmental Stages ◽

Zygotic Embryos ◽

Scale Production ◽

Cedrus Libani ◽

Hormone Levels ◽

Significant Difference

Somatic embryogenesis offers a great potential for large-scale production of Cedrus libani, which is important not only as a forest tree, but also for the development of a timber industry. In an attempt to optimize conditions for embryogenic callus induction, we used zygotic embryos at different developmental stages as explants, compared different media, and used several hormone levels and combinations. Results indicated that post-cotyledonary immature embryos had highest induction efficiency. Four different media namely 1/2 MS, Durzan, Litvay's, and Von Arnold supplemented with similar hormone levels showed no significant difference in efficiency of callus induction. Induction frequencies of embryogenic callus from explants subjected to different hormone levels and combinations were dependent on the developmental stage of the explant.

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Production of Gymnemic Acid Depends on Medium, Explants, PGRs, Color Lights, Temperature, Photoperiod, and Sucrose Sources in Batch Culture ofGymnema sylvestre

The Scientific World JOURNAL ◽

10.1100/2012/897867 ◽

2012 ◽

Vol 2012 ◽

pp. 1-11 ◽

Cited By ~ 8

Author(s):

A. Bakrudeen Ali Ahmed ◽

A. S. Rao ◽

M. V. Rao ◽

Rosna Mat Taha

Keyword(s):

Batch Culture ◽

Callus Induction ◽

Large Scale ◽

Time Factors ◽

Callus Cultures ◽

Scale Production ◽

Gymnemic Acid ◽

Plant Parts ◽

Culture Optimization

Gymnema sylvestre(R.Br.) is an important diabetic medicinal plant which yields pharmaceutically active compounds called gymnemic acid (GA). The present study describes callus induction and the subsequent batch culture optimization and GA quantification determined by linearity, precision, accuracy, and recovery. Best callus induction of GA was noticed in MS medium combined with 2,4-D (1.5 mg/L) and KN (0.5 mg/L). Evaluation and isolation of GA from the calluses derived from different plant parts, namely, leaf, stem and petioles have been done in the present case for the first time. Factors such as light, temperature, sucrose, and photoperiod were studied to observe their effect on GA production. Temperature conditions completely inhibited GA production. Out of the different sucrose concentrations tested, the highest yield (35.4 mg/g d.w) was found at 5% sucrose followed by 12 h photoperiod (26.86 mg/g d.w). Maximum GA production (58.28 mg/g d.w) was observed in blue light. The results showed that physical and chemical factors greatly influence the production of GA in callus cultures ofG. sylvestre. The factors optimized forin vitroproduction of GA during the present study can successfully be employed for their large-scale production in bioreactors.

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Micropropagation of Environmental Stress Tolerant Local Potato (Solanum tuberosum L.) Varieties of Bangladesh

Plant Tissue Culture and Biotechnology ◽

10.3329/ptcb.v24i1.19218 ◽

2014 ◽

Vol 24 (1) ◽

pp. 101-109 ◽

Cited By ~ 1

Author(s):

Fazlima Parveen ◽

Mahmuda Khatun ◽

Aparna Islam

Keyword(s):

Shoot Apex ◽

Large Scale ◽

Solanum Tuberosum L ◽

In Vitro Regeneration ◽

Nodal Explants ◽

Direct Regeneration ◽

Nodal Segment ◽

Scale Production ◽

Potato Varieties

In vitro response of four explants namely, leaf, shoot apex, nodal and internodal, in three stress tolerant Bangladeshi potato varieties, viz. Zaubilati, Shadaguti and Challisha were tested. Of all the varieties, Shadaguti responded best for all the explants. Among the explants nodal segment responded best, followed by shoot apex. For all these explants and varieties, shoot regeneration response was tested in response to two cytokines, BAP and Kn. When compared between BAP and Kn supplementation, Kn treatment performed better than that of BAP for nodal and shoot apex, while opposite was observed for remaining explants. Interestingly, hormone free basal PROP medium was found to be best for nodal explants of both Zaubilati and Challisha varieties. While nodal explants of Shadaguti showed the best result in 0.5 mg/l Kn. Shoot apex performed best in 0.5 mg/l Kn for all the varieties. As an explant both internode and leaf did not perform well. Direct regeneration from these explants was found best in PROP + 0.2 mg/l GA3 + 0.5 mg/l IAA; 1.0 mg/l BAP, 1.5 mg/l BAP and 2.0 mg/l BAP for Zaubilai, Challisha and Shadaguti, respectively. For rooting, of the in vitro grown shoots half strength of PROP medium supplemented with 0.5 mg/l IBA was found to be best. Cent percent survival of transplanted plantlets was recorded. The successful protocol for in vitro regeneration was developed which can be used for large scale production of these abiotic stress tolerant potato varieties. Plant Tissue Cult. & Biotech. 24(1): 101-109, 2014 (June) D. O. I. http://dx.doi.org/10.3329/ptcb.v24i1.19218

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In vitro Regeneration of Vitex negundo L. - A Multipurpose Woody Aromatic Medicinal Shrub

Plant Tissue Culture and Biotechnology ◽

10.3329/ptcb.v18i1.3263 ◽

1970 ◽

Vol 18 (1) ◽

pp. 37-42 ◽

Cited By ~ 6

Author(s):

