In vitro callus induction and regeneration of multiple shoots through callus derived from leaf and epicotyl explants in pigeon pea (Cajanus cajan L.)

2017 ◽  
Author(s):  
Padmavathi A.V. Thangella ◽  
B. Fakrudin
Keyword(s):  
Callus Induction ◽  
Leaf Shape ◽  
Leaf Explants ◽  
Multiple Shoots ◽  
Pigeon Pea ◽  
Callus Cultures ◽  
Natural Conditions ◽  
Cotyledonary Leaf ◽  
Higher Doses

An efficient in vitro protocol was developed for callus induction, high frequency plant regeneration through callus cultures derived from cotyledonary leaf and epicotyl explants, rooting of shoots derived from callus and establishment onto the natural conditions in two cultivars of pigeon pea; ICPL 87119 and ICPL 8863. Cotyledonary leaf and epicotyl explants were tested for callus induction across 48 different combinations and concentrations of auxins and cytokinins in MS medium, wherein, higher doses of auxins (15 mg/1 NAA) in combination with lower doses of cytokinins (0.5 mg/l kinetin) induced regenerable callus from leaf explants while lower doses of auxins (0.2 mg/1 NAA) in combination with higher doses of cytokinins (8 mg/1 kinetin) induced regenerable callus from epicotyl explants in both the genotypes. Plantlet regeneration from leaf and epicotyl derived callus was optimized at 0.05 mg/l TDZ in both genotypes. Rooting was optimized on ½ MS + 0.5 mg/1 IBA media in both genotypes. Well-rooted plants were acclimatized and established successfully into natural conditions in potting mixture-containing soil: FYM in 1:1 ratio resulting in 48.01 per cent survivability. Regenerated plants were uniform morphologically with normal leaf shape and growth. This protocol finds its significance in rapid multiplication of transgenic plants.

scholarly journals In Vitro plant regeneration from leaf explants of Tagetes erecta L.

2021 ◽  
Vol 56 (2) ◽  
pp. 69-74
Author(s):  
JL Munshi ◽  
R Baksha ◽  
MZ Rahaman ◽  
NN Huque ◽  
EA Zinat ◽  
...  
Keyword(s):  
Callus Induction ◽  
Callus Formation ◽  
Leaf Shape ◽  
Leaf Explants ◽  
Multiple Shoots ◽  
Tagetes Erecta ◽  
Ms Medium ◽  
Shoot Bud ◽  
Tagetes Erecta L

Regeneration of multiple shoots via callus induction and organogenesis was obtained from young leaf explants of the field grown marigold (Tagetes erecta L.). Callus induction and shoot regeneration at various frequencies were observed using different concentrations and combinations of growth regulators. Highest percentage (90%) of callus formation was observed within two weeks on MS medium supplemented with 5.0 mg/l BAP with 2.5 mg/l NAA. The maximum percentage (80%) of shoot bud formation (10±0.5/callus) was obtained from MS medium containing 1.0 mg/l BAP with 0.5 mg/l kinetin. The regenerated shoots developed highest percentages (90%) of roots on half strength MS medium supplemented with 1.0 mg/l IBA. The plantlets when transferred into potsoil 80% survived. Regenerated plants were morphologically uniform with normal leaf shape and growth pattern. Bangladesh J. Sci. Ind. Res.56(2), 69-74, 2021

scholarly journals In vitro Regeneration and Agrobacterium-mediated Genetic Transformation of Tomato (Lycopersicon esculentum Mill.)

1970 ◽  
Vol 19 (1) ◽  
pp. 101-111 ◽  
Author(s):  
Rakha Hari Sarker ◽  
Khaleda Islam ◽  
M.I. Hoque
Keyword(s):  
Lycopersicon Esculentum ◽  
Genetic Transformation ◽  
In Vitro Regeneration ◽  
Leaf Explants ◽  
Multiple Shoots ◽  
Multiple Shoot ◽  
Root Induction ◽  
Cotyledonary Leaf ◽  
Lycopersicon Esculentum Mill

