Tissue Culture Laboratory Research Articles

In vitro Propagation and Ex vitro Acclimatization of Potato (Solanum tuberosum L.) Using Nodal Cutting Explants

Potato (Solanum tuberosum L.) is an economic tuberous crop cultivated worldwide in the temperate, tropical and subtropical zones. It occupies the fourth largest food crop following wheat, rice and maize. The aim of this study is to establish a protocol for in vitro initiation, multiplication, rooting and ex vitro acclimatization of potato plants (Solanum tuberosum L.) cultivars Lady Balfour and Bellini. This study was carried out in the plant tissue culture laboratory, the Faculty of Agriculture, Saba basha, Alexandria University, Egypt during the period from 2013 to 2016. An efficient and reliable protocol for in vitro propagation and ex vitro acclimatization of potato (Solanum tuberosum L.) was optimized. Nodal cutting explants were inoculated on various initiation or establishment media with different combinations of NAA and KIN and the neoformed shoots were cultured on proliferation (multiplication) media with different combinations of NAA and BAP for the development of multiple shoots, and the elongation media to elongate of the neoformed shoot. The subsequent elongated shoots were rooted, and acclimatized ex vitro, successfully. The best medium for shoot initiation was MS medium supplemented with 1.0 mg/l KIN. The favorable medium for multiplication was the tested medium augmented with 2.0 mg/l BA and 0.250 mg/l NAA. In addition, the most effective medium for elongation was the used medium enriched with 0.250 mg/l NAA. Furthermore, in vitro the shoots showed healthy root development when the tested medium was supplemented with combination of 1.0 mg/l IBA and 0.50 mg/l NAA (rooting stage).The combination of sand:perlite:peatmoss (1:3:3, v:v:v) was used as substrates for the hardening of the in vitro plantlets, as a potting mix, was the best suited mix for the acclimatization of plantlets ex vitro.

Awaken Your Senses at Spring’s Height at Atlanta’s Botanical Garden

Back to table of contents Previous article Next article Annual MeetingFull AccessAwaken Your Senses at Spring’s Height at Atlanta’s Botanical GardenAaron LevinAaron LevinSearch for more papers by this authorPublished Online:18 Feb 2016https://doi.org/10.1176/appi.pn.2016.2b2AbstractA short cab ride away from the APA Annual Meeting but well worth the side trip lies the home of thousands of extraordinary flowers, plants, trees, and even a few familiar vegetables.The Atlanta Botanical Garden, at 1345 Piedmont Avenue, features many collections of plants from around the world—from the bright orchids and neotropical blueberries of the cloud forests of the Andes Mountains in South America to succulent collections native to Madagascar.Among much real greenery, a friendly copper frog by artist Beau Smith invites visitors to the Atlanta Botanical Garden to take a bit of a breather beside him on the bench.Aaron LevinThe Tropical Rotunda includes garden habitats from lowland tropical forests in Central America, Central Africa, Southern Mexico, and Southeast Asia. There are also selections of idiosyncratic island environments from the Seychelles, New Zealand, New Guinea, New Caledonia, and the Comoros Islands.The garden features some truly exotic plants, like the nepenthes, originally from South Asia and Australia. This carnivorous plant dines on insects and the occasional inattentive small rodent who slip into the large cup that characterizes the flower after sipping nectar produced by the plant. There, they are digested.In addition to offering visitors a chance to see flowers from around the world, the Botanical Garden works to conserve and restore rare and threatened plant species native to the Southeastern United States. The garden’s Tissue Culture Lab uses cloning and other techniques to propagate plants like native orchids and trillium, which can later be replanted in their native habitats. Efforts by garden staff and volunteers, for example, have helped repopulate the nearly vanished Cigar Orchid in Florida’s Fakahatchee Strand State Preserve. The Nepenthes Chandelier by glass artist Dale Chihuly hanging by the entrance of the Atlanta Botanical Garden was inspired by the insect-eating plant whose name it bears. Many more of Chihuly’s brightly colored glass creations will be on display in May.Aaron LevinNot all plants are simply a feast for the eyes. The Botanical Garden’s Edible Garden reminds visitors that what looks good might also taste good. The adjacent Outdoor Kitchen features Atlanta’s top chefs, who offer cooking classes on Saturdays and Sundays at noon, 1 p.m., and 2 p.m., showcasing seasonal recipes using ingredients harvested from the Edible Garden. Any harvested food not used in educational programs is donated to local charities. Lunch is available at the Cafȳ at Linton’s in the Garden, supervised by Chef Linton Hopkins, a James Beard Award winner and popular Atlanta restaurateur. The menu is based on fresh, local ingredients inspired by the Garden. Beyond its own leafy wonders, the Botanical Garden will feature a major exhibit of the works of renowned American glassblower Dale Chihuly during the APA Annual Meeting. Abstract but clearly reminiscent of the natural world, some of these large glass sculptures have already been installed, but many more will be in place throughout the gardens in May.To give visitors an opportunity to explore the 20 installations in the light of evening, the Botanical Garden will offer extended hours on Wednesdays through Sundays in May. ■For more information, visit the Atlanta Botanical Garden website. ISSUES NewArchived

EFFECT OF LOW COST GELLING AGENTS AND SOME GROWTH REGULATORS ON MICROPROPAGATION OF Philodendron selloum

This study was carried out in Plant Tissue Culture Laboratory of the Faculty of Environ. Agric. Sci. El-Arish, North Sinai, Suez CanalUniversity during the period from 2012 to 2015. The present studies were undertaken to look for additional cheap and easily available substitutes of agar to be used in tissue culture media. The obtained results show that MS medium supplemented with BA at 8 mgl-1 resulted in significant increase for all Philodendron growth parameters i.e, number of adventitious shoots/explant, adventitious shoot length (cm), main shoot length (cm) and number of leaves/shoot respectively (4.94, 8.09, 7.60 and 5.33, respectively) after six weeks from culture date comparing with control (MS medium free). However, 12 weeks period was not largely effective in enhancing growth parameters so that, for economical feasibility plants should not be cultured further than 6 weeks. While, in multiplication stage data present that MS medium supplemented with 8mgl-1 with 0.4 NAA mgl-1 produced the highest growth values. Moreover, the combination between agar at 4gl-1 and locust bean (LB) at 4gl-1 as gelling agentproduced the maximum growth values. For rooting stage, data clear that number of roots increased and recorded the maximum number (31.67) when shoots of Philodendron were cultured on half MS medium supplemented with 1.0 mgl-1 IBA on the absence of activated charcoal. Finally, Rooted plantlets were acclimatized successively and the highest survivability percentage (100%) was achieved by using the media mixture of vermiculite: peat (1:1) and plants irrigated with quarter MS strength after six weeks.

Effect of Banana Extract on Growth and Development of Protocorm Like Bodies in <i>Dendrobium</i> sp. orchid

An experiment was carried out in Plant Tissue Culture Laboratory, Department of Crop Botany, Bangladesh Agricultural University, Mymensingh to investigate the effect of banana extract on micropropagation of <i>Dendrobium</i> sp. var. Sonia orchid through PLBs. The experiment was conducted during July 2012 to October 2013. Half-Murashige and Skoog (1/2MS) medium were used as basal medium and the medium was supplemented with banana extract at 12.5, 25, 50, 100, and 200 ml L-1 with a control, where no banana extract was supplemented. The cultures were done in 100 ml conical flasks and maintain at 25°C with 30µ mol m-2 S-1 lighting provided by florescent tubes for 16 hours per day. Banana extract showed significant effect on growth and development of PLBs. Among the treatments, 100 ml L-1 banana extract enhanced new PLBs regeneration from explanted PLBs and growth and development of PLBs. Present research indicated that nutrient requirement for PLBs multiplication and plantlets growth of Dendrobium orchid is quantitatively different in vitro. Finally, 100 ml L-1 and 25 ml L-1 of banana extract may be recommended as supplement into 1/2MS medium for PLBs multiplication and plantlet regeneration, respectively in vitro.The Agriculturists 2015; 13(1) 101-108

Determination of hormonal combination for increased multiplication of tissue culture potato plantlets

Use of plant growth hormones either singly or in combination is vital for rapid multiplication of virus-free in-vitro potato ( Solanum tuberosum L. ) plantlets for the production of clean seed potato. This study was carried out at Kachwekano Zonal Agricultural Research and Development Institute, Kabale district in south-western Uganda to identify a suitable hormonal combination and optimum concentrations for production of a high number of In-vitro plantlets for three farmer-preferred varieties Kachpot-1, Kinigi, and Victoria. Eight hormonal combinations were formulated and tested using a completely randomized design with three replicates in the tissue culture laboratory. Ten shoot tips from in-vitro raised plantlets were excised and transferred to each of these hormonal combinations. The effect of hormonal combinations was variety dependant. N3 produced the highest mean number of nodes (6.42), buds (4.32) and tallest plantlets (7.38) for Kinigi. N2 produced the highest number of buds in Kachpot 1. N4 produced the tallest plantlets (7.19), highest mean number of buds (5.88) and nodes (7.66) for Victoria. Therefore, N3, N2 and N4 should be used for rapid In-vitro propagation of Kinigi, Kachpot 1 and Victoria respectively. Key words : In-vitro propagation, Solanum tuberosum

Differentiation of human umbilical cord mesenchymal stem cells using 5-azacytidine to generate a cardiac patch on thermo-responsive polymer.

