Search results for: plant tissue culture

Authors: Sakinah Abdullah, Rosna Mat Taha

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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: Dioscorea alata, tissue culture, antioxidant, in vivo, in vitro, DPPH

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

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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: crepis capillaris, 2B chromosome, tissue culture, polyploidy

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

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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 20 g/l sucrose, 7 g/l agar and different plant growth regulators. Seeds of Taraxacum kok-saghyz were cultured on media containing different levels of BA and 2,4-D (0,5 and 1,0 and 3,0 mg/L) to direct shoot regeneration study. Leaf explants were cultured in different combination of BA (at three levels: 0.5, 1.0 and 3.0 mg/L) and zeatin (at two levels: 0.5 and 1.0 mg/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 3.0 mg/L) and NAA at two levels (0.5 and 1.0 mg/L) in all possible combinations were used for shoot regeneration from callus. The results showed that the treatment containing 1.0 mg/L 2,4-D in combination with 1.0 mg/L BA was found to be the best one for shoot regeneration from seeds. The treatment with 1.0 mg/L BA in combination with 1.0 mg/L zeatin were found to be suitable treatments for callus production from leaf explants, as well. Moreover, 0.5 mg/L BA alone or in combination with 1.0 mg/L NAA were found to be the best treatments for shoot regeneration from callus.

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

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Authors: Maged El-Sayed Mohamed

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Plant tissue culture practices are presented nowadays as the most promising substitute to a whole plant in the terms of secondary metabolites production. They offer the advantages of high production, tunability and they have less effect on plant ecosystems. Lantana camara is a weed, which is common all over the world as an ornamental plant. Weeds can adapt to any type of soil and climate due to their rich cellular machinery for secondary metabolites’ production. This characteristic is found in Lantana camara as a plant of very rich diversity of secondary metabolites with no dominant class of compounds. Aim: This trait has encouraged the author to develop tissue culture experiments for Lantana camara to be a platform for production and manipulation of secondary metabolites through biotransformation. Methodology: The plant was collected in its flowering stage in September 2014, from which explants were prepared from shoot tip, auxiliary bud and leaf. Different types of culture media were tried as well as four phytohormones and their combinations; NAA, 2,4-D, BAP and kinetin. Explants were grown in dark or in 12 hours dark and light cycles at 25°C. A metabolic profile for the produced callus was made and then compared to the whole plant profile. The metabolic profile was made using GC-MS for volatile constituents (extracted by n-hexane) and by HPLC-MS and capillary electrophoresis-mass spectrometry (CE-MS) for non-volatile constituents (extracted by ethanol and water). Results: The best conditions for the callus induction was achieved using MS media supplied with 30 gm sucrose and NAA/BAP (1:0.2 mg/L). Initiation of callus was favoured by incubation in dark for 20 day. The callus produced under these conditions showed yellow colour, which changed to brownish after 30 days. The rate of callus growth was high, expressed in the callus diameter, which reached to 1.15±0.2 cm in 30 days; however, the induction of callus delayed for 15 days. The metabolic profile for both volatile and non-volatile constituents of callus showed more simple background metabolites than the whole plant with two new (unresolved) peaks in the callus’ nonvolatile constituents’ chromatogram. Conclusion: Lantana camara callus production can be itself a source of new secondary metabolites and could be used for biotransformation studies due to its simple metabolic background, which allow easy identification of newly formed metabolites. The callus production gathered the simple metabolic background with the rich cellular secondary metabolite machinery of the plant, which could be elicited to produce valuable medicinally active products.

