In Vitro Callus Development on Immature Leaf Explants of Liberica Coffee (Coffea liberica L. cv. Liberika Tungkal Komposit) by the Application of 2.4-D and BAP

ABSTRACTThis study was aimed to obtain an appropriate medium composition with various combinations of Picloram + BAP for the proliferation of embryogenic callus from immature leaf explants of durian. The experiment was carried out at the Plant Biotechnology Laboratory, Faculty of Agriculture , the University of Jambi from January through to November 2012. Five levels of Picloram (1.0, 2.0, 3.0, 4.0, 5.0 mg L-1) in combination with three levels of BAP (0, 0.5, 1.5 mg L-1) were tested. Therefore, there were 15 treatment combinations with 4 replicates resulting in 60 experimental unit. Each unit consisted of 4 culture flasks containing one immature leaf explant. Cultures were kept in culture room with 16 h photoperiod and 1000 lux light intensity. The results showed that: 1) callus proliferation on immature leaf explants of durian cv. Selat was dependent upon the level of Picloram + BAP added to culture medium, 2) the addition of 3.0 - 5.0 mg L-1Picloram without BAP was found to be effective in promoting callus proliferation on the majority of cultured explants, 3) all regenerated callus showed similar characteristics, but embryogenic properties was not seen yet, and 4) the application of tissue culture technique in the propagation of durian cv. Selat needs further comprehensive investigation, particularly on factors directly affecting culture development and inducing somatic embryogenesis.Key words: tissue culture, in vitro culture, micropropagation, plant hormones, auxin, cytokinin, fruit crops. ABSTRAKPenelitian ini bertujuan untuk mendapatkan komposisi media yang tepat dari kombinasi Picloram + BAP untuk proliferasi kalus embriogenik dari eksplan daun dewasa durian. Penelitian dilakukan di Laboratorium Bioteknologi Tanaman, Fakultas Pertanian, Universitas Jambi dari Januari hingga November 2012. Perlakuan kombinasi media zat pengatur tumbuh adalah lima taraf Picloram (1.0, 2.0, 3.0, 4.0, 5.0 mg L-1) dengan kombinasi tiga taraf perlakuan BAP (0, 0.5, 1.5 mg L-1). Terdapat 15 kombinasi media perlakuan dengan 4 ulangan, sehingga terdapat 60 kombinasi satuan percobaan. Setiap unit percobaan terdapat 4 botol kultur dengan satu eksplan daun. Kultur disimpan di ruang kultur selama 16 jam penyinaran dan intensitas cahaya 1000 lux. Hasil penelitian menunjukkan bahwa: 1) Proliferasi kalus dari eksplan daun muda buah durian tergantung pada taraf kombinasi picloram + BAP yang ditambahkan ke media kultur; 2) Penambahan 3.0-5.0 mg L-1Picloram tanpa BAP memberikan hasil yang efektif untuk menginduksi proliferasi kalus pada sebagian besar eksplan; 3) Regenerasi kalus menunjukkan karakteristik serupa, tetapi embriogenik kalus tidak muncul, dan 4) Perbanyakan eksplan daun durian dengan tehnik kultur jaringan membutuhkan penelitian lebih lanjut, terutama pada faktor yang berpengaruh langsung pada induksi embriogenesis somatik.Kata kunci: auksin, buah, kultur jaringan, kultur in vitro, mikropropagasi, sitokinin, zat pengaturtumbuh,

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FACTORS AFFECTING THE REGENERATION OF PEPPER (CAPSICUM ANNUUM L.)

HortScience ◽

10.21273/hortsci.25.9.1120 ◽

1990 ◽

Vol 25 (9) ◽

pp. 1120G-1120

Author(s):

J. L. Jacobs ◽

C. T. Stephens

Keyword(s):

Growth Hormone ◽

Silver Nitrate ◽

Callus Formation ◽

Leaf Explants ◽

Regeneration System ◽

Factors Affecting ◽

Nitrate Concentrations ◽

Leaf Differentiation ◽

Whole Plants

Several growth hormone combinations and silver nitrate concentrations were examined for their effect on regeneration of different pepper genotypes. Primary leaf explants from in vitro seedlings were cultured on a revised Murashige and Skoog medium supplemented with auxin, cytokinin and 1.6% glucose. Combinations of different concentrations of indole-3-acetic acid (IAA), 0-5 mg/l, and 6-benzylaminopurine (BAP), 0-5 mg/l, were tested to determine the most effective medium for shoot primordium formation. Experiments with IAA and BAP did not result in a specific growth hormone combination appropriate for regeneration of all genotypes tested. Of the silver nitrate concentrations tested, 10 mg/l resulted in the best shoot and leaf differentiation and reduced callus formation. Differences in organogenic response of individual genotypes were evaluated on a single regeneration medium. Whole plants were regenerated from 11 of 63 genotypes examined. Based on these experiments, a reproducible regeneration system for pepper was developed with a total of 500 plants regenerated to date.

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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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In vitro shoot regeneration of boldo from leaf explants

Ciência Rural ◽

10.1590/s0103-84782010005000173 ◽

2010 ◽

Vol 40 (10) ◽

pp. 2210-2213

Author(s):

Monalize Salete Mota ◽

Juliana de Magalhães Bandeira ◽

Eugenia Jacira Bolacel Braga ◽

Valmor João Bianchi ◽

José Antonio Peters

Keyword(s):

Shoot Regeneration ◽

Leaf Explants ◽

Shoot Development ◽

High Potential ◽

Naphthaleneacetic Acid ◽

Regeneration System ◽

Bud Induction ◽

Molecular Studies ◽

In Vitro Shoot Regeneration

A shoot regeneration system for Plectranthus neochilus was studied from leaf explants. Leaves developed under in vitro conditions were cultured on Wood Plant Medium supplemented with 0.2mg dm-3 α-naphthaleneacetic acid (NAA) and different 6-benzilaminopurine (BAP) or thidiazuron (TDZ) concentrations (0, 1.5, 3.0, 4.5 and 6.0mg dm-3). An increase in percentage of responsive explants (85.3%) and in the number of shoots developed per explant (3.2) was observed when the explants were treated with 5.3 and 4.7mg dm-3 BAP, respectively. The leaf explants cultured on media supplemented with TDZ became vitreous and did not form buds. The regeneration system used is efficient for boldo bud induction and shoot development, showing high potential for advanced cellular and molecular studies.

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Production of bergenin, an active chemical constituent in the callus of Bergenia ciliata (Haw.) Sternb.

Botanica Orientalis Journal of Plant Science ◽

10.3126/botor.v8i0.5557 ◽

2012 ◽

Vol 8 ◽

pp. 40-44 ◽

Cited By ~ 3

Author(s):

Umesh Krishna Shrestha ◽

Bijaya Pant

Keyword(s):

Leaf Explants ◽

Chemical Constituent ◽

Wild Plants ◽

Plant Science ◽

Naphthalene Acetic Acid ◽

Active Chemical ◽

In The Wild ◽

The Media ◽

Benzyl Amino Purine

In vitro culture of Bergenia ciliata (Haw.) Sternb. was carried out for the examination of bergenin content. Leaf explants were cultured in MS (Murashige and Skoog) basal media supplemented with or without phytohormones. The hormonal series maintained were in the range of 0-2 mg l-1 for BAP (6-benzyl amino purine) and 0-1.5 mg l-1 for NAA (α-naphthalene acetic acid). Bergenin content of in vitro grown tissues of B. ciliata was compared with that of wild plants collected from three different localities of Nepal. The best growth of callus and plantlets occurred in the media containing BAP 1.0 mg l-1 + NAA 1.0 mg l-1 and BAP 1.5 mg l-1 + NAA 1.0 mg l-1. Production of bergenin was high in the media supplemented with 1.0 mg l-1 BAP + 1.5 mg l-1 NAA (3.40 μg g-1) and 2.0 mg l-1 BAP + 1.5 mg l-1 NAA (3.05 μg g-1) under experimental condition. The bergenin content in the wild plants collected from Langtang, Jumla and Godawari was found to be 4.28 μg g-1, 4.53 μg g-1 and 3.64 μg g-1 respectively. This study shows that the in vitro cultured callus of B. ciliata is capable of synthesizing bergenin in quantity comparable to that of the wild plant.doi: http://dx.doi.org/10.3126/botor.v8i0.5557 Botanica Orientalis – Journal of Plant Science (2011) 8: 40-44

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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 Propagation of Digitalis trojana Ivanina., an Endemic Medicinal Plant of Turkey

10.5772/intechopen.94378 ◽

2020 ◽

Author(s):

Nurşen Çördük ◽

Cüneyt Aki

Keyword(s):

Acetic Acid ◽

Medicinal Plant ◽

In Vitro Propagation ◽

Leaf Explants ◽

Naphthalene Acetic Acid ◽

Ms Medium ◽

Northwestern Turkey ◽

Helen Of Troy ◽

Endemic Medicinal Plant

Digitalis trojana Ivanina is a member of the Plantaginaceae family and known by its common name, Helen of Troy foxglove. It is perennial endemic to Çanakkale and Balıkesir, northwestern Turkey. In order to develop an efficient shoot regeneration protocol, the leaf explants of D. trojana were cultured on Murashige and Skoog (MS) medium containing 6-benzyl adenine (0.1, 0.5, 1.0, 3.0, 5.0 mg/L) and α-naphthalene acetic acid (0.1, 0.5, 1.0 mg/L), 3% (w/v) sucrose and 0.8% (w/v) agar. The highest number of regenerated shoots was obtained from leaf explants that were cultured on MS medium with 3.0 mg/L BA+0.1 mg/L NAA. Regenerated shoots were rooted on MS medium without plant growth regulators. Rooted plants (2–3 cm) were separately transferred to pots containing a mixture of peat and perlite (2:1 v/v) and acclimatized successfully in a growth chamber.

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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 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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