BAP Medium Research Articles

In vitro propagation of black cumin (Nıgella satıva L.) plants

This study was carried out to determine in vitro development using Black cumin leaf and stem explants. ?ameli black cumin variety was used as plant material. Five different nutrient mediums (1. LS2.5, 2. MS, 3. MS + 0.5 mg.l-1 IAA, 4. MS + 0.5 mg.l-1 BAP, 5. MS + 0.5 mg.l-1 IAA + 0.5 mg.l-1 BAP) containing 30 g sugar were used in this study. As a result of the research, 100% callus formation was detected in the stem explants cultured in the number 1 and number 5 mediums. These were followed by stem explants cultured in medium 4 with a success rate of 96%. Of this rate, 66% was shoot formation, and 30% was callus formation. Direct shoot regeneration was performed only on stem explants cultured in mediums 4 and 3, with a 66% success rate in medium four and a 36% success rate in medium 3. The highest plant regenerations from calluses were gained from stem explants (273.3%) in medium 4, followed by calluses gained from leaf explants (262.5%) in the same medium. These were followed by cultures in medium 3, with calluses derived from stem explants (255%) and leaf explants (150%). No plant regeneration was determined from calluses gained in the medium 1. Thus it is evident that high auxin content and auxin-cytokinin balanced mediums encouraged callus formation in the black cumin plants. The addition of only IAA or BAP to the medium promoted shoot formation in the stem explants, but direct shoot regeneration was not thereby achieved from the leaf explants. These results show that, for in vitro clonal propagation studies done on black cumin plants, a high auxin containing medium is preferable if the aim is callus formation. If the aim is direct shoot regeneration, BAP or other cytokinin-containing medium is preferred.

KARDELENİN (Galanthus Woronowoii ) İN VİTRO REJENERASYONU

The study was conducted in Tissue Culture Laboratory of the Field Crops Department of Agricultural Faculty of the Ege University in 2019. In the study, the bulbs of Snowdrop (Galanthus woronowii), is one of the geophytes of Turkey, were used as genetic material and were investigated for in vitro regeneration. The effects of different levels of MS+BAP (0.1; 0.5; 1.0; 2.0; 3.0 and 4.0 mg/l) and MS (control) were cultured in vitro. When the results were evaluated; the highest mean for bulb regeneration was obtained MS+4.0 mg/l BAP medium for shoot number (3.3) and shoot length (1.3 cm). Shoots of growing were sub cultured in media containing MS+0.5 mg/l IBA and MS+1.0 mg/l IBA. Medium of MS+ 1.0 IBA had the highest mean for root number (6.0) and root length (4.5 cm). As a result, Galanthus woronowii is to be suitable for in vitro production and can be evaluated for commercial production.

Classification of Intestinal Microorganisms Cultured from the Feces of Pet Dogs Fed Commercial Feed

The purpose of this study was to investigate the microorganisms found in the intestines of dogs fed commercial feed though the aerobic culture of dog excrement. 16 specimens of pet dog feces were cultured employing BAP, MaC, and EMB medium. Strains of Escherichia coli were detected in all the cultured specimens, along with a few other bacteria of significance. All 16 samples were identified as containing multiple E. coli strains using a API20E kit. Two samples showed a swarming phenomenon in the BAP medium that was identified as a Proteus mirabilis strain. In 10 samples, only E. coli species were identified, while E. coli species and P. mirabilis species were identified in 2 samples. A small number of E. coli and gram-positive bacteria were detected in 3 specimens, and a small number of E. coli and gram-positive rods were detected in one specimen. It has been shown that E. coli is predominantly distributed in the feces of dogs fed with commercial feed. The fact that E. coli is distributed in the feces of dogs is of concern as it can cause serious problems such as water pollution and food poisoning.

Bioactive compound and bioactivity fidelitous micropropagation method of Blumea lacera (Burm. f.) DC.: A large scale production potential

