Optimizing the concentrations of plant growth regulators for in vitro shoot cultures, callus induction and shoot regeneration from calluses of grapes

Aim: To optimize the concentrations of growth regulators in the media for the proficient micropropagation of grapevine (Vitis vinifera L.) cv. King’s Ruby.Methods and results: Apical meristems of the grape cultivar were used to establish in vitro shoot cultures. Nodal explants, each containing an axillary bud, taken from in vitro grown shoots were inoculated in shoot proliferation medium, i.e., half strength Murashige and Skoog (MS) medium supplemented with benzyl aminopurine (BAP), kinetin, glycine and gibberellic acid (GA3). A higher number of shoots (5.33) with greater shoot length (2.75 cm) was produced in the medium supplemented with 1.0 mg L-1 BAP and 0.1 mg L-1 GA3. Calluses were induced from leaf explants taken from in vitro grown shoots. Callus induction was greater (73.00%) on the medium containing 2.0 mg L-1 2,4-dichlorophenoxyacetic acid (2,4-D), 0.3 mg L-1 BAP and 0.2 mg L-1 α-naphthaleneacetic acid (NAA). The maximum frequency of shoot regeneration (53.33%) was achieved on the medium supplemented with 1.5 mg L-1 BAP and 0.5 mg L-1 NAA, and the regenerated shoots successfully formed roots on growth regulator-free half strength MS medium.Conclusion: Optimizing the concentration of BAP and GA3 and omitting the glycine and kinetin in the culture medium increased the number and length of shoots. Similarly, for inducing the callus of the leaf explants, taken from in vitro grown shoots, it is recommended to adjust the medium with the higher concentration of 2,4-D and lower concentrations of BAP. Moreover, the maximum number of shoots was regenerated on a medium supplemented with relatively high levels of both BAP and NAA (1.5 and 0.5 mg L-1, respectively). Finally, we suggest the half strength MS medium that is free from growth regulators for the root formation of the regenerated shoots.Significance and impact of the study: Optimizing the concentration of growth regulators is crucial for the efficient micropropagation of a grape cultivar. Knowing the specific balance between the growth regulators is necessary to establish in vitro shoot cultures, callus induction and shoot regeneration and, hence, to propagate disease-free true to type grape cultivars in a short time.

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Influence of growth regulators and explants on shoot regeneration in carnation

Horticultural Science ◽

10.17221/1/2009-hortsci ◽

2009 ◽

Vol 36 (No. 4) ◽

pp. 140-146 ◽

Cited By ~ 4

Author(s):

J.K. Kanwar ◽

S. Kumar

Keyword(s):

Acetic Acid ◽

Shoot Regeneration ◽

Growth Regulators ◽

Dianthus Caryophyllus ◽

Naphthalene Acetic Acid ◽

Ms Medium ◽

Shoot Bud ◽

Dichlorophenoxy Acetic Acid ◽

Number Of Shoots

The influence of growth regulators, explants and their interactions on in vitro shoot bud formation from callus was studied in Dianthus caryophyllus L. The leaf and internode explants were cultured on Murashige and Skoog (MS) medium containing different concentrations of growth regulators. The highest callus induction was observed with 2 mg/l 2,4-dichlorophenoxy acetic acid (2,4-D) and 1 mg/l benzyl adenine (BA). Out of twenty seven shoot regeneration media tested, only 2 mg/l thidiazuron (TDZ) and zeatin alone or in combination with naphthalene acetic acid (NAA) and/or indole acetic acid (IAA) could differentiate calli. The highest average number of shoots was observed with 2 mg/l TDZ and 1 mg/l IAA. Significant differences were observed in calli producing shoots and number of shoots per callus in the explants of leaf and internode. The shoots were elongated and multiplied on MS medium supplemented with 1 mg/l BA and solidified with 1% agar. The shoots were rooted and hardened with 76% survival success in pots after six weeks of transfer to the pots.

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IN VITRO REGENERATION AND MASS PROPAGATION OF AZIMA TETRACANTHA [LAM.] FROM THE LEAF EXPLANTS THROUGH CALLUS CULTURE

Kongunadu Research Journal ◽

10.26524/krj202 ◽

2017 ◽

Vol 4 (2) ◽

pp. 52-56

Author(s):

Mallika Devi T

Keyword(s):

Callus Induction ◽

In Vitro Regeneration ◽

Leaf Explants ◽

Shoot Formation ◽

Ms Medium ◽

Apical Leaf ◽

Half Strength ◽

Regenerated Plantlets ◽

Azima Tetracantha

In the present study the protocol for callus induction and regeneration in Azima tetracantha has been developed in culture medium. The young apical leaf explants were used for callus induction on MS medium containing BAP and NAA at 1.0 and 0.4mgl-1 respectively showed maximum callus induction (73%). The amount of callus responded for shoot formation (74%) was obtained in the MS medium containing BAP (1.5 mgl-1) and NAA (0.3mgl-1).The elongated shoots were rooted on half strength medium supplemented with IBA (1.5 mgl-1) and Kn (0.4 mgl-1) for shoots rooted. Regenerated plantlets were successfully acclimatized and hardened off inside the culture and then transferred to green house with better survival rate.

