Micropropagation and Polyploid Induction of Acer platanoides ‘Crimson Sentry

Protocols were developed for micropropagation and induction of autopolyploids in a fastigiate cultivar of Norway maple (A. platanoides L. ‘Crimson Sentry’). Murashige and Skoog (MS) medium, woody plant medium (WPM), and Quoirin and Lepoivre medium were supplemented with 2 μM 6-benzylaminopurine (BA), meta-Topolin, 6-(γ,γ-dimethylallylamino) purine, kinetin, or thidiazuron to evaluate microshoot proliferation. Murashige and Skoog medium with 2 μM BA yielded the most microshoots (3.2) and longest microshoots (30.6 mm) per subsample after 5 weeks. The influence of BA concentration on proliferation was evaluated at 0, 2, 4, 8, or 16 μM. Optimal multiplication rate was achieved at 2 or 4 μM BA producing approximately 2.8 microcuttings per subsample after 5 weeks. To induce in vitro rooting, half-strength WPM was supplemented with 0, 5, 10, 20, 40, or 80 μM indole-3-butyric acid (IBA). Optimal in vitro rooting (70%), number of roots (2.5), and root length (15 mm) per subsample were achieved with 10 μM IBA after 8 weeks. To induce polyploidy, microcuttings were pretreated for 7 days on MS medium with 4 μM BA alone or combined with 1 μM IBA, indole-3-acetic acid (IAA), or 1-naphthaleneacetic acid prior to treatment in liquid MS medium containing 15 μM oryzalin for 3 days. Homogenous tetraploids were only obtained from shoots pretreated with IAA. This research provides optimized protocols for micropropagation and autopolyploid induction of A. platanoides ‘Crimson Sentry’ and demonstrates the development of tetraploid lines for use in future improvement programs.

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Micropropagation of Acacia chundra (Roxb.) DC.

Horticultural Science ◽

10.17221/648-hortsci ◽

2008 ◽

Vol 35 (No. 1) ◽

pp. 22-26 ◽

Cited By ~ 7

Author(s):

R. Rout G ◽

K. Senapati S ◽

S. Aparajeta

Keyword(s):

Acetic Acid ◽

Growth Regulators ◽

Basal Medium ◽

Multiplication Rate ◽

Ms Medium ◽

Leguminous Tree ◽

Half Strength ◽

Basal Salts ◽

Indole 3 Acetic Acid

An in vitro propagation of an economic leguminous tree, Acacia chundra, has been standardized. Induction of bud sprout was obtained from shoot tip and nodal explants derived from in vitro grown plants of A. chundra on the Murashige and Skoog (MS) basal medium supplemented with 6-benzylaminopurine (BA) (1.0 mg/l) and 20 mg/l adenine sulfate (Ads). The rate of multiplication was obtained on MS medium supplemented with 1.5 mg/l BA, 0.01 to 0.05 mg/l (indole-3-acetic acid) IAA and 50 mg/l Ads. The multiplication rate varied from 3 to 6 shoots depending on the growth regulators used. Excised shoots were rooted on half-strength MS basal salts supplemented with 0.25 mg/l indole-3-butyric acid (IBA) or IAA and 20 g/l (w/v) sucrose after 10 to 12 days of culture. The micropropagated plantlets have been acclimatized and successfully transferred to soil.

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Effect of Sucrose Concentrations and Incubation Periods on in Vitro Rooting of Moris Pineapple (Ananas comosus)

AL-MUKHTAR JOURNAL OF SCIENCES ◽

10.54172/mjsc.v34i4.145 ◽

2019 ◽

Vol 34 (4) ◽

pp. 230-242

Author(s):

Abdelhamid M Hamad

Keyword(s):

