Tissue Culture Laboratory Research Articles

Effect of different combinations of 6-benzyl amino purine and naphthalene acetic acid on multiple shoot proliferation of plantain ( Musa spp.) cv. Matoke from meristem derived explants

This study was conducted at the Amhara Region Agricultural Research Institute Tissue culture Laboratory, Ethiopia from April to June 2012 to investigate the effect of different concentrations of 6-benzyl amino purine (BAP) and naphthalene acetic acid (NAA) on virus free plant regeneration and shoot multiplication. The culture meristem grew into callus after 28 days then adventitious plantlets were developed. Among the different concentrations, 5 mg/l BAP + 1.0 mg/l NAA showed highest shoot proliferation (1.00, 1.67, 1.75 and 3.08 shoots per clump) at 10, 20, 30 and 60 days after induction (DAI), respectively. Good numbers of shoots were achieved at 5 mg/l BAP + 0.5 mg/l NAA at 60 DAI (3.08). The longest shoot was produced by the concentration of 5 mg/l BAP + 1.0 mg/l NAA showed (0.43, 2.42, 2.63 and 3.42 shoots per plantlet) at 10, 20, 30 and 60 DAI, respectively. The maximum number of leaves (1.67, 2.67, 3.67 and 4.33 leaves per explant) at 10, 20, 30 and 60 DAI produced on the medium supplemented with 5.0 mg/l BAP and 0.50 mg/l NAA. The lowest number of leaves was obtained from the control treatment. The longest leaves were produced by the concentration of 5.0 mg/l BAP + 1.0 mg/l NAA (1.52, 2.27, 2.70 and 3.13 cm) at 10, 20, 30 and 60 DAI, respectively. Keywords : Plantain, regeneration, micro propagation African Journal of Biotechnology Vol. 12(7), pp. 709-719

Pertumbuhan dan Perkembangan Cabai Keriting (Capsicum annuum L.) secara In Vitro pada beberapa Konsentrasi BAP dan IAA

<p style="text-align: justify;">Curly red pepper (Capsicum annuumL.) is one of the crops that are important and widely cultivated in Indonesia. The purpose of this research was to determine the effect of the use of plant growth regulators BAP and IAA in various doses on the growth and development of curly red peppers in vitro. This research was conducted on March until October 2011 in Tissue Culture Laboratory Department of Agronomy and Horticulture, Faculty of Agriculture, Bogor Agricultural University. This research using Randomized Complete Design with combination of plant growth reguators as the factor. The results showed that combinations of growth regulators significantly influenced root induction, high growth, number of leaves, callus and shoot multiplication. Plant growth regulator combination of 0 mg L-1 BAP + 1 mg L-1 IAA was very good treatment in inducing roots. Combination of 2 mg L-1 BAP + 0.2 mg L-1 IAA effect high growth and shoot induction. Combination of 1 mg L-1 BAP + 0 mgL-1 IAA was the best treatment in increasing number of leaves. While for callus and multiplication, the best treatment was combination of 4 mg L-1 BAP + 0 mg L-1 IAA.</p><p>Keyword: induction, growth regulators, pepper plant</p>

ORGANOGENESIS TANAMAN ANGGUR HIJAU (Vitis vinifera L.) PADA MEDIUM MSDENGAN PENAMBAHAN IAA (Indole Acetid Acid) DANBERBAGAI KONSENTRASI BAP (Benzil Amino Purin)

Research onorganogenesisof Grape (Vitis viniferaL.) on MS medium supplemented with IAA and various sconcentrations of BAP, was conducted at the Tissue Culture Laboratory, Faculty of Forestry Untad Palu, during periode of Januari and April 2012.This research was aimed to obtain the best combination ofIAA and BA Pin MS medium that stimulate organogenesis of Grape. This experiment was arranged in completely randomized design with 4 treatments and 3 replications.The treatmentswere: MS medium+ 0.1ppm IAA+ 0.1 ppm BAP(B1), MS medium+ 0.1 ppm IAA +0.3 ppm BAP (B2), MS medium + 0.1ppm IAA + 0.5ppm BAP(B3) and MS medium+ 0.1 ppm IAA+0.7BAP(B4). Parameters observed in this study were the days appear of shoots, the days appear of leaves, percentage of explant producing shoots (%), and the presence or absence of roots. The results showedthat MS medium+ 0.1 ppm IAA+0.7ppm BAP(B4) gived the best results in speedupof shootsandleaves,in 7 days after induction.

Induksi Kalus Tanaman Kakao (Theobroma cacao L.) Pada Media MS Dengan Penambahan 2,4-D, BAP Dan Air Kelapa

Study on callus induction of cacao ( Theobroma cacao L . ) was conducted at the Tissue Culture laboratory, Forestry faculty, Tadulako University, Palu, during periode of January until April 2012. This research was aimed to obtain the best medium formulation for induction and growth of cacao callus. Calli was induced from embryo on MS based medium containing of 2,4-D, BAP and coconut water. The intact plant as explant was taken from Local Farming. This experiment was arranged in completely randomized design with 4 treatments and 3 replications. The treatments were : MS medium + 2 ppm 2,4-D + 15% coconut water (N1), MS medium + 2 ppm 2,4-D + 0,2 ppm BAP, 15% coconut water (N2), MS medium + 2 ppm 2,4-D + 0,2 ppm BAP (N3), and MS medium + 2 ppm 2,4-D (N4). Parameters observed in this study were the days appear of callus, percentage of explant producing callus (%), also observation on morphology and the cell of callus. The best medium for callus induction was MS medium + 2 ppm 2,4-D + 15% coconut water (N1). This medium produces healthy-compac callus which active in cell proliferation, in 6 days after induction.

Perbanyakan Tanaman Melon (Cucumis melo L.) Secara In Vitro Pada Medium Ms Dengan Penambahan Indole Acetic Acid (IAA) Dan Benzil Amino Purin (BAP)

The research on organogenesis of melon ( Cucumis melo L.) was done at Tissue Culture Laboratory Forestry Faculty UNTAD Palu, during January until May 2012. The aim of this research was to determine the best concentration of IAA and BAP hormones in inducing organs of this plant. Sterile seedling of melon was used as explants in this work. The experiment was arranged in Completely Randomized Design (CRD) with 4 treatments and 4 replications. The treatments were MS0 + 0,1 ppm IAA + 0,5 ppm BAP (M1), MS0 + 0,1 ppm IAA + 0,7 ppm BAP (M2), MS0 + 0,1 ppm IAA + 0,9 ppm BAP (M3) and MS0 + 0,1 ppm IAA + 1 ppm BAP (M4). Parameters observed on this study were the day appear of shoot and leaves, the number of shoots and leaves and also the present of callus and root. The result showed that the best medium for organs induction was MS0 + 0,1 ppm IAA + 1 ppm BAP, which shoot and leaf emerged in the day of 3,75 and 5,75 after induction, respectively. This media was also induced the number of shoots and leaves, i.e 4,75 and 8,75, respectively. Nevertheles this media was not suitable enough in inducing root formation.

Microbial contaminants of date palm (Phoenix dactylifera L.) in Iraqi tissue<br>culture laboratories

The date palm is one of the most important economic species of the palm family, grown mainly for its fruits (dates). Nowadays there is increased demand for date palm fruits around the world. To meet this demand, several propagation methods have been utilized, among them micropropagation which has been used in Iraq and many other countries for large-scale multiplication of date palm. Micropropagation faces several constraints; one is microbial contamination which represents a major challenge to the initiation and maintenance of date palm micropropagation laboratories. In recent years, two major groups of contaminants have been identified and isolated from different date palm tissue culture laboratories in Iraq. The first group is fungi. Several fungal species have been isolated and identified as contaminants; most predominant are: Aspergillus niger, Alternaria alternataand Penicilliumspp. The second group is bacteria; predominantly of the genera Bacillus, Staphylococcusand Proteus.

Knowledge and Awareness of Routine Biosafety Measures and Proper Waste Disposal Practices among Healthcare Workers in Karachi, Pakistan

This study assessed the knowledge and awareness regarding routine biosafety measures and proper waste disposal practices among healthcare workers in Karachi, Pakistan, and also highlighted areas in which training was needed. To increase knowledge of biosafety and waste disposal requirements, the Virology & Tissue Culture Laboratory, Department of Microbiology, Jinnah University for Women, Karachi, Pakistan in association with the Biosafety Association of Pakistan (BSAP) executed a collaborative project entitled “Training Workshop on Proper Waste Disposal from Hospitals & Pathological Laboratories in Karachi, Pakistan,” with financial support from the Biosecurity Engagement Program (BEP), the United States Department of State (DOS), and Civilian Research & Development Foundation (CRDF) for health-related issues in developing countries. A 50-question pre-workshop survey was administered to better understand the current biosafety and biosecurity measures in place at 250 state and private-sector clinical facilities (hospital and pathological laboratories) in the city. Participants from the 250 facilities were selected on the basis of first-come, first-served, but with preference for participants without prior knowledge of biosafety and biosecurity. After the training workshop, an evaluation survey was conducted with 28 participants. Results of that survey indicate that healthcare workers (HCWs) need to be offered training and refresher courses, in Urdu, to increase their level of knowledge and awareness of biosafety and waste disposal protocols to reduce both occupational exposures and the spread of infectious diseases in the community through improper disposal of infectious waste.

Improvement of Selected Induction Culture Media on Callus Induction in Anther Culture of Anthurium and a Histological Study on its Callus Formation

Improvement of selected induction culture media on callus induction in anther culture of anthurium and a histologicalstudy on its callus formation were studied at the tissue culture laboratory of the Indonesian Ornamental CropsResearch Institute from February to October 2008. The objectives of the study were to optimize selected media forcallus formation, reveal cell origin of callus derived from anther culture and shoot formation process. Selectedmedia improved in the study were 1) MMS-TBN containing 0,5 mg/l TDZ, 1,0 mg/l BAP and 0,01 mg/l NAA (Winartomedium, WM) and 2) MMS III supplemented with 1,5 mg/l TDZ, 0,75 mg/l BAP and 0,02 mg/l NAA (Winarto andRachmawati medium, WRM). Improvement treatments were carried out by omission and application of 2,4-D in 0.5mg/l and reduction of medium strength of full, half, quarter, one eighth, one sixteenth, and zero strength. Afactorial experiment was arranged using a randomized complete block design with four replications. Results ofthis study indicated that the highest callus induction was clearly established in WRM. The medium stimulatedpotential growth of anther (PGA) up to 81% with 49% of percentage of anther regeneration (PAR) and 2.7 number ofcallus formed per replication (NCF). Significant improvement in callus formation was also recorded by reduction ofmedium strength of WRM to one eighth compared to others. The reduction induced PGA up to 58% with 29% of PARand 1.8 NCF. From histological studies it was well recognized that regenerated callus on half anthers cultured wasoriginated from middle layer cells of anther wall. The morphogenic response of anther wall cells caused primarilyon no androgenesis effect in microspore cells.

SOMATIC EMBRYOGENESIS AND ADVENTITIOUS SHOOT REGENERATION FROM LEAF EXPLANTS AND STEM NODES OF

The present study was carried out in Plant Tissue Culture Laboratory Department of Horticulture , College of Agriculture & Forestry , Mosul University to propagate lilium plants Lilium longiflorium cv. Aster lily a white flowers in vitro by using nodes from plants growing in field in 5 May after flowering and cultured in MS medium supplemented with different concentrations of Kinetin or BA and rooting the shoots produced in vitro , as well as to explain the effect of sucrose on bulblet produced on rooted shoots , also medium or basal parts of leaves were cultured on MS medium supplemented with different concentrations of BA interacted with different concentrations of 2,4-D or on MS medium supplemented with BA and NAA , nodes taken from plants grown in field also cultured in 10 July before plant dormancy on MS medium supplemented with different concentrations of 2,4-D . Data refers that highest number of shoots 8.1 shoot/explant obtained from nodes cultured on MS medium supplemented with 0.5 mg/l BA after eight weeks from planting date , 100% rooting and highest number of roots 11.8 root/explant was obtained from cultured shoots produced in vitro on MS medium supplemented with 0.5 mg/l IBA , highest weight and volume 0.997gm , 0.43 cm3 for bulblet were obtained from cultured rooted shoots on MS medium supplemented with 120 gm/l sucrose , callus produced from culturing nodes three time on MS medium with 0.2 mg/l 2,4-D produced 50% shoots with average 5 shoot /explant after cultured on MS medium without growth regulators and stages of somatic embryogenesis were observed (globular, heart, torpedo) on callus, shoots initiated directly on basal leaf parts cultured on medium supplemented with 0.1 mg/l 2,4-D , otherwise callus initiated on basal leaf parts cultured on medium supplemented with 0.5 mg/l BA and 0.1mg/l NAA and when recultured these callus somatic embryogenesis were observed on it , all plantlets produced in vitro acclimated in laboratory and transferred to plastic house and then to field with 100% survival.

