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.

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 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.

scholarly journals Combining Medicinal Plant In Vitro Culture with Machine Learning Technologies for Maximizing the Production of Phenolic Compounds

Antioxidants ◽  
2020 ◽  
Vol 9 (3) ◽  
pp. 210 ◽  
Author(s):  
Pascual García-Pérez ◽  
Eva Lozano-Milo ◽  
Mariana Landín ◽  
Pedro Pablo Gallego
Keyword(s):  
Machine Learning ◽  
Phenolic Compounds ◽  
In Vitro Culture ◽  
Culture Media ◽  
Radical Scavenging Activity ◽  
Radical Scavenging ◽  
Total Phenolic ◽  
Starting Point ◽  
Plant In Vitro Culture

We combined machine learning and plant in vitro culture methodologies as a novel approach for unraveling the phytochemical potential of unexploited medicinal plants. In order to induce phenolic compound biosynthesis, the in vitro culture of three different species of Bryophyllum under nutritional stress was established. To optimize phenolic extraction, four solvents with different MeOH proportions were used, and total phenolic content (TPC), flavonoid content (FC) and radical-scavenging activity (RSA) were determined. All results were subjected to data modeling with the application of artificial neural networks to provide insight into the significant factors that influence such multifactorial processes. Our findings suggest that aerial parts accumulate a higher proportion of phenolic compounds and flavonoids in comparison to roots. TPC was increased under ammonium concentrations below 15 mM, and their extraction was maximum when using solvents with intermediate methanol proportions (55–85%). The same behavior was reported for RSA, and, conversely, FC was independent of culture media composition, and their extraction was enhanced using solvents with high methanol proportions (>85%). These findings confer a wide perspective about the relationship between abiotic stress and secondary metabolism and could serve as the starting point for the optimization of bioactive compound production at a biotechnological scale.

474 Effect of four assisted hatching techniques and two in vitro culture media on embryo hatching rate

2017 ◽  
Vol 95 (suppl_4) ◽  
pp. 231-231
Author(s):  
N. C. Negota ◽  
L. P. Nethenzheni ◽  
N. R. Serota
Keyword(s):  
In Vitro Culture ◽  
Culture Media ◽  
Assisted Hatching ◽  
Hatching Rate

160 COMPARATIVE STUDY OF IN VITRO CULTURE MEDIA AND ASSISTED HATCHING TECHNIQUES ON MOUSE EMBRYOS

2017 ◽  
Vol 29 (1) ◽  
pp. 188
Author(s):  
N. C. Negota ◽  
L. P. Nethenzheni ◽  
M. L. Mphaphathi ◽  
D. M. Barry ◽  
T. L. Nedambale
Keyword(s):  
In Vitro Culture ◽  
Chorionic Gonadotropin ◽  
Culture Medium ◽  
Culture Media ◽  
Assisted Hatching ◽  
Hatching Rate ◽  
Significant Difference ◽  
Equine Chorionic Gonadotropin ◽  
F1 Generation

The in vitro culture media and assisted hatching techniques remain challenging obstacles to be utilised widely. Mechanical, chemical, enzymatic thinning, and laser-assisted techniques have been used previously but information is still lacking on its application in livestock. The aim of this study was to compare the effect of 2 in vitro culture media (Hamster F10 and TMC-199) and 4 (mechanical, chemical, enzymatic, and laser) assisted hatching techniques on blastocyst formation and hatching rate using murine embryos as a model. The C57/b and Balb/c breeds were raised until they reached maturity and bred naturally to produce F1 generation. The light in the breeding house was controlled at 14 h light and 10 h dark. Feed and water were provided ad libitum for the mice. Superovulation of females were stimulated using equine chorionic gonadotropin and human chorionic gonadotropin. The F1 generation was used for the collection of the 400 blastocysts and randomly allocated into 4 assisted hatching techniques. Blastocysts were paired into a group of 10 and replicated 4 times for each assisted hatching technique. The general linear model of SAS version 9.4 (SAS Institute Inc., Cary, NC, USA) was used to analyse the data. Assisted hatching techniques of laser, mechanical, enzymatic, and chemical yielded 46.9 ± 37.1, 51.1 ± 40.2, 39.1 ± 35.8, and 33.3 ± 4.5%, respectively, under in vitro culture of Hamster F10. The TCM-199, laser, mechanical, enzymatic, and chemical assisted hatching techniques yielded 56.3 ± 43.3, 52.6 ± 35.5, 49.2 ± 37.5, and 33.9 ± 35.5%, respectively, with a significant difference. There was no significant difference observed in assisted hatching techniques and Hamster F10 culture medium. However, the hatching rate of embryos for all techniques was higher when in vitro cultured in TCM than cultured in Hamster F10. Hatching rate of blastocysts increased from chemical, enzymatic, mechanical, and laser with response to Hamster F10 and TCM; thus, laser is a suitable assisted hatching technique with TCM-199.

In vitro culture of pointed gourd (Trichosanthes dioica Roxb.)

