Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 9

Tissue Culture Related Abstracts

9 Surface Sterilization of Aquatic Plant, Cryptopcoryne affinis by Using Clorox and Mercury Chloride

Authors: Sridevi Devadas

Abstract:

This study was aimed to examine the combination efficiency of Clorox (5.25% Sodium Hypochlorite) and mercury chloride (HgCl2) as reagent for surface sterilization process of aquatic plant, Cryptocoryne affinis (C. affinis). The treatment applied 10% of the Clorox and 0.1 ppm of mercury chloride. The maximum exposure time for Clorox and mercury chloride was 10 min and 60 sec respectively. After exposed to the treatments protocols (T1-T15) the explants were transferred to culture room under control temperature at 25°C ± 2°C and subjected to 16 hours fluorescence light (2000 lumens) for 30 days. The both sterilizing agents were not applied on control specimens. Upon analysis, the result indicates all of the treatments protocols produced sterile explants at range of minimum 1.5 ± 0.7 (30%) to maximum 5.0 ± 0.0 (100%). Meanwhile, maximum 1.0 ± 0.7 numbers of leaves and 1.4 ± 0.6 numbers of roots have been produced. The optimized exposure time was 0 to 15 min for Clorox and 30 sec for HgCl2 whereby 90% to 100% sterilization was archived at this condition.

Keywords: Tissue Culture, Cryptocoryne affinis, surface sterilization, clorox, mercury chloride

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8 Surface Sterilization Of Aquatic Plant, Cryptocoryne affinis by Using Clorox and Mercury Chloride

Authors: Sridevi Devadas

Abstract:

This study was aimed to examine the combination efficiency of Clorox (5.25% Sodium Hypochlorite) and mercury chloride (HgCl2) as a reagent for surface sterilization process of aquatic plant and cryptocoryne affinis (C. affinis). The treatment applied 10% of the Clorox and 0.1ppm of mercury chloride. The maximum exposure time for clorox and mercury chloride was 10min and 60sec respectively. After exposed to the treatments protocols (T1-T15) the explants were transferred to culture room under control temperature at 25°C ± 2°C and subjected to 16 hours fluorescence light (2000 lumens) for 30 days. The both sterilizing agents were not applied on control specimens. Upon analysis, The result indicates all of the treatments protocols produced sterile explants at range of minimum 1.5 ± 0.7 (30%) to maximum 5.0 ± 0.0 (100%). Meanwhile, maximum 1.0 ± 0.7 numbers of leaves and 1.4 ± 0.6 numbers of roots have been produced. The optimized exposure time was 0 to 15 min for Clorox and 30 sec for HgCl2 whereby 90% to 100% sterilization was archived at this condition.

Keywords: Tissue Culture, Cryptocoryne affinis, surface sterilization, clorox, mercury chloride

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7 Empirical Measures to Enhance Germination Potential and Control Browning of Tissue Cultures of Andrographis paniculata

Authors: Nidhi Jindal, Ashok Chaudhury, Manisha Mangal

Abstract:

Andrographis paniculata, (Burm f.) Wallich ex. Nees (Family Acanthaceae) popularly known as King of Bitters, is an important medicinal herb. It has an astonishingly wide range of medicinal properties such as anti-inflammatory,antidiarrhoeal, antiviral, antimalarial, hepatoprotective, cardiovascular, anticancer, and immunostimulatory activities. It is widely cultivated in southern Asia. Though propagation of this herb generally occurs through seeds, it has many germination problems which intrigued scientists to work out on the alternative techniques for its mass production. The potential of tissue culture techniques as an alternative tool for AP multiplication was found to be promising. However, the high mortality rate of explants caused by phenolic browning of explants is one of the difficulties reported. Low multiplication rates were reported in the proliferation phase, as well as cultures decline characterized by leaf fall and loss of overall vigor. In view of above problems, a study was undertaken to overcome seed dormancy to improve germination potential and to investigate further on the possible means for successful proliferation of cultures via preventive approaches to overcome failures caused by phenolic browning. Experiments were conducted to improve germination potential and among all the chemical and mechanical trials, scarification of seeds with sand paper proved to be the best method to enhance the germination potential (82.44%) within 7 days. Similarly, several pretreatments and media combinations were tried to overcome browning of explants leading to the conclusion that addition of 0.1% citric acid and 0.2% of ascorbic acid in the media followed by rapid sub culturing of explants controlled browning and decline of explants by 67.45%.

