The technique of planttissueculture has been well accepted and applied in the mass propagation of planting materials in various crops and plants. In India numerous micropropagation units are producing millions of plantlets catering the needs for the increasing demand of quality planting materials. The advantages of this technique lie in the production of plantlets that are disease free and genetically identical to the elite mother plants. Application of planttissueculturetechnique is the only viable means for the large scale production of banana planting materials which is not possible through conventional propagation. The article discusses the strategies of the mass pro- duction of commercially important banana in Mizoram using planttissueculture techniques.
lack of good healthy planting materials (Das et al; 2013). The conventional methods are slow and not adequate for rapid multiplication. Further, the yield is drastically reduced by viral and nematode infections etc. and also due to ruthless exploitation of medicinal herbs form natural habitats. Not only these but some of the members revive naturally through seeds, but however their cultivation rate and poor seed dormancy are crucial factors for its propagation. As a result of which many of the valuable useful plant members are getting extinct or endangered. Planttissueculturetechnique would be useful for conservation of rare and endangered plants for the production of industrially important phytochemicals.
Identification and characterization of bacterial and fungal contaminants: Microbial contaminants were isolated from 211 invitro cultureplant-lets at planttissueculture laboratory, University of Maiduguri. Bacterial isolates were aseptically streaked onto sterile nutrient agar (NA) medium and the culture were incubated at 37 o c for 24 hours. All the isolated contaminants were purified by serial dilution technique (Collins and Lyne 1984). Whereas, fungal isolates were aseptically transferred onto Petri dishes containing potato dextrose agar (PDA) growth medium and the cultures were incubated at 24 o c for 7 days. The purified isolates were stained for morphological characterization based on vegetative cell shape, gram reaction and presence of spores. Below are pictures of the two fungi isolated potatoes dextrose agar. Fig.1 and Fig.2.
The similarities of the effects induced by the stress in the plant cultured in vitro and in vivo conditions suggest that the in vitro system can be used as an alternative to field evaluations for studying the general effect of water-stress on plant growth and development. The tissueculturetechnique is a rapid method to propagate plants in vitro, especially, medicinal plants. The present study is carried out at biology Lab, Biology Department, University College in ALKhafji, University of HAFR ALBATIN, to study the effect of MS salts strength, sucrose concentration on micro propagation and anti-oxidant
In the present study, protocol for callus induction and regeneration for the medicinal plant species, Cryptolepis grandiflora has been developed by employing tissueculture technology. Young leaf explant inoculated on MS medium containing the growth regulators, BAP and NAA at 2.0 and 1.2 mgl -1 respectively showed higher callus induction (81%). The amount of callus responded for shoot formation greater was (78%) when subcultured on to the MS medium containing BAP (2.0 mg/L) and GA 3 (0.5 mg/L). The elongated shoots were rooted well on half
Study and exploration of alfalfa transformation using different types of explant materials not only provide options to meet different needs and situations, but can also provide possibility for a short time frame and feasibility for transfer of new genes into previously transformed alfalfa plants for multiple trait improve- ment. Besides using ordinary organs for explant transformation, somatic embryos derived from tissueculture can also be used for transformation in alfalfa. Use of somatic embryos as explants in alfalfa genetic transformation can be conducted via direct and indirect somatic embryogenesis     . In direct embryogenesis, somatic embryos were first infected by Agrobacterium . The second- ary embryogenesis was induced through direct and spontaneous new embryo- genesis from primary somatic embryos without callus phase. This method ap- peared to be effective for alfalfa transformation. However, molecular analysis, such as Southern blotting, which is essential to confirm transformation, was not conducted in recovered plants   . The concern is that selection through direct embryogenesis may be less effective and may result in escapes . In addition, spontaneous embryogenesis appeared to be very genotype spe- cific and thus the technique may be restricted to unique clones and genotypes. Liu et al .  report a transformation system which also used somatic embryos as explants. The system was based on cell dedifferentiation or cell reprogramming of somatic embryos induced by plant growth regulators, subsequent induction and development of embryogenic calli, and somatic embryo formation from the calli cultured on plant growth regulator-free medium. The biological and rege- neration pathway of secondary embryogenesis reported by Liu et al .  is dif- ferent from direct and repetitive embryogenesis   . The transforma- tion reported by Liu  could be applied to different genotypes and no escapes were observed. Introduction of multiple genes of interest into previously trans- formed plants to confer plants with different traits is a direction of wide applica- tion of transgenic technology . Use of somatic embryos as explants from trans- genic plants to initiate transformation to introduce new genes of interest can re- duce the time for development of plants that contain multiple new traits. In ad- dition, the system may be an option for those cultivars in which use of other or- gans for transformation is difficult.
culture. Contamination from different sources, such as bacteria and fungi, reduces the productivity and can completely prevent successful culture (Colgecen et al., 2009). The growth media in which the planttissue is inoculated is also a good source of nutrients for microbial growth. These microbes compete with planttissue for nutrients and some of them produce phytotoxins, which result in culture mortality, tissue necrosis, and reduced shoot proliferation and rooting (Kane, 2003). The diversity and range of tolerance of various microorganisms mostly bacteria and fungi makes it quite challenging to eradicate the same through traditional approaches. The effective management of the microorganisms in tissueculture needs greater interpretation and research. Internal and external contamination of the microorganisms, mostly bacteria and fungi pose problems due to their greater growth rate than the plant cell, which drains all the nutrients and creates an unfavourable environment for the explants to grow (Cassells, 1991). Surface sterilization of explants may sometime fail to establish the aseptic condition of the culture medium due to systemic infection of various microorganisms and latent contamination can be observed which spoils the whole experimental setup. Inside the plant they have very little microbial competition and remain mostly latent and hardly show any symptoms in plants (Peñalver et al., 1994; Hallman et al., 1997) and therefore they cannot be sorted by visual observation. By surface sterilization, most of the epiphytic microorganisms are eradicated except the systemic or the endogenous one (Habiba et al., 2002). Therefore, in various cases an interdisciplinary approach with other branches of biological science needs importance. Although varieties of techniques and protocols are formulated and optimized to minimize the microbial contamination, designing more efficient methods and techniques to eliminate the contamination so as to prevent the labour intensive and expensive experiments seems necessary (Mahna et al., 2013).
processes and interactions, to quantify the impact of single variables on the performance of the systems. In the field of engineering reactors, it is noted today a growing deployment bioreactor temporary immersion and/or permanent, which promote the production of tissues, cells or plant organs on a large scale (Konstas and Kinzios, 2003; Xie et al., 2003; Rodrigues et al. (2006) describe the development of new temporary immersion prototypes and or permanent, and the study of its operation in different cultures is essential to reduce costs, increase productivity and maintain of plant material subjected to this technique. RechFilho (2004) describes the in vitro culture systems of plant cells in bioreactors were initially used in the United States, Cuba and France. Currently many countries have begun in order to produce seedlings on a large scale that agricultural interest, especially ornamental, fruit and forest species, with greater speed and efficiency compared to other conventional methods used in parallel, there is a reduction in process costs (Soccol et al., 2008;Scheidt et INTERNATIONAL JOURNAL
culture and study the development of small, isolated segments of plant tissues or isolated cells. Around the mid twentieth century, the notion that plants could be regenerated or multiplied from either callus or organ culture was widely accepted and practical application in the plant propagation industry ensued. The technique was heralded as the universal mass clonal plant propagation system for the future and the term ‘micropropagation’ was introduced to describe more accurately the processes. In the present study callus cultures of B. monosperma were raised from nodal segments of this plant. The observations of present investigation are in agreement with Thorpe and Patel (1984) that tissue or organs used as source of explants can also be determinant for the success of planttissueculture (Akbas et al., 2008). It has been observed that juvenile and actively dividing plant responded effectively in vitro condition due to vigorous vegetative development stage and absence of reproductive structure formation. Even in juvenile stage, tissue and organ regeneration is reported to be more with the younger and actively dividing tissues (Endress, 1994).