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In vitro callus induction of gerbera from leaf explant

International Journal of Advanced Geosciences ◽

10.14419/ijag.v8i1.30000 ◽

2020 ◽

Vol 8 (1) ◽

pp. 1

Author(s):

Sadia Afrin Jui ◽

Md. Mijanur Rahman Rajib ◽

M. Mofazzal Hossain ◽

Sharmila Rani Mallik ◽

Iffat Jahan Nur ◽

...

Keyword(s):

Acetic Acid ◽

Growth Regulators ◽

Callus Induction ◽

Leaf Explants ◽

Leaf Explant ◽

Naphthalene Acetic Acid ◽

Dichlorophenoxyacetic Acid ◽

Ms Medium ◽

Indole 3 Acetic Acid

The experiment was designed to evaluate the effect of growth regulators on leaf explant of Gerbera for callus induction. Various kinds of plant growth regulators such as 6-Benzylaminopurine (BAP), α-Naphthalene acetic acid (NAA), 2, 4-Dichlorophenoxyacetic acid (2, 4-D), Indole-3-acetic acid (IAA) were used to initiate cultures. These were added to Murashige and Skoog medium in different combinations and concentrations. Leaf explants cultured on MS medium supplemented with BAP+ 2, 4-D+ IAA in T4 treatment & BAP+ 2,4-D in T5 treatment showed the best results for callus induction. On the other hand callus was induced early in the combination of BA+ 2,4-D + IAA hormone in T5, T9 & T8 treatment respectively. The rate of callus induction was very low in BA + NAA combinations but it was much earlier.

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Haploid plantlet regeneration through anther culture in oilseed Brassica species

Bangladesh Journal of Agricultural Research ◽

10.3329/bjar.v34i4.5844 ◽

1970 ◽

Vol 34 (4) ◽

pp. 693-703 ◽

Cited By ~ 4

Author(s):

MA Alam ◽

MA Haque ◽

MR Hossain ◽

SC Sarker ◽

R Afroz

Keyword(s):

Plant Regeneration ◽

Anther Culture ◽

Growth Regulators ◽

Callus Induction ◽

Brassica Species ◽

Plantlet Regeneration ◽

Root Induction ◽

Ms Medium ◽

Half Strength

Anther of five varieties of Brassica species, namely BARI Shariaha-7, Tori-7, Agrani, Daulat and Safal were cultured in vitro to observe their regeneration potentiality. Different concentrations and combinations of growth regulators were supplemented in MS medium. The range of callus induction was 12.50-87.50 %. Maximum callus induction (75.00%) was observed on MS +4 mg/L 2, 4-D + 1.0 mg/L BAP. Among the genotypes, BARI Sharisha-7 showed the highest percentage of callus induction (60.42%). Among the treatments, highest percentage of shoot regeneration (75.00%) was observed on MS + 4 mg/L BAP + 1.0 mg/L NAA. BARI Sharisha-7 also showed the highest rate of plant regeneration (66.67%). Root induction was highest (75%) on half strength MS medium supplemented with 1.0 mg/L IBA and 0.5 mg/L NAA. The plantlets with sufficient roots thus obtained were transferred successfully to plastic pots and subsequently to the field. BARI Sharisha-7 and Tori-7 survived easily in the pots as well as in the field but Safal was very poor in survivability both in the pots and in the field. Key Words: Brassica; haploid; anther culture; in vitro regeneration.DOI: 10.3329/bjar.v34i4.5844Bangladesh J. Agril. Res. 34(4) : 693-703, December 2009

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An Efficient Protocol for High-frequency Direct Multiple Shoot Regeneration from Internodes of Peppermint (Mentha x piperita)

Natural Product Communications ◽

10.1177/1934578x1000501223 ◽

2010 ◽

Vol 5 (12) ◽

pp. 1934578X1000501

Author(s):

Sanjog T. Thul ◽

Arun K. Kukreja

Keyword(s):

Shoot Regeneration ◽

Multiple Shoots ◽

Shoot Elongation ◽

Shoot Formation ◽

Multiple Shoot ◽

Ms Medium ◽

Mentha X Piperita ◽

Half Strength ◽

Multiple Shoot Regeneration

A simple, repeatable and efficient protocol for direct multiple shoot regeneration from internodal explants has been defined in peppermint ( Mentha x piperita var. Indus). In vitro regenerated shoots of peppermint were excised into 4 to 8 mm long internodes and cultured on Murashige and Skoog's medium supplemented with different cytokinins. In the hormonal assay, 3.0 mg L-l zeatin or 6-isopentenyl adenine independently supplemented to half strength MS medium exhibited multiple shoot regeneration, while thiaduzorn (0.1-3.0 mg L−1) showed no morphogenetic effect. A maximum of 85% in vitro cultured explants showed multiple shoot formation with an average of 7 shoots per explant on MS medium supplemented with zeatin. Multiple shoots were initiated within three weeks of cultivation. Internodes with regenerated multiple shoots were transferred to half - strength MS medium without supplementing with any plant growth hormone for shoot elongation and rhizogenesis. Rooted plants acclimatized and grew to maturity under glasshouse conditions. The plantlets developed were phenotypically identical to the parent plant and exhibited 96 % survival.

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In vitro propagation of six selected sugarcane mutant clones through leaf explants

IOP Conference Series Earth and Environmental Science ◽

10.1088/1755-1315/883/1/012075 ◽

2021 ◽

Vol 883 (1) ◽

pp. 012075

Author(s):

R Purnamaningsih ◽

D Sukmadjaja ◽

S Suhesti ◽

S Rahayu

Keyword(s):

Callus Induction ◽

Casein Hydrolysate ◽

Leaf Explants ◽

Shoot Proliferation ◽

Medium Composition ◽

Culture Techniques ◽

High Productivity ◽

Media Formulation ◽

Number Of Shoots

Abstract Six mutant clones of sugarcane with high productivity have been produced through tissue culture techniques combined with mutations using gamma-ray irradiation and Ethyl Methane Sulfonate. The six mutant clones have been tested for stability in the field. They are proven to have high productivity and yields, so that they are very potential to be developed as superior varieties. To support the planting material sufficiency of these clones, an efficient propagation method was needed. Media formulations with different physical properties and composition of growth regulators were tested to obtain high seedling propagation rates. The media formulation for callus induction was Murashige dan Skoog (MS) + 3 mg/l 2,4-D + 3 g/l casein hydrolysate + 3% sucrose and for shoot regeneration was MS + 0,5 mg/l BA + 0,1 mg/l IBA + 100 mg/l PVP and 2% sucrose. Shoot proliferation was carried out on MS liquid (1, ½) + (0.3; 0.5 mg/l) BA + 0.1 mg/l IBA + 1 mg/l Kinetin + (0; 0.5 mg/l) GA3+ sucrose 2%. The results showed that callus induction, callus regeneration, and shoot proliferation of sugarcane mutant clones were influenced by the genotype and medium composition. The fastest callus induction was obtained from the MSP-4 clone (5.82 days), and the longest was MSB-7 (8.82 days). The largest callus diameter was obtained from MSB-6 clone on MS medium containing 1 mg/l BA, 100 mg/l PVP, and 2% sucrose. The highest number of shoots was obtained from the MSB-6 clone, while the least number of shoots conducted from the MSB-8 clone. The MSB-8 clones were more difficult to regenerate compared to the others. The best media formulation for shoot proliferation was ½ MS containing 0.5 mg/l BA, 1 mg/l Kinetin, and 0.1 mg/l IBA, while the best formulation for rooting was ½ MS.

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In vitro regeneration performance of Corchorus olitorius

Journal of the Bangladesh Agricultural University ◽

10.3329/jbau.v8i1.6390 ◽

1970 ◽

Vol 8 (1) ◽

pp. 1-6 ◽

Cited By ~ 1

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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Callus Induction in Baru (Dipteryx alata Vog.) Explants

Notulae Botanicae Horti Agrobotanici Cluj-Napoca ◽

10.15835/nbha47111366 ◽

2018 ◽

Vol 47 (2) ◽

pp. 538-543

Author(s):

Rodrigo Kelson S. REZENDE ◽

Ana Maria N. SCOTON ◽

Maílson V. JESUS ◽

Zeva V. PEREIRA ◽

Fernanda PINTO

Keyword(s):