Incubation Period ◽

In Vitro Rooting ◽

Ananas Comosus ◽

Ms Medium ◽

Incubation Periods ◽

Half Strength

The effect of 6 sucrose concentrations (5, 10, 15, 20, 25, 30 g/l) over 4 incubation periods (30, 45, 60, 75 days) on in vitro rooting of Moris pineapple cultured in liquid half strength MS medium enriched with IBA at 2.0 mg/l was investigated. At all incubation periods, all shoots in medium enriched with sucrose at 5 g/l failed to root, and no roots formed within the first 30 days in medium enriched with sucrose at 10 g/l. After 30 days of incubation, the highest rooting percentage (66 %), tallest plantlets (23 mm tall), highest (3.4 roots) and longest (5.3 mm) root per shoot were obtained in medium enriched with sucrose at 25, 10, 15, 15 g/l respectively, while after 45 days, the highest of all rooting aspects (75 %, 32.3 mm tall, 3.7 roots, 7 mm long), were obtained in medium enriched with sucrose at 15 g/l. After 60 days, the highest rooting percentage (91.7 %) and tallest plantlets (36.7 mm tall) were obtained in medium enriched with sucrose at 20 g/l while highest roots per shoot (3.7 roots) and longest root (10.7 mm) were obtained in medium enriched with sucrose at 15 g/l. After 75 days, all shoots rooted (100 %) in medium enriched with sucrose at 10 and 20 g/l, while sucrose at 25 g/l resulted in tallest plantlets (46.3 mm tall) and at 20 g/l resulted in highest (4.7 roots) and longest roots (27.3 mm). At each incubation period, there was a different optimum sucrose enrichment for different rooting parameters.

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

Glasnik Sumarskog fakulteta ◽

10.2298/gsf1511099m ◽

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.

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IN VITRO RESPONSE BY Terminalia arjuna GENOTYPES DURING MICROPROPAGATION

Journal of Experimental Biology and Agricultural Sciences ◽

10.18006/2021.9(1).44.50 ◽

2021 ◽

Vol 9 (1) ◽

pp. 44-50

Author(s):

Meena Choudhary ◽

◽

Ashok Gehlot ◽

Sarita Arya ◽

Inder Dev Arya ◽

...

Keyword(s):

Shoot Proliferation ◽

In Vitro Rooting ◽

Terminalia Arjuna ◽

Ms Medium ◽

Demand And Supply ◽

Half Strength ◽

Modified Ms Medium ◽

In Vitro Response ◽

Different Doses

Terminalia arjuna is an important tree of the medicinal and sericulture industry, commonly known as Arjun. It’s bark is rich in secondary metabolites makes this plant highly valuable in medicine industry to treat cardiovascular disease. Overexploitation due to high demand in medicine, low seed germination, limitations of the conventional method of propagation push this plant towards being endangered. To conserve germplasm of such tree species and meet the requirement in medicinal industry, some non-conventional propagation method like micropropagation has been developed. The present work highlighted the effect of three genotypes (G-1, G-2, and G-3) on tissue culture of T. arjuna situated at Jodhpur, Rajasthan, India. In vitro shoot proliferation was achieved on a modified MS medium enriched with BAP + additives. Among the tested genotypes, Genotype -1 showed maximum bud break response (100%) followed by G-3 (93.33 %) and G-2 (86.66%). Further multiplication of these shoots on modified MS medium containing BAP + NAA + additives gave 11.38±0.26 (G-1), 9.44±0.21 (G-2) and 10.22±0.32 (G-3) shoots. In vitro rooting was done by pulse treatment with IBA for 10 min prior to transfer on hormone free half strength MS medium containing 0.1% activated charcoal. Maximum in vitro rooting was obtained in G-1 (80%) followed by G-3 (71.11%) and G-2 (68.88%). In the present study, it was observed that optimum growth in all three genotypes required different doses of Plant Growth Regulator. Thus, by identifying and multiplying the best performing genotypes the gap between demand and supply of such medicinal plant can be fulfilled.