Event Abstract Back to Event Differentiation of human umbilical cord mesenchymal stem cells using 5-azacytidine to generate a cardiac patch on thermo-responsive polymer. Lakshmi R. Nair1 and Kumary T. V1 1 Sree Chitra Tirunal Institute for Medical Sciences and Technology, Tissue Culture Laboratory, India Introduction Human heart, the non-tirelessly working; pumping organ. Incidence and prevalence of cardiovascular diseases (CVDs) are increasing worldwide. Current treatment strategies mitigate only the symptoms, with no role in damaged tissue regeneration. This calls for myocardial tissue engineering (MTE) with scaffolds and cells. A bioengineered cardiac patch with native extracellular matrix can overrule these limitations. Cell sheet technology thus helps in developing transplantable scaffold and suture free in vitro tissue constructs. For this, choice of cell source is also important. Hence hUMSCs which are least researched upon for MTE, but reported to be better source of seed cells for generating cardiomyocytes were used (1). Materials and Methods HiFi Wharton's jellyMesenchymalStemCellswere used for the study. Cells were cultured, characterised by immunostaining and flow cytometry for CD90, CD105, Stro-1 and CD34. Cells were grown on NGMA, the in-house developed thermo-responsive polymer and differentiated using 5µM 5-azacytidine. After 24 hours, the cells were washed twice with PBS and the media was changed to normal LG-DMEM and studied for 21 days with media change at alternate days. The differentiated cells were characterised for cardiac markers Troponin T, Myosin heavy chain, Alpha cardiac sarcomeric actinin and Connexin 43 by immunostaining and by RT PCR for Connexin-43, Cardiac myosin heavy chain, Cardiac troponin-T, GATA4, Nkx2.5 and house keeping gene GAPDH. The sheet was retrieved by lowering the temperature. FDA – PI live dead staining was performed to ensure the viability of the retrieved cell sheet. Results and Discussion The hUCMSCs are being looked upon with great enthusiasm, as they have proved to be highly competent to beat the bone marrow derived MSCs; one of the most extensively researched cell source; in availability, proliferation and even immuno-modulation. The risk of viral contamination is also lower for hUCMSCs (2). In spite of being better seed cells for for generating cardiomyocytes, they have never been advocated as a potential cardiac patch differentiated using 5-azacytidine alone as inducer, except for the study by Zhao et al., 2011 which depicts sphingosine -1 - phosphate in combination with cardiac myocytes conditioned medium for cardiac differentiation(3). Cells were found to express CD90 (Fig. 1), CD105 and Stro-1 (Fig 2) by immunostaining and flow cytometry, confirming stemness and homogeneity. Differentiated cells aligned parallel to one another and showed morphological features similar to cardiomyocytes. Molecular expression of sarcomeric-alpha-actinin, Nkx 2.5 and Connexin 43 (Fig 3), the molecules which are major to cardiac cells for cell contraction and electrical impulse propagation to other myocardial cells in the syncytium, when transplanted were observed. Cells differentiated to myocardial lineage were also retrieved as intact viable cell sheet which is highly recommended for suture free transplantations. Conclusion Differentiated hUCMSCs cells have been retrieved as a viable cardiac patch for suture free transplantation with minimal cell loss and effective nutrient diffusion. Also, our study, illustrates more economical method by adding small amount of inducer to cells, for a short period. Lakshmi R Nair acknowledges University Grants Commission, under the Ministry of Human Resources and Development, Government of India for the fellowships offered under CSIR/UGC/NET