Agrobacterium-mediated genetic transformation system has been developed for two tomato (Lycopersicon esculentum Mill.) varieties, namely Pusa Ruby (PR) and BARI Tomato-3 (BT-3). Prior to the establishment of transformation protocol cotyledonary leaf explants from the two varieties were cultured to obtain genotype independent in vitro regeneration. Healthy multiple shoot regeneration was obtained from the cut ends of cotyledonary leaf segments for both the varieties on MS containing 1.0 mg/l BAP and 0.1 mg/l IAA. The maximum root induction from the regenerated shoots was achieved on half the strength of MS medium supplemented with 0.2 mg/l IAA. The in vitro grown plantlets were successfully transplanted into soil where they flowered and produced fruits identical to those developed by control plants. Transformation ability of cotyledonary leaf explants was tested with Agrobacterium tumefaciens strain LBA4404 harboring binary plasmid pBI121, containing GUS and npt II genes. Transformed cotyledonary leaf explants were found to produce multiple shoots on MS containing 1.0 mg/l BAP and 0.1 mg/l IAA. Selection of the transformed shoots was carried out by gradually increasing the concentration of kanamycin to 200 mg/l since kanamycin resistant gene was used for transformation experiments. Shoots that survived under selection pressure were subjected to rooting. Transformed rooted plantlets were transferred to soil. Stable expression of GUS gene was detected in the various tissues from putatively transformed plantlets using GUS histochemical assay.  Key words: In vitro regeneration, transformation, tomato D.O.I. 10.3329/ptcb.v19i1.5004 Plant Tissue Cult. & Biotech. 19(1): 101-111, 2009 (June)

scholarly journals Callus Induction and Organogenesis in Sugarcane (Saccharum officinarum L.) var 93v297

2015 ◽  
Vol 48 ◽  
pp. 14-22
Author(s):  
Arjun ◽  
Rao Srinath
Keyword(s):  
Sugar Cane ◽  
Callus Induction ◽  
Leaf Explants ◽  
Multiple Shoots ◽  
Saccharum Officinarum ◽  
Ms Medium ◽  
Sand Soil ◽  
Maximum Root Length ◽  
Maximum Root

An efficient protocol for induction of callus and regeneration of a sugar cane var 93v297 has been developed and reported here. Callus induction from immature young leaf explants derived from 2-3-month-old plants was achieved on Murashige and Skoog’s (MS) medium supplemented with different auxins viz, 2,4-D, NAA and IAA. Among different auxins, 2, 4-D at 3.5mg/l + 0.5mg/l BAP was found favourable in inducing callus. Addition of coconut milk and BAP further enhanced the growth of callus maximum being on MS medium supplemented with 0.5mg/l BAP (3602.33±0.88mg). Calli were further evaluated for regeneration. MS medium supplemented with 1.0 mg/l BAP was found suitable where 100% calli regenerated with maximum number of multiple shoots per callus mass (41.40±0.89). Highest number of root emergence (28.33±1.16) and maximum root length (3.40±0.67cm) was achieved on MS medium supplemented with 3mgl/l NAA. The in vitro grown plants were transferred to polycups containing a mixture of sterilized sand, soil and cocopeet (1:1:1) for hardening. The hardened plants were transferred to green-house conditions where they survived with 90% frequency.

scholarly journals In vitro Propagation of Ionidium suffruticosum Ging. ? A Seasonal Multipotent Medicinal Herb.

1970 ◽  
Vol 19 (2) ◽  
pp. 143-150
Author(s):  
Arunkumar B. Sonappanavar ◽  
M. Jayaraj ◽  
Asha N. Bagadekar ◽  
Anant V. Bhandarkar
Keyword(s):  
Key Words ◽  
Shoot Regeneration ◽  
Callus Induction ◽  
Plant Tissue ◽  
In Vitro Propagation ◽  
Leaf Explants ◽  
Callus Cultures ◽  
Medicinal Herb ◽  
In Vitro Shoots

Indirect regeneration of plant was obtained through organogenesis in leaf callus cultures of Ionidium suffruticosum. Leaf explants were found to be best suited for callus induction on MS with 2, 4-D (0.5 and 1.0 mg/l). Maximum shoot regeneration was obtained in MS supplemented with Kn (4.0 mg/l) alone and NAA (0.4 mg/l) with Kn (2.0 m/l).  The in vitro shoots thus obtained were successfully rooted in MS supplemented with Kn (4.0 mg/l) alone and with NAA (2.0 mg/l) and Kn (0.2  mg/l).  Seventy per cent of the rooted plants survived and they were successfully acclimated in soil. Key words: Ionidium suffruticosum, micropropagation, Medicinal herb D.O.I. 10.3329/ptcb.v19i2.5431 Plant Tissue Cult. & Biotech. 19(2): 143-150, 2009 (December)

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

2015 ◽  
Vol 25 (1) ◽  
pp. 1-12 ◽  
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)

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

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.

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

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.

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

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.

scholarly journals Callus Induction in Baru (Dipteryx alata Vog.) Explants

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.

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

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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