scholarly journals Problems Encountered during In vitro Culture Establishment in Terminalia arjuna

2020 ◽  
pp. 154-160
Author(s):  
Meena Choudhary ◽  
Inder Dev Arya ◽  
Sarita Arya
Keyword(s):  
In Vitro Culture ◽  
Culture Media ◽  
Terminalia Arjuna ◽  
Culture Initiation ◽  
Surface Sterilization ◽  
Initial Stage ◽  
In Vitro Shoots ◽  
Half Strength ◽  
Bud Growth

The main aim of present study was to overcome the problems associated with the in vitro culture initiation in Terminalia arjuna. The micropropagation of tree species is not easy as shrubs and herbs. Many problems encountered from explant collection to in vitro culture establishment. The problems that have been occurred during T. arjuna micropropagation were culture contamination, phenolic exudation, bud growth inhibition, shoots yellowing and leaf fall. All these problems have been solved by applying certain treatments prior to explant collection and inoculation. The mother tree was lopped in November months (six months prior to explant collection) to remove any inhibitory substance and release bud growth. Different sterilizing agents were used to minimize the bacterial and fungal contamination. Some modification in culture media (use of different concentration of NH4NO3 and KNO3 salts and adenine sulphate) was done. Surface sterilization of nodal explants collected from lopped branches with 0.1% HgCl2 for 8 min., treatment with chilled antioxidant solution (Ascorbic acid, Citric acid and PVP) and half strength of NH4NO3 and KNO3 salts of MS medium supported 100% bud break response with proliferation of green and healthy in vitro shoots. Removing these hurdles already in the initial stage of micropropagation is very important and maximize mass in vitro propagation of this medicinally important Arjun tree. 

scholarly journals Micropropagation of Isoplexis chalcantha Svent, O´Shanahan from Mature Plants

1970 ◽  
Vol 18 (2) ◽  
pp. 131-137 ◽  
Author(s):  
S. Mederos-Molina
Keyword(s):  
Culture Medium ◽  
Culture Media ◽  
Nodal Segments ◽  
Axillary Shoots ◽  
In Vitro Shoots ◽  
Half Strength ◽  
Modified Ms Medium ◽  
High Degree ◽  
First Time

Culture medium requirements for micropropagation of Isoplexis chalcantha was achieved for the first time after high degree of contamination and phenolic exudates were detected and solved. Cultures were established from axillary shoots using juvenile branches collected from this medicinal plant. Most satisfying results were obtained using a solidified and a modified MS medium (NO3- : NH4+ ratios) enriched with ascorbic acid or soluble PVP plus GA3, BAP and NAA. Explants (nodal segments) were used for in vitro shoots multiplication and best results were achieved with modified MS plus BAP and auxins. Vigorous shoots rooted without symptoms in the half-strength modified MS enriched with low concentration of IBA. Key words: Isoplexis chalcantha, axillary shoots, contamination, phenolic exudates, culture media, NO3- : NH4+ ratios D.O.I. 10.3329/ptcb.v18i2.3395 Plant Tissue Cult. & Biotech. 18(2): 131-137, 2008 (December)

scholarly journals Mass propagation of Bambusa bambos (L.) Voss through in vitro culture

2017 ◽  
Vol 5 (2) ◽  
pp. 15-26 ◽  
Author(s):  
Raihan I Raju ◽  
Shyamal K Roy
Keyword(s):  
In Vitro Culture ◽  
Basal Medium ◽  
Nodal Segments ◽  
Garden Soil ◽  
Root Induction ◽  
Ms Medium ◽  
Mass Propagation ◽  
Surface Sterilization ◽  
Half Strength