Electrosprayed calcium phosphate nano-carriers as therapeutic agents

Event Abstract Back to Event Electrosprayed calcium phosphate nano-carriers as therapeutic agents Yogeshwar Chakrapani1, 2, Sampath T. Kumar1, Kumary T. V2 and Deepa K. Raj2 1 Indian Institute of Technology Madras, Medical Materials Laboratory, India 2 Sree Chitra Tirunal Institute for Medical Sciences and Technology, Tissue Culture Laboratory, India Introduction: Calcium phosphate particles intended as bone fillers and drug delivery agents are gaining interest worldwide. Size of the particles becomes an important factor when they are intended to act as site specific delivery agents, for therapeutic purposes. Cellular uptake, velocity of drug carrier in physiological pathways and specificity towards binding of therapeutic molecules are some key parameters that are greatly influenced by particle size[1]. The aim of the current study was to produce calcium phosphate particles through a modified electrospraying process[2]; characterize them physicochemically, evaluate their cytocompatibility and to analyse the efficacy of the particles to act as drug delivery systems. Materials and Methods: Calcium nitrate tetrahydrate (CNt) and triethyl phosphite (TEP) were used as the calcium and phosphorous precursors with an aqueous solvent. Sols with the required Ca/P ratio (1.67 & 1.61) were prepared by adding CNt solution drop wise to the hydrolysed TEP solution. The obtained sols were aged and subsequently blended with a polyvinylalcohol solution. The solution was loaded into a 10ml syringe fitted with an 18G blunted needle. The tip to collector distance, flow rate and voltage were optimised for electrospraying. The as sprayed particles were dried overnight and then subjected to a controlled heating upto 500˚C. The particles obtained on heating 1day and 5days aged sols of 1.67 and 1.61 Ca/P ratio will be referred to as CaP167-1d, CaP167-5d, CaP161-1d and CaP161-5d respectively. Phase confirmation was done through X-ray diffraction (XRD). Stability of phases was tested by heating upto 900˚C. The particles size and morphology were analysed using transmission electron microscopy (TEM). In vitro biological evaluation of the particles were performed using L-929 and HOS cell lines. The drug delivering capacities of the particles were assessed using tetracycline as model drug. Results and Discussion: Figure 1 shows the results of XRD and TEM analysis. XRD patterns were matched with that of hydroxyapatite (JCPDS file no. 9-432). At 500˚C the patterns of all the particles are similar whereas on heating to 900˚C, CaP167-1d and 5d are the same while CaP161-1d and 5d develop peaks of β-tricalcium phosphate. From TEM imaging it is evident that increasing the aging time of sols has lead to an increase in the electrosprayed particle size. MTT assay confirms the non cytotoxic nature of the particles (Fig. 2a). Cell adhesion and proliferation on the surface of pelletized particles confirms the biocompatibility (Fig. 2b). Cellular uptake studies confirm the potential of the particles to act as therapeutic delivery agents. The loading and release profiles reveal that particles with a CaP ratio of 1.61 show a higher loading but a lesser release when compared to those with a CaP ratio of 1.67. Also smaller particles show a better loading and release profiles than their larger size counterparts. Conclusion: In conclusion, a modified electrospraying process has been used to produce HA and CDHA particles in different size regimes. The particles do not elicit any toxicity and are readily taken up by the cells. Thus,electrosprayed CaP particles with controllable sizes are valid candidates for targeted delivery of therapeutic agents.The process opens up avenues for producing other ceramic particles as well. Dr. H. K. Varma, Bioceramics Laboratory, SCTIMST

Induksi PLB Anggrek Vanda sumatrana Schltr. Liar Pada Media MS dengan Penambahan BAP dan NAA serta Ploidisasi dengan Kolkisin

The study about PLB induction of wild Vanda sumatrana Schltr. on MS media suplement with BAP and NAA and ploidisation by colchicine treatment was conducted from December 2014 until November 2015 at the Laboratory of Genetics and Cell Biology and Laboratory of Plant Physiology and Tissue Culture, Biology department, Faculty of Mathematic and Natural Science, Andalas University, Padang. The study aimed to 1) knowing the best concentration of 6-Benzyl amino purin (BAP) and α-Naphtalene acetic acid (NAA) for Protocorm Like Bodies (PLB) induction from shoot tip of V. sumatrana, 2) knowing the PLB response of V. sumatrana to concentrations and soak period of colchicine and 3) find the effective concentrations and soak period of colchicine to induce tetraploid on PLB of V. sumatrana. Shoot tips from in-vitro cultured of V. sumatrana were subcultured on Murashinge and Skoog (MS) medium supplement with 3 mg/l BAP + 0,5 mg/l NAA, 3 mg/l BAP and 1,5 mg/l BAP. PLB of diploid V. sumatrana from the best treatment were soaked in 0.05% and 0.1% colchicine for 24 and 48 hours respectively in MS liquid medium, as control were set PLB without colchicine treatment. The results showed that MS medium supplemented with 1.5 mg/l BAP was the best formula to induce PLB. The highest percentage of survival rate of PLB and percentage of survived PLB regenerated shoot was obtained from 0.05% colchicine with 24 hours soak period treatment. The effective treatment to induce tetraploid on PLB of V. sumatrana Schltr. was obtained from 0.05% colchicine solution for 24 hours soak period.