Ascorbic Acid ◽

Callus Induction ◽

Callus Formation ◽

Leaf Explants ◽

Naphthalene Acetic Acid ◽

Ms Medium ◽

Alternative Technique ◽

Propagation Methods ◽

Dipteryx Alata

Baru (Dipteryx alata Vog.) is a species with great economic and environmental potential; it has popular acceptance, besides being a very productive species. Alternative propagation methods are important for species maintenance and exploration. Thus, micropropagation emerged as an alternative technique, providing genetic stability and the production of a large number of seedlings. The aim of the present investigation was to develop a callus induction protocol for in vitro baru explants. The tested explants were nodal, internodal and foliar segments. The explants were disinfected for 30 seconds in 70% alcohol (v/v) and 2 minutes in sodium hypochlorite (1.25% active chlorine). This was followed by triple washing. The inoculation was carried out in test tubes containing 15 mL MS medium (30 g L-1 sucrose, 6 g L-1 agar and 100 mg L-1 ascorbic acid) supplemented with 2.0 mg L-1 naphthalene acetic acid (NAA). The solution also contained 0.0, 2.5 or 5.0 mg L-1 of 6-benzylaminopurine (BAP) with the pH adjusted to 5.8. In the incubation phase, the explants were cultured for seven days in the dark and then subjected to a photoperiod of 16 hours (43 µmol m-2 s-1) at 25 ± 2 °C. The treatments were studied with 2.5, 5.0, 7.5 or 10.0 mg L-1 BAP additions to the MS. Callus formation, contamination and oxidation evaluations were undertaken. The results obtained when using 2.0 mg L-1 NAA concluded that such a treatment should be used to induce callogenesis from nodal explants, while for the tested baru leaf explants, the best results for callus formation were given by the combination of 2.0 mg L-1 NAA with 2.5 mg L-1 of BAP to.

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In vitro Regeneration and Rapid Multiplication of Dendrobium bensoniae, an Indigenous Ornamental Orchid

The Agriculturists ◽

10.3329/agric.v14i2.31341 ◽

2017 ◽

Vol 14 (2) ◽

pp. 24-31 ◽

Cited By ~ 1

Author(s):

S S Riva ◽

A Islam ◽

M E Hoque

Keyword(s):

Shoot Regeneration ◽

In Vitro Regeneration ◽

Ms Medium ◽

Shoot Induction ◽

Rapid Multiplication ◽

Number Of Shoots

An experiment was conducted on in vitro regeneration and multiplication of Dendrobium bensoniae. Different concentrations of BA and IBA alone or combination of both hormones were used as treatment for regeneration. It was revealed that shoot regeneration from node was the best at 2.0 mg/l BA supplemented to MS medium. It gave better responses than all other concentrations and combinations of BA and BA+IBA, used in the present study. The highest number of shoots and leaves were found when 1.0 mg/l BA with 1.5 mg/l IBA was supplemented into MS medium. For rooting, 0.5 mg/l BA with 1.0 mg/l IBA was found to be the most effective. The well-rooted plantlets were successfully acclimatized under 70-80% humidity and planted in pots and transferred to the shade house for establishment. Around 85% of plantlets survived in the field. From the present result, it may be recommended that MS medium supplemented with 2.0 mg/l BA may be used for rapid shoot induction and regeneration of D. bensoniae.The Agriculturists 2016; 14(2) 24-31

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Development of an in vitro Callus Induction Protocol and Shoot Proliferation for Selected Jatropha (Jatropha curcas) Accessions

Plant Tissue Culture and Biotechnology ◽

10.3329/ptcb.v30i1.47798 ◽

2020 ◽

Vol 30 (1) ◽

pp. 131-141

Author(s):

Hundessa Fufa ◽

Jiregna Daksa

Keyword(s):

Shoot Regeneration ◽

Callus Induction ◽

Jatropha Curcas ◽

Plant Tissue ◽

Leaf Explants ◽

Shoot Proliferation ◽

Adventitious Shoot ◽

Regeneration Medium ◽

Adventitious Shoot Buds

The present study was undertaken to establish a protocol for in vitro callusing of three Jatropha accessions, namely Metema, Adami Tulu and Shewa Robit from leaf explants. The medium supplemented with combination of 4.44 μM BAP and 4.52 μM 2,4-D resulted in maximum percentage of callus (100%) formed for all accessions. The maximum shoot regeneration (66.67%) from callus with 10.13 number of shoot was obtained from Shewa Robit in MS medum fortified with TDZ (2.27 μM ) and IBA (0.49 μM ). The presence of TDZ in the shoot regeneration medium has greater influence on the induction of adventitious shoot buds, whereas MS supplemented with BAP alone and combination with IBA did not induce shoot regeneration from callus culture. The results obtained in the present study would facilitate the high callus induction and regeneration responses in Jatropha for its improvement using biotechnological tools. Plant Tissue Cult. & Biotech. 30(1): 131-141, 2020 (June)

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