scholarly journals Modulation of the anatomical and physiological responses of in vitro grown Alcantarea imperialis induced by NAA and residual effects of BAP

2020 ◽  
Vol 26 (2) ◽  
pp. 283-297
Author(s):  
João Paulo Rodrigues Martins ◽  
Luiz Carlos de Almeida Rodrigues ◽  
Thayna dos Santos Silva ◽  
Andreia Barcelos Passos Lima Gontijo ◽  
Antelmo Ralph Falqueto
Keyword(s):  
In Vitro Propagation ◽  
Growth Traits ◽  
Adventitious Shoots ◽  
Residual Effects ◽  
Physiological Status ◽  
Naphthaleneacetic Acid ◽  
Negative Effects ◽  
Direct Exposure

Abstract During in vitro propagation, cytokinins (CKs) and auxins (AUXs), such as 6-benzylaminopurine (BAP) and 1-naphthaleneacetic acid (NAA), are often used to induce adventitious shoots and roots, respectively. However, it is not clear how CKs affect plants over a long period of in vitro propagation as well as the synergy of direct exposure to AUX with previous CK treatments. The aim was to assess the physiological and anatomical responses of Alcantarea imperialis in function of the interaction of both previous BAP treatments and direct NAA exposure during in vitro propagation. Plants previously grown in vitro were transferred to media containing 0, 5, 10 or 15 μM BAP. After 60 days, the adventitious shoots from each previous BAP treatment were subcultured in media with 0, 2 or 4 μM NAA. Pigment content, anatomical and growth traits were assessed in the plants from each treatment. Both previous BAP treatments and direct NAA exposure altered the anatomy and pigment contents of plants as well as their growth traits. BAP induced negative effects over the long term on physiological status as well as changed the plants’ anatomy. NAA supplementation in the medium can partially reverse the negative effects induced by BAP. The application of 2 μM NAA during in vitro rooting improved the plants’ quality.

In vitro propagation of Crambe maritima

1987 ◽  
Vol 65 (1) ◽  
pp. 72-75 ◽  
Author(s):  
J. Y. Peron ◽  
E. Regnier
Keyword(s):  
In Vitro Propagation ◽  
Indoleacetic Acid ◽  
Adventitious Shoots ◽  
Basal Medium ◽  
Naphthaleneacetic Acid ◽  
Petiole Explants ◽  
Acclimatization Period

A method for rapid micropropagation of sea kale (Crambe maritima L.) was developed. Petiole explants placed in vitro on a medium containing 0.5 mg/L indoleacetic acid (IAA), 6.0 mg/L kinetin, and 1.5 mg/L benzylaminopurine developed callus within 15 days and shoots within 28 days. Nearly four adventitious shoots could be developed within 3 weeks by placing the initial shoot on media without IAA. To develop roots, the shoots were then transferred to the basal medium containing 0.1 to 1.0 mg/L indolbutyric or α-naphthaleneacetic acid. Rooted plantlets were obtained within 2 or 3 weeks. After an acclimatization period of 6 weeks in a greenhouse in unsterilized medium, the plantlets could be set outdoors.

scholarly journals 108 Shoot Regeneration from Ovaries of Various Cultivars of Hosta

HortScience ◽  
1999 ◽  
Vol 34 (3) ◽  
pp. 460B-460
Author(s):  
David J. Williams ◽  
Karim H. Al-Juboory
Keyword(s):  
Shoot Regeneration ◽  
Adventitious Shoots ◽  
Shoot Tips ◽  
Effective Alternative ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Slow Process ◽  
Number Of Shoots

The objective of this study was to evaluate the ability of various cultivars of Hosta ovary explants to generate adventitious shoots and obtain variegated plants in vitro. Immature inflorescences along with 8 to 10 cm of scape were harvested from Hosta cultivars. The ovaries were prepared for culture by cutting immature florets before anthesis. The florets were first cut just above the top of the immature ovary to remove the sigma, style, corolla, and anther. Then the calyx and filament bases were also removed. Ovaries were transversely cut into halves and transferred to baby jars containing Hosta initiation medium supplemented with naphthaleneacetic acid (NAA) at 0.5 mg/L and 6-benzylamino purine (BA). The explants produced adventitious shoots from ovary base via organogenesis. The number of shoots regenerated from shoot tips and callus increased linearly with repeatedf subculturing on MS medium. This method would provide an effective alternative to conventional propagation crown division of Hosta, an expensive and slow process. The long-term goal of this project is to improve Hosta.

scholarly journals Efficient Regeneration of Hedychium coronarium through Protocorm-Like Bodies

Agronomy ◽  
2020 ◽  
Vol 10 (8) ◽  
pp. 1068
Author(s):  
Xiu Hu ◽  
Jiachuan Tan ◽  
Jianjun Chen ◽  
Yongquan Li ◽  
Jiaqi Huang
Keyword(s):  
In Vitro Culture ◽  
In Vitro Propagation ◽  
Adventitious Shoots ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Culture Methods ◽  
Hedychium Coronarium ◽  
Mature Node ◽  
First Time

Hedychium coronarium J. Koenig is a multipurpose plant with significant economic value, but it has been overexploited and listed as a vulnerable, near threatened or endangered species. In vitro culture methods have been used for propagating disease-free propagules for its conservation and production. However, explant contamination has been a bottleneck in in vitro propagation due to the use of rhizomes as the explant source. Plants in the family Zingiberaceae have pseudostems that support inflorescences, while rhizomes are considered true stems. The present study, for the first time, reported that the pseudostem bears nodes and vegetative buds and could actually be true stems. The evaluation of different sources of explants showed that mature node explants derived from the stem were the most suitable ones for in vitro culture because of the lowest contamination and the highest bud break rates. Culture of mature node explants on MS medium supplemented with 13.32, 17.76, and 22.20 μM 6-benzylaminopurine (BA), each in combination with 9.08 μM thidiazurin (TDZ) and 0.05 μM α-naphthaleneacetic acid (NAA) induced the conversion of buds to micro-rhizomes in six weeks. More than 96% of the micro-rhizomes cultured on MS medium supplemented with 17.76 μM BA, 6.81 μM TDZ, and 2.46 μM indole-3-butyric acid (IBA) were converted to globular-shaped clumps with protocorm-like bodies (PLBs). Further culture of a piece of the clumps induced more than 15 adventitious shoots. Adventitious roots were produced at the base of adventitious shoots, and plantlets were readily transplanted to a substrate for acclimatization in a shaded greenhouse. The survival rate of the plants in the greenhouse was up to 90%. Plants grew vigorously, and there were no off-types from the regenerated 11,100 plants. Our study also, for the first time, shows that H. coronarium can be regenerated via PLBs, which may represent a new way of the in vitro propagation of H. coronarium. The established protocol could be used for the increased propagation of H. coronarium for conservation or commercial production.

scholarly journals In vitro Propagation and Assessment of Genetic Relationships of Citrus Rootstocks Using ISSR Molecular Markers

2017 ◽  
Vol 45 (2) ◽  
pp. 383-391 ◽  
Author(s):  
Constantinos SALIS ◽  
Ioannis E. PAPADAKIS ◽  
Spyridon KINTZIOS ◽  
Marianna HAGIDIMITRIOU
Keyword(s):  
Molecular Markers ◽  
Genetic Variability ◽  
Root Length ◽  
In Vitro Propagation ◽  
Genetic Relationships ◽  
Shoot Proliferation ◽  
Poncirus Trifoliata ◽  
Naphthaleneacetic Acid ◽  
Issr Molecular Markers

The behavior of six citrus rootstocks, Volkameriana, Citrumelo ‘Swingle’, Citrange ‘Carrizo’, Poncirus trifoliata ‘Serra’, Poncirus trifoliata ‘Rubidoux’ and Poncirus trifoliata ‘Flying Dragon’, in in vitro propagation was studied and compared for shoot proliferation and rooting. In addition, the genetic relationships among the rootstocks studied and other Citrus species, using the Inter-Simple Sequence Repeats (ISSR) molecular markers, were investigated. Nodal explants of three months old shoots were used in Murashige and Skoog medium supplemented with N6-benzyladenine (BA) for shoot proliferation and with naphthaleneacetic acid (NAA) for rooting. The rootstock Volkameriana showed a statistically significant higher number of shoots (1.81), shoot length (15.14 mm) and number of leaves per explant (5.81), while all three Poncirus trifoliata rootstocks showed the lowest numbers. The number of roots and root length per explant were evaluated at the end of the rooting phase. The rootstock ‘Swingle’ showed a higher number of roots per explant (4.2) followed by ‘Flying Dragon’ (3.93) and ‘Carrizo’ (3.23) rootstocks. The rootstocks ‘Swingle’ (140.8 mm), Volkameriana (148 mm) and ‘Flying Dragon’ (131.12 mm) had significantly higher root length per explant compared to ‘Carrizo’ (31 mm) and ‘Rubidoux’ (34.5 mm). The ISSR molecular marker technique used in the present study grouped successfully the different species, varieties and rootstocks studied, revealing their genetic variability. The genetic variability observed among the rootstocks ranged between 0.29 (Poncirus trifoliata ‘Serra’ and Citrumelo ‘Swingle’) and 0.60 (Volkameriana and Citrumelo ‘Swingle’). The response of the rootstocks studied in in vitro propagation however is not related to their genetic affinity.

scholarly journals A Novel Antiinflammatory Maintains Glucocorticoid Efficacy with Reduced Side Effects

2003 ◽  
Vol 17 (5) ◽  
pp. 860-869 ◽  
Author(s):  
Michael J. Coghlan ◽  
Peer B. Jacobson ◽  
Ben Lane ◽  
Masaki Nakane ◽  
Chun Wei Lin ◽  
...  
Keyword(s):  
Side Effects ◽  
Inflammatory Disease ◽  
Target Genes ◽  
Negative Effects ◽  
Interacting Protein ◽  
Altered Gene ◽  
Antiinflammatory Effects

Abstract Glucocorticoids (GCs) are commonly used to treat inflammatory disease; unfortunately, the long-term use of these steroids leads to a large number of debilitating side effects. The antiinflammatory effects of GCs are a result of GC receptor (GR)-mediated inhibition of expression of proinflammatory genes as well as GR-mediated activation of antiinflammatory genes. Similarly, side effects are most likely due to both activated and repressed GR target genes in affected tissues. An as yet unachieved pharmaceutical goal is the development of a compound capable of separating detrimental side effects from antiinflammatory activity. We describe the discovery and characterization of AL-438, a GR ligand that exhibits an altered gene regulation profile, able to repress and activate only a subset of the genes normally regulated by GCs. When tested in vivo, AL-438 retains full antiinflammatory efficacy and potency comparable to steroids but its negative effects on bone metabolism and glucose control are reduced at equivalently antiinflammatory doses. The mechanism underlying this selective in vitro and in vivo activity may be the result of differential cofactor recruitment in response to ligand. AL-438 reduces the interaction between GR and peroxisomal proliferator-activated receptor γ coactivator-1, a cofactor critical for steroid-mediated glucose up-regulation, while maintaining normal interactions with GR-interacting protein 1. This compound serves as a prototype for a unique, nonsteroidal alternative to conventional GCs in treating inflammatory disease.

scholarly journals ITGA6+ Human Testicular Cell Populations Acquire a Mesenchymal Rather than Germ Cell Transcriptional Signature during Long-Term Culture

2020 ◽  
Vol 21 (21) ◽  
pp. 8269
Author(s):  
Robert B. Struijk ◽  
Callista L. Mulder ◽  
Saskia K. M. van Daalen ◽  
Cindy M. de Winter-Korver ◽  
Aldo Jongejan ◽  
...  
Keyword(s):  
Germ Cell ◽  
Cell Sorting ◽  
In Vitro Propagation ◽  
Cell Type ◽  
Testicular Cell ◽  
Testicular Cells ◽  
Cell Type Composition ◽  
Type Composition

Autologous spermatogonial stem cell transplantation is an experimental technique aimed at restoring fertility in infertile men. Although effective in animal models, in vitro propagation of human spermatogonia prior to transplantation has proven to be difficult. A major limiting factor is endogenous somatic testicular cell overgrowth during long-term culture. This makes the culture both inefficient and necessitates highly specific cell sorting strategies in order to enrich cultured germ cell fractions prior to transplantation. Here, we employed RNA-Seq to determine cell type composition in sorted integrin alpha-6 (ITGA6+) primary human testicular cells (n = 4 donors) cultured for up to two months, using differential gene expression and cell deconvolution analyses. Our data and analyses reveal that long-term cultured ITGA6+ testicular cells are composed mainly of cells expressing markers of peritubular myoid cells, (progenitor) Leydig cells, fibroblasts and mesenchymal stromal cells and only a limited percentage of spermatogonial cells as compared to their uncultured counterparts. These findings provide valuable insights into the cell type composition of cultured human ITGA6+ testicular cells during in vitro propagation and may serve as a basis for optimizing future cell sorting strategies as well as optimizing the current human testicular cell culture system for clinical use.

A comparison of genetic stability in tea [Camellia sinensis (L.) Kuntze] plantlets derived from callus with plantlets from long-term in vitro propagation

2019 ◽  
Vol 138 (3) ◽  
pp. 467-474 ◽  
Author(s):  
Lidiia Samarina ◽  
Maya Gvasaliya ◽  
Natalia Koninskaya ◽  
Ruslan Rakhmangulov ◽  
Alexander Efremov ◽  
...  
Keyword(s):  
Camellia Sinensis ◽  
Genetic Stability ◽  
In Vitro Propagation

scholarly journals Micropropagation of Rare Scutellaria Havanensis Jacq. And Preliminary Studies on Antioxidant Capacity and Anti-cancer Potential

2021 ◽  
Author(s):  
Lani Irvin ◽  
Yarelia Zavala Ortiz ◽  
Kamila Rivera Rivera ◽  
Brajesh Nanda Vaidya ◽  
Samantha H Sherman ◽  
...  
Keyword(s):  
Antioxidant Capacity ◽  
In Vitro Propagation ◽  
Adventitious Shoots ◽  
Colorectal Cancer Cell Line ◽  
Dependent Manner ◽  
Flavonoid Content ◽  
Leaf Extracts ◽  
Total Polyphenol ◽  
Bud Induction

We report the development of in vitro propagation protocols through adventitious shoot induction pathway for a rare and medicinal Scutellaria havanensis. In vitro propagation studies using nodal explants showed MS medium supplemented with 10µM 6-Benzylaminopurine induced highest number of adventitious shoots in a time dependent manner. A ten - day incubation was optimum for shoot bud induction as longer exposures resulted in hyperhydricity of the explants and shoots induced. We also report preliminary evidence of Agrobacterium tumefaciens EHA105 - mediated gene transfer transiently expressing of green fluorescent protein in this species. Transformation studies exhibited amenability of various explant tissues, internode being the most receptive. As the plant has medicinal value, research was carried out to evaluate its potential antioxidant capacity and the efficacy of methanolic leaf extracts in curbing the viability of human colorectal cancer cell line HCT116. Comparative total polyphenol, and flavonoid content measurement of fresh and air dried leaf extract revealed that the fresh leaf extracts contain higher total polyphenol and flavonoid content. The HCT 116 cell viability assessed by colorimetric assay using a 3-(4, 5-dimethyl-thiazol-2-yl)-2, 5-diphenyltetrazolium bromide, showed a steady growth inhibition after 24 hours of incubation. Scanning Electron Microscopy of leaf surface revealed high density of glandular and non-glandular trichomes. This research provides basis for the conservation of this rare plant and future phytochemical screening and clinical research.

scholarly journals 889 PB 469 IN VITRO SELECTION FOR DWARFISM IN APPLE

HortScience ◽  
1994 ◽  
Vol 29 (5) ◽  
pp. 561c-561
Author(s):  
Mushtaq Sarwar ◽  
Robert M. Skirvin
Keyword(s):  
In Vitro Selection ◽  
Adventitious Shoots ◽  
Shoot Formation ◽  
Leaf Segment ◽  
Adult Growth ◽  
Napthaleneacetic Acid ◽  
Selection For ◽  
Cut Surface

Adventitious shoots were regenerated from apple (`Wijcik', `McIntosh', `Macspur', `M-26' and `Mutsu') by excising leaves from in vitro-grown shoots, cutting them into three sections, and plating them onto regeneration media. Cultures were kept in the dark for 1 to 4 weeks and then moved to light for further shoot development. MS medium supplemented with thiadiazuron (2-3 μM) and napthaleneacetic acid (5 μM) produced the highest number of shoots per leaf segment. `Wijcik' and `M-26' regenerated best from big leaves, whereas `McIntosh' and `Macspur' regenerated best from small leaves. Shoot formation was enhanced by 3 to 4 weeks of dark treatment and by placing the leaf on medium with its abaxial surface uppermost. The cut surface of leaf segments produced the most regeneration sites. In vitro adventitious shoots were transferred to various concentrations of BA (5, 10, 15, 20, 25 and 30 μM to screen them for BA tolerance and to predict their adult growth habit. These shoots will be rooted and transferred to greenhouse and field conditions for long-term evaluations.

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