1970 ◽  
Vol 35 (1) ◽  
pp. 135-142 ◽  
Author(s):  
MA Malek ◽  
D Khanam ◽  
M Khatun ◽  
MH Molla ◽  
MA Mannan
Keyword(s):  
Plant Regeneration ◽  
In Vitro Culture ◽  
Culture Media ◽  
Culture Conditions ◽  
Root Induction ◽  
Ms Medium ◽  
Trichosanthes Dioica ◽  
Pointed Gourd ◽  
Regenerated Plantlets

An experiment was conducted to study the in vitro culture of pointed gourd. Cotyledon rescued from physiologically matured seeds (PMS) and immatured seeds (IMS) of pointed gourd were used as explants. Cotyledon excised from PMS responded very well in all culture conditions. Plant regenerated from cotyledon of PMS ranged from 38 to 96% in different hormonal formulations of culture media. Highest percentage of shoot regeneration was observed in MS + 1.0 mg/l BAP and lowest in MS + 2.5 mg/l BAP. No plant regeneration was observed in cotyledon from immatured seeds. The highest percentage of root induction (99%) was achieved in half MS medium supplemented with 0.5 mg/l NAA. The regenerated plantlets were successfully established in earthen pot. Keywords: Cotyledon; in vitro; pointed gourd. DOI: 10.3329/bjar.v35i1.5874Bangladesh J. Agril. Res. 35(1) : 135-142, March 2010

scholarly journals 146COMPARISON OF EMBRYO CULTURE MEDIA FOR BLASTOCYST DEVELOPMENT AFTER INTRACYTOPLASMIC SPERM INJECTION OF IN VITRO-MATURED EQUINE OOCYTES

2004 ◽  
Vol 16 (2) ◽  
pp. 195
Author(s):  
Y.H. Choi ◽  
D.D. Varner ◽  
K. Hinrichs
Keyword(s):  
In Vitro Culture ◽  
Embryo Culture ◽  
Intracytoplasmic Sperm Injection ◽  
Culture Media ◽  
Polar Body ◽  
Developmental Competence ◽  
Blastocyst Development ◽  
Vitro Fertilization ◽  
Significant Difference

Research on in vitro culture of equine embryos has been scant, due to failure of equine in vitro fertilization to be repeatably successful. We have recently obtained high fertilization rates of equine oocytes via intracytoplasmic sperm injection (ICSI) using a piezo drill (Choi et al., 2002 Reproduction 123, 455–465). Culture of presumptive zygotes in G1.2/2.2 medium resulted in 63% cleavage and an average of 15 cells at 4d, but only 2 to 9% blastocyst development at 7 days (Choi et al., 2003 Theriogenology 59, 1219–1229). In the present study, we evaluated the effect of two different culture media, G1.3/G2.3 v. DMEM/F-12, with or without FBS, on blastocyst development after ICSI. Oocytes were collected from slaughterhouse-derived ovaries by follicular scraping and were matured in vitro for 24h in M199 with 10% FBS and 5μUmL−1 FSH. After culture, oocytes having a polar body (198/305; 65%) were fertilized by ICSI with frozen-thawed equine sperm using a piezo drill. Presumptive zygotes were cultured in 1 of 4 media: G1.3/G2.3 (which includes 0.8% BSA) with or without 10% FBS, or in DMEM/F-12 with 0.5% BSA, with or without 10% FBS. Culture was performed in microdroplets at 5μL/zygote under oil at 38.2°C in an atmosphere of 5% CO2, 5% O2 and 90% N2 for 7.5 days. In G1.3/2.3 treatments, G1.3 media were completely refreshed at 48h, zygotes were transferred to G2.3 (with or without FBS as per the first stage) at 96h, and were completely refreshed with the same media at 144h. In DMEM/F-12 treatments, media were completely refreshed every other day. Three to 5 replicates were performed in each treatment, and data were analyzed by chi-square test. There were no significant differences in cleavage rates (59–64%) among treatments. The rate of development to blastocyst, per oocyte injected, in G1.3/G2.3/BSA (1/49, 2%) was significantly lower (P&lt;0.05) than that for the other three treatments: G1.3/2.3/BSA/FBS (9/49, 18%), DMEM/F-12/BSA (9/50, 18%), or DMEM/F-12/BSA/FBS (10/50, 20%). There was no significant difference in blastocyst development among the latter three treatments. These findings indicate that G1.3/2.3 media with BSA only do not adequately support growth of equine embryos. Development of up to 20% of injected oocytes to the blastocyst stage in G media supplemented with FBS, in DMEM/F-12/BSA or in DMEM/F-12/BSA/FBS represents the highest in vitro equine blastocyst rate in medium alone (i.e. without co-culture) yet reported. The success of DMEM/F-12 as an embryo culture medium may provide a relatively simple basis for equine in vitro culture programs. To determine whether this medium was able to support further developmental competence, we cultured equine embryos resulting from nuclear transfer of in vitro-matured oocytes in DMEM/F-12+10% FBS (without BSA). We transferred 4 resulting blastocysts to recipient mares by transcervical transfer; one pregnancy is ongoing at 230d gestation at the time of this writing. This work was supported by the Link Equine Research Endowment Fund, Texas A&amp;M University.

scholarly journals Experiencia práctica: socialización de conceptos, aplicaciones y beneficios de la biotecnología en Costa Rica experience: socializing the concepts, applications and benefits of biotechnology in Costa Rica

2019 ◽  
Vol 67 (2SUPL) ◽  
pp. S26-S35
Author(s):  
Emanuel García-Jiménez ◽  
Andrés-M. Gatica-Arias ◽  
Frank Solano-Campos ◽  
Ana Abdelnour-Esquivel
Keyword(s):  
Costa Rica ◽  
In Vitro Culture ◽  
New Technologies ◽  
Human Diet ◽  
Culture Techniques ◽  
Biotechnological Processes ◽  
Growing Population

Biotechnological processes have accompanied humanity since the beginning of civilization; proof of this is the use of yeasts for the preparation of bread, wine and beer. Biotechnology has also been fundamental in the improvement of plants and animals that are part of the human diet; and in vitro culture techniques have accelerated the processes for obtaining better crops  for our growing population. The project “Biotechnology for everyone: Socialization of concepts, applications and benefits” socializes the concepts, applications and benefits of biotechnology among educators, opinion-forming groups, and producers in the agricultural and food sectors. The project has successfully used new technologies to reach its goals.

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. 

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