Keywords: capillary electrophoresis-mass spectrometry, gas chromatography, metabolic profile, plant tissue culture

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Authors: Gayatri Nahak, Satyajit Kanungo, Rajani Kanta Sahu

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Holarrhena antidysenterica Linn. (Apocynaceae) is a typical Indian medicinal plant popularly known as “Indrajav”. Traditionally the plant has been considered a popular remedy for the treatment of dysentery, diarrhea, intestinal worms and the seeds of this plant are also used as an anti-diabetic remedy. In the present study axillary shoot multiplication, callus induction and shoot regeneration from callus culture were obtained on Murashige and Skoog (MS) medium supplemented with different concentrations and combinations of plant growth regulators. Then in vivo and in vitro grown healthy plants were selected for study of antioxidant activity through DPPH and OH methods. Significantly higher antioxidant activity and phenol contents were observed in vitro raised plant in comparison to in vivo plants. The findings indicated the greater amount of phenolic compounds leads to more potent radical scavenging effect as shown in in vitro raised plant in comparison to in vivo plants which showed the ability to utilize tissue culture techniques towards development of desired bioactive metabolites from in vitro culture as an alternative way to avoid using endangered plants in pharmaceutical purposes.

Keywords: Holarrhena antidysenterica, in vitro, in vivo, antioxidant activity

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Authors: Totik Sri Mariani, Sukrasno Isna, Tet Fatt Chia

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Purwoceng (Pimpinella pruatjan Molk) is a legend plant used for increasing stamina by Kings in Java Island, Indonesia. Purpose of this study was to perform meristem culture and detected its stigmasterol by thin layer chromatography (TLC). Our result show that meristem culture could be propagated and grew into plantlet. After extracting intact acclimatized plant derived from meristem culture by hexane, we could detected stigmasterol by TLC. For suggestion, our extraction and TLC method could be used for detecting stigmasterol in others plant.

Keywords: purwoceng (pimpinella pruatjan), meristem culture, extraction, thin layer chromatography

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Authors: Ekramul Hoque, Zinat Ara Eakut Zarin, Ershad Ali

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The growth and development of explants is governed by the effect of nutrient medium. Ammonium nitrate (NH4NO3) as a major salt of stock solution-1 for the preparation of tissue culture medium. But, it has several demerits on human civilization. It is use for the preparation of bomb and other destructive activities. Hence, it is totally ban in our country. A new chemical was identified as a substitute of ammonium nitrate. The concentrations of the other ingredients of major and minor salt were modified from the MS medium. The formulation of new medium is totally different from the MS nutrient composition. The most widely use MS medium composition was used as first check treatment and MS powder (Duchefa Biocheme, The Netherland) was used as second check treatment. The experiments were carried out at the Department of Biotechnology, Sher-e-Bangla Agricultural University, Dhaka, Bangladesh. Two potato varieties viz. Diamant and Asterix were used as experimental materials. The regeneration potentiality of potato onto new medium was best as compare with the two check treatments. The traits -node number, leaf number, shoot length, root lengths were highest in new medium. The plantlets were healthy, robust and strong as compare to plantlets regenerated from check treatments. Three subsequent sub-cultures were made in the new medium to observe the growth pattern of plantlet. It was also showed the best performance in all the parameter under studied. The regenerated plantlet produced good quality minituber under field condition. Hence, it is concluded that, a new plant tissue culture medium as discovered from the Department of Biotechnology, Sher-e-Bangla Agricultural University, Dhaka, Bangladesh under the leadership of Professor Dr. Md. Ekramul Hoque.

Keywords: new medium, potato, regeneration, ammonium nitrate

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Authors: Zelikha Labbani

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Many biotechnology methods have been used in plant breeding programs. The in vitro isolated microspore culture is the one of these methods. For durum wheat, the use of this technology has been limited for a long time due to the low number of embryos produced and also most regeneration plants are albina. The objective of this paper is to show that using isolated microspores culture on durum wheat is possible due to the development of the new methods using the new pretreatment of the microspores before their isolation and cultivation.

Keywords: isolated microspore culture, pretreatments, in vitro embryogenesis, plant breeding program

