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

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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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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In vitro Regeneration of Vitex negundo L. - A Multipurpose Woody Aromatic Medicinal Shrub

Plant Tissue Culture and Biotechnology ◽

10.3329/ptcb.v18i1.3263 ◽

1970 ◽

Vol 18 (1) ◽

pp. 37-42 ◽

Cited By ~ 6

Author(s):

M. Jawahar ◽

S. Ravipaul ◽

M. Jeyaseelan

Keyword(s):

Callus Induction ◽

In Vitro Regeneration ◽

Leaf Explants ◽

Shoot Proliferation ◽

Optimal Level ◽

Vitex Negundo ◽

Indirect Organogenesis ◽

Shoot Bud ◽

Bud Initiation

A rapid and efficient protocol was developed for inducing indirect organogenesis using leaf explants of Vitex negundo L. Explants were cultured on MS with different concentrations of 2,4-D and IAA in combination with BAP for callus induction. The frequency of callus induction increased with increasing concentration of IAA (0.3 mg/l) and BAP (0.3 mg/l) at optimal level. The shoot buds appeared emerging as green coloured protuberances on the callus. The high frequency of shoot bud initiation and shoot proliferation was observed on MS containing 0.3 mg/l IAA and 0.3 mg/l BAP. The regenerated shoots were successfully rooted on MS supplemented with 0.5 mg/l IBA. Rooted plants were transferred to pots containing sand, soil and manure in the ratio of 1 : 1 : 1. Nearly 90% survival of in vitro plants were recorded. Key words : Vitex negundo, In vitro, Leaf, Callus, Regeneration D.O.I. 10.3329/ptcb.v18i1.3263 Plant Tissue Cult. & Biotech. 18(1): 37-42, 2008 (June)

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

OENO One ◽

10.20870/oeno-one.2015.49.1.95 ◽

2015 ◽

Vol 49 (1) ◽

pp. 37 ◽

Cited By ~ 6

Author(s):

Nadra Khan ◽

Maqsood Ahmed ◽

Ishfaq Hafiz ◽

Nadeem Abbasi ◽

Shaghef Ejaz ◽

...

Keyword(s):

Shoot Regeneration ◽

Growth Regulators ◽

Callus Induction ◽

Leaf Explants ◽

Shoot Cultures ◽

Ms Medium ◽

Grape Cultivar ◽

Half Strength ◽

Number Of Shoots

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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Efficient Plant Regeneration via Indirect Organogenesis in Carnation (Dianthus Caryophyllus Semperflorens flore pleno) Cultivars

Journal of Horticultural Research ◽

10.2478/johr-2021-0020 ◽

2022 ◽

Vol 0 (0) ◽

Author(s):

Hamid Reza SABAGHI ◽

Gholamreza SHARIFI-SIRCHI ◽

Pejman AZADI ◽

Mohammad Hossein AZIMI

Keyword(s):

Plant Regeneration ◽

Callus Induction ◽

Dianthus Caryophyllus ◽

Casein Hydrolysate ◽

Leaf Explants ◽

Naphthaleneacetic Acid ◽

Ms Medium ◽

Regeneration Rate ◽

Shoot Number

ABSTRACT Callus induction and plant regeneration are important steps of in vitro plant breeding of ornamental plants. In this study, the effects of different combinations of plant growth regulators (PGRs), promoters, and minerals on callus induction and plant regeneration in different carnation cultivars were studied in a completely randomized design with three replications. For callus induction, 16 different combinations of 2,4-dichlorophenoxyacetic acid (2,4-D), 6-benzylaminopurine (BA), 1-naphthaleneacetic acid (NAA), and casein hydrolysate (CH) were studied using in vitro leaf explants. The Murashige and Skoog (MS) medium supplemented with 0.2 mg·dm-3 of 2,4-D and 200 mg·dm-3 of CH showed the highest frequency of callus induction. Among the cultivars, ‘Noblesse’ showed the highest rate of callus induction (91.67%). Regarding regeneration, BA, NAA, silver nitrate (AgNO3), and adenine hemisulfate (As) were used in ten different combinations. The ‘Cameron’, ‘Tabasco’, and ‘Noblesse’ cultivars with 95.24% regeneration percentage showed the highest rate of plant regeneration. Generally, in most cultivars, the highest regeneration rate and shoot number per explant were found in the MS medium supplemented with 3 mg·dm-3 of BA, 0.6 mg·dm-3 of NAA, 5 mg·dm-3 of AgNO3, and 40 mg·dm-3 of As. According to the results, the highest regeneration frequency was obtained when 40 mg·dm-3 of As was added to the medium. Finally, the flow cytometry analysis indicated that there were no significant differences between in vitro regenerated and control plants in terms of DNA ratios.

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Seed Germination and in vitro Plant Regeneration through Callus Culture of Two Lychnis Species

Plant Tissue Culture and Biotechnology ◽

10.3329/ptcb.v25i1.24110 ◽

2015 ◽

Vol 25 (1) ◽

pp. 1-12 ◽

Cited By ~ 1

Author(s):

Kee Hwa Bae ◽

Eui Soo Yoon

Keyword(s):

Plant Regeneration ◽

Seed Germination ◽

Callus Induction ◽

In Vitro Propagation ◽

Ornamental Plants ◽

Leaf Explants ◽

Adventitious Shoot ◽

Temperature Treatment ◽

In Vitro Plant Regeneration

Lychnis cognate Maxim and Lychnis fulgens Fish. Ex Spreng are two valued ornamental plants in Korea. Soaking of seeds in GA3 solution remarkably promoted germination up to 60%, but the control (0 mg/l) was not effective (> 5%). To select an adequate temperature for seed germination, seeds, previously soaked in a 1000 mg/l GA3 for 24 hrs, were incubated at 15, 20, 25, and 30°C. Seed germination of over 20% was obtained at 15, 20, and 25°C, but only 10% at 30°C. These results indicate that the seeds of L. cognate and L. fulgens are in a such dormant state that they hardly germinate even by dormancy breaker (GA3) and low (15 ? 25°C) temperature treatment. The highest callus induction was observed in the leaf explants of the seedlings on MS containing specific concentrations of 3.0 mg/l BA and 1.0 mg/l NAA. The adventitious shoot was formed < 90% of calli on 1/2 WPM medium. The height of in vitro propagated plantlet was no different media used for regeneration. This in vitro propagation protocol should be useful for conservation of endangered and ornamental plant.Plant Tissue Cult. & Biotech. 25(1): 1-12, 2015 (June)

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Optimization of medium composition for in vitro shoot proliferation and growth of date palm cv. Mejhoul

3 Biotech ◽

10.1007/s13205-016-0430-x ◽

2016 ◽

Vol 6 (1) ◽

Cited By ~ 10

Author(s):

Mouaad Amine Mazri ◽

Reda Meziani ◽

Jamal El Fadile ◽

Az-eddine Ezzinbi

Keyword(s):

Date Palm ◽

Shoot Proliferation ◽

Medium Composition ◽

In Vitro Shoot Proliferation

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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 somatic embryogenesis of superior clones of robusta coffee from Lampung, Indonesia: Effect of genotypes and callus induction media

Biodiversitas Journal of Biological Diversity ◽

10.13057/biodiv/d210849 ◽

2020 ◽

Vol 21 (8) ◽

Author(s):

Dwi Hapsoro ◽

Rahmadyah Hamiranti ◽

Yusnita Yusnita

Keyword(s):

Somatic Embryogenesis ◽

Callus Induction ◽

Leaf Explants ◽

Regeneration Medium ◽

Embryogenic Calli ◽

Robusta Coffee ◽

Primary Callus ◽

Ascorbic Acids ◽

Superior Clones

Abstract. Hapsoro D, Hamiranti R, Yusnita Y. 2020. In vitro somatic embryogenesis of superior clones of robusta coffee from Lampung, Indonesia: Effect of genotypes and callus induction media. Biodiversitas 21: 3811-3817. This study aimed to investigate the effects of genotypes and primary callus induction media on somatic embryogenesis of superior robusta coffee clones of Lampung. Leaf explants of clones Tugusari, Komari, Tugino, and Wanto were cultured on two types of primary callus induction media (PCIM). PCIM1 consisted of half-strength MS salts, 30 gL-1 sucrose, added with (mgL-1) 0.1 thiamine-HCl, 0.5 nicotinic acids, 0.5 pyridoxine-HCl, 100 Myo-inositol, 200 ascorbic acids, 150 citric acids, and 1 benzyl adenine. PCIM2 consisted of NPCM salts, 30 gL-1 sucrose, added with (mgL-1) 15 thiamine-HCl, 1 nicotinic acid, 1 pyridoxine-HCl, 2 glycines, 130 Myo-inositol, 200 ascorbic acids, 150 citric acids, 1 2,4-dichlorophenoxyacetic acid, and 2 thidiazuron. The highest percentage (100%) of primary callus formation was found in Komari and Wanto clones. PCIM2 resulted in more primary calli than PCIM1. When subcultured to embryogenic callus induction medium, primary calli of clone Komari and Wanto developed into a high percentage of embryogenic calli, while those of the other two turned brown and died. PCIM2-derived primary calli developed into more embryogenic calli. When subcultured on somatic embryo (SE) regeneration medium, these calli underwent the formation of SE of various stages. When subcultured to plant regeneration medium, these SEs developed into plantlets.

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Micropropagation of Tarenaya rosea (Cleomaceae) from leaf explants

Rodriguésia ◽

10.1590/2175-7860202172020 ◽

2021 ◽

Vol 72 ◽

Author(s):

Claudia Simões-Gurgel ◽

Tatiana Carvalho de Castro ◽

Cátia Henriques Callado ◽

Lívia da Silva Cordeiro ◽

Norma Albarello

Keyword(s):

Large Scale ◽

Plant Conservation ◽

Leaf Explants ◽

Plant Production ◽

Anthocyanin Content ◽

Alternative Source ◽

Regeneration Rate ◽

Culture Techniques ◽

Regenerated Plants

Abstract In vitro culture techniques are recognized as efficient strategies for large-scale plant production, as well as providing alternatives for plant conservation. In this study the micropropagation of Tarenaya rosea was established using petiole and foliar blade segments cultivated on MS medium with 6-benzyladenine (BA) and/or 6-furfurylaminopurine (KIN). The regeneration rate from explants was evaluated after 30-days in culture, as well as the proliferation rate from explant-derived shoots, reached after four subcultures performed at 30-days in culture. In vitro propagation occurred by both direct (DO) and indirect (IO) organogenesis. The highest regeneration rates by DO (50% to 100%) were reached on media containing only BA, while morphogenic calluses (IO) were mainly formed with BA+KIN. Explants on media with BA showed the presence of small black nodules on their surface, and histological analysis revealed the presence of trichomes with anthocyanin content. Elongation and rooting were reached on growth regulator-free MS. Acclimatization rates around 80% were achieved and the in vitro-regenerated plants were successfully maintained under field conditions. Results show significant morphogenetic potential of T. rosea from leaf explants, mainly when cultivated in the presence of 4.4 µM BA, providing a new alternative source of plant material for biotechnological and in vitro conservation studies.

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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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