Blumea lacera (Burm. f.) DC. is an annually grown infallible medicinal plant that is used in many popular systems of traditional medicine but fresh plant material for medicinal uses and research purposes is critically unavailable year-round due to its very short life cycle. A micropropagation method of this medicinal plant ensuring fidelity of the bioactive compounds and bioactivity has thus been developed in order to maintain its year-round supply chain. In this regard, axillary multiple shoot regeneration from field grown nodal explant was established where the highest frequency (79.03 ± 0.04 %) induction of axillary multiple shoot bud organogenesis with the maximum number (13.60 ± 0.17 shoots/explant) of induced shoot bud was observed on MS + 3.0 mg/L BAP + 1.0 mg/L Kinetin medium. The induced axillary shoot bud exhibited the highest frequency (94.66 ± 0.33 %) of proliferation and elongation with the maximum number (32.16 ± 2.35) and length (7.28 ± 2.63 cm) of developed shoots/explant when sub-cultured in the same medium composition among tested media. The highest frequency (98.36 ± 0.85 %) of rooting with the maximum number (35.53 ± 1.53) and length (4.66 ± 1.72 cm) of root achieved on half-strength MS + 3.0 mg/L IAA medium. Explant suitability study found that in vitro grown leaf explants were superior over nodal explants for multiple shoot induction and proliferation. Leaf explants were collected from shoots that developed on MS + 1.5 mg/L BAP medium and cultured on MS + 3.0 mg/L BAP + 1.0 mg/L Kinetin medium. The cultures were maintained in the same medium composition following a subculture passages method for adventitious multiple shoot regeneration. A successive passage of subcultures achieved nearly 300 developed shoots/explant within 20 weeks. Regenerated plantlets retained all tested bioactive compounds and bioactivity properties like the donor plant thus maintaining medicinal fidelity.

Regeneration Rate of Eggplant Somatic Embryogenic In Various Maturation Media

Ralstonia solanacearum is one of the most important pathogen that causes bacterial wilt disease in eggplant and inhibits eggplant production. Improvement of eggplant varieties resistant to bacterial wilt can be accomplished through genetic manipulation. Regeneration of in vitro plants isone of the important tools to supports plant improvementthrough biotechnology. This study was aimed to determine the rate of eggplant regeneration in various maturation media, and to find the best medium for eggplant regeneration based on maturation rate and the number of cotyledon produced. We used resistant eggplant (accession 032) as the material to produce somatic embryogenic.There were 7 types of regeneration media used in this research. MS medium was supplemented with a certain concentration of plant growt regulators , such as: 1 mg / L + BAP 1 mg / L, NAA 4mg / L, TDZ 0.005 mg / L, TDZ 0.001 mg / L, CuSO4 2mM + BAP 1 mg / L, CuSO4 2mM + BAP 2 mg / L and Kinetin 1 mg / L + CuSO4 2mM. Three clumps of callus per plate with three replications were transferred to MS suplemented medium. The parameters observed were the color of callus before and after they were transfered to regeneration medium, the day of formation of globular, heart-shaped, tubular and cotyledonary phase, and the number of cotyledons formed. The results obtained showed the somatic embryogenic color of the 032 genotype was white with friable structure before being transferred to regeneration medium and was turned to yellowish white after being transferred to the regeneration medium. On the day sixth, friable embryogenic somatic of eggplant was developed into nodule on medium MS + NAA 4 mg / L, MS + CuSO4 2mM + BAP medium 1 mg / L, and MS + CuSO4 2mM + BAP 2 mg / L. Somatic embryogenic callus of accession 032 were able to pass complete globular, heart-shaped, tubular and cotyledonary phase. The most responsive medium for somatic embryogenic callus regeneration, based on the days of the callus phases formation and the number of early-phase cotyledons obtained, were MS medium suplemented with CuSO4 2mM + BAP,and CuSO4 2 mM + BAP 2 mg / L.Keywords: eggplant, Ralstonia solanacearum, regeneration, cotyledonary, clump, BAP

An efficient protocol for in vitro regeneration and conservation of Shirui lily (Lilium mackliniae Sealy): a lab-to-land approach to save the rare endangered Asiatic lily species

An efficient protocol for direct and indirect shoot regeneration and proliferation from bulb scales of Shirui lily (Lilium mackliniae Sealy), an endangered Asiatic lily species endemic to the Shirui hill peak, Manipur, India, has been developed. Bulb scales were isolated from mature bulbs and cultured on Murashige and Skoog (MS) basal medium supplemented with different concentrations of 6-benzylaminopurine (BAP), kinetin (KIN), or thidiazuron (TDZ). For direct shoot regeneration from bulb scale explants, 0.5 mg L−1 BAP yielded the highest shoot induction (3.5 shoots per scale; a 96.7% response). For indirect de novo organogenesis, optimum callus induction was achieved with 2.0 mg L−1 2,4-dichlorophenoxyacetic acid (2,4-D), and shoot organogenesis was higher (16.2) when subcultured onto 0.5 mg L−1 BAP medium. Multiple shoot regeneration and pseudo-bulb formation protocols were assessed; the highest shoot proliferation (10.1) occurred with 0.5 mg L−1 BAP and 1.0 mg L−1 gibberellic acid (GA3). Rooting response was 96% with 0.5 mg L−1 1-naphthalene acetic acid (NAA), with multiple roots per shootlet. Plantlet survival was increased to 92.5% during the hardening-off process by using hydroponics with Hoagland’s solution in a mist chamber. Clonal fidelity was assessed through random amplified polymorphic DNA (RAPD) analysis comparing the mother plant and regenerated plantlets. After confirming genetic uniformity, the pseudo-bulblets with four to six leaves and three to four roots were successfully established at the Shirui hills peak. This in vitro regeneration and ex vitro conservation approach could be helpful to save this rare endangered species in a sustainable way.