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Authors: Sushil Pradhan

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Biotechnology contributes to sustainable development in several ways such as biofertilizer production, biopesticide production and management of environmental pollution, tissue culture and biodiversity conservation in vitro, in vivo and in situ, Insectivorous medicinal plant Drosera burmannii Vahl belongs to the Family-Droseraceae under Order-Caryophyllales, Dicotyledoneae, Angiospermeae which has 31 (thirty one) living genera and 194 species besides 7 (seven) extinct (fossil) genera. Locally it is known as “Patkanduri” in Odia. Its Hindi name is “Mukhajali” and its English name is “Sundew”. The earliest species of Drosera was first reported in 1753 by Carolous Linnaeus called Drosera indica L (Indian Sundew). The latest species of Drosera reported by Fleisch A, Robinson, AS, McPherson S, Heinrich V, Gironella E and Madulida D.A. (2011) is Drosera ultramafica from Malaysia. More than 50 % species of Drosera have been reported from Australia and next to Australia is South Africa. India harbours only 3 species such as D. indica L, Drosera burmannii Vahl and D. peltata L. From our Odisha only D. burmannii Vahl is being reported for the first time from the district of Subarnapur near Sonepur (Arjunpur Reserve Forest Area). Drosera plant is autotrophic but to supplement its Nitrogen (N2) requirement it adopts heterotrophic mode of nutrition (insectivorous/carnivorous) as well. The colour of plant in mostly red and about 20-30cm in height with beautiful pink or white pentamerous flowers. Plants grow luxuriantly during November to February in shady and moist places near small water bodies of running water stream. Medicinally it is a popular herb in the locality for the treatment of cold and cough in children in rainy season by the local Doctors (Kabiraj and Baidya). In the present field investigation an attempt has been made to understand the unique reproductive phase and life cycle of the plant thereby planning for its conservation and propagation through various techniques of tissue culture and biotechnology. More importantly besides morphological and anatomical studies, cytological investigation is being carried out to find out the number of chromosomes in the cell and its genomics as there is no such report as yet for Drosera burmannii Vahl. The ecological significance and biodiversity conservation of Drosera with special reference to energy, environmental and chemical engineering has been discussed in the research paper presentation.

Keywords: insectivorous, medicinal, drosera, biotechnology, chromosome, genome

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Authors: Nidhi Jindal, Ashok Chaudhury, Manisha Mangal

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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: plant tissue culture, empirical measure, germination, tissue culture

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Authors: Naira Sahakyan, Margarit Petrosyan, Armen Trchounian

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One of the tasks in contemporary biotechnology, pharmacology and other fields of human activities is to obtain biologically active substances from plants. They are very essential in the treatment of many diseases due to their actually high therapeutic value without visible side effects. However, sometimes the possibility of obtaining the metabolites is limited due to the reduction of wild-growing plants. That is why the plant cell cultures are of great interest as alternative sources of biologically active substances. Besides, during the monitored cultivation, it is possible to obtain substances that are not synthesized by plants in nature. Isolated culture of Ajuga genevensis with high growth activity and ability of regeneration was obtained using MS nutrient medium. The agar-diffusion method showed that aqueous extracts of callus culture revealed high antimicrobial activity towards various gram-positive (Bacillus subtilis A1WT; B. mesentericus WDCM 1873; Staphylococcus aureus WDCM 5233; Staph. citreus WT) and gram-negative (Escherichia coli WKPM M-17; Salmonella typhimurium TA 100) microorganisms. The broth dilution method revealed that the minimal and half maximal inhibitory concentration values against E. coli corresponded to the 70 μg/mL and 140 μg/mL concentration of the extract respectively. According to the photochemiluminescent analysis, callus tissue extracts of leaf and root origin showed higher antioxidant activity than the same quantity of A. genevensis intact plant extract. A. genevensis intact plant and callus culture extracts showed no cytotoxic effect on K-562 suspension cell line of human chronic myeloid leukemia. The GC-MS analysis showed deep differences between the qualitative and quantitative composition of callus culture and intact plant extracts. Hexacosane (11.17%); n-hexadecanoic acid (9.33%); and 2-methoxy-4-vinylphenol (4.28%) were the main components of intact plant extracts. 10-Methylnonadecane (57.0%); methoxyacetic acid, 2-tetradecyl ester (17.75%) and 1-Bromopentadecane (14.55%) were the main components of A. genevensis callus culture extracts. Obtained data indicate that callus culture of A. genevensis can be used as an alternative source of biologically active substances.

