scholarly journals Effect of different sucrose and nitrogen salt levels in the medium and temperature on in vitro propagation of Helleborus niger L.

2015 ◽  
Vol 32 (2) ◽  
pp. 161-171 ◽  
Author(s):  
Eleonora Anna Gabryszewska
Keyword(s):  
Developmental Stages ◽  
Leaf Shape ◽  
Morphological Observation ◽  
Multiplication Rate ◽  
Ms Medium ◽  
Ex Vitro ◽  
Axillary Shoots ◽  
The Media ◽  
Helleborus Niger

<em>Helleborus niger</em> L. is a rhizomatous, herbaceous perennial with overwintering, divided, basal leaves. The objective of the study was to investigate the influence of different levels of sucrose (10, 20, 30, 40, 50, 60, 70, and 80 g l<sup>−1</sup>) and nitrogen salts (25%, 50%, and 100% according to MS medium) as well as temperature (15°C, 20°C) on in vitro multiplication and rooting and ex vitro acclimatization of <em>H. niger</em>. The growth and multiplication of axillary shoots were performed on modified MS medium supplemented with various growth regulators (2iP, BAP and kinetin – each at a concentration of 1.0 mg l<sup>−1</sup>, GA<span><sub>3</sub></span> 2.5 mg l<sup>−1</sup>). For the induction of roots, the medium was supplemented with IBA 1 mg l<sup>−1</sup> and NAA 0.1 mg l<sup>−1</sup>. Rooted plants were transplanted in a peat–perlite substrate (4:1) in a heated greenhouse for ex vitro acclimatization. The multiplication rate of <em>H. niger</em> shoots, in vitro rooting, and ex vitro acclimatization were strongly dependent on the sucrose/nitrogen salt relationship in the medium. The highest multiplication rate of axillary shoots (3.7) was found at a temperature of 15°C or 20°C, on the medium with cytokinins and GA<span><sub>3</sub></span> supplemented with sucrose 20–30 g l<sup>−1</sup> and nitrogen salts at 50%. Sucrose at a concentration of 50 g l<sup>−1</sup> strongly stimulated the number of roots per microplant (5.8–6.0) on the media with a reduced level of nitrogen salts (25% and 50%) when the temperatures were 20°C and 15°C, respectively. The plants rooted on the media with a high sucrose/nitrogen salt ratio showed acclimatization rates which ranged from 82% to 100%. Morphological observation of plantlets revealed obvious differences in leaf shape and size and the architecture of the root system as well as differences in the developmental stages of shoots grown on media with different sucrose and nitrogen salt concentrations.

scholarly journals Optimization of Micropropagation Protocol for Goji Berry (Lycium barbarum L.)

2016 ◽  
Vol 73 (2) ◽  
pp. 141 ◽  
Author(s):  
Alexandru Fira ◽  
Nirmal Joshee ◽  
Victoria Cristea ◽  
Manuela Simu ◽  
Monica Harta ◽  
...  
Keyword(s):  
Shoot Proliferation ◽  
Optimal Number ◽  
Wheat Starch ◽  
Ms Medium ◽  
Ex Vitro ◽  
Benzyl Adenine ◽  
Lycium Barbarum ◽  
Floating Cell ◽  
The Media

Micropropagation of Lycium barbarum cv. 'Ningxia N1' was achieved. The cultures were by initiated by axenical seed germination. The highest shoot proliferation was obtained on the MS media with 1.33 or 2.22 µM benzyl adenine, gelled with wheat starch as an agar alternative. The treatments with 2.22 µM benzyl adenine ensured proliferation rates superior to the ones with 1.33 μM benzyl adenine, but the latter provided longer and more robust shoots. Use of large microcuttings as an explant onto the multiplication media ensured higher in vitro explant survival, higher number of shoots regeneration and more vigorous plantlets. The microcuttings inserted vertically into the media yielded superior growth and multiplication as compared to the microcuttings placed horizontally. The non-rooted, elongated shoots from the treatment 1.33 μM benzyl adenine were either rooted in vitro on a hormone-free MS medium with starch or used for direct ex vitro rooting and acclimatization. The optimal number of microcuttings/vessel for in vitro rooting was 40 and the rooted plantlets were efficiently acclimatized ex vitro by three methods: float hydroculture in floating cell trays, floating perlite, and in Jiffy7 pellets.

scholarly journals Micropropagation of phytoplasma-affected Limonium sinuatum Mill. plants

2014 ◽  
Vol 69 (2) ◽  
pp. 109-113 ◽  
Author(s):  
Eleonora Gabryszewska ◽  
Maria Kamińska ◽  
Małgorzata Korbin ◽  
Anna Rudzińska-Langwald
Keyword(s):  
Tissue Culture ◽  
Electron Microscope ◽  
Multiplication Rate ◽  
Fresh Weight ◽  
Axillary Shoots ◽  
The Media

Healthy and AY-affected plants of <em>L.sinuatum</em> have been propagated in vitro for 12 months on the media with and without cytokinins. In the contrary to the healthy plants the phytoplasma affected statice showed abnormal proliferation of the axillary shoots, shortening of the internodes, smaller leaves and severe chlorosis. On the medium without cytokinins, diseased plants proliferated and formed 7.0 axillary shoots per explant but the healthy ones only formed 2.3 shoots; however, the fresh weight of them was similar. On the media with cytokinins, the multiplication rate and fresh weight of healthy shoots greatly in-creased, but of the diseased plants were on the same laevel or decreased. During tissue culture phytoplasma could be detected in symptomatic plants by PCR as well as electron microscope however, the phytoplasmas showed the symptoms of degeneration.

scholarly journals THE INFLUENCE OF GROWTH REGULATORS ON DAHLIA PROPAGATION IN TISSUE CULTURE AND ACCLIMATIZATION OF PLANTS IN ex vitro CONDITIONS

2019 ◽  
Vol 18 (5) ◽  
pp. 49-61
Author(s):  
Barbara Marcinek ◽  
Marzena Parzymies ◽  
Monika Poniewozik ◽  
Danuta Kozak ◽  
Wojciech Durlak
Keyword(s):  
Shoot Multiplication ◽  
Multiplication Rate ◽  
Ex Vitro ◽  
Fresh Weight ◽  
Flower Stalk ◽  
Axillary Shoots ◽  
First Order ◽  
Isopentenyl Adenine ◽  
Consequent Effect

The aim of work was to evaluate the influence of cytokinins on dahlia propagated in vitro and their consequent effect on acclimatization. Plant material were shoot tips and nodes. From three cytokinins, benzyladenine, kinetin and 2-isopentenyl-adenine, only BA effectively stimulated shoot multiplication from axiliary buds. The highest multiplication rate was obtained from nodes in presence of 0.25–0.5 mg·dm–3 BA. Higher concentrations shortened internodes and decreased leaf blades and growth of callus. 1 mg·dm–3 of KIN and 2iP positively influenced shoots growth and size of leaves. Gibberellic acid (GA3) used with BA increased the number of axillary shoots. The best quality shoots and the highest multiplication rate were obtained when 2 mg·dm–3 BA was used with 5 mg·dm–3 GA3. Cytokinins affected rooting and acclimatization ex vitro. Dahlias shoots multiplicated in presence of 1 mg·dm–3 KIN or 2iP rooted faster in the soil and 100% survived in field, while those from 1 mg·dm–3 BA media rooted slowly, had shorter shoots and only 60% survived. Plants bloomed after 11–12 weeks in the field. Dahlia plants that had been multiplicated in presence of KIN had a bigger diameter and fresh weight in the field. BA and 2iP positively influenced flowers diameter, length of flower stalk and a number of first order shoots.

scholarly journals IN VITRO PROPAGATION OF PATUMMA (CURCUMA ALISMATIFOLIA GAGNEP.)

HortScience ◽  
1992 ◽  
Vol 27 (6) ◽  
pp. 585d-585
Author(s):  
Surawit Wannakrairoj
Keyword(s):  
Survival Rate ◽  
In Vitro Propagation ◽  
Culture Vessel ◽  
Multiplication Rate ◽  
Ex Vitro ◽  
Lateral Bud ◽  
The Media ◽  
Pre Treatment ◽  
Better Than