M. Jawahar ◽

S. Ravipaul ◽

M. Jeyaseelan

Keyword(s):

Callus Induction ◽

In Vitro Regeneration ◽

Leaf Explants ◽

Shoot Proliferation ◽

Optimal Level ◽

Vitex Negundo ◽

Indirect Organogenesis ◽

Shoot Bud ◽

Bud Initiation

A rapid and efficient protocol was developed for inducing indirect organogenesis using leaf explants of Vitex negundo L. Explants were cultured on MS with different concentrations of 2,4-D and IAA in combination with BAP for callus induction. The frequency of callus induction increased with increasing concentration of IAA (0.3 mg/l) and BAP (0.3 mg/l) at optimal level. The shoot buds appeared emerging as green coloured protuberances on the callus. The high frequency of shoot bud initiation and shoot proliferation was observed on MS containing 0.3 mg/l IAA and 0.3 mg/l BAP. The regenerated shoots were successfully rooted on MS supplemented with 0.5 mg/l IBA. Rooted plants were transferred to pots containing sand, soil and manure in the ratio of 1 : 1 : 1. Nearly 90% survival of in vitro plants were recorded. Key words : Vitex negundo, In vitro, Leaf, Callus, Regeneration D.O.I. 10.3329/ptcb.v18i1.3263 Plant Tissue Cult. & Biotech. 18(1): 37-42, 2008 (June)

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New Basal Media for Protocorm-Like Body and Callus Induction of Hybrid Cymbidium

Journal of Fruit and Ornamental Plant Research ◽

10.2478/v10290-012-0022-8 ◽

2012 ◽

Vol 20 (2) ◽

pp. 127-133 ◽

Cited By ~ 12

Author(s):

Jaime A. Teixeira da Silva

Keyword(s):

Callus Induction ◽

High Frequency ◽

Callus Formation ◽

Basal Medium ◽

Naphthaleneacetic Acid ◽

Control Medium ◽

Industry Standard ◽

Basal Media ◽

Bacto Agar

Abstract High frequency protocorm-like body (PLB) production from hybrid Cymbidium Twilight Moon ‘Day Light’ has been developed through a new medium, Teixeira Cymbidium (TC) medium. Two new TC media containing variable amounts of macroand micronutrients and other additives, inspired by Winarto and Teixeira (WT) medium for Anthurium and Murashige and Skoog (MS) basal medium were used to induce PLBs and callus. Control medium was research- and industry-standard Vacin and Went (VW) medium. The first TC medium, TCPLB, could induce significantly more PLBs than on VW while high levels of macronutrients in the second TC medium, TCCALLUS, and MS were required to induce callus. All PLB induction media contained 0.1 mg/l α-naphthaleneacetic acid (NAA) and 0.1 mg/l kinetin (KIN), 2 g/l tryptone and 20 g/l sucrose, and solidified with 8 g/l Bacto agar while callusinduction media were identical, except that KIN was substituted by thidiazuron (TDZ). Basal medium had a significant effect on PLB and callus formation. This protocol could be used to induce PLBs and callus from other Cymbidium species or cultivars.

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Effect of the Carbon Source and Plant Growth Regulators (PGRs) in the Induction and Maintenance of an In Vitro Callus Culture of Taraxacum officinale (L) Weber Ex F.H. Wigg

Agronomy ◽

10.3390/agronomy11061181 ◽

2021 ◽

Vol 11 (6) ◽

pp. 1181

Author(s):

María Eugenia Martinez ◽

Lorena Jorquera ◽

Paola Poirrier ◽

Katy Díaz ◽

Rolando Chamy

Keyword(s):

Carbon Source ◽

Plant Growth ◽

Plant Growth Regulators ◽

Callus Culture ◽

Growth Regulators ◽

Callus Induction ◽

Large Scale ◽

Taraxacum Officinale ◽

Naphthaleneacetic Acid

Taraxacum officinale (L.) Weber ex F.H. Wigg, commonly known as dandelion, is a cosmopolitan and perennial weed, which has medicinal properties. In vitro propagation methods are widely used on plants that have difficulties in cultivation and, consequently, low extraction yields of active metabolites. Thus, callus culture has been considered to be useful for the accumulation of several metabolites. In this study, we aimed to establish an efficient protocol for callus induction and maintenance of T. officinale, for which explant type, carbon source, light conditions, and nine different combinations of plant growth regulators (PGRs), such as 1-naphthaleneacetic acid (NAA) (from 0.05 to 0.5 mg/L) and 6-benzylaminopurine acid (BAP) (from 0.5 to 3.0 mg/L), were evaluated. The results showed that hypocotyls and roots from sterile seedlings are the best sources for callus induction, with 100% of callogenesis at every condition tested, and more than 95% of viability and friability. Complete darkness and a medium supplemented with sucrose at 2.3% (w/v) and 0.5 mg/L of NAA and 0.5 mg/L of BAP were the best conditions for callus induction, showing callus with low organogenesis and high friability. This study provides a basis for future studies on improving large-scale callus propagation and further establishment of suspension culture systems for commercial purposes.

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