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

Search results for: tissue culture

9 21st Century Biotechnological Research and Development Advancements for Industrial Development in India

Authors: Monisha Isaac

Abstract:

Biotechnology is a discipline which explains the use of living organisms and systems to construct a product, or we can define it as an application or technology developed to use biological systems and organisms processes for a specific use. Particularly, it includes cells and its components use for new technologies and inventions. The tools developed can be further used in diverse fields such as agriculture, industry, research and hospitals etc. The 21st century has seen a drastic development and advancement in biotechnology in India. Significant increase in Government of India’s outlays for biotechnology over the past decade has been observed. A sectoral break up of biotechnology-based companies in India shows that most of the companies are agriculture-based companies having interests ranging from tissue culture to biopesticides. Major attention has been given by the companies in health related activities and in environmental biotechnology. The biopharmaceutical, which comprises of vaccines, diagnostic, and recombinant products is the most reliable and largest segment of the Indian Biotech industry. India has developed its vaccine markets and supplies them to various countries. Then there are the bio-services, which mainly comprise of contract researches and manufacturing services. India has made noticeable developments in the field of bio industries including manufacturing of enzymes, biofuels and biopolymers. Biotechnology is also playing a crucial and significant role in the field of agriculture. Traditional methods have been replaced by new technologies that mainly focus on GM crops, marker assisted technologies and the use of biotechnological tools to improve the quality of fertilizers and soil. It may only be a small contributor but has shown to have huge potential for growth. Bioinformatics is a computational method which helps to store, manage, arrange and design tools to interpret the extensive data gathered through experimental trials, making it important in the design of drugs.

Keywords: Biotechnology, Agriculture, advancement, bio-services, bio-industries, bio-pharmaceuticals

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

Authors: R. M. Taha, S. Mohajer, 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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7 An Effect of Organic Supplements on Stimulating Growth of Vanda and Mokara Seedlings in Tissue Culture

Authors: CHOCKPISIT THEPSITHAR, Kullanart Obsuwan

Abstract:

This study aimed to investigate effect of different organic supplements on growth of Vanda and Mokara seedlings. Vanda and Mokara seedlings approximately 0.2 and 0.3 cm. in height were sub-cultured onto VW supplemented with 150 ml/L coconut water, 100 g/L potato extract, 100 g/L ‘Gros Michel’ banana (AAA group) and 100 g/L ‘Namwa’ banana (ABB group). The explants were sub-cultured onto the same medium every month for 3 months. The best medium increased stem height to 0.52 and 0.44 Cm. in Vanda and Mokara respectively was supplemented with coconut water. The maximum fresh weight of Vanda (0.59 g) was found on medium supplemented with ‘Gros Michel’ banana while Mokara cultured on medium supplemented with Potato extract had the maximum fresh weight (0.27 g) and number of roots (5.20 roots/shoot) statistically different (p≤ 0.05) to other treatments. However, Vanda cultured on medium supplemented with ‘Namwa’ banana had the maximum number of roots (3.80 roots/shoot). Our results suggested that growth of different orchid genera was responded diversely to different organic supplements. 

 

Keywords: orchid, in vitro propagation, fresh weight, plant height

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6 Molecular Analysis of Somaclonal Variation in Tissue Culture Derived Bananas Using MSAP and SSR Markers

Authors: Emma K. Sales, Nilda G. Butardo

Abstract:

The project was undertaken to determine the effects of modified tissue culture protocols e.g. age of culture and hormone levels (2,4-D) in generating somaclonal variation. Moreover, the utility of molecular markers (SSR and MSAP) in sorting off types/somaclones were investigated.

Results show that somaclonal variation is in effect due to prolonged subculture and high 2,4-D concentration. The resultant variation was observed to be due to high level of methylation events specifically cytosine methylation either at the internal or external cytosine and was identified by methylation sensitive amplification polymorphism (MSAP).Simple sequence repeats (SSR) on the other hand, was able to associate a marker to a trait of interest.

These therefore, show that molecular markers can be an important tool in sorting out variation/mutants at an early stage.

Keywords: methylation, MSAP, somaclones, SSR, subculture

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5 Recovering Taraxacum kok-saghyz Rodin. via Seed and Callus Culture

Authors: K. Uteulin, S. Mukhambetzhanov, I. Rakhimbaev

Abstract:

This experiment was performed to optimize the medium for tissue culture of Taraxacum kok-saghyz Rodin. Different tissue culture approaches such as shoot regeneration from seed, callus formation from leaf explants and plant regeneration from callus were investigated in this study. All the explants were cultured on MS basal medium supplemented with 20g/l sucrose, 7g/l agar and different plant growth regulators. Seeds of Taraxacum kok-saghyzwere cultured on media containing different levels of BA and 2,4-D (0.5, 1.0 and 3.0mg/L) to direct shoot regeneration study. Leaf explants were cultured in different combination of BA (at three levels: 0.5, 1.0 and 3.0mg/L) and zeatin (at two levels: 0.5 and 1.0mg/L) to examine callus formation. After the callus formation the formed calli were cultured on different combinations of BA and NAA for shoot regeneration. BA at three levels (0.5 and 1.0 and 3.0mg/L) and NAA at two levels (0.5 and 1.0mg/L) in all possible combinations were used for shoot regeneration from callus. The results showed that the treatment containing 1.0mg/L 2,4-D in combination with 1.0mg/L BA was found to be the best one for shoot regeneration from seeds. The treatment with 1.0mg/L BA in combination with 1.0mg/L zeatin were found to be suitable treatments for callus production from leaf explants, as well. Moreover, 0.5mg/L BA alone or in combination with 1.0mg/L NAA were found to be the best treatments for shoot regeneration from callus.

Keywords: callus, Taraxacum kok-saghyz Rodin, shoot regeneration

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4 In vitro Culture Medium Sterilization by Chemicals and Essential Oils without Autoclaving and Growth of Chrysanthemum Nodes

Authors: CHOCKPISIT THEPSITHAR, Aree Thongpukdee, Wittaya Deein

Abstract:

Plant tissue culture is an important in vitro technology applied for agricultural and industrial production. A sterile condition of culture medium is one of the main aspects. The alternative technique for medium sterilization to replace autoclaving was carried out. For sterilization of plant tissue culture medium without autoclaving, ten commercial pure essential oils and 5 disinfectants were tested. Each essential oil or disinfectant was added to a 20-mL Murashige and Skoog (MS) medium before medium was solidified in a 120-mL container, kept for 2 weeks before evaluating sterile conditions. Treated media, supplemented with essential oils or disinfectants, were compared to control medium, autoclaved at 121 degree Celsius for 15 min. Sterile conditions of MS medium were found 100% from betel oil or clove oil (18 mL/20 mL medium), cinnamon oil (36 mL/20 mL medium), lavender oil or holy basil oil (108 mL/20 mL medium), and lemon oil or tea tree oil or turmeric oil (252 mL/20 mL medium), compared to 100% sterile condition from autoclaved medium. For disinfectants, 2% iodine + 2.4% potassium iodide, 2% merbromine solution, 10% povidone-iodine, 6% sodium hypochlorite or 0.1% thimerosal at 36 mL/20 mL medium provided 100% sterile conditions. Furthermore, growth of new shoots from chrysanthemum node explants on treated media (fresh weight, shoot length, root length and number of node) were also reported and discussed in the comparison of those on autoclaved medium.

Keywords: Disinfectants, essential oils, chrysanthemum, sterilizing agents, MS medium, sterilization of medium without autoclaving

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3 Prevention of Biofilm Formation in Urinary Catheter by Coating Enzymes/ Gentamycin/ EDTA

Authors: Niraj A. Ghanwate, P V Thakare, P R Bhise, Ashish Dhanke, Shubhangi Apotikar

Abstract:

Urinary Tract Infections (UTI) account for an estimated 25-40% nosocomial infection, out of which 90% are associated with urinary catheter, called Catheter associated urinary tract infection (CAUTI). The microbial populations within CAUTI frequently develop as biofilms. In the present study, microbial contamination of indwelling urinary catheters was investigated. Biofilm forming ability of the isolates was determined by tissue culture plate method. Prevention of biofilm formation in the urinary catheter by Pseudomonas aeruginosa was also determined by coating the catheter with some enzymes, gentamycin and EDTA. It was found that 64% of the urinary catheters get contaminated during the course of catheterization. Of the total 6 isolates, biofilm formation was seen in 100% Pseudomonas aeruginosa and E. coli, 90% in Enterococci, 80% in Klebsiella and 66% in S. aureus. It was noted that the biofilm production by Pseudomonas was prolonged by 7 days in amylase, 8 days in protease, 6 days in lysozyme, 7days in gentamycin and 5 days in EDTA treated catheter.