Patumma, a native of Thailand, is a new ornamental crop. Its increased demand for export make the species vulnerable to extinction. Cloning methodology is thus needed for the production of both existing and newly developed clone. Young inflorescent segment and lateral bud from rhizome can be used as explants. For decontamination of the lateral bud, the success was depended both on pre-treatment and disintestation procedure. The bud from dried rhizome was better than one from fresh rhizome. Prior to disinfestation with sodium hypochlorite, pre-treatment of rhizome in 52°C water for 5 minutes could replaced the use of antibiotic. Plantlet were placed on modified MS media with 0, 6.67, 13.32, 19.98 and 26.64 μmol/l benzyladenine (BA) or 0.19, 0.56, 1.67 and 5 μmol/l kinetin. The best multiplication rate of 4.83 fold was obtained when longitudinally-divided rhizome was on the medium with 13.32 μmol/l BA. The result also showed that wild-collected and selected clones responded to the media similarly. When the MS media modified with 13.32, 15.54, 17.76 and 19.98 μmol/l BA in combination with 15, 30 and 45 g/l sucrose were tested, the multiplication rate of non-divided plantlets were all the same. Acclimatization by open the culture vessel for 3 day improved plantlet survival rate ex vitro.

scholarly journals In vitro multiplication, micromorphological studies and ex vitro rooting of Hybanthus enneaspermus (L.) F. Muell. – a rare medicinal plant

2018 ◽  
Vol 77 (1) ◽  
pp. 80-87 ◽  
Author(s):  
Mahipal S. Shekhawat ◽  
M. Manokari
Keyword(s):  
Medicinal Plant ◽  
Large Scale ◽  
Nodal Segments ◽  
Ms Medium ◽  
Ex Vitro ◽  
Culture Bottle ◽  
Hybanthus Enneaspermus ◽  
Half Strength

AbstractHybanthus enneaspermusis a rare medicinal plant. We defined a protocol for micropropagation,ex vitrorooting of cloned shoots and their acclimatization. Surface-sterilized nodal segments were cultured on Murashige and Skoog (MS) medium with different concentrations of 6-benzylaminopurine (BAP) and kinetin (Kin). Medium supplemented with 1.5 mg L−1BAP was found optimum for shoot induction from the explants and 6.4±0.69 shoots were regenerated from each node with 97% response. Shoots were further proliferated maximally (228±10.3 shoots per culture bottle with 7.5±0.43 cm length) on MS medium augmented with 1.0 mg L−1each of BAP and Kin within 4–5 weeks. The shoots were rootedin vitroon half strength MS medium containing 2.0 mg L−1indole-3 butyric acid (IBA). The cloned shoots were pulse-treated with 300 mg L–1 of IBA and cultured on soilrite® in a greenhouse. About 96% of the IBA-pulsed shoots rootedex vitroin soilrite®, each shoot producing 12.5±0.54 roots with 5.1±0.62 cm length. Theex vitrorooted plantlets showed a better rate of survival (92%) in a field study thanin vitrorooted plantlets (86%). A comparative foliar micromorphological study ofH. enneaspermuswas conducted to understand the micromorphological changes during plant developmental processes fromin vitrotoin vivoconditions in terms of variations in stomata, vein structures and spacing, and trichomes. This is the first report onex vitrorooting inH. enneaspermusand the protocol can be exploited for conservation and large-scale propagation of this rare and medicinally important plant.

scholarly journals Shoot multiplication of Pogostemon cablin var. Sidikalang and patchouli oil profile

2019 ◽  
Vol 11 (2) ◽  
Author(s):  
POPY HARTATIE HARDJO ◽  
DANNY PUTRA SENTOSA SUSANTO ◽  
WINA DIAN SAVITRI ◽  
MARIA GORETTI MARIANTI PURWANTO
Keyword(s):  
Growth Index ◽  
Shoot Multiplication ◽  
High Price ◽  
Ms Medium ◽  
Ex Vitro ◽  
Average Growth ◽  
In Vitro Multiplication ◽  
Patchouli Alcohol ◽  
Pogostemon Cablin