Keywords: Ajuga genevensis, antibacterial activity, antioxidant activity, callus cultures

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Authors: Rashmi Ranade, Neelu Joshi

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Barleria prionitis L. is a well explored Indian medicinal plant valued for its stem and leaf which forms an important ingredient of many Ayurvedic formulations. It is used for the treatment of various disorders like toothache, bleeding gums, strengthening gums, whooping cough, inflammation, arthritis, enlargement of scrotum and sciatica etc. The plant is propagated vegetatively through stem cuttings. Frequent harvesting of this plant has led to the shortage of planting material, and it has acquired the status of vulnerable plant species. Plant tissue culture technology offers a very good alternative for propagation and conservation of such plant species. The present investigation was undertaken to develop in vitro regeneration protocol for B. prionitis L. via callus organogenesis pathway. Stem and leaf explants were used for this purpose. Different media and plant growth regulators were optimized to develop the protocol. The problem of phenol secretion and browning and in vitro cultures at the establishment phase was successfully curbed with the usage of antibrowning agents such as ascorbic acid and activated charcoal. Optimum shoot multiplication was achieved by the use of liquid media and incorporation of silver nitrate and TIBA (triiodobenzoic acid) into the media. High percent rooting (76%) was observed on WPM media supplemented with IBA (2.0 mg/l), IAA (0.5 mg/l), GA3(0.5) and activated charcoal(500 mg/l). The rooted plantlets were subjected to in vitro hardening on sterile potting mix (soil:farmyard manure:compost; 1:2:1) and acclimatized under greenhouse conditions. Around 85% survival of plantlets was recorded upon acclimatization. This lab scale protocol would be tested for in vitro scaling up production of B. prionitis L.

Keywords: explant browning, liquid culture, micropropagation, shoot multiplication, phenolic secretion

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Authors: Sun Il Yu, Jung Hyun Joo, Hwa Sung Shin

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Astrocytes are known as dominant cells in CNS and play a role as a supporter of CNS activity and regeneration. Recently, three-dimensional culture of astrocytes were actively applied to understand in vivo astrocyte works. Electrospun nanofibers are attractive for 3D cell culture system because they have a high surface to volume ratio and porous structure, and have already been used for 3D astrocyte cultures. In this research, the stiffness of cellulose acetate (CA) nanofiber was controlled by heat treatment. As stiffness increased, astrocyte cell viability and adhesion increased. Reactivity of astrocyte was also upregulated in stiffer CA nanofiber in terms of GFAP, an intermediate filament protein. Finally, we demonstrated that stiffness-controllable CA is attractive for astrocyte tissue engineering.

Keywords: astrocyte, cellulose acetate, nanofiber, tissue scaffold

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Authors: Sridevi Devadas

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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: Cryptocoryne affinis, surface sterilization, tissue culture, clorox, mercury chloride

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Authors: Sridevi Devadas

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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: Cryptocoryne affinis, surface sterilization, tissue culture, clorox, mercury chloride

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Authors: Muhammad Aasim, Mehmet Karataş, Fatih Erci, Şeyma Bakırcı, Ecenur Korkmaz, Burak Kahveci

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Water hyssop (Bacopa monnieri L. Wettst.) is an important medicinal aquatic/semi aquatic plant native to India where it is used in traditional medicinal system. The plant contains bioactive compounds mainly Bacosides which are the main ingridient of commercial drug available as memory enhancer tonic. The local name of water hyssop is Brahmi and brahmi based drugs are available against for curing chronic diseases and disorders Alzheimer’s disease, anxiety, asthma, cancer, mental illness, respiratory ailments, and stomach ulcers. The plant is not a cultivated plant and collection of plant from nature make palnt threatened to endangered. On the other hand, low seed viability and availability make it difficult to propagate plant through traditional techniques. In recent years, plant tissue culture techniques have been employed to propagate plant for its conservation and production for continuous availability of secondary metabolites. On the other hand, application of nanoparticles has been reported for increasing biomass, in vitro regeneration and secondary metabolites production. In this study, silver nanoparticles (AgNPs) were applied at the rate of 2, 4, 6, 8 and 10 ppm to Murashihe and Skoog (MS) medium supplemented with 1.0 mg/l Benzylaminopurine (BAP), 3.0% sucrose and 0.7% agar. Leaf explants of water hyssop were cultured on AgNPs containing medium. Shoot induction from leaf explants were relatively slow compared to medium without AgNPs. Multiple shoot induction was recorded after 3-4 weeks of culture comapred to control that occured within 10 days. Regenerated shoots were rooted successfully on MS medium supplemented with 1.0 mg/l IBA and acclimatized in the aquariums for further studies.