Fluorescent Pseudomonas sp. is emerging as largest and potentially most promising group of PGPR (plant growth promoting rhizobacteria) that are involved in plant growth enhancement. Plant growth regulators viz., auxins, cytokinins and gibberellins help in plant growth and development. In present study, fluorescent Pseudomonas strains isolated from rhizospheric soil from temperate fruit zone of Himachal Pradesh were investigated for plant growth regulator production i.e. auxins, gibberellins and cytokinins in nutrient broth. All the strains tested produced plant growth regulators in concentrations auxins (1.83-21μg/ml), gibberellins (116.1-485.8μg/ml) and cytokinins (45.4-295.4μg/ml). Two strains (An-1-kul and An-13-kul) were selected on the basis of over all PGPR activities for production of growth regulators. Molecular characterization of best selected Pseudomonas strains were done by 16S-rRNA technique. Plant growth regulators produced by best isolates were further studied to observe their effect on growth of callus formation, shoot formation of broccoli and root elongation of cabbage
extension is the slow propagation rate through the formal method, which requires several years . Many reasons have been reported for low production of sugarcane from them an important is inability of rapid multiplication of see practice. In this procedure favorable clone is can be clear, which takes about 6 to 7 years normally for getting effective quality of better planting material. This high time consuming process can be because of a large bottleneck in effective breeding program . Planttissueculture is an assemblage of procedures employed to sustain or cultureplant groups, tissues otherwise appendages below sterilized situations in a nutrient culture average of identified concerto. Today, the technique of planttissueculture has become a potent tool to study, solve basic problems and applied in plant biotechnology [3-5] conducted initial trials to regenerate plants through in vitro techniques. Many writers have developed procedures for the in vitro regeneration of sugarcane during corn cultivation, axillary blossom, furthermore shoot pour culture [6-8]. The growth of tissueculture technology for the rapid generation of disease-free planting material has been an important step towards sufficient seed production, faithful to the character and quality of sugarcane . Thus, the application of planttissueculture techniques provides an alternative method for the propagation and improvement of sugarcane . Planttissueculture offers the best methodology through the micropropagation of sugarcane for planting quality and planting material at a quicker pace in a shorter period. Tissueculture preserve add to the spread probable near 20-35 moments .
Mutagenesis is one of the powerful tools for breeding biotechnology. Chemical mutagenesis is an easy tool, which doesn’t require special tools and provide high mutation rate. Effects of different cytokines (BA, 2ip and Kim at 0.4 mg/l) on in vitro shootlet proliferation of Eustoma grandiflorum were studied. Different concentrations of colchicine (30, 60, 120 and 240 mg/l) and sodium azide (5, 10, 15 and 20 mg/l) were added to free- hormone MS medium. In vitro shootlets, of Eustoma grandiflorum, were culture for 7, 14 and 28 days. After each treatment period, the shootlets were transferred to fresh MS medium free (without the chemical mutagenesis). At acclimatization stage, most of morphological parameters and anthocyanin pigment contents in flower recorded decline by most treatments of sodium azide. All colchicine treatments reached to morphological stage and formed bud initiation and death before flowering. Contrary, highest survival % of acclimatized plants (70%) and highest values of number of branches, branches length (cm), leaf area (cm2), most of floral parameters, photosynthetic pigments, carotenoids and anatomical structure were obtained from ( 5,10 mg/l sodium azide for 28, 7 days) and (60, 30mg colchicine for 28 days).
Mutations are important sources needed by plant breeders as a source of genetic variations that can result in plants superior to their origins. Mutations can occur in sexual cells that are easily transmitted to subsequent generations. If the mutations occur in Somatic cells, to the next generation that have been cultivated, and since the rate of occurrence of genetic mutations very little in nature so it became necessary to use different techniques to develop mutations in both genomes in vivo or in vitro. Mutagenesis in vitro is one of the technologies that can be used in this field. Through this technique, millions of cells can be exposed to stress, whether physical such as emulsions or chemical, such as Ethel methane sulphonate (EMS) Selection of mutant cells and replanting of plants. This technique can be used for breeding purposes to obtain plants with specific specifications such as NaCl salt tolerance or pesticide and toxin resistance by adding such media to the food media used in the development of cells showing resistance to this stresses (5).
Previous studies found that chitosan does not improve the production of BGSL hydrolysis product compared to elicitation using methyl jasmonate (Al-Gendy & Lockwood, 2005). The highest PEGSL content in treated plant is 1.59 ± 0.01 µmol/g FW at 20 % and BGSL content is 0.66 ± 0.001 µmol/g at 5 % of coconut water. Both identified glucosinolates respond differently on coconut water treatment. PEGSL tend to increase at higher concentration of coconut water while BGSL tend to increase at low concentration of coconut water. Plant growth hormone which used in tissueculture are naturally present in coconut (Agampodi & Jayawardena, 2006) which affect the growth of the in vitro plant.