INDOOR MICROBIAL CONTAMINATION AND ITS RELATION TO PHYSICAL INDOOR AIR QUALITY (IAQ) CHARACTERISTICS AT DIFFERENT LABORATORY CONDITIONS

Laboratory usually refers to a room or building equipped with chemicals and biological agents for scientific experimentation and research. Due to its own indoor hazards and its cause of health implications, it is an urge to assess and to understand the physical indoor air quality (IAQ) characteristics in the laboratory and the variables affecting the degree of exposure to occupants. The main objectives of this study was aimed to assess and to compare the physical IAQ characteristics and airborne pollutants including particulate matters (PM) and gaseous pollutants between laboratories, to identify microbial contaminants via bacterial counts as well as scientific bacterial-kits species identification and to correlate the relationship of physical IAQ characteristics, airborne pollutants and microbial contaminants between different laboratory settings within the same building which are Natural Product (NP) laboratory, Plant Tissue Culture (PTC) laboratory, and Microbiology laboratory. The physical IAQ characteristics and airborne PM was measured using VelociCalc multi-function ventilation meter 9565 and DustMate environmental dust detector respectively. Surface Air System Indoor Air Quality (SAS IAQ) was used to capture the microbial contaminants and after that bacterial counting and identification were done. The scientific method protocol and standard reference limits were compared based on Industrial Code of Practice on Indoor Air Quality (ICOP) (2010) regulated by the Department of Occupational Safety and Health (DOSH). The temperature of PTC laboratory, velocity of NP laboratory and Microbiology laboratory, the respirable particulate matter (PM) of all three laboratories and Colony Forming Unit (CFU) count of PTC and NP laboratory exceeded the standard limit regulated by DOSH. This study demonstrated that Gemella morbillorum is the common bacterial species available in the environment with poor IAQ and there is a significant relationship between physical characteristics, airborne pollutants and microbial contaminants between the different types of laboratory settings. In conclusion, priority should be given to NP laboratory as it is exposed to poor IAQ conditions and immediate action should be taken to eliminate the problems.

Pengaruh Pemberian N 2,4-D Terhadap Pertumbuhan Dan Metabolisme Kalus Kedelai Pada Kondisi Hipoksida Secara Invitro

Soybean demand continues to increase as the number of people , but the production has not be enable to meet the needs of national soybean. One effort to increase production is to indentificat soybean acreage of inundation. The aims of the research was to determine about Identification callus of soybean toward inundation by in vitro. This research was carried out in The Tissue Culture Laboratory, Agriculture’s Faculty Of North Sumatera University from August to March 2015. Completely Randomized Design with two factors was used, first factor was auxin 2,4-D concentration consist of three leevels : 0 ppm; 2 ppm; 4 ppm. The second factor was Inundation and without Inundation. The parameters measured were are visualization of callus, percentage of growth callus, percentage of weight callus , the amount of chlorophyll a and b, and concentration of protein. The results showed that 2,4-D concentration and Inundation give significant effect on visualization of callus, percentage of growth callus, percentage of weight callus , the amount of chlorophyll a and b, and concentration of protein , but it have no significantly effect on the formation of chlorophyll a in the phase of inundation and interactions between as given significantly effect on all parameters. Keywords : 2,4-D, Hipoksida, In vitro Soybean demand continues to increase as the number of people , but the production has not be enable to meet the needs of national soybean. One effort to increase production is to indentificat soybean acreage of inundation. The aims of the research was to determine about Identification callus of soybean toward inundation by in vitro. This research was carried out in The Tissue Culture Laboratory, Agriculture’s Faculty Of North Sumatera University from August to March 2015. Completely Randomized Design with two factors was used, first factor was auxin 2,4-D concentration consist of three leevels : 0 ppm; 2 ppm; 4 ppm. The second factor was Inundation and without Inundation. The parameters measured were are visualization of callus, percentage of growth callus, percentage of weight callus , the amount of chlorophyll a and b, and concentration of protein. The results showed that 2,4-D concentration and Inundation give significant effect on visualization of callus, percentage of growth callus, percentage of weight callus , the amount of chlorophyll a and b, and concentration of protein , but it have no significantly effect on the formation of chlorophyll a in the phase of inundation and interactions between as given significantly effect on all parameters. Keywords : 2,4-D, Hipoksida, In vitro