Indirect Organogenesis through Seedling-Derived Leaf Segments of Ficus Religiosa - a Multipurpose Woody Medicinal Plant

The aim of this study is to introduce the suitable protocol for indirect regeneration from seedling-derived leaf segment of Ficus religiosa. The leaf explant successfully produced callus on MS medium containing various concentrations of auxin in combination with BAP. The maximum callus induction (100%) was achieved in MS medium containing 0.5 mg/l 2,4-D plus 0.05 mg/l BAP and MS medium containing 1.5 mg/l NAA plus 0.15 mg/l BAP as well. MS medium consisting of 2,4-D produced yellow-brownish and friable callus (type I) while the yellowish and compact calli (type II) were obtained in MS medium consisting of NAA. On the other hand, MS medium supplemented with IBA formed greenish and compact calli (type Ш). The regeneration rate in type II callus was less than the type I, and there was no shoot induction observed on type Ш calli. MS medium supplemented with 1.5 mg/l BAP in combination with 0.15 mg/l IBA had the highest regeneration frequency (100%) and maximum shoot numbers (5.16) as well as shoot length (2.56 cm) in type I callus. A maximum of 93.33% root induction was observed in MS medium supplemented with 2.0 mg/l IBA plus 0.1mg/l NAA. The plantlets were successfully transferred to the greenhouse. This system could be utilized for large-scale multiplication of Ficus religiosa.

English

A reproducible in vitro shoot regeneration protocol was developed for pigeonpea variety ICPL 87 through proliferation of axillary buds from cotyledonary nodes. The seedlings were cultured on MS + Kin (2 mgL-1) + BAP (5 mgL-1) medium and de-capitated on 7th day of culturing, which resulted in the induction of multiple shoots from cotyledonary node region after 15 days of culturing. Miniature shoots were elongated on MS + adenine suphate (30 mgL-1) medium; the elongated shoots were rooted ex vitro using a rooting powder ROOTEXTM...

Morphological and Biochemical Responses to Gibberellic Acid in Magnolia × ‘Spectrum’ in Vitro

AbstractThe total soluble sugar content and antioxidant enzyme activities were studied for the first time during axillary shoot formation inMagnolia× ‘Spectrum’ in vitro in response to BAP (0.3 mg l−1), different levels of gibberellic acid (GA3; 0.0, 0.1, 0.5, 1.0 mg l−1), sucrose (20 and 30 g l−1) and nitrogen salts (KNO3/NH4NO3; 100/100% and 75/50% relative to MS medium). Among various GA3and sucrose/nitrogen salts ratios, the most effective axillary multiplication (5.9 shoots/explant) and leaf formation (25.7 leaves per multiplied clumps) were obtained after addition of GA3at 0.1 mg l−1to a BAP medium containing 20 g l−1sucrose and reduced levels of nitrogen salts (75% KNO3and 50% NH4NO3). The addition of GA3to the BAP medium enhanced shoot formation by 36% and leaf formation by 27%. The highest shoot formation capacity ofM.× ‘Spectrum’ in vitro coincided with enhanced levels of soluble sugar and peroxidase (POD) activity. Increasing GA3concentration from 0.1 to 1.0 mg l−1in the above medium resulted in inhibition of shoot and leaf formation and a decrease in the soluble sugar content. The influence of GA3on the activities of catalase (CAT) and POD depended on its concentration and the levels of sucrose and nitrogen salts in the medium. The highest increase in CAT and POD activities, that coincided with the enhanced shoot formation capacity ofM.× ‘Spectrum’ in vitro, was observed after addition of GA3to the medium containing high levels of sucrose and nitrogen salts.

Establishment of in vitro regeneration protocol for Adhatoda vasica Nees

An in vitro regeneration protocol of Adhatoda vasica has been developed using excised nodal segments and juvenile leaves for multiple shoots regeneration directly or through callus induction. Explants were cultured on MS medium with different concentrations of IAA, NAA, BAP, GA3 and Kn singly or in combinations. MS medium supplemented with BAP (10.0 mg/l) was found best for multiple shoot formation, in which 93.33% explants produced multiple shoots. After two months, maximum number of multiple shoots were 10.6 ± 1.82, highest length of plantlets was 5.2 ± 2.20 cm. 100% calli formation were observed on MS medium supplemented with IAA (0.05 mg/l) + NAA (0.05 mg/l) + BAP (1.0 mg/l). Callus initiation started after 14 days and gave light green colored callus. Best callus mediated shoot regeneration was found on MS+10.0 mg/l BAP medium. Root induction of in vitro raised shoots was best on ½ MS + IBA (1.0 mg/l). Well rooted plantlets were transferred to plastic pots containing garden soil and compost in a ratio of 2:1 for hardening. The ultimate survival rate under natural condition was about 80%.Bangladesh J. Sci. Ind. Res. 51(1), 75-80, 2016

Multiple shoot induction and jasmonic versus salicylic acid driven elicitation for enhanced andrographolide production in Andrographis paniculata

Multiple shoot induction at an average of (15.25 ± 2.128 and 16.16 ± 3.851 shoots/explant) was obtained from Andrographis paniculata cotyledonary node explants on Murashige and Skoog’s (MS) medium supplemented with 2.0 mg/L 6-Benzylaminopurine (BAP) and Zeatin, respectively after 8 weeks of culture. Further proliferation of cultures in MS + BAP medium at a maximum of 150 ± 18.40 multiple shoots was obtained from single cotyledonary node explants after three subculture (84 days) in vitro passage. Elicitation as yield enhancement strategy, a protocol was developed using multiple shoots to treat the cultures with jasmonic acid (JA) and salicylic acid (SA). Various concentrations of JA (1, 5, 10, 25, 50, 75 and 100 µM) and SA (10, 20, 50 and 100 µM) were used for elicitation. Treatment of JA at 1.0 µM concentration resulted in 1.322 % dry weight (DW) with 2.6-fold increase in andrographolide production after fifth week compared to 0.508 % DW in untreated control. However, JA treatment at 25 and 50 µM promoted 3.3, 3.0-fold enhancement in andrographolide production (1.624 and 1.481 % DW), respectively; after eighth week compared to control. Treatment of 10, 20 and 50 µM SA resulted in 3.0, 3.4 and 3.1-fold andrographolide content (1.479, 1.654 and 1.483 % DW), increase after eighth week, respectively; compared to control (0.478 % DW). This is the first report on elicitation of A. paniculata multiple shoot cultures using signal molecules (JA and SA). The present findings may be helpful for in vitro manipulation and enhanced production of andrographolides.

In Vitro Growth and Rooting of Mangosteen (Garcinia mangostana L.) on Medium with Different Concentrations of Plant Growth Regulator

Propagation of mangosteen is challenging for many reasons, including limited seed set, slow rate of seedling growth, and difficulty with root formations. The objective of this research was to find the best combination of medium and plant growth regulator for in vitro growth and rooting of mangosteeen seed. Various types of explant (a whole seed; seed divided into 2, 3, and 4 cross sections; seed divided into 2, 3, and 4 longitudinal sections) were treated with five concentrations of benzyl amino purine (BAP; 0, 2.5, 5, 7.5, 10 mg/L) for shoot induction in ½ Nitrogen (N) Murashige and Skoog (MS) medium. The shoots were rooted on MS and woody plant medium (WPM) media with several combinations of indole butyric acid (IBA) and naphtalene acetic acid (NAA). Treatments for root induction were applied as follows: (i) low dose, given during induction of rooting, (ii) soaking the base of the shoots in medium treated with a high dose of auxin for 5 days, and then growing the shoots in MS ½ N with 1 mg/L NAA + 1 mg/L BAP medium. Our result show that BAP positively affected mangosteen bud growth. The best medium for mangosteen shoot regeneration was found to be MS ½ N + 5 mg/L BAP. This medium induced the highest number of shoots from the seed explant cut into four cross sections. We found the best medium to induce in vitro rooting of mangosteen shoot was MS ½ N + 3 mg/L indole butiric acid (IBA) + 4 mg/L NAA medium. Some treatment negatively affected growth. Soaking the mangosteen shoot base in a medium with an overly high dose of auxin seemed to disrupt and inhibit growth of the mangosteen shoot.