Plant somatic embryogenesis (SE) research has been in- vestigated in planttissueculture in recent years. The de- finition of SE is that the plant somatic cell develops into a new plant with the similar progress of zygotic embryo development . The origination of embryogenesis is di- fferent between SE and zygotic embryogenesis; however, both embryogenesis are close in structure and bio-chemi- cal properties . Plant SE is the expression of plant cell totipotency, and the successful SE of different plants re- quires specific cultural environments . The SE research progress of fruit trees is relatively slow compared with the rapid development of vegetables and flowers. Whereas, due to the nutrient and economic value of fruit trees in agricultural production, SE of fruit trees has made rapid progress in the past decade years. The author reviews the factors that affect SE and its application to the field of horticulture science in recent years, providing the refer- ence for future embryogenesis research.
Rootable shoots were excised from lavishly multiplying shoot clusters of both basmati varieties (Ran Bas and Bas 370) and were then transferred singly to culture tubes. Rooting medium was MS supplemented with different concentrations of NAA (Napthalene acetic acid) (0.1mg/l, 0.2mg/l and 0.3mg/l). The percentage of root induction was recorded after 30 days of culture. After the roots were well developed, the rooted plants were taken out of culture tubes, washed gently to remove agar and then transferred to the pots with a mixture of sand and soil in the ratio of 2:1. The plantlets in the pots were covered with jars to maintain the humidity. After 2 weeks, the jars were removed and the established plants were then transferred to soil in the field conditions and their survival rate was observed.
The aim of this study employ technology of tissueculture in the possibility of increasing secondary metabolites of plant marigold is made by developing callus from explants, then treated subcultured callus by some elicitors, which might help to increase the secondary metabolites of Calendula officinalis L., which had highly medical importance specially in the pharmaceutical industries, and compared the secondary metabolites resulting from treatment of the callus by elicitors with the secondary metabolites from the mother plant without treatments.Calendula officinalis L. (Asteraceae), known as calendula or marigold, is an annual specie widely used around the world as a medicinal plant. It is native to the area surrounding the Mediterranean, it is today and has been historically grown much more widely, throughout many temperate zones (Ao, 2007). C. officinalis L. is grown for medicinal herbal (Mohammad and Kashani, 2012), anti-tumor (Matic et al.,
The culture vessel design depicted in Fig. 2 was devel- oped to be compatible with both semi-solid culture and liquid based rocker systems [24–28], as well as being a suitable size to integrate commercially available RGB LED strips. One of the major limitations of FDM 3D printing with respect to planttissueculture is that most materials currently used have relatively low melting points and are not suited to heat sterilization or autoclaving. However, while polycarbonate (PC) is not commonly used for 3D printing it is amenable to heat sterilization and has good optical clarity so was used in this study. Problems were encountered with this material related to warping, poor adhesion to the print bed, delamination between layers, and achieving water-tight prints. Warping and delami- nation were related in large part to poor adhesion to the build plate, and parts that did not stick well would inevi- tably fail. To improve build plate adhesion, several mate- rials and adhesives were evaluated and the most effective combination was printing onto PolyEthylene terephtha- late (PET) tape treated with a thin layer of disappearing purple glue stick (Elhmer’s Products, OH, USA). Another factor that was critical for successful printing with PC was accurate bed leveling and optimizing the height of the first layer. Warping was further reduced by printing with a fully enclosed 3D printer that helped create more uniform temperatures and even cooling of the molten plastic. Once these factors and the slicing parameters in the software were optimized (see Table 1), PC vessels
Copper is one of the eight essential micronutrients for all higher plants, being a component of the protein structure of a range of enzymes involved in electron transport and redox reactions in mitochondria, chloroplasts, cell walls and the cytoplasm of plant cells (Marschner, 1 986; Welch, 1 995). A common characteristic of heavy metals in general, regardless of whether they are biologically essential or not, is that they may already exert toxic effects in low concentrations compared with macro-nutrients (Verkleij and Schat, 1 989). In the case of Cu, it is toxic for most plant species, at concentrations higher than 20-50 mg kg-1 dry matter (DM) (Asher, 199 1 ; Gartrell, 198 1 ; Robson and Reuter, 1 98 1 ; Welch, 1 995). Timperley et al. ( 1 970) concluded that the Cu concentrations of most plants tend to be internally rather than externally regulated, so that Cu concentrations in plants tend to be relatively constant, generally between 5-20 mg kg-1 DM, irrespective of the available Cu concentration in the rooting media. The concentrations of Cu in soil solutions, however, are much lower (5-200 ppb). These concentrations would not test internal plant regulation of Cu (Jarvis and Whitehead, 1 98 1 , 1 983 ; Welch, 1 995; Whitehead, 1987).