Keywords: Water hyssop, Silver nanoparticles, In vitro, Regeneration, Secondary metabolites

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Authors: Emma K. Sales, Nilda G. Butardo

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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, 2, 4-D

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Authors: Münüre Tanur Erkoyuncu, Mustafa Yorgancılar

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Crop improvement through genetic engineering depends on effective and reproducible plant regeneration systems. Immature embryos are the most widely used explant source for in vitro regeneration in barley (Hordeum vulgare L.). However, immature embryos require the continuous growth of donor plants and the suitable stage for their culture is also certainly limited. On the other hand, mature embryos can be procured and stored easily; they can be studied throughout the year. In this study, an effective callus induction and plant regeneration were aimed to develop from mature embryos of different barley genotypes. The effect of medium (MS₁ and MS₂), auxin type (2,4-D, dicamba, picloram and 2,4,5-T) and concentrations (2, 4, 6 mg/l) on callus formation and effect of cytokinin type (TDZ, BAP) and concentrations (0.2, 0.5, 1.0 mg/l) on green plant regeneration were evaluated in mature embryo culture of barley. Callus and shoot formation was successful for all genotypes. By depending on genotype, MS₁ is the best medium, 4 mg/l dicamba is the best growth regulator in the callus induction and MS₁ is the best medium, 1 mg/l BAP is the best growth regulator in the shoot formation were determined.

Keywords: barley, callus, embryo culture, mature embryo

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Authors: Anna Vildová, H. Hendrychová, J. Kubeš, L. Tůmová

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The abiotic elicitation is one of the methods for increasing the secondary metabolites production in plant tissue cultures and it seems to be more effective than traditional strategies. This study verified the use of silver nitrate as elicitor to enhance flavonolignans and flavonoid taxifolin production in suspension culture of Sylibum marianum (L.) Gaertn. Silver nitrate in various concentrations (5.887.10-3 mol/L, 5.887.10-4 mol/L, 5.887.10-5 mol/L) was used as elicitor. The content of secondary metabolites in cell suspension cultures was determined by high performance liquid chromatography. The samples were taken after 6, 12, 24, 48, 72 and 168 hours of treatment. The highest content of taxifolin production (2.2 mg.g-1) in cell suspension culture of Silybum marianum (L.) Gaertn. was detected after silver nitrate (5.887.10-4 mol/L) treatment and 72 h application. Flavonolignans such as silybinA, silybin B, silydianin, silychristin, isosilybin A, isosilybin B were not produced by cell suspension culture of S. marianum after elicitor treatment. Our results show that the secondarymetabolites could be released from S. marianum cells into the nutrient medium by changed permeability of cell wall.

Keywords: Silybum marianum (L.) Gaertn., elicitation, silver nitrate, taxifolin

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Authors: Szilvia Várallyay, Szilvia Veres, Éva Bódi, Farzaneh Garousi, Béla Kovács

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The agricultural environment is contaminated with heavy metals and other toxic elements, which means more and more threats. One of the most important toxic element is the arsenic. Consequences of arsenic toxicity in the plant organism is decreases the weight of the roots, and causes discoloration and necrosis of leaves. The toxicity of arsenic depends on the quality and quantity of the arsenic specialization. The arsenic in the soil and in the plant presents as a most hazardous specialization. A dicotyledon plant were chosen for the experiment, namely sunflower. The sunflower plants were grown in nutrient solution in different As(III) levels. The content of As, P, Fe were measured from experimental plants, using by ICP-MS.Negative correlation was observed between the higher concentration of As(V) and As(III) in the nutrition solution and the content of P in the sunflower tissue. The amount of Fe was decreasing if we used a higher concentration of arsenic (30 mg kg-1). We can tell the conclusion that the arsenic had a negative effect on the sunflower tissue P and Fe content.