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Micropropagation and in vitro conservation of Alcantarea nahoumii (Bromeliaceae), an endemic and endangered species of the Brazilian Atlantic Forest

Acta Scientiarum Biological Sciences ◽

10.4025/actascibiolsci.v42i1.52940 ◽

2020 ◽

Vol 42 ◽

pp. e52940

Author(s):

Simone Sacramento dos Santos Silva ◽

Everton Hilo de Souza ◽

Fernanda Vidigal Duarte Souza ◽

Cristina Ferreira Nepomuceno ◽

Maria Angélica Pereira de Carvalho Costa

Keyword(s):

Atlantic Forest ◽

Culture Media ◽

In Vitro Conservation ◽

Naphthaleneacetic Acid ◽

High Survival ◽

Multiplication Rate ◽

Ms Medium ◽

Mature Seeds ◽

Stem Segments

Alcantarea nahoumii (Leme) J. R. Grant is a species native to the Atlantic Forest that stands out for ornamental purposes. The objective of this work was to evaluate the in vitro germination of A. nahoumii seeds and establish a micropropagation protocol for production of seedlings so as to minimize the effects of predatory extractivism and develop an in vitro conservation system. Mature seeds were disinfested, established in three culture media (MS, MS½ and MS⅓) and incubated at four temperatures (20, 25, 30 and 35ºC) in a germination chamber. In the micropropagation experiment, stem segments were introduced in MS medium supplemented with 0.5 μM of 1-naphthaleneacetic acid (NAA) and 0.0, 2.2, 4.4 and 6.6 μM of 6-benzylaminopurine (BAP). For the in vitro conservation, plantlets were established in MS or MS½ medium supplemented with 15 g L-1 or 30 g L-1 of sucrose. The plants were acclimated with commercial substrate. The highest seed germination percentages were promoted by temperature conditions of 20 and 25ºC, with MS culture medium. The highest multiplication rate of shoots was obtained from the treatment without addition of the growth regulator or when combined with 2.2 μM of BAP + 0.5 μM of NAA. The acclimation of the plants occurred with high survival rate. The species can be conserved in vitro under slow growth condition for 24 months when incubated in MS medium supplemented with 30 g L-1 of sucrose.

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Pomegranate Micropropagation, Callogenesis and Genetic Integrity Assessment Using Simple Sequence Repeat Markers

Journal of Agricultural Science ◽

10.5539/jas.v11n1p237 ◽

2018 ◽

Vol 11 (1) ◽

pp. 237

Author(s):

Tebogo Stimela ◽

Remmy W. Kasili ◽

Edward G. Mamati

Keyword(s):

Acetic Acid ◽

Simple Sequence Repeat ◽

Naphthaleneacetic Acid ◽

Genetic Integrity ◽

Sequence Repeat ◽

Simple Sequence Repeat Markers ◽

Half Strength ◽

Indole 3 Acetic Acid ◽

Simple Sequence

In recent years, the awareness of pomegranate health benefits has grown exponentially; nonetheless the existing propagation methods remain a challenge to supply adequate suitable planting materials needed for commercial production. Micropropagation can lead to mass production of plantlets and callus-mediated in vitro regeneration can open avenues for the use of genetic engineering to improve this crop. The aim of this study was to evaluate appropriate conditions for pomegranate micropropagation, callogenesis and use Simple Sequence Repeat markers to screen for somaclonal variation. Cytokinins (Benzylaminopurine, Kinetin and Thiadiazol-5ylurea) were tested for shoot induction from nodal explants while auxins (1-Naphthaleneacetic acid, Indole-3-butyric acid and Indole-3-acetic acid) were tested for root induction of in vitro regenerated shoots. 1-Naphthaleneacetic acid combined with Benzylaminopurine was assessed for their ability to induce callus from cotyledon and leaf explants. Genetic integrity between mother plant, callus and in vitro regenerated shoots were assessed using eight Simple Sequence Repeat markers. Maximum number of shoots and leaves were obtained on full strength Murashige and Skoog media with 6.9 µM kinetin. The highest number of roots was achieved on half strength Murashige and Skoog media with 4.9 µM Indole-3-butyric acid and the longest root was got on half strength Murashige and Skoog media with 5.3 µM Indole-3-acetic acid. Leaves and cotyledons demonstrated to be potential explants for callus formation at all hormonal combination levels tested. Eight out of 13 amplified alleles were polymorphic. A wider genetic variation was found with similarity coefficient range of 0.46-0.92. More somaclonal variation was in regenerated shoots compared to callus.