PRODUCTION CORDYLINE FRUTICOSA, L. BY USING TISSUE CULTURE

Under in vitro conduction this experiment was carried out at Tissue Culture laboratory at ,Horticulture Research Institute, Agriculture Research Center, during 2009 – 2010 year to substitute chemical hormones with natural compound at establishing stage with studying proliferation ,rooting and acclimatization of Cordyline fruticosa, explants. The results emphasized that using 20% Clorox for 20 min for tips' and 10-20 min, for node with immersion in 70% ethyl alcohol for five seconds gave the best results (100%) survival and free contamination of this explants. Pollen grains collected by hand or bee at 4.00g/l induced significantly increased in callus production, explants development, survival% ,length of shoots ,number of formed leaves but not reached to the level of significance in increasing number of formed shoots for both types of explants tips and one node .One node cutting failed in rooting by addition the two types of pollen grains to cultured medium. Furthermore the use of1.0 mg/l for IBA , NAA or IAA were the best concentrations for callus formation, explants development and survival% .Using MS medium contained 1.0 mg/l NAA and 2.0 mg/l BA or KI significantly increased number and length of shoots, BA was better than KI. The shoots of Cordyline fruticosa, successfully rooted when they cultured in MS medium supplemented with 2.0 mg/l IBA or NAA or IAA. Plantlets after root development formulation of the agricultural media of sand and peat moss at a ratio of 1:3 were effective in increasing survival %,and acclimatization of plant parameters expressed by plant height, shoot thickness and number of leaves. So, it could be recommend to use pollen grains for substitute chemical hormones but it's not commercial. Cultured explants on MS medium contained 1.0mg/l.(NAA or IBA or IAA)and 2.0mg/lBA for producing the best cordyline production. In addition ,using the mixture of sand and peatmoss at a ratio 1-3 for the best acclimatization of Cordyline fruticosa,plant .

Influence of Different Media Variety and Growth Regulator on Mycelial Colony Proliferation of Mushroom

An experiment was conducted at the Mushroom growth house and Tissue Culture Laboratory, Horticulture Demonstration and Training Centre (HDTC), Kewatkhali, Mymensingh during February to May, 2006 to investigate the mycelial colony proliferation of different mushroom species (Oyster (Pleurotus florida), Milky (Calocybe indica) and Button (Agaricus biporus)) in different media (viz. PDA, YPDA and MEA) and different combinations and concentrations ( 0, 1, 5, 10 and 20 ppm) of different growth regulators (IAA and NAA). As a media, the best mycelial growth of 9.34 cm was found in YPDA (Yeast Potato Dextrose Agar) at 21 DAI. As a variety, the best mycelial growth of 11.90 cm was performed by Oyster at 21 DAI. In case of combined effect of different media and variety for mycelial growth, the best growth (12.57 cm) was found in Oyster with YPDA media at 21 DAI. In case of growth regulator, the best mycelial growth of 5.50 cm was found at 21 DAI with 10 ppm IAA and 4.03 cm was found at 18 DAI with 5 ppm NAA and 5.38 cm at 21 DAI with control. In case of combined effect of IAA and NAA, the best mycelial colony proliferation (6.72 cm) was found with 10 ppm IAA + 5 ppm NAA at 21 DAI. It may be concluded that among three varieties and media, Oyster was the best variety and YPDA was the superior media for mushroom cultivation. Malt extract supplemented with 10 ppm IAA+ 5 ppm NAA as found to be the best for mycelial colony proliferation of Button mushroom DOI: http://dx.doi.org/10.3329/jesnr.v5i1.11586 J. Environ. Sci. & Natural Resources, 5(1): 223 – 227, 2012

