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

1970 ◽  
Vol 18 (1) ◽  
pp. 37-42 ◽  
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)

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 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)

scholarly journals Sterilization procedure and callus regeneration in black turmeric (Curcuma caesia)

2019 ◽  
Vol 6 (49) ◽  
Author(s):  
A. S. Abubakar ◽  
R. N. Pudake
Keyword(s):  
Acetic Acid ◽  
Callus Induction ◽  
In Vitro Regeneration ◽  
Leaf Explants ◽  
Surface Sterilization ◽  
To Come ◽  
Dichlorophenoxy Acetic Acid ◽  
And Control ◽  
Rhizome Explants

Sterilization procedure, media composition, explants selection and control of physical environment are critical for successful cultures and callus induction with surface sterilization being very challenging in most plants. Five different sterilization methods were evaluated to come up with the best for subsequent use to establish an in vitro regeneration method for the induction of callus in Curcuma caesia using excised leaf and rhizome explants. Murashige and Skoog (MS) media supplemented with various concentration of 2,4-Dichlorophenoxy acetic acid (2,4-D)/Indole-3-acetic acid (IAA) (0.5- 5.0mg/L), singly or in combination with Benzyl aminopurine (BAP)/Kinetin (KIN) (0.1-5.0mg/L), 0.3% sucrose and 0.08% agar were used. The result of the sterilization procedures showed 15% NaHClO3 (5min) + 70% Ethanol (30s) + 0.1% HgCl2 (5min) to be the most effective in controlling contamination in C. caesia among all the treatments tested. The response to callus induction was found to depend on the type of explants used and growth regulators combination. Leaf explants gave the highest percentage of callus induction. Highest percentage of callus induction (66.70%) was obtained in the growth regulator combination of 2, 4-D (0.5mg/L) + BAP (0.1mg/L) and least (14.29%) in IAA (2.0mg/L) + BAP (0.5mg/L). Equal and higher concentration of 2, 4-D + BAP of 5.0mg/L each also provided better result (40.00%). No callus was obtained in all the single concentration of 2, 4-D used.

scholarly journals High Frequency Regeneration of Plantlets from Immature Male Floral Explants of Musa paradisica cv. Puttabale - AB Genome

2011 ◽  
Vol 21 (2) ◽  
pp. 199-205 ◽  
Author(s):  
K. Girish Kumar ◽  
V. Krishna ◽  
Venkatesh Venkatesh ◽  
K. Pradeep
Keyword(s):  
Callus Induction ◽  
High Frequency ◽  
In Vitro Regeneration ◽  
Floral Part ◽  
Bud Formation ◽  
Shoot Bud ◽  
Immature Male ◽  
Floral Explants ◽  
High Frequency Induction

High frequency in vitro regeneration for mass multiplication from immature male floral explants of Musa paradisica cv. Puttabale on MS supplemented with adenine sulfate (160 mg/l), tyrosine (100 mg/l), sucrose (40 g/l) and gelled with 0.8 g/l agar was attempted.  For callus induction the combinations of 2, 4-D and BAP were tested at 1.0 - 10.0 mg/l and 0.5 - 5.0 mg/l, respectively. For shoot bud formation combinations of BAP and TDZ were also tested at 1.0 - 5.0 mg/l and 0.1 - 0.5 mg/l, respectively.  Luxuriant proliferation and high frequency induction (97.0%) of  callus  was  noticed from the  accessory  floral  part  of  the  explant  at 7.0 mg/l 2, 4-D and 1.0 mg/l BAP, later it preceded towards the gynoecium. Interaction of BAP (2.0 – 5.0 mg/l) and TDZ (0.2 - 0.5 mg/l) would provoke high frequency shoot bud differentiation from the floral calli and a mean of 29.40 ± 6.10 shootlets per callus was obtained at 4 mg/l BAP and 0.4 mg/l TDZ. Rooting of the microshoots was achieved on MS containing 0.6 mg/l NAA and 0.2% activated charcoal.     Key words: Musa pardisica, Puttabale, Regeneration, Male floral explants.   D. O. I. 10.3329/ptcb.v21i2.10243   Plant Tissue Cult. & Biotech. 21(2): 199-205, 2011 (December)

scholarly journals Efficient Regeneration of Tobacco (Nicotiana tabacum L.) Plantlets from Cotyledon, Hypocotyl and Leaf Explants: An Excellent Model Plant for Gene Function Analysis

2020 ◽  
pp. 1-9
Author(s):  
Md. Shoyeb ◽  
Kanis Fatema ◽  
Md. Abdur Rauf Sarkar ◽  
Atikur Rahman ◽  
Shaikh Mizanur Rahman
Keyword(s):  
Gene Function ◽  
Callus Induction ◽  
Function Analysis ◽  
In Vitro Regeneration ◽  
Leaf Explants ◽  
Leaf Explant ◽  
Regeneration Ability ◽  
Ms Medium ◽  
Gene Function Analysis