In-vitro Callus Induction and Rosmarinic Acid Quantification in Callus Culture of Satureja khuzistanica Jamzad (Lamiaceae).

In the present study, an efficient protocol has been developed for callus induction and production of RA in callus culture of Satureja khuzistanica for the first time. In-vitro callus induction was achieved from young shoot tip explants cultured on MS and B5 media supplemented with different concentrations of IBA (0.1, 1.0, 2.0 and 5.0 mgL(-1)) solely or in combination with cytokinins BAP and KIN (1.0, 2.0 and 5.0 mgL(-1)). B5 medium supplemented with 1.0 mgL(-1) IBA plus 5.0 mgL(-1) BAP and MS medium fortified with 2.0 mgL(-1) IBA and 2.0 mgL(-1) BAP were the most favorable media for callus formation with the highest induction rate (96%). Maximum growth index (2.89 and 2.63) and maximum callus biomass (2.34 and 2.33 g fresh weight) were obtained from the callus cultured on B5 medium supplemented with 1.0 mgL(-1) IBA plus 5.0 mgL(-1) BAP and MS medium fortified with 1.0 mgL(-1) IBA plus 1.0 mgL(-1) KIN, respectively. Determination and quantification of RA in cultured calli were performed by HPLC UV/MS analysis. Calli induced from the plant and maintained on supplements of IBA and BAP in the absence of light produced RA 7.5% based on dry weight (DW). No differentiation was observed in any callus during the course of this study.

Micropropagation of Urginea maritima (L.) Baker s. str.

A method of micropropagation of <i>Urginea maritima</i> (L.) Baker s.str. (<i>Liliaceae</i>) by adventitious shoot formation was developed. Bulb scales and leaf fragments were used as primary and secondary explants, respectively. The most favourable for shoot regeneration were media: MS supplemented with BAP or kinetin (bulb scales) and medium C containing NAA along with BAP (leaf explants). No difficulties in rooting and adapting of plants to greenhouse conditions were observed.

Effects of cytokinins on antioxidant enzymes in in vitro grown shoots of Pelargonium hortorum L. H. Bayley

<p>The aim of this study was to determine the influence of <em>meta</em>-topolin (<em>m</em>T) and 6-benzyl-aminopurine (BAP) on the hydrogen peroxide (H<sub>2</sub>O<sub>2</sub>) level and antioxidant enzymes activities in relation to the shoot formation and senescence process in <em>Pelargonium hortorum</em> cultivars, which differ in their susceptibility to leaf yellowing under <em>in vitro</em> conditions.</p><p>In an early senescing cultivar ‘Grand Prix’, the addition of an aromatic cytokinin <em>m</em>T to abscisic acid (ABA)-enriched Murashige and Skoog (MS) basal medium more efficiently inhibited leaf yellowing than BAP. In both genotypes, <em>meta</em>-topolin was also the most effective in shoot formation. It was found that <em>Pelargonium</em> species varying in their susceptibility to senescence differ in H<sub>2</sub>O<sub>2</sub> production and antioxidant enzymes activities. Generally, <em>meta</em>-topolin more effectively enhanced H<sub>2</sub>O<sub>2 </sub>production and POD activity than BAP and control medium, but its effect depended on genotype. The highest H<sub>2</sub>O<sub>2</sub> production stimulated by <em>m</em>T was observed on day 5 of subculture in late senescing cv. ‘Bergpalais’. In both geranium genotypes, superoxide dismutase (SOD) and catalase (CAT) levels were highest at the beginning of the subculture period, during the initiation of shoot formation. SOD showed the highest activity on day 5 of subculture on the medium without cytokinin and generally being higher in cv. ‘Bergpalais’ than in cv. ‘Grand Prix’. CAT activity was positively regulated by both cytokinins. POD activity was most effectively enhanced by <em>m</em>T, but on different days of subculture - on the 2<sup>nd</sup> day of subculture in cv. ‘Bergpalais’ and on the 22<sup>nd</sup> day of subculture in cv. ‘Grand Prix’. The enhanced activity of POD in the presence of <em>m</em>T, 4-fold higher than on control medium, at the end of subculture in <em>P. hortorum </em>‘Grand Prix’ coincided with the inhibition of leaf senescence.</p>

Callus Induction and Plant Regeneration from Node Explants of Ziziphus Spina- Christi