Keywords: Pseudomonas, Biofilm, Enzymes, EDTA, CAUTI

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2 Screening and Evaluation of in vivo and in vitro Generated Insulin Plant (Vernonia divergens) for Antimicrobial and Anticancer Activities

Authors: Santosh KUMAR, KANAK SINHA, Anand Prakash, Anita K Verma

Abstract:

Vernonia divergens Benth., commonly known as “Insulin Plant” (Fam: Asteraceae) is a potent sugar killer. Locally the leaves of the plant, boiled in water are successfully administered to a large number of diabetic patients. The present study evaluates the putative anti-diabetic ingredients, isolated from the in vivo and in vitro grown plantlets of V. divergens for their antimicrobial and anticancer activities. Sterilized explants of nodal segments were cultured on MS (Musashige and Skoog, 1962) medium in presence of different combinations of hormones. Multiple shoots along with bunch of roots were regenerated at 1mg l-1 BAP and 0.5 mg l-1 NAA. Micro-plantlets were separated and sub-cultured on the double strength (2X) of the above combination of hormones leading to increased length of roots and shoots. These plantlets were successfully transferred to soil and survived well in nature. The ethanol extract of plantlets from both in vivo & in vitro sources were prepared in soxhlet extractor and then concentrated to dryness under reduced pressure in rotary evaporator. Thus obtainedconcentrated extracts showed significant inhibitory activity against gram negative bacteria like Escherichia coli and Pseudomonas aeruginosa but no inhibition was found against gram positive bacteria. Further, these ethanol extracts were screened for in vitro percentage cytotoxicity at different time periods (24 h, 48 h and 72 h) of different dilutions. The in vivo plant extract inhibited the growth of EAC mouse cell lines in the range of 65, 66, 78, and 88% at 100, 50, 25 & 12.5μg mL-1 but at 72 h of treatment. In case of the extract of in vitro origin, the inhibition was found against EAC cell lines even at 48h. During spectrophotometric scanning, the extracts exhibited different maxima (ʎ) - four peaks in in vitro extracts as against single in in vivo preparation suggesting the possible change in the nature of ingredients during micropropagation through tissue culture techniques.

Keywords: Tissue Culture, anti-cancer, Anti-microbial, EAC mouse cell, Vernonia divergens

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1 Effect of Various Concentrations of Humic Acid on Growth and Development of Eggplant Seedlings in Tissue Cultures at Low Nutrient Level

Authors: Natdhera Sanmanee, Kullanart Obsuwan, Kamolchanok Panishkan, Suluck Namchote, Sirichai Dharmvanij

Abstract:

Humic acids (HAs) have been shown to activate some ion uptakes along with stimulating the lateral roots at effective concentration of micronutrients. However, the effects of HA on ion adsorption by plant roots are not easily explainable due to the varieties of HAs that differ from origins. Therefore, this study was aimed to investigate the effect of various concentrations of HA obtained from the compost derived from mix manures and some agricultural wastes on the growth of eggplant seedlings (Solanum melongena L. cv. Chao Praya) in tissue cultures at low nutrient level. Egg plant seeds were surfaced sterilized and germinated in ½ Murashige and Skoog medium (MS) without HA added or in ¼ MS supplemented with 0, 25, 50, 75 and 100 ppm of HAs. Then, they were cultured for 4 weeks under the controlled environment. The results showed that seedlings grown on ¼MS supplemented with HAs at the concentration of 25 and 50 ppm had the average plant heights (2.49 and 2.28 cm, respectively) higher than the other treatments. Both treatments also significantly showed the maximum average fresh and dry weights (p<0.05). Also the later yielded the highest average number of leaves and the longest average root length (p<0.05). However, there was no statistically different in the number of roots among treatments (p>0.05). This suggested that HAs at the concentration of 25 and 50 ppm could improve the growth of egg plant seedlings in tissue cultures at low nutrient level (¼ MS).

Keywords: Tissue Culture, Growth, Humic Acid , seedling, dry weight, fresh weght

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