Keywords: arsenic, sunflower, ICP-MS, toxicity

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Authors: Rosna Mat Taha, Sakinah Abdullah, Sadegh Mohajer, Asmah Awal

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Lycium barbarum L. (Goji) belongs to Solanaceae family and native to some areas of China. Ethnobotanical studies have shown that this plant has been consumed by the Chinese since ancient times. It has been used as medicine in providing excellent effects on cardiovascular system and cholesterol level, besides contains high antioxidant and antidiabetic properties. In the present study, some tissue culture work has been carried out to induce callus, in vitro regeneration from various explants of Goji and also some acclimatization protocols were followed to transfer the regenerated plants to soil. The main aims being to establish high efficient regeneration system for mass production and commercialization for future uses, since the growth of this species is very limited in Malaysia. The optimum hormonal regime and the most suitable and responsive explants were identified. It was found that leaves and stems gave good responses. Murashige and Skoog’s (MS) medium supplemented with 2.0 mg/L NAA and 0.5 mg/L BAP was the best for callus induction and MS media fortified with 1.0 mg/L NAA and 1.0 mg/L BAP was optimum for in vitro regeneration. The survival rates of plantlets after acclimatization was 63±1.5 % on black soil and 50±1.3 % on mixed soil (combination of black and red soil at a ratio of 2 to 1), respectively.

Keywords: callus, acclimatization, in vitro culture, regeneration

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Authors: Weiliang Wen, Xinyu Guo, Ying Zhang, Jianjun Du, Boxiang Xiao

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Reconstructing and analyzing three-dimensional (3D) models from situ measured data is important for a number of researches and applications in plant science, including plant phenotyping, functional-structural plant modeling (FSPM), plant germplasm resources protection, agricultural technology popularization. It has many scales like cell, tissue, organ, plant and canopy from micro to macroscopic. The techniques currently used for data capture, feature analysis, and 3D reconstruction are quite different of different scales. In this context, morphological data acquisition, 3D analysis and modeling of plants on different scales are introduced systematically. The commonly used data capture equipment for these multiscale is introduced. Then hot issues and difficulties of different scales are described respectively. Some examples are also given, such as Micron-scale phenotyping quantification and 3D microstructure reconstruction of vascular bundles within maize stalks based on micro-CT scanning, 3D reconstruction of leaf surfaces and feature extraction from point cloud acquired by using 3D handheld scanner, plant modeling by combining parameter driven 3D organ templates. Several application examples by using the 3D models and analysis results of plants are also introduced. A 3D maize canopy was constructed, and light distribution was simulated within the canopy, which was used for the designation of ideal plant type. A grape tree model was constructed from 3D digital and point cloud data, which was used for the production of science content of 11th international conference on grapevine breeding and genetics. By using the tissue models of plants, a Google glass was used to look around visually inside the plant to understand the internal structure of plants. With the development of information technology, 3D data acquisition, and data processing techniques will play a greater role in plant science.

Keywords: plant, three dimensional modeling, multi-scale, plant phenotyping, three dimensional data acquisition

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Authors: Shackira Am, Jos T. Puthur

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The phytostabilization potential of a halophyte Acanthus ilicifolius L. has been evaluated with special attention to the nutritional as well as anatomical adaptations developed by the plant. Distribution of essential elements influenced by the excess Zn²⁺ ions in the root tissue was studied by FEG-SEM EDX microanalysis. Significant variations were observed in the uptake and allocation of mineral elements like Mg, P, K, S, Na, Si and Al in the root of A. ilicifolius. The increase in S is in correlation with the increased synthesis of glutathione which might be involved in the biosynthesis of phytochelatins. This in turn might be aiding the plant to tolerate the adverse environmental conditions by stabilizing the excess Zn in the root tissue itself. Moreover it is revealed that most of the Zn were accumulated towards the central region near the vascular tissue. Treatment with ZnSO₄ in A. ilicifolius caused significant increase in the number of glandular trichomes on the adaxial leaf surface as compared to the leaves of control plants. In addition to this, A. ilicifolius when treated with ZnSO₄, exhibited a deeply stained layer of cells immediate to the endodermis, forming more or less a ring like structure around the xylem vessels. Phloem cells in these plants were crushed/reduced in numbers. There were no such deeply stained cells forming a ring around the xylem vessels in the control plants. These adaptive responses make the plant a suitable candidate for the phytostabilization of Zn. In addition the nutritional adjustment of the plant equips them for a better survival under increased concentration of Zn²⁺.