Ziziphus spina -christi (L.) ecologically and economically important due to its tolerance to drought and salinity. The purpose of this study was developing a standardized protocol to induce a high frequency of callus in Z. spina -christi for breeding programs with the final objective of plantlet regeneration from node explants. Node explants of aseptic seedlings, were cultured on MS medium containing the hormone, such as 2, 4-D, NAA, TDZ and BAP. For regeneration, different levels of the PGRs including BAP, TDZ and Kin were applied. Also rooting of seedlings obtained from 1/2MS medium, were performed in the presence of IBA and NAA. Results showed that culture medium containing 2, 4-D and TDZ at concentrations of 0.5 and 1 mg/l ( ) has the highest amount of callus induction. Most of the node explants cultured on BAP medium were producing direct shoot at callusing phase. 5 mg/l of Kin and 1 mg/l of TDZ respectively, had the highest average shoot with numerical average of 84.8, 95.8 but the shoot produced by TDZ had an unusual shape but applying Kin, direct normal shoots were produced. The value of root produced by the hormone IBA more than NAA at all concentrations. KeywordsCallus Induction; Growth Regulators; Node Explants; Ziziphus Spina -Christi

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.

Orchid Micropropagation: Regeneration Competence of Anther Culture

The possibility of using anthers for initiating in vitro cultures of Rhynchostylis retusa (Orchidaceae), has been positively tested. The study was designed to study the effects of physiological status, stage of microspore development and pre-cold treatment on androgenic response in R.retusa. The anthers from open flowers (2 days of anthesis) in tetrad stage failed to respond despite variations in the chemical regime; whereas those from unopened buds (1.25-1.35cm long) with an intact operculum & pre-cold treatment with microspores in early & late-uninucleate decussate regenerated provided their nutritional complexities were satisfied through an exogeneous supply of Peptone (2mg/l) in the BAP (10mg/l) and NAA enriched Sharma medium. The best response was obtained when a 24hrs cold treatment was employed at 4°C in darkness. The complete plantlet (2-3 leaves &1-2 roots) was formed in 24wks.The plantlet was accilimatized & the survival rate is 70%.

Micropropagação de Dioscorea multiflora Grised

Dioscorea multiflora uma planta nativa do Sul do Brasil produz a diosgenina como metabólito secundário majoritário, uma substância potencialmente usada pela indústria farmacêutica para a produção de cortisona e substâncias com ação contraceptiva. Objetivou-se, neste trabalho otimizar o protocolo de micropropagação de D. multiflora, visando a produção de mudas em escala comercial. Segmentos nodais subcultivados em meio MS sólido foram transferidos para multiplicação em meio MS suplementado com BAP (0,01; 0,1; 0,5; 1,0 e 3,0 mg L-1)e meio MS suplementado com 0,1 mg L-1 ou 0,5 mg L-1 de BAP acrescido de diferentes concentrações de sacarose (2, 4, 6, 8 e 10%). Para o enraizamento, as brotações foram cultivadas em meio MS suplementado com AIB (0,1; 0,5; 1,0 e 3,0 mg L-1) e meio MS suplementado com ANA (0,1; 0,5; 1,0 e 3,0 mg L-1). Os experimentos in vitro foram instalados em delineamento experimental inteiramente casualizado e cada tratamento constituiu-se de 3 repetições e 10 cubetas/parcela. Plântulas com e sem raízes foram aclimatizadas em casa de vegetação. Melhores resultados de multiplicação e enraizamento foram obtidos em meio MS + 0,1 mg L-1 de BAP (80%) e em meio MS + 1,0 mg L-1 de AIB (42,6%), respectivamente. Não houve diferença quanto à porcentagem de sobrevivência das plântulas in vitro e ex vitro durante a aclimatização (75%). O protocolo de micropropagação para D. Multiflora é efetivo e pode ser usado para a produção em escala comercial.

Isolation and characterization of non-Frankia actinobacteria from root nodules of Alnus glutinosa, Casuarina glauca and Elaeagnus angustifolia

Actinobacterial isolates randomly obtained on nitrogen-free BAP medium from surface sterilized root nodules of Alnus glutinosa, Casuarina glauca and Elaeagnus angustifolia sampled from fields were reported. They were assigned on the basis of partial 16S rRNA sequences to Micromonospora, Nocardia and Streptomyces genera. The isolates have been screened for hydrolytic activities, indole acetic acid (IAA) and siderophores production, phosphate solubilization and antagonistic activities. Results suggest putative traits as plant growth promoting bacteria proprieties of the isolates that occur in unique association in root nodules of the three analysed actinorhizal host species.