USING TISSUE CULTURE TECHNIQUE IN MICROPROPAGATION OF SWEET POTATO (ipomoea batatas)

The current study was conducted at tissue culture laboratory of Vegetable and Floriculture Department, Faculty of Agriculture, Mansoura University during the period from 2010 – 2012. A protocol is described for rapid and large-scale propagation of the sweet potato (Ipomoea batatas) by in vitro culture of shoot tips or nodal segments as explants. The best sterilization method for the explants was observed by using Hg Cl2 at 0.1 % for 14minutes and70%ethanol for 30 seconds. Nodal segments were found to be more efficient than shoot tips for sweet potato shoots regeneration on MS medium (Murashig and skoog, 1962) supplemented with 2ip at 4 mg/l + 0.5 mg/l GA3. Adding activated charcoal (AC) to the culture medium at 2 mg/l was the most effective treatment to avoid of browning phenomenon occurrence (0.00%). Among the five different cytokinins Thidiazuron (TDZ), kinetin (kin), 6-benzyl aminopurine (BAP), - N6-(2-isopentenyl) adenine 2ip and phloroglycenol (PG) and four concentrations of each, TDZ at 1.00 mg/l gave the best results for shoots multiplication. Concerning the rooting stage, Indole-3-acetic acid (IAA) at 2.0 mg/l clearly in henced roots development. Mixture of peat moss :vermiculite : perlite (1:1:1 v/v/v ) recorded the highest results for survived plantlets in terms of survival percentage,

Tissue Culture Laboratory of Institute of Neurosurgery is 50 years

Tissue Culture Laboratory of Institute of Neurosurgery is 50 years

Rapid cell sheet detachment using spin-coated pNIPAAm films retained on surfaces by an aminopropyltriethoxysilane network

The ability to harvest cell sheets grown on thermoresponsive polymers, such as poly(N-isopropylacrylamide) (pNIPAAm), has been widely studied for use in tissue engineering applications. pNIPAAm is of special interest because of the phase change that it undergoes in a physiologically relevant temperature range. Two primary approaches have been adopted to graft pNIPAAm chains covalently onto tissue culture polystyrene dishes: electron beam irradiation and plasma polymerization. These approaches often involve non-easily accessible (e.g. e-beam) facilities and complicated procedures that have hindered most tissue culture laboratories in adopting this technology for their specific applications. In this study, we developed a simple and cost-effective approach to create thermoresponsive surfaces using commercially available pNIPAAm. Using a simple spin-coating technique, thermoresponsive thin films were deposited on glass slides or silicon wafers using pNIPAAm blended with a small amount of 3-aminopropyltriethoxysilane (APTES), which enhances the retention of pNIPAAm on the surface. We found that the thermoresponsive films created using our method support cell attachment and proliferation without additional adhesive proteins as well as cell sheet detachment within minutes.

In vitro conservation and production of vigorous and desiccate tolerant synthetic seeds in Stevia rebaudiana

Artificial (Synthetic) seed production along with microprpagation can solve many problems with regeneration of Stevia rebaudiana, which produce tiny and non-viable seeds. Shoot tips and axillary buds nodal segments were used from in vitro cultures for encapsulation. The explants were taken from green houses and tissue culture laboratory of AgriBiotech Research Farms, Lahore, Pakistan. Different concentrations of sodium alginate and calcium chloride affect the shape and germination of artificial seeds. A gelling matrix of 3% sodium alginate and 100 mM calcium chloride was found most suitable for the formation of firm, clear and isodiametric ideal beads. No effect of the duration of sodium alginate treatment on bead formation was observed. However, timing of CaCl2 treatment proved to be crucial with 100 mM CaCl2, treatment time of 15 min was suitable for isodiametric bead formation. The synthetic seeds pre-treated with KN03 germinated earlier than seeds without pretreatment with KNO3. Root formation took place after 15 days of shoot formation without addition of any auxin. In liquid medium, the frequency of conversion was high. Frequency to plantlet conversion of synthetic seeds decreased gradually as the storage duration at 4°C increased. After 15, 30, 45 and 60 days of storage, frequencies to plantlet conversion from encapsulated and non-encapsulated segments were 86, 63, 50, 23.3 60.3, 40, 20.6 and 10%, respectively. Key words: Stevia rebaudiana, synthetic seed, in vitro conservation, Na-Alginate, encapsulation.