Abstract. Hardjo PH, Susanto DPS, Savitri WD, Purwanto MGM. 2019. Shoot multiplication of Pogostemon cablin var. Sidikalang and patchouli oil profile. Nusantara Bioscience 11: 123-127. Pogostemon cablin Benth. is a plant producing patchouli oil, which mostly consists of patchouli alcohol compound. Patchouli oil has great potentials in the world market because of its stability and high price. In this study, in vitro multiplication of Sidikalang variety of Acehnese patchouli shoots was done on solid and liquid Murashige & Skoog (MS) medium. This study aimed to determine the effect of cytokinins in various combinations of shoot multiplication and to compare the patchouli oil yield of in vitro and ex vitro culture. In vitro multiplication of Acehnese patchouli shoots by using solid MS medium with addition of 0.2 ppm benzyl aminopurine (BAP) and 0.2 ppm Kinetin resulted in shoot explants with an average growth index of 82.198 ± 0.690. Patchouli oil extraction was done on 7 weeks old in vitro shoot explants cultured on solid MS medium + 0.2 ppm BAP + 0.2 ppm Kinetin using water distillation method. In vitro shoots yielded 2.5% patchouli oil and contained ± 35% patchouli alcohol compound, whereas ex vitro shoots produced 4% patchouli oil and contained ± 25% patchouli alcohol compound. The qualitative analysis by using thin layer chromatography (TLC) showed that there were similarities in the number of spot and Rf value for each spot of ex vitro and in vitro patchouli oil.

scholarly journals STUDY ON THE IN VITRO MORPHOGENETIC RESPONSE FROM SHOOT AND CALLUS CULTURES OF Blepharis maderaspatensis (L.) Heyne ex. Roth

2020 ◽  
Vol 1 (40) ◽  
pp. 28-38
Author(s):  
Trang Phuong Nguyen Thi ◽  
Quang Minh Bui ◽  
Hai Duc Le ◽  
Linh Quoc Nguyen
Keyword(s):  
High Efficiency ◽  
Raw Materials ◽  
Callus Cultures ◽  
Culture Conditions ◽  
Ms Medium ◽  
Axillary Shoots ◽  
Scientific Database ◽  
Medicinal Value ◽  
Shoot Growth Rate

Blepharis maderaspatensis (L.) Heyne ex. Roth is a short-term plant which contains many important secondarycompounds with high medicinal value. Currently, most of the researches focus on chemical composition and pharmacological activity, but the source of raw materials is very limited. In this study, the first step is transferring the samples from nature into in vitro culture conditions to understand the effects of the factors related to shooting and callus morphogenesis was performed, the first node from shoots apical meristem was isolated and sterilized with 1.5% NaOCl for 20 minutes to achieve high efficiency with 86.11% sterile samples and 85.56% shoot growth rate after 2 weeks of culture on MS medium. The shoot generation from axillary shoots was continued to be investigated with the highest number of shoots formed on MS medium supplemented with BA (1 mg / l) showed 1.53 shoots/implant which the height and the number of leavesare 3.65cm and 6.67, respectively. Besides, the formation of callus from leaves of MS medium supplemented with 2.4 - D (0.25 mg / l) achieved the rate of 66.67% of cultured samples, forming good callus after 4 weeks of culture. The results of the study not only contribute importantly to understanding morphogenesis for micropropagation purposes but also serve as the scientific database for further studies at the cellular and molecular levels of this plant.

scholarly journals Micropropagation of Achillea millefolium L. on half-strength ms medium and direct rooting and acclimatization of microshoots in hydroponic culture

2015 ◽  
pp. 99-112
Author(s):  
Marija Markovic ◽  
Dragana Skocajic ◽  
Mihailo Grbic ◽  
Matilda Djukic ◽  
Dragica Obratov-Petkovic ◽  
...  
Keyword(s):  
Medicinal Plant ◽  
Nutrient Solution ◽  
Ph Value ◽  
Ex Vitro Rooting ◽  
In Vitro Rooting ◽  
Efficient Production ◽  
Ms Medium ◽  
Ex Vitro ◽  
Half Strength

The aim of this study was to determine the possibility of micropropagation of the medicinal plant A. millefolium on half-strength MS medium and ex vitro rooting and acclimatization of the obtained microshoots in hydroculture in order to establish an efficient production method. Two explant types were used: basal and terminal cuttings, and better results were achieved when terminal cuttings were used. The development of shoots in the multiplication phase was successful with a regeneration percentage of 100%. Ex vitro rooting in a modified Hoagland nutrient solution was successful (83%), but the percentage of in vitro rooting on half-strength MS medium without hormones was higher (95%). However, bearing in mind that mass production of A. millefolium is more efficient when the phase of in vitro rooting is excluded, this method could be recommended for commercial propagation of this medicinal plant. It is necessary to conduct additional research in order to optimize the composition, EC and pH value of the hydroponic nutrient solution.