Protocol for mass propagation of Bambusa bamboos (L.) Voss was developed through in vitro culture. Nodal segments containing pre-existing axillary bud, after surface sterilization, were inoculated on liquid Murashige and Skoog’s (MS) basal medium containing different concentrations and combinations of cytokinins (BAP, TDZ and Kn). The highest direct shoot induction (90%) was obtained in the MS liquid medium supplemented with 2.0 mg/l BAP and 1.0 mg/l TDZ with maximum average number of shoots (3.14 ± 0.06) per explant. Highest shoot multiplication (16.58 ± 0.24 shoots per culture) with highest average shoot length (9.21 ± 0.13 cm) was obtained when in vitro raised shoots were cultured in gelrite gelled MS medium in conjunction with 2.0 mg/l BAP and 1.0 mg/l TDZ. Incorporation of 10% coconut water with 4% sucrose in the above mentioned medium resulted satisfactory shoot growth and development with an average 26.7 ± 0.60 shoots per culture. For root induction, in vitro raised shoots were divided into clumps of 4-5 shoots in each clump and transferred onto both liquid and gelled half-strength MS medium containing different concentrations and combinations of auxins (IBA and NAA). Maximum rooting (86.67%) was achieved in half-strength of MS medium fortified with 2.5 mg/l IBA and 2.5 mg/l NAA with an average 8.72 ± 0.42 root per shoot. The rooted plantlets were then transferred to polybags containing garden soil, sand and compost mixture with 1:1:1 ratio. After a month the hardened plantlets were then transferred to the larger pots containing garden soil and compost with 1:1 ratio for sufficient growth and finally transplanted to the field. In this process, the highest 100% survivability was recorded from well-established rooted plantlets. The regenerated plants showed well developed root and shoot systems in field condition.Jahangirnagar University J. Biol. Sci. 5(2): 15-26, 2016 (December)

Rhizome Buds Disinfection for Preparation of Red Ginger (Zingiber officinale Roxb. var. rubrum Rosc.) In Vitro Culture

2020 ◽  
Vol 3 (1) ◽  
pp. 19-26
Author(s):  
Popy Hartatie Hardjo ◽  
Alfian Hendra Krisnawan
Keyword(s):  
In Vitro Culture ◽  
Axenic Culture ◽  
Zingiber Officinale ◽  
Culture Initiation ◽  
Surface Sterilization ◽  
Sterilization Method ◽  
Ginger Rhizome ◽  
Red Ginger ◽  
Rhizome Buds

The success of culture initiation depends on explant surface sterilization techniques. Suitable concentration, combinations, and duration of exposure of sterilizing agents are important to raise in vitro culture successfully. The aim of this work is to obtain the suitable sterilization method for explant buds of red ginger rhizome to get the axenic culture. Four sterilizing agents, fungicide, bactericide, Cefotaxime antibiotic, and NaOCl were tested for sterilization by various concentration and duration of exposure. The results showed that sterilizing agents 200 mg/L Cefotaxime and 100 mg/L Benomyl combined with NaOCl decreased the contamination of explants, and achieved 20% axenic culture.

scholarly journals Rheological and Microstructural Features of Plant Culture Media Doped with Biopolymers: Influence on the Growth and Physiological Responses of In Vitro-Grown Shoots of Thymus lotocephalus

2021 ◽  
Vol 2 (2) ◽  
pp. 538-553
Author(s):  
Natacha Coelho ◽  
Alexandra Filipe ◽  
Bruno Medronho ◽  
Solange Magalhães ◽  
Carla Vitorino ◽  
...  
Keyword(s):  
In Vitro Culture ◽  
Culture Medium ◽  
Culture Media ◽  
Average Pore Size ◽  
Physiological Responses ◽  
Gum Arabic ◽  
Shoot Elongation ◽  
Hydroxyethyl Cellulose ◽  
Plant Culture