PENGARUH PEMBERIAN JENIS DAN KONSENTRASI AUKSIN TERHADAP INDUKSI PERAKARAN PADA TUNAS Dendrobium sp SECARA IN VITRO

ABSTRACT
 Dendrobium orchids are the most widely grown because it has the advantages of easy to maintain and most easily grown. Induction of rooting in bud of Dendrobium sp by giving the different types and concentrations of auxin is one of many ways to get plantlets in large quantities and relatively quick time. The purpose of this research is to obtain the types and concentrations of auxin is the best in inducing rooting in bud of Dendrobium sp. This research was conducted at the Tissue Culture Laboratory, Department of Biology, Faculty of Mathematics and Science, State University of Jakarta on February to June 2013. The method used was description and experimental, the design of a factorial with two groups consisting of one control group and one experimental group for the provision of the types and concentrations of auxin on root induction in Dendrobium sp bud with each group consists three replications. Analysis of the data using three-way anova. Based on this research, Dendrobium sp bud induced by the different types and concentrations of auxin showed a distinct influence on the growth of bud in qualitative and quantitative characters. Qualitative data on root induction, shows that there is a root that grows one and two roots and shoots are green, white and brown. The results based on quantitative data showed that the bud of Dendrobium sp induced by auxin type 2,4-D and concentration of 0.75 ppm had an average number higher root length compared with the type of auxin IAA and NAA. Dendrobium sp bud are planted in the medium by the addition of auxin type of IAA and NAA at a concentration of 1.25 ppm has an average number of roots better than the auxin type 2,4-D.
 
 Keywords: Dendrobium sp, Types and Concentrations of Auxin, Rooting Induction

In Vitro Propagation and Ex Vitro Acclimatization of Magnolia (Magnolia grandiflora, Linn) Trees

This study was carried out in the tissue culture laboratory, Faculty of Agriculture, Saba basha, Alexandria University, Egypt during the period from 2013 to 2015. An efficient and reliable protocol for in vitro propagation of Magnolia grandiflora, Linnwas optimized. However, nodal explants from field grown of magnolia were used during in vitro culture study for induction of multiple shoots. Nodal explants were effectively surface sterilized with 30% Clorox (sodium hypochlorite) as commercial bleaches for 20 min plus 1.5mg/l mercuric chloride for 5 min with few drops of Tween-20, also. Nodal explants were inoculated on various initiation or establishment media with different combinations of IBA and KIN and the neoformed shoots were cultured on proliferation (multiplication) media for the development of multiple shoots, and the elongation media to elongate of the neoformed shoot. The subsequent elongated shoots were rooted, successfully. The best medium for shoot initiation was Woody Plant Medium (WPM) supplemented with 2.0 mg/l KIN and 1.00 IBA. The favourable medium for multiplication was the tested medium augmented with 5.0 mg/l KIN and 1.00 mg/l IBA.. Furthermore, the in vitro shoots showed healthy root development when the tested medium was supplemented with combination of 1.00 mg/l IBA and NAA ,each in turn (rooting stage). The shoots of Magnolia grandiflora multiplication and rooted successfully when they cultured in WP medium supplemented with 1.00mg/l charcoal. The combination of sand: compost (1:3) was used as substratum for the hardening of the in vitro plantlets, as a potting mix, was the best suited mix for the acclimatization of plantlets.

PENGARUH KONSENTRASI NAA DAN AIR KELAPA TERHADAP MULTIPLIKASI TEMULAWAK (CURCUMA XANTHORRIZHA ROXB.) SECARA IN VITRO

<p><em>Java turmeric is a medicinal plant which in top ranks in its uses. Many people choose herbal treatments for their health. This study aims to investigate the response of the multiplication of java turmeric against granting PGR NAA and young coconut water in vitro. This research was conducted at the Laboratory of Biotechnology and Tissue Culture, Sebelas Maret University, Surakarta. The research was arranged in a Fully Randomized Design with 16 treatments and 3 replications.</em><em> </em><em>The results showed that giving kombination treatment of 0</em><em>,5</em><em> ppm NAA and 60% young coconut water was able to increase root amount with by an average of 20,33. NAA 1,5 ppm can increase buds amount at 2 buds. Giving 20% young coconut water treatment were also capable of increase average length buds, root length in sequence 7,37 cm and 7,37 cm.</em></p>

TINGKAT KOLONISASI PERAKARAN BIBIT PISANG JANTAN YANG DIINOKULASI DENGAN BEBERAPA DOSIS INOKULAN FUNGI MIKORIZA ARBUSKULA SERTA LAMA PEMBERIAN FOSFAT

Male banana is one of the varieties of bananas that becomes a main commodity of West Sumatra. As a main commodity, then growth, quality and productivity need to be maintained and improved. Soil microbial technology application, such as the development of biological agents of Fungi Mycorrhizae Fungi (AMF), is one of the strategies need to be tested and developed to support the development of a national program of healthy banana seedlings as well as issues of pressing environmental preservation based fertilizers and chemical pesticides. This research was conducted at the Laboratory of Plant Physiology and tissue culture and greenhouse of Faculty of Agriculture, University of Andalas, Padang. This research used an experimental method arranged in a factorial completely-randomized design consisting of two factors with three replications. Factor A: Dose mycorrhizae (A0 = no inoculation, A1 = 5 g inoculant FMA PU10, A2 = 10 g inoculant FMA PU10, A3 = 15 g inoculant PU10, A4 = 20 g inoculant PU10. Factor B: Timing of phosphate (B0 = 0 HSI inoculant FMA PU10 B1 = 10 HSI inoculant FMA PU10, B2 = 20 HSI inoculant FMA PU10, B3 = 30 HSI inoculant FMA PU10, B4 = 40 HSI inoculant FMA PU10). The research shows that inoculation of inoculant FMA on different doses give a significant effect on the percentage of male banana seedling of root colonization. Where dosing 10 grams (A2) shows that the highest root colonization percentage compared with other treatments equals to 53. 33%. The provision of phosphate on 30th day of HSI Mycorrhizae shows that the highest percentage of colonization is 45.55%.

English

The present attempt aimed to optimize micropropagation protocols supplemented with different concentrations and combinations of benzylaminopurine (BAP) (0, 2, 4, 6 mg L-1) and indole acetic acid (IAA) (0.5 and 1.0 mg L-1). Exotic banana (Musa spp) genotypes GCTCV-215 (AAA), ‘Yangambi’ Yangambi Km-5 (AAA) and FHIA-23 (AAAA) were used in research work. Experiments were conducted at Plant Tissue Culture Laboratory of Nuclear Institute of Agriculture (NIA), Tando Jam. Data collected for in vitro shoot consists of the following parameters: days for bud initiation, rate of shoot proliferation (%), number of multiple shoots, shoot length (cm) and fresh mass of shoot (g). Significant (p≤ 0.05) variations were observed for varieties, treatments and varieties x treatment for all the parameters. Synergistic effects of BAP and IAA were observed in GCTCV-215 and Yangambi Km-5. Out of various treatments, best concentration for multiple shoot in short period of time for GCTCV-215 and Yangambi Km-5 was found in 4.0 mg /l BAP + 0.5 mg L-1 IAA. Maximum fresh mass of shoot observed at same concentration and combination of BAP and IAA and for shoot length combination of 4.0 mg L-1 BAP with 1.0 mg L-1 IAA was found to be most suitable for GCTCV-215 and Yangambi Km-5. FHIA-23, show better performance in MS medium supplemented with only BAP at concentration 4.0 mg/L-1. After development of root, in vitro plantlets were shifted from growth room to green house in polythene bags containing garden soil and humus mixture in ratio (1:1). Key words: Micropropagation efficiency, exotic musa genotype, growth regulators.

Effect of mutagens on regeneration and growth of in vitro grown epicotyl segments of rough lemon seedlings (Citrus jambhiri Lush.)

The present study on the effect of mutagens on regeneration and growth of in vitro grown epicotyls segments of rough lemon seedlings (Citrus jambhiri Lush.) was carried out during the years 2009- 2010 and 2010- 2011 in the Tissue Culture Laboratory, Department of Fruit Science, Punjab Agricultural University, Ludhiana. Developmental characteristics of the in vitro grown epicotyls segments on regeneration media were treated employing gamma radiation in Gray(Gy) at 0, 5,10,15, 20,25,30,35,40 and 45 Gy; the alkylating agent ethyl methane sulfonate (EMS) and methyl methane sulfonate (MMS) each at 0, 0.1, 0.2, 0.3, 0.4,0.5 and 0.6% (v/v) were evaluated. Epicotyl segments from one month old in vitro grown seedling were cultured in regeneration medium (MS+BAP 1.0 mglit-1) under controlled laboratory conditions (25±2? C, 16 hr photoperiod, 2000 lux light). LD50, the dose required to kill half of the tested population corresponded to 35Gy for gamma radiation, 0.3% each for EMS and MMS treatments. Number of days taken for regeneration increased with increasing dose of gamma irradiation, EMS and MMS. Percent regeneration, number of buds, number of shoots, shoot length, number of leaves, internodal length, primary root length and number of secondary roots decreased with increasing dose of gamma radiation , EMS and MMS. The study would be beneficial to induce desirable variations in plant growth characteristics of rough lemon by the use of mutagens treatment.

In vitro PROPAGATION OF SOME CITRUS ROOTSTOCKS. 1- EFFECT OF EXPLANT SOURCE AND GROWING MEDIA GROWTH REGULATORS CONTENT.