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In Vitro Long-Term Cultures of Papaya (Carica Papaya L.).

10.21203/rs.3.rs-262638/v1 ◽

2021 ◽

Author(s):

Jose Javier Regalado González ◽

Manuel López Granero ◽

Carlos Lopez Encina

Keyword(s):

Morphological Characteristics ◽

Basal Medium ◽

Multiplication Rate ◽

Ms Medium ◽

High Quality ◽

Poor Growth ◽

Average Multiplication ◽

Half Strength

Abstract We present the data on proliferation corresponding to 10 years of continuous incubation in vitro of papaya shoots, and propose a reliable method for long-term micropropagation for papaya, using two types of explants: Microshoots from somatic embryos, and from axillary buds of papaya. Three different media were assayed. The proliferation medium (PPRM) allowed to maintain papaya shoots under continuous proliferation during 20 years, maintaining a consistent behaviour. Most of the shoots developed in PPRM rooted during the incubation, and after acclimated easily, maintaining the morphological characteristics of the parental plants, flowering and setting fruits normally. The PPRM medium consist in MS medium supplemented with NAA (0.1 mg l-1), BA (0.5 mg l-1), GA3 (0.5 mg l-1) and Adenine sulphate (40 mg l-1). The average multiplication rate was higher than 20 shoots per explant along the long-term assay. The elongation medium (PELM), was designed to recover shoots with a poor growth, and allowed the development of high quality shoots ready for rooting, and consist in a MS basal medium supplemented with NAA (0.1 mg l-1), Kin (0.5 mg l-1) and GA3 (1 mg l-1). The rooting medium (PROM) was designed to induce high quality roots from non-rooted shoots and consist in a half strength MS medium plus IBA (1mg l-1). On PROM, agar can be exchanged for expanded vermiculite. Acclimation took place inside an acclimatization tunnel under progressive hydric stress. After 4 weeks, the plant recovery rate was 90% for plants maintained under continuous proliferation during ten years.

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In vitro Propagation of Solanecio biafrae and Determination of Genetic Stability of Plantlets Using RAPD and ISSR Markers

Journal of Horticultural Research ◽

10.1515/johr-2016-0004 ◽

2016 ◽

Vol 24 (1) ◽

pp. 29-36 ◽

Cited By ~ 1

Author(s):

Jelili Opabode ◽

Oluyemisi Akinyemiju

Keyword(s):

Random Amplified Polymorphic Dna ◽

Issr Markers ◽

Naphthaleneacetic Acid ◽

Ms Medium ◽

Germplasm Exchange ◽

Indole Butyric Acid ◽

Half Strength ◽

Micropropagated Plants

Abstract An efficient and reproducible micropropagation protocol of Solanecio biafrae (Oliv. & Hiern) C. Jeffrey has been developed from nodal stem segments. Shoot development was obtained on Murashige and Skoog (MS) medium supplemented with benzylaminopurine (BAP) alone and in combination with zeatin and 1-naphthaleneacetic acid (NAA). Elongated shoots were rooted in the presence of zeatin or 3-indole-butyric acid (IBA) alone or in combinations. The highest number of explants forming shoots (100%) as well as the highest number of shoots per explant (3.4) and the longest shoots (22 mm) were recorded on medium containing 4.0 mg·dm−3 BAP, 2.0 mg·dm−3 NAA, and 1.0 mg·dm−3 zeatin. About 76% of shoots formed roots on half-strength MS medium free of plant growth regulators. The best root formation (approximately 88%) was recorded on the medium containing 1.0-1.5 mg·dm−3 IBA. The micropropagated shoots with well-developed roots were efficiently acclimatized under greenhouse conditions. The random amplified polymorphic DNA (RAPD) and intersimple sequence repeat (ISSR) amplification products were monomorphic in micropropagated plants and similar to those of mother plant showing their genetic uniformity. This is the first report of micropropagation of S. biafrae, which will facilitate in vitro mass propagation, conservation, and germplasm exchange of this endangered African vegetable.