Keywords: Acanthus ilicifolius, mineral elements, phytostabilization, zinc

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Authors: Ennus Tajuddin Tamboli, Madhukar Garg, Mohd. Mujeeb, Sayeed Ahmad

Abstract:

An efficient protocol for simultaneous production of forskolin and rosmarinic acid in in vitro callus derived from the leaves of Coleus forskohlii Briq. has been developed. MS media was used for the establishment of cultures and NAA + 6-BA (1.0 ppm) was found best for callus growth. The callus was further subjected to treatment with various elicitor/precursors viz. chitosan, thidiazuron and methyl jasmonate to observe their effect on production of biomass and accumulation of secondary metabolites. The content of forskolin and rosmarinic acid were estimated by HPTLC, in comparison to natural explant which showed 2 fold and 10 fold rise in forskolin and rosmarinic acid content, respectively. Methy1 jasmonate 50 µM was found best for production of forskolin, whereas thidiazuron showed best results in the yield of rosmarinic acid, separately in static culture. However, combined treatment in suspension culture showed moderated effect for increase in secondary metabolites but the biomass increased significantly as compared to static culture.

Keywords: plant tissue culture, secondary metabolites, coleus, forskolin, rosmarinic acid, HPTLC

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Authors: Kumalo F. I., Malimabe M. A., Gumede T. P., Mosoabisane M. F. T.

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This study investigates the fabrication and characterization of bio-nanocomposites consisting of poly (butylene succinate) (PBS) and poly (ԑ-caprolactone) (PCL), reinforced with cellulose extracted from Eucomis autumnalis, a medicinal plant. Bio-nanocomposite films were prepared using the solvent casting method, with cellulose content ranging from 1 to 3 wt%. Comprehensive analysis was conducted using FTIR, SEM, TEM, DSC, TGA, and XRD, to assess morphological, thermal, and structural properties. The results indicated significant improvements in the thermal stability and morphological properties with increasing cellulose content, showcasing the potential of these materials for tissue engineering applications. The use of cellulose extracted from a medicinal plant highlight the potential for sustainable and biocompatible materials in biomedical applications.

Keywords: Bionanocomposites, poly(butylene succinate), poly(caprolactone), eucomis autumnalis, medicinal plant

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Authors: Neeraj Malhotra

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Background: Bioreactors in tissue engineering are used as devices that apply mechanical means to influence biological processes. They are commonly employed for stem cell culturing, growth and expansion as well as in 3D tissue culture. Contemporarily there use is well established and is tested extensively in the medical sciences, for tissue-regeneration and tissue engineering of organs like bone, cartilage, blood vessels, skin grafts, cardiac muscle etc. Methodology: Literature search, both electronic and hand search, was done using the following MeSH and keywords: bioreactors, bioreactors and dentistry, bioreactors & dental tissue engineering, bioreactors and regenerative dentistry. Articles published only in English language were included for review. Results: Bioreactors like, spinner flask-, rotating wall-, flow perfusion-, and micro-bioreactors and in-vivo bioreactor have been employed and tested for the regeneration of dental and like-tissues. These include gingival tissue, periodontal ligament, alveolar bone, mucosa, cementum and blood vessels. Based on their working dynamics they can be customized in future for regeneration of pulp tissue and whole tooth regeneration. Apart from this, they have been successfully used in testing the clinical efficacy and biological safety of dental biomaterials. Conclusion: Bioreactors have potential use in testing dental biomaterials and tissue engineering approaches aimed at regenerative dentistry.