This investigation was carried out during 2013 to 2015 seasons in tissue culture laboratory of Horticulture Department, Agriculture Faculty, Mansoura University, to make comparssion between shoot tips and epicotyls explants in vitro propagation of five Citrus rootstocks namely, Sour orange (Citrus aurantium, L.), Volkamer lemon (Citrus volkmeriana, Ten.). Cleopatra mandarin (Citrus reshni, Blanco), Rangpur lime and Troyer citrange . Our results showed that, epicotyl of Rangur rootstock, Sour orange and Volkamariana which cultured on MS medium containing BAP+Kin (BAP at 1 mg/L, Kin at 1.5 mg/L) gave distinct significant increase of length of shoots/plantlet from epicotyl explants comparing with another tested treatment in this study. Also, explants of epicotyl that took from Volkamariana, Sour orange and Rangpur on MS medium supplemented with IBA+NAA(IBA at 2 mg/L, NAA at 2mg/L) in vitro had significant increase of length of roots/explant comparing with other two citrus rootstocks in this respect.

Response of Durum Wheat Seedlings to Salinity

Salinity is an important source of abiotic stress, limiting crop performance in most arid and semi-arid areas of the world. This research was conducted to determine the effects of salinity on physiological parameters of durum wheat (Triticum durum Desf.) genotypes. The research was conducted in the tissue culture laboratory at the Agriculture Faculty of Dicle University. The study consisted of one durum wheat commercial cultivar, five local cultivars and four advanced genotypes. There were three replications in a split-plot experimental design. Genotypes were germinated in four NaCl concentrations (0, 50, 100, 150 mM) in plastic boxes. There were statistically assured significant differences among the genotypes for all salt concentrations and all observed parameters (coleoptile length, seedling length, root length, seedling fresh weight, root fresh weight, seedling dry weight, root dry weight, germination rate and seedling vigor). There was significant decrease in all examined parameters depending on the increase of salt concentration. The ‘Sorgul’ genotype was most tolerant to salinity, in terms of root length and root dry weight, whereas ‘Altintoprak 98’ was most tolerant as measured by the impact of salinity on coleoptile length, seedling fresh weight, germination rate and seedling vigour. The ‘Beyaziye’ genotype was the most sensitive to salinity-induced stress. The results from this study demonstrated differences among durum wheat genotypes for seedling parameters measured in the presence of salinity stress.

Response of Durum Wheat Seedlings to Salinity

Salinity is an important source of abiotic stress, limiting crop performance in most arid and semi-arid areas of the world. This research was conducted to determine the effects of salinity on physiological parameters of durum wheat (Triticum durum Desf.) genotypes. The research was conducted in the tissue culture laboratory at the Agriculture Faculty of Dicle University. The study consisted of one durum wheat commercial cultivar, five local cultivars and four advanced genotypes. There were three replications in a split-plot experimental design. Genotypes were germinated in four NaCl concentrations (0, 50, 100, 150 mM) in plastic boxes. There were statistically assured significant differences among the genotypes for all salt concentrations and all observed parameters (coleoptile length, seedling length, root length, seedling fresh weight, root fresh weight, seedling dry weight, root dry weight, germination rate and seedling vigor). There was significant decrease in all examined parameters depending on the increase of salt concentration. The ‘Sorgul’ genotype was most tolerant to salinity, in terms of root length and root dry weight, whereas ‘Altintoprak 98’ was most tolerant as measured by the impact of salinity on coleoptile length, seedling fresh weight, germination rate and seedling vigour. The ‘Beyaziye’ genotype was the most sensitive to salinity-induced stress. The results from this study demonstrated differences among durum wheat genotypes for seedling parameters measured in the presence of salinity stress.

Kandungan Klorofil Daun Planlet Anggrek Bulan (Phalaenopsis amabilis (L.) Bl.) Hasil Seleksi dengan Asam Salisilat secara In vitro

Phalaenopsis amabilis is one of Indonesias national flowers. P.amabilis has beautifulflowers and exciting and high economic value. However, there are some constraints in its development, one of which is Fusarium wilt caused byFusarium oxysporum, for this is not maximized in handling. One of effective procedure to control the disease is to use salicylic acid as a selective agent resistance to the disease. Study of the effect of the addition of salicylic acid(concentrations 15, 30, 45 and 60 ppm) have been studied in vitro in mediumVW(Vacint and Went) on the content ofchlorophyll a, b, and total leaf plantlets P.amabilis. The research was implemented in tissue culture laboratory, Department of Biology, Faculty of MIPA, University of Lampung from February to March 2015. This study used a completely randomized design with 3 replications. Analysis of variance and LSD test performed at 5% significance level. Calculation and extraction of chlorophyll was conducted by Harbourne method(1987). The absorbance was measured with a (Shimadzu UV 800)spectrophotometer at wavelength of 663 and 646 nm. The results showed that the content of chlorophyll a, b, and total plantlets of P. amabilis leaves significantly increased at the concentration of salicylic acid in the medium VW 15, 30, 45 and 60 ppm compared with control(0 ppm).Key words : Phalaenopsis amabilis plantlets, chlolophyll, salicylic acid, VW medium.