OA01.43. Phytochemical And Tissue Culture Studies With Terminalia Arjuna

Purpose: There is genetic variability in Terminalia species which is a very tall tree ranging from 5 to 30 meters. Government of India has established a tissue culture laboratory and the callus formation and differentiation tests were conducted at Ranchi, and it was found that this plant can be used for pharmacognosy and fibre. Method: In pharmacognosy, through tissue culture, anti-oxidative properties of ethanol extract of Terminalia bark was tested against sodium flouride induced oxidative stress in heart. The activities of various antioxidant enzymes, levels of cellular metabolites reduced carbonyl contents were already determined in the cardiac tissue. Tissue culture is a specialised area of production which should be exploited by Ayurvedic researches. Terminalia Arjuna was cultured on nutrient media supplemented with different concentration of phyto-hormones along with auxins and cytokinins implemented with coconut water. Result: Adenine Sulphate at the rate 25 mg/lt were added for nodal and auxiliary explants, and these nodal explants and shoot tips of T. arjuna were cultured again to have shoot proliferation. This can be used for other herbal plants for exploitation in medicinal and other useful purposes. In Ayurveda it can help in cost reduction and efficacy enhancement. Conclusion: Culture the new variety of Terminalia species can be developed with the help of tissue culture from Ayurvedic point of view, which can reduce the level of lipid profile, and angiotensin activity should be compared with others.

Influence of Taraxacum officinale Wigy and Malva parvifloral L. on the active ingredient of tomato in vitro

Research was done in College of Science for Women/ Biology Department/ Plant Tissue Culture Lab. from 2009 to 2010 Results showed that the best plant growth regulator combination for the highest callus quantity production was 1 mg/l Naphthalene acetic acid (NAA) with 2 mg/l Benzyl adenine (BA) so it was fixed in all the preceding experiments. Alcoholic extract for the two plants were added to full Murashigue & Skooge (MS.) and half MS. (modified) with concentration (2,4) mg/l for both as a replacement for vitamins and salts. These extracts showed an effect on the increasing of Ascorbic acid (vit. C) in full MS. medium with 4 mg/l in induced callus of tomato with peak area 68.859 by using Malva parviflora L. and 48.478 by using Taraxacum officinale Wigy and they effect in less degree on Nicotinic acid (vit. B5) with T. officinale Wigy extract effect only at 4 mg/l. with peak area 47.871. They effect on Caffaic acid were nearly abcent.

PERFORMANCE OF COCONUT EMBRYO CULTURE ACCESSIONS INTRODUCED AT THE INTERNATIONAL COCONUT GENEBANK FOR LATIN AMERICA AND THE CARIBBEAN (ICG-LAC)