Keywords: bioreactors, biological process, mechanical stimulation, regenerative dentistry, stem cells

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Authors: Hoo Cheol Lee, Dae Seung Kim, Sang Myung Jung, Gwang Heum Yoon, Hwa Sung Shin

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Loss of bone tissue can occur due to a bone tissue disease and aging or fracture. Renewable formation of bone is mainly made by its differentiation and metabolism. For this reason, osteoblasts have been studied for regeneration of bone tissue. So, tissue engineering has attracted attention as a recovery means. In tissue engineering, a particularly important factor is a scaffold that supports cell growth. For osteoblast scaffold, we used the cellulose nanowhisker (CNW) extracted from marine organism. CNW is one of an abundant material obtained from a number of plants and animals. CNW is polymer consisting of monomer cellulose and this composition offers biodegradability and biocompatibility to CNW. Mechanical strength of CNW is superior to the existing natural polymers. In addition, substances of marine origin have a low risk of secondary infection by bacteria and pathogen in contrast with those of land-derived. For evaluating its suitability as an osteoblast scaffold, we fabricate CNW film for osteoblast culture and performed the MTT assay and ALP assay to confirm its cytotoxicity and effect on differentiation. Taking together these results, we assessed CNW is a potential candidate of a material for bone tissue regeneration.

Keywords: bone regeneration, cellulose nanowhisker, marine derived material, osteoblast

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Authors: Krasimir I. Ivanov, Penka S. Zapryanova, Stefan V. Krustev, Violina R. Angelova

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Three commonly used digestion methods (dry ashing, acid digestion, and microwave digestion) in different variants were compared for digestion of tobacco leaves. Three main macroelements (K, Ca and Mg) were analysed using AAS Spectrometer Spectra АА 220, Varian, Australia. The accuracy and precision of the measurements were evaluated by using Polish reference material CTR-VTL-2 (Virginia tobacco leaves). To elucidate the problems with elemental recovery X-Ray and SEM–EDS analysis of all residues after digestion were performed. The X-ray investigation showed a formation of KClO₄ when HClO4 was used as a part of the acids mixture. The use of HF at Ca and Mg determination led to the formation of CaF₂ and MgF₂. The results were confirmed by energy dispersive X-ray microanalysis. SPSS program for Windows was used for statistical data processing.

Keywords: digestion methods, plant tissue, determination of macroelements, K, Ca, Mg

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Authors: Sunil Dehipawala, Aregama Sirisumana, stephan Smith, P. Schneider, G. Tremberger Jr, D. Lieberman, Todd Holden, T. Cheung

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The arsenic and iron environments in different growth stages have been studied with EXAFS and XANES using Brookhaven Synchrotron Light Source. Collard Greens plants were grown and tissue samples were harvested. The project studied the EXAFS and XANES of tissue samples using As and Fe K-edges. The Fe absorption and the Fourier transform bond length information were used as a control comparison. The Fourier transform of the XAFS data revealed the coexistence of As (III) and As (V) in the As bonding environment inside the studied plant tissue samples, although the soil only had As (III). The data suggests that Collard Greens has a novel pathway to handle arsenic absorption in soil.

Keywords: EXAFS, fourier transform, metalloproteins, XANES

Procedia PDF Downloads 328

Authors: Arash Sepehri Bonab

Abstract:

Techniques to switch cells between development and differentiation, which tend to be commonly exclusive, are utilized in arrange to supply an expansive cell mass that can perform particular separated capacities required for the tissue to develop. Approaches to tissue engineering center on the have to give signals to cell populaces to advance cell multiplication and separation. Current tissue regenerative procedures depend primarily on tissue repair by transplantation of synthetic/natural inserts. In any case, restrictions on the existing procedures have expanded the request for tissue designing approaches. Tissue engineering innovation and stem cell investigation based on tissue building have made awesome advances in overcoming the issues of tissue and organ damage, useful loss, and surgical complications. Bone tissue has the capability to recover itself; in any case, surrenders of a basic estimate anticipate the bone from recovering and require extra support. The advancement of bone tissue building has been utilized to form useful options to recover the bone. This paper primarily portrays current advances in tissue engineering in different fields of bone and talks about the long-term trend of tissue designing innovation in the treatment of complex diseases.

Keywords: tissue engineering, bone, technologies, treatment

Procedia PDF Downloads 97