scholarly journals Diazotrophic bacteria in the growth of micropropagated ornamental pineapple

2019 ◽  
Vol 13 (2) ◽  
pp. 269-278
Author(s):  
Adriano Bortolotti Silva ◽  
Ligiane Aparecida Florentino ◽  
Dalvana De Sousa Pereira ◽  
Paulo Roberto Correa Landgraf ◽  
Ana Carolina Rodrigues Alves ◽  
...  
Keyword(s):  
Shoot Growth ◽  
Ex Vivo ◽  
Control Treatment ◽  
In Vitro Cultivation ◽  
Ms Medium ◽  
Ex Vitro ◽  
Diazotrophic Bacteria ◽  
Bacterial Strains ◽  
Indole 3 Acetic Acid

Ornamental pineapple is a hardy plant with significant landscaping value. Tissue culture of plants is viable for producing plants with a high phytosanitary quality. However, one of the difficulties with this cultivar is the acclimatization process, which is slow and can cause losses. The objective of the present study was to verify the potential of inoculation with diazotrophic bacteria for in vitro and ex vivo growth of ornamental pineapple. A group of diazotrophic bacterial strains selected at the Universidade José do Rosário Vellano (UNIFENAS) was prioritized in this study, and the treatments included bacterial strains UNIFENAS (100-13, 100-60, 100-68, 100-153, 100-167 and 100-198). These strains were evaluated in terms of their capacity to produce indole 3-acetic acid. Subsequently, plants were cultivated in a medium composed of MS medium salts (1/4), adding 1 mL of the bacterial strain. In the control treatment, the plants were maintained in 2 mL of MS medium. 7 days after inoculation, the plants were transplanted into the MS, where they were maintained for 30 days. After in vitro cultivation, the plants were transferred to pots containing commercial Plantmax® substrate and maintained under these conditions for 60 days. The diazotrophic bacteria were able to synthesize auxins, and their inoculation promoted greater growth in vitro and ex vitro in the plants. In the acclimatization phase, the plants inoculated with UNIFENAS strains (100-60, 100-68 and 100-153) promoted a higher shoot growth, chlorophyll content and nitrate reductase enzyme activity.

scholarly journals In vitro regeneration performance of Corchorus olitorius

1970 ◽  
Vol 8 (1) ◽  
pp. 1-6 ◽  
Author(s):  
M Hoque ◽  
KM Nasiruddin ◽  
GKMN Haque ◽  
GC Biswas
Keyword(s):  
Shoot Regeneration ◽  
Callus Induction ◽  
Root Formation ◽  
Plantlet Regeneration ◽  
Ms Medium ◽  
Corchorus Olitorius ◽  
The Media ◽  
Regenerated Plantlets

The experiment was conducted during May to December 2008 in the Biotechnology Laboratory of Bangladesh Agricultural University, Mymensingh to observe the callus induction, regeneration potentiality and to establish a suitable in vitro plantlet regeneration protocol of Corchorus olitorius. MS medium supplemented with different phytohormone concentrations and combinations were used to observe the callus induction, shoot regeneration and root formation ability of the cotyledon with attached petiole derived explant of three genotypes viz. O-9897, O-72 and OM-1. The highest callus induction (92.85%) was observed in O-9897 followed by O-72 (82.14%) in the MS media supplemented with 2.5 mg/L BAP + 0.5 mg/L IAA. Genotype O-9897 in MS media supplemented with 2.5 mg/L BAP + 0.5 mg/L IAA produced the highest percentage of shoot regenerants (83.33%) followed by O-72 (75.00%) in the media supplemented with 2.5 mg/L BAP + 0.5 mg/L IAA. The root formation from regenerants was the best on halfstrength of MS media supplemented with 0.6 mg/L IBA in genotype O-9897 (45.00%). The in vitro regenerated plantlets from the genotypes O-9897 could be established in the field. Therefore, the genotypes O-9897 of C. olitorius in MS media supplemented with 2.5 mg/L BAP + 0.5 mg/L IAA could be used for callus induction and shoot regeneration. Keywords: Regeneration; Phytohormone; Corchorus olitorius DOI: 10.3329/jbau.v8i1.6390J. Bangladesh Agril. Univ. 8(1): 1-6, 2010

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