Keywords: Tissue Culture, Plant Tissue Culture, Germination, empirical measure

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6 Comparative Study of Antioxidant Activity in in vivo and in vitro Samples of Purple Greater Yam (Dioscorea alata L).

Authors: Sakinah Abdullah, Rosna Mat Taha

Abstract:

Antioxidants are compounds that protect cells against the damaging effects of reactive oxygen species such as singlet oxygen, superoxide, peroxyl radicals, and peroxynitrite which result in oxidative stress leading to cellular damage. Natural antioxidant are in high demand because of their potential in health promotion and disease prevention and their improved safety and consumer acceptability. Plants are rich sources of natural antioxidant. Dioscorea alata L. known as 'ubi badak' in Malaysia were well known for their antioxidant content, but this plant was seasonal. Thus, tissue culture technique was used to mass propagate this plant. In the present work, a comparative study between in vitro (from tissue culture) and in vivo (from intact plant) samples of Dioscorea alata L. for their antioxidant potential by 2,2-diphenil -1- picrylhydrazyl (DPPH) radical scavenging activity method and their total phenolic and flavonoid contents were carried out. All samples had better radical scavenging activity but in vivo samples had the strongest radical scavenging activity compared to in vitro samples. Furthermore, tubers from in vivo samples showed the greatest free radical scavenging effect and comparatively greater phenolic content than in vitro samples.

Keywords: Tissue Culture, in vivo, In vitro, DPPH, antioxidant, Dioscorea alata

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5 In vitro Environmental Factors Controlling Root Morphological Traits of Pineapple (Ananas comosus L. Merr)

Authors: S. Mohajer , R. M. Taha, M. Adel

Abstract:

Developing our knowledge of when pineapple roots grow can lead to improved water, fertilizer applications, and more precise culture management. This paper presents current understanding of morphological traits in pineapple roots, highlighting studies using incubation periods and various solid MS media treated with different sucrose concentrations and pH, which directly assess in vitro environmental factors. Rooting parameters had different optimal sucrose concentrations and incubation periods. All shoots failed to root in medium supplemented with sucrose at 5 g/L and no roots formed within the first 45 days in medium enriched with sucrose at 10 g/L. After 75 days, all shoots rooted in medium enriched with 10 and 20 g/L sucrose. Moreover, MS medium supplied with 20 g/L sucrose resulted in the longest and the highest number of roots with 27.3 mm and 4.7, respectively. Root function, such as capacity for P and N uptake, declined rapidly with root length. As a result, the longer the incubation period, the better the rooting responses would be.

Keywords: Tissue Culture, Environmental Factors, pineapple, in vitro rooting

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4 Effect of Interaction between Different Concentrations of Colchicine, Time Duration and Two Verities of Crepis capillaris on Chromosome Polyploidy in vitro Culture

Authors: Mosleh M. S. Duhoky, Payman A. A. Zibari

Abstract:

These experiments were conducted at Tissue Culture Laboratory/ Faculty of Agriculture and Forestry/ University of Duhok during the period from January 2011 to May 2013. The objectives of this study were to study the effects of interaction between three different factors on percentage of polyploidy of Crepis capillaris by using Tissue culture technology. Concerning the data it is obvious that shaking of Crepis capillaris with 2B chromosome with 0.15 mM for ten days inscribed a high percentage of polyploidy within most fifteen passages.

Keywords: Tissue Culture, Polyploidy, crepis capillaris

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3 Influence of Elicitors on Callus Growth and Active Ingredient in Echinacea purpurea

Authors: Mohamed Abdelfattah Meawad Hamza, H. A. Bosila, M. A. Zewil

Abstract:

This research aims to study the effect of different sources of elicitors for increase growth and active ingredients in callus of Echinacea purpurea plant. Callus that have been obtained from leaf explant, was used to conduct the following studies. A study of the impact of both the phenylalanine and tyrosine (50, 100,150 and 200 mg/l.) individually and casein hydrolysate (100, 200 and 300 mg/l.) supplemented to MS medium. Results show that Casein hydrolysate 100 mg/l. has achieved the better results in both callus fresh weight 1.881 g/explant after 8 weeks of the incubation period and callus growth rate 0.398 g/explant after 6 weeks of the incubation period, while gave add 200 mg/l. The best results in total carbohydrate 2.444 mg/ 100 mg dry weight. Phenylalanine 150 mg/l. has achieved the best results in callus dry weight 0.156 g/explant after 8 weeks of incubation period. Tyrosine 200 mg/l. recorded the best result for positive production of caffeic acid 0.460 mg/ 100 mg dry weight after 4 weeks incubation period.

Keywords: Tissue Culture, casein, echinacea, tyrosine

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2 Enhancement of Growth Regulators to Callus Formation and Silymarin Content from Different Explants of Silybum marianum Seedling

Authors: M. A. Hamza, H. A. Bosila, M. A. Zewil, I. M. Harridy

Abstract:

Silymarin is one active component extracted from milk thistle Silybum marianum; it is flavonoid recognized for its ability to benefit people with liver disorders and as a protective compound against liver damaging agents. For this reason, this research aims to study the effect of growth regulators (BA+NAA) and explant type (cotyledon, hypocotyl, and root) to increase the growth and active ingredients (silymarin) in callus of S. mariaum plant. The results showed that cotyledon explant which have been cultured in MS medium supplemented with BA 0.4 mg/l. +NAA 0.25 mg/l. Led to obtain the best results in callus fresh weight (1.847a) and callus dry weight (0.155a). On the other hand, the same explant (cotyledon) cultured in MS medium supplemented with BA 1.6 mg/l. + NAA 0.5 mg/l. The suitable condition to silymarin content (0.132 mg/100 mg dry weight). And also, it turned out, lack of importance of the use of hypocotyl and root in the production of callus and silymarin compared to cotyledon.

Keywords: Tissue Culture, callus, silybum, cotyledon

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1 Influence of Genotype, Explant, and Hormone Treatment on Agrobacterium-Transformation Success in Salix Callus Culture

Authors: Lukas J. Evans, Danilo D. Fernando

Abstract:

Shrub willows (Salix spp.) have many characteristics which make them suitable for a variety of applications such as riparian zone buffers, environmental contaminant sequestration, living snow fences, and biofuel production. In some cases, these functions are limited due to physical or financial obstacles associated with the number of individuals needed to reasonably satisfy that purpose. One way to increase the efficiency of willows is to bioengineer them with the genetic improvements suitable for the desired use. To accomplish this goal, an optimized in vitro transformation protocol via Agrobacterium tumefaciens is necessary to reliably express genes of interest. Therefore, the aim of this study is to observe the influence of tissue culture with different willow cultivars, hormones, and explants on the percentage of calli expressing reporter gene green florescent protein (GFP) to find ideal transformation conditions. Each callus was produced from 1 month old open-pollinated seedlings of three Salix miyabeana cultivars (‘SX61’, ‘WT1’, and ‘WT2’) from three different explants (lamina, petiole, and internodes). Explants were cultured for 1 month on an MS media with different concentrations of 6-Benzylaminopurine (BAP) and 1-Naphthaleneacetic acid (NAA) (No hormones, 1 mg⁻¹L BAP only, 3 mg⁻¹L NAA only, 1 mg⁻¹L BAP and 3 mg⁻¹L NAA, and 3 mg⁻¹L BAP and 1 mg⁻¹L NAA) to produce a callus. Samples were then treated with Agrobacterium tumefaciens at an OD600 of 0.6-0.8 to insert the transgene GFP for 30 minutes, co-cultivated for 72 hours, and selected on the same media type they were cultured on with added 7.5 mg⁻¹L of Hygromycin for 1 week before GFP visualization under a UV dissecting scope. Percentage of GFP expressing calli as well as the average number of fluorescing GFP units per callus were recorded and results were evaluated through an ANOVA test (α = 0.05). The WT1 internode-derived calli on media with 3 mg-1L NAA+1 mg⁻¹L BAP and mg⁻¹L BAP alone produced a significantly higher percentage of GFP expressing calli than each other group (19.1% and 19.4%, respectively). Additionally, The WT1 internode group cultured with 3 mg⁻¹L NAA+1 mg⁻¹L BAP produced an average of 2.89 GFP units per callus while the group cultivated with 1 mg⁻¹L BAP produced an average of 0.84 GFP units per callus. In conclusion, genotype, explant choice, and hormones all play a significant role in increasing successful transformation in willows. Future studies to produce whole callus GFP expression and subsequent plantlet regeneration are necessary for a complete willow transformation protocol.

Keywords: Tissue Culture, Agrobacterium, callus, Salix

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