The present work is a partial report for the introduction of eleven accessions of dwarf coconut and one accession of giant coconut tree into the International Coconut Genebank for Latin America and the Caribbean (ICG-LAC), a partnership between Embrapa Coastal Tablelands (CPATC) and Bioversity International, coordinated by Coconut Genetic Resources Network (COGENT). These accessions, originated from the International Coconut Genebank for Asia, India and Oceania (ICG-AIO), situated at Ivory Coast, were introduced in Brazil through zygotic embryos in November 2008. The mature fruits and endosperm cylinders with embryos were submitted at Ivory Coast to a phytosanitary treatment process according to international technical recommendations for safe movement of vegetal germplasm. At the Laboratory of Plant Tissue Culture of CPATC, the embryos were excised, surface sterilized and inoculated in Y3 culture medium. The first evaluation occurred after nine days of inoculation, and it was verified that 1.38% of the embryos were contaminated by fungi and 15.30% by bacteria. After 12 months of inoculation variation of in vitro development was observed within and among accessions and a high percentage of non-germinated and oxidated embryos. The normal germination percentage varied from 3.33% for ‘Niu Leka Dwarf’ to 33.3% for ‘Malayan Green Dwarf’ accessions. The non-germinated embryos percentage varied from 18.47% for ‘Tahitian Red Dwarf’ to 47.72% for ‘Sri Lanka Green Dwarf’ accessions. After 12 months, the contamination was higher for ‘Niu Leka Dwarf’ (70%). The percentage of non-germinated and oxidated embryos varied from 18.47% (‘Tahitian Red Dwarf’) to 47.72% (‘Sri Lanka Green Dwarf’). The great number of collected cylinders with exposed embryos (not protected by endosperm) and cracked endosperm, in addition to the long storage time (7 to 11 days) from collection to inoculation, favored bacterial proliferation. The non uniform stage of fruits/embryos maturation contributed to high variation of in vitro development.

Effect of interaction of 6-benzyl aminopurine (BA) and sucrose for efficient microtuberization of two elite potato (Solanum tuberosum L.) cultivars, Desiree and Cardina

Single node and multinode explants of two potato cvs., Desiree and Cardinal were tested for in vitro microtuber production. Explants were taken from tissue culture laboratory of Seed Centre, University of the Punjab, Lahore, Pakistan in 2004. Experiments were designed in completely randomized pattern. 34 media treatments of Murashige and Skoog (1962) with varying concentrations of sucrose (4, 6, 8, 10 and 12%) either alone or in combination (5-8% sucrose) with 6-benzyl amino purine (BA) were studied. In the case of cv. Desiree, medium MB10 (BA 6 mg/l, sucrose 6%) and for cv. Cardinal, medium MK20 (BA 5 mg/l, sucrose 8%) in term of induction, mean number and mean fresh weight per single node explant were optimized. In comparison, cv. Desiree genotypically was observed to be slightly slow in response to in vitro microtuber induction and development than cv. Cardinal. Key words: Potato, microtuberproduction, 6-benzyl aminopurine (BA), sucrose.

Regenerasi dan Pertumbuhan Beberapa Varietas Tebu (Saccharum officinarum L.) secara In Vitro

<p>(Saccharum officinarum L.) through In Vitro Culture.<br />Deden Sukmadjaja and Ade Mulyana. The research was<br />conducted at the Laboratory of Tissue Culture The Biology of<br />Cell and Tissue Researcher Group ICABIOGRAD, Bogor from<br />June to November 2009 to studied growth and regenerations<br />response some varieties of sugarcane through in vitro<br />culture. The research activities have been carried out in<br />three steps, i.e., callus formation, regeneration of shoots and<br />roots regeneration. The type of explants used in the study<br />was in vitro planlet explants of both sugarcane varieties.<br />Seven media formulations were used for the callus induction<br />and regeneration of shoots, while five media formulations<br />were used for the roots regeneration. The results<br />showed that the highest respond for calluses induction was<br />Bulu Lawang varieties at media formulation MS + 2.4-D 2<br />mg.l-1 + BAP 0.4 mg.l-1 + CH 2000 mg.l-1 and PS 951 varieties<br />at media formulation MS + 2.4-D 1 mg.l-1 + BAP 0.4 mg.l-1.<br />While the highest respond for regeneration of shoots was<br />Bulu Lawang varieties at media formulation MS0 (control<br />MS) dan PS 951 varieties at media formulation MS + BAP 1<br />mg.l-1 + kinetin 1 mg.l-1 + NAA 0.5 mg.l-1 + GA3 0.5 mg.l-1.<br />The highest respond of roots regeneration was Bulu Lawang<br />and PS 951 varieties at media formulation MS + IBA 1 mg.l-1.<br />Acclimatization of plantlets produced were grew successfully<br />about 90-100% in greenhouse.</p>