In vitro culture is an important biotechnological tool in plant research and an appropriate culture media is a key for a successful plant development under in vitro conditions. The use of natural compounds to improve culture media has been growing and biopolymers are interesting alternatives to synthetic compounds due to their low toxicity, biodegradability, renewability, and availability. In the present study, different culture media containing one biopolymer (chitosan, gum arabic) or a biopolymer derivative [hydroxyethyl cellulose (HEC), carboxymethyl cellulose (CMC)], at 100 or 1000 mg L−1, were tested regarding their influence on the growth and physiological responses of Thymus lotocephalus in vitro culture. Cellulose-based biopolymers (HEC and CMC) and gum arabic were used for the first time in plant culture media. The results showed that CMC at 100 mg L−1 significantly improved shoot elongation while chitosan, at the highest concentration, was detrimental to T. lotocephalus. Concerning only the evaluated physiological parameters, all tested biopolymers and biopolymer derivatives are safe to plants as there was no evidence of stress-induced changes on T. lotocephalus. The rheological and microstructural features of the culture media were assessed to understand how the biopolymers and biopolymer derivatives added to the culture medium could influence shoot growth. As expected, all media presented a gel-like behaviour with minor differences in the complex viscosity at the beginning of the culture period. Most media showed increased viscosity overtime. The surface area increased with the addition of biopolymers and biopolymer derivatives to the culture media and the average pore size was considerably lower for CMC at 100 mg L−1. The smaller pores of this medium might be related to a more efficient nutrients and water uptake by T. lotocephalus shoots, leading to a significant improvement in shoot elongation. In short, this study demonstrated that the different types of biopolymers and biopolymer derivatives added to culture medium can modify their microstructure and at the right concentrations, are harmless to T. lotocephalus shoots growing in vitro, and that CMC improves shoot length.

scholarly journals In vitro culture as an aid to conservation of indigenous ferns: Diplazium proliferum

2015 ◽  
Vol 6 (1) ◽  
Author(s):  
Zubeir M. Golamaully ◽  
Vishwakalyan Bhoyroo ◽  
Nadeem Nazurally ◽  
Vineshwar Gopal
Keyword(s):  
Growth Rate ◽  
In Vitro Culture ◽  
Mercuric Chloride ◽  
Rare Species ◽  
Vascular Plants ◽  
Culture Media ◽  
Fungal Contamination ◽  
Increase Growth Rate ◽  
Growing Population

With the ever growing population and economic needs of Mauritius, the flora of Mauritius has never been in more danger and one group of vascular plants is even more in peril; ferns.<em> Diplazium proliferum</em> is indigenous to the Mascarene region and is considered as a rare species in Mauritius. The need to develop a tested <em>in vitro</em> propagation protocol is a must to protect the biodiversity of Mauritius. This experiment was geared towards the establishment of a proper sterilization technique and the effect of 6-benzylaminopurine (BAP) and light on <em>in vitro</em> culture of this fern. Sterilization with 0.05% Mercuric chloride was effective to eliminate fungal contamination and allow germination of spores. Culture media supplemented with BAP did not significantly increase growth rate of both gametophytes and sporophytes of<em> D. proliferum</em>. Present results suggest efficient sterilization methods to be a crucial stage for successful<em> in vitro r</em>egeneration of ferns. The established protocol will be used as an optimized baseline protocol for the propagation of other indigenous ferns.

Influence of Safeners on the in vivo and in vitro Metabolism of Bentazon and Metolachlor by Grain Sorghum Shoots: a Preliminary Report

1991 ◽  
Vol 46 (9-10) ◽  
pp. 906-914 ◽  
Author(s):  
Donald E. Moreland ◽  
Frederick T. Corbin
Keyword(s):  
Grain Sorghum ◽  
In Vitro Studies ◽  
Naphthalic Anhydride ◽  
Surface Sterilization ◽  
Polar Components ◽  
In Vitro Shoots ◽  
Treated Seed ◽  
Cytochrome P 450

Abstract Metabolism of bentazon and metolachlor by excised shoots and a microsomal fraction iso­lated from the shoots, of 3-day-old, dark-grown, grain sorghum (Sorghum bicolor cv. Funk G 522 DR) seedlings was studied. The effects of seed treatments, on the subsequent metabo­lism of the herbicides, with the safeners naphthalic anhydride, oxabetrinil, and CGA 133205 were compared against surface-sterilization and Captan-treatments. Bentazon was aryl hydroxylated in both in vivo and in vitro studies with the hydroxylated derivative undergoing glycosylation only under in vivo conditions. Both shoots and microsomes isolated from shoots of safener-treated seed showed enhanced metabolism of bentazon relative to the controls. In­ hibition by tetcyclacis, a potent inhibitor of plant cytochrome P-450 monooxygenases, in both the in vivo and in vitro studies, and a requirement for NADPH in the in vitro studies suggested that the formation of hydroxybentazon was mediated by a cytochrome P-450 monooxygenase. Metolachlor was metabolized to polar material and O-desmethylmetolachlor under in vivo conditions. Only the demethylated product was formed in vitro. Shoots isolated from safener-treated seed showed enhanced formation of polar com pounds which were assumed to have arisen from conjugation with glutathione. Tetcyclacis did not affect the formation of the polar components. However, the formation of O-desmethylmetolachlor was depressed in the shoots excised from safener-treated seed under both in vivo and in vitro conditions. Tetcyclacis completely prevented formation of the demethylated metabolite. Hence, formation of this meta­bolite is considered to be P-450 mediated. The differential response obtained with the safeners, i.e., stimulation of aryl hydroxylation of bentazon and depression of metolachlor demethylation, suggests that the reactions are probably catalyzed by different cytochrome P-450 mono­oxygenases.

scholarly journals Research on in vitro micropropagation of Lilium brownii F.E. Brown

2016 ◽  
Vol 14 (1) ◽  
pp. 121-129
Author(s):  
Vũ Hoài Sâm ◽  
Bùi Đức Quỳnh ◽  
Nguyễn Thị Hương ◽  
Nguyễn Văn Khiêm
Keyword(s):  
Subtropical Climate ◽  
Ms Medium ◽  
In Vitro Plant Regeneration ◽  
Medicinal Species ◽  
Surface Sterilization ◽  
The North ◽  
In Vitro Micropropagation ◽  
Half Strength ◽  
Over Exploitation

Lilium brownii Brown belonging Lilium genus and Liliaceae family is well-known as a popular medicinal species, as well as food source and beautiful ornamental flowers. The specie has unique and ornamental floral characteristics such as light and elegant fragrance and perianth color rapidly changing from yellowish cream to white during anthesis. In traditional medicine, it is used for treatment cough, sedation diuretic, bronchitis... In nature, it can be found in subtropical climate moutainous areas in the North such as Sa Pa, Bat Xat, Mu Cang Chai; Sin Ho and Phong Tho, Quang Ba and Dong Van. In recent years, this species has been listed in the Red List for medicinal plants in Vietnam due to over-exploitation. The only effective strategy for sustaible conservation this species is in vitro micropropagation. In this study, in vitro plant regeneration and micropropagation of L. brownii was established from bubles and stem nodes. After surface sterilization with 0.1% HgCl­2 in 10 minutes, healthy young shoots were obtained from initial bubles and stem nodes on MS medium supplemented with 0.5 mg/l BAP or 0.5 mg/l NAA, respectively.  Bulblets also were formed from young shoot on MS supplemented with 0.5 mg/l NAA. The highest number of 4.5 bulblets per an explant was recorded from longitude-divided bubbles on MS medium containing 0.5 mg/l NAA and 0.2 mg/l BAP after 60 days in culture. The regererated plants produced quality roots on half strength MS supplemented with the combination of 1.0 mg/ l NAA and 0.2 mg / l BAP. More than 90% of rooted plants in vitro were survival on artificial soil TN1 in the nursery.

scholarly journals Adjustment of Mineral Elements in the Culture Medium for the Micropropagation of Three Vriesea Bromeliads from the Brazilian Atlantic Forest: The Importance of Calcium

HortScience ◽  
2009 ◽  
Vol 44 (1) ◽  
pp. 106-112 ◽  
Author(s):  
Alice Noemí Aranda-Peres ◽  
Lázaro Eustáquio Pereira Peres ◽  
Edson Namita Higashi ◽  
Adriana Pinheiro Martinelli
Keyword(s):  
New Media ◽  
Mineral Composition ◽  
Culture Media ◽  
Dry Weight ◽  
Defined Medium ◽  
Ms Medium ◽  
Media Formulation ◽  
Half Strength

Many different species of Bromeliaceae are endangered and their conservation requires specific knowledge of their growth habits and propagation. In vitro culture of bromeliads is an important method for efficient clonal propagation and in vitro seed germination can be used to maintain genetic variability. The present work aims to evaluate the in vitro growth and nutrient concentration in leaves of the epiphyte bromeliads Vriesea friburguensis Mez, Vriesea hieroglyphica (Carrière) E. Morren, and Vriesea unilateralis Mez, which exhibit slow rates of growth in vivo and in vitro. Initially, we compared the endogenous mineral composition of bromeliad plantlets grown in half-strength Murashige and Skoog (MS) medium and the mineral composition considered adequate in the literature. This approach suggested that calcium (Ca) is a critical nutrient and this was considered for new media formulation. Three new culture media were defined in which the main changes to half-strength MS medium were an increase in Ca, magnesium, sulfur, copper, and chloride and a decrease in iron, maintaining the nitrate:ammonium rate at ≈2:1. The main difference among the three new media formulated was Ca concentration, which varied from 1.5 mm in half-strength MS to 3.0, 6.0, and 12 mm in M2, M3, and M4 media, respectively. Consistently, all three species exhibited significantly higher fresh and dry weight on M4, the newly defined medium with the highest level of Ca (12 mm). Leaf nitrogen, potassium, zinc, magnesium, and boron concentrations increased as Ca concentration in the medium increased from 1.5 to 12 mm.

scholarly journals Effective medium for in vitro sprouting of the buds and multiplication of the plantlets, and identification of the fungal contaminant associated with the explants of Gnetum

2021 ◽  
Vol 16 (2) ◽  
pp. 001-013
Author(s):  
Abwe Mercy Ngone ◽  
Lawrence Monah Ndam ◽  
Rita Mungfu Njilar ◽  
Doungous Oumar ◽  
Thomas Eku Njock
Keyword(s):  
Culture Medium ◽  
Culture Media ◽  
Fungal Growth ◽  
Growth Media ◽  
Fungal Contamination ◽  
Surface Sterilization ◽  
Pure Cultures ◽  
Nodal Cuttings ◽  
Baseline Information

Plant tissue culture requires the optimization of growth media. Gnetum, known locally in Cameroon as “Eru” is an indigenous gymnospermous vegetable with diverse medicinal, nutritional, cultural and socio-economic values. This resource is over-exploited and expected to neighboring countries, resulting to increased scarcity in the forest. Preliminary work on the in vitro culture of nodal cuttings was faced by the problem of fungal contamination. It was therefore necessary to isolate and identify the fungal contaminant, optimize the surface sterilization of field material and compose an appropriate medium for sprouting. Pure cultures of the fungus were obtained and grown on Potato Dextrose Agar (PDA) and Sabouraud Dextrose Agar (SDA). The identification was based on the appearance of the fungal growth on plates and also on the microscopic view. This was affected by the use of keys. Gnetum explants were disinfected with the various concentrations of disinfectants, preceded in some instances by pre-treatments, as well as incorporating fungicides in the culture medium. Two different culture media were employed: the Woody Plant Medium (WPM) and the Murashige and Skoog (MS) based establishment medium (Y-1). Gnetum was found to live in association with a complex of Microsporum species. The level of contamination of cultures was reduced from 100% to 40% when pre-treated before disinfection and even lower to 10% by incorporating fungicides in the medium. Sprouting was observed in WPM. This study provides baseline information on the in vitro propagation of Gnetum and thus opens up avenues for more research to be carried out in this field.

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