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Non-symbiotic Seed Germination and In vitro Plant Development of Pholidota articulata

Nepalese Horticulture ◽

10.3126/nh.v15i0.36648 ◽

2021 ◽

Vol 15 ◽

pp. 44-51

Author(s):

R. Devendra Prasad ◽

Shreeti Pradan ◽

Mukti Ram Poudel ◽

Bijaya Pant

Keyword(s):

Acetic Acid ◽

Seed Germination ◽

Plant Production ◽

Sustainable Utilization ◽

Ms Medium ◽

Natural Habitats ◽

Half Strength ◽

Indole 3 Acetic Acid ◽

Symbiotic Seed Germination

Pholidota articulata is an epiphytic orchid mostly used in ornamental cut/pot flower and in traditional medicine. As it has high ornamental and medicinal values, its population from natural habitats is decreasing, therefore, it is listed in the Appendix-II of Convention on International Trade in Endangered Species (CITES). The objective of the present study is to obtain the in vitro plants of P. articulata from seed culture to conserve its germplasm. The in vitro seed germination was carried out in different strengths of Murashige and Skoog (MS) and Knudson C (KnC) medium supplemented with various plant hormones. On the half-strength of MS medium, seeds were started to germinate after 4 weeks of primary culture and they were developed into protocorms with first leaf primordium earlier than on the other medium. Therefore, in vitro developed protocorms were sub-cultured on the half-strength of MS medium supplemented with different concentrations of 6-benzylaminopurine (BAP), gibberellic acid (GA3) and α-naphtalene acetic acid (NAA). They were successfully developed into shoots on the 1.5 mg/l BAP supplemented half-strength of MS medium. Later, they were inoculated on the half-strength of MS medium supplemented with different concentration of α-napthalene acetic acid (NAA), indole-3-acetic acid (IAA) and indole-3-butyric acid (IBA) for the root formation, where IBA supplemented medium was found effective for the development of roots. Thus, this study provides a reliable protocol for non-symbiotic seed germination and plant production, and reveals that seed-derived protocorms are good explants for the in vitro mass propagation for conservation and sustainable utilization in horticulture.

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ENHANCED REGENERATION OF SHOOTS FROM PEACH CELLS INFECTED WITH A SHOOTY MUTANT STRAIN OF AGROBACTERIUM.

HortScience ◽

10.21273/hortsci.25.9.1150d ◽

1990 ◽

Vol 25 (9) ◽

pp. 1150d-1150

Author(s):

A. Smigocki ◽

F. Hammerschlag

Keyword(s):

Prunus Persica ◽

Northern Analysis ◽

Dichlorophenoxyacetic Acid ◽

Naphthaleneacetic Acid ◽

Ms Medium ◽

Gene Transcripts ◽

Half Strength ◽

Low Levels ◽

2,4 Dichlorophenoxyacetic Acid

Immature `Redhaven' peach (Prunus persica L. Batsch) embryos were infected with Agrobacterium tumefaciens strain tms328::Tn5 carrying the functional cytokinin gene. Shoots were regenerated from callus grown on MS medium without added phytohormones and subsequently rooted on half-strength MS medium with 2.8 -naphthaleneacetic acid. These plants exhibited an increased frequency of branching in vitro. Low levels of cytokinin gene transcripts were detected in these cells by Northern analysis, and using an ELISA assay, the cytokinins zeatin and zeatinriboside were determined to be on the average 30-fold higher. From these results, the expression of the cytokinin gene appears to promote growth of cells in the absence of phytohormones thus serving as a marker for transformation and a promoter of morphogenesis without a 2,4-dichlorophenoxyacetic acid inductive step.

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