Tobacco has been widely used as a model plant for stable and non-stable gene function analysis. Successful Agrobacterium-mediated transformation mainly depends on in vitro regeneration of tobacco plant. However, a reliable and standard regeneration protocol of tobacco using multiple explants is limited. In this study, we established a reliable and reproducible regeneration protocol of tobacco using three different explants i.e. cotyledon, hypocotyl and leaf. Preliminary, surface sterilized tobacco seeds were germinated on growth regulator free MS medium. Thereafter, in vitro germinated explants were inoculated into Murashige and Skoog [1] media supplemented with different combination and types of growth regulators for callus induction and subsequent regeneration of plantlets. It was revealed that, regeneration ability of explants is greatly influenced by type and nature of the explant. Among the three explants, higher callus induction (95%) was obtained in MS medium supplemented with 2.0 mg l-1 kinetin + 2.0 mg l-1 IAA from leaf explant. Also, leaf explant exhibited much higher regeneration ability (95%) than hypocotyl (60%) and cotyledon (45%) explants. Significantly highest number of shoots (8.0) were regenerated from leaf explants cultured on MS medium supplemented with 3.0 mg l-1 Kinetin+1.0 mg l-1 IAA compared to the other hormone combinations. Regenerated mature shoots were showed normal root after transferred onto ½ MS medium containing 0.3 mg l-1 IBA. This study will provide valuable information related to in vitro regeneration of tobacco plantlets using cotyledon, hypocotyl and leaf explants and will be used as a standard protocol for Agrobacterium-mediated transformation for gene function analysis.

scholarly journals In vitro Regeneration of Japanese Burdock (Arctium lappa L.) from Cotyledon and Leaf Derived Explants

2022 ◽  
Vol 31 (2) ◽  
pp. 123-134
Author(s):  
Mustafa Abul Kalam Azad ◽  
Md Arifuzzaman ◽  
Md Mobarok Hossain ◽  
Md Sohel Arman ◽  
Muhammad Nurul Amin
Keyword(s):  
In Vitro Regeneration ◽  
Adventitious Shoots ◽  
Leaf Explants ◽  
Direct Shoot Regeneration ◽  
Indirect Organogenesis ◽  
Arctium Lappa ◽  
Sand Mixture ◽  
The Third ◽  
Rooting Medium

Considering the vegetable and medicinal values, a micropropagation protocol has been established for Japanese Burdock (Arctium lappa L.) by culturing the explants of cotyledon and leaf obtained from in vitro grown seedlings. Direct shoot regeneration was achieved from cotyledon and leaf explants on MS fortified with 4.0 μM BAP and 2.0 μM IBA or NAA after 5 weeks of culture. In addition, both the explants also formed callus from their cut margins within 6 weeks of cultivation on medium complemented with 6.0 μM BAP and 4.0 μM IBA or NAA. Adventitious shoots were also redeveloped through indirect organogenesis from the cotyledon and leaf-derived callus within 10 weeks of culture on MS containing 4.0 μM BAP and 2.0 μM IBA or NAA. The highest rate of shoot reproduction was attained at the third subculture, and more than 12.6 shoots were formed per callus clump. Within 4 weeks of transfer to the rooting medium on MS containing 6.0 μM IBA, the cultured micro-shoots produced highest 5.3 roots per cultured shoot. Rooted plantlets were successfully established on a soil-composed-sand mixture under natural condition with 93.3% survival rate Plant Tissue Cult. & Biotech. 31(2): 123-134, 2021 (December)

scholarly journals In vitro Regeneration of Alstroemeria cv. ‘Balance’ by Indirect Organogenesis

2017 ◽  
Vol 14 (2) ◽  
pp. 607-614
Author(s):  
Hossein Nazarian ◽  
Maryam Beigi Harchegani ◽  
Mahmoud Otroshy ◽  
Ali Motamedi
Keyword(s):  
Callus Induction ◽  
Callus Formation ◽  
In Vitro Regeneration ◽  
Culture Technique ◽  
Stem Explants ◽  
Indirect Organogenesis ◽  
High Concentrations ◽  
Two Phases ◽  
Positive Effect

ABSTRACT: This study was designed in order to optimize the indirect organogenesis (during callus induction and regeneration) of Alstroemeria cv. ‘Balance’ through tissue culture technique in two phases; the first stage: callus induction by rhizome segments, leaf and nodal stem which in the start, callus formation media were examined using two types of auxins; 2,4-D and NAA and a cytokinin; BAP in four different experimentations. In the second stage, calli derived from rhizome segments and nodal stem explants were transferred to regeneration media. The results revealed that 2,4-D in combination with BAP in the rhizome segments and nodal stem explants were efficient as compared to NAA. The highest yield of callus formation was also obtained in the rhizome segments explants. According to the results, it can be suggested that NAA as auxin, does not have direct positive effect on cell division in Alstroemeria. The 2,4-D is toxic at high concentrations and may bring about cell death. Eventually, the composition of 0.5 mg/l NAA with 3 mg/l BAP and callus derived from nodal stem explants may be introduced as the best combination for regeneration. These results indicate the necessity of the BAP cytokinin presence for regeneration. In addition, the maximum length of the shoot was obtained from combination of BAP with nodal stem explants, without the presence of NAA.

scholarly journals Plant Regeneration from Leaf-derived Callus Cultures of Primrose (Primula vulgaris)

HortScience ◽  
2016 ◽  
Vol 51 (5) ◽  
pp. 558-562 ◽  
Author(s):  
Sadiye Hayta ◽  
Mark A. Smedley ◽  
Jinhong Li ◽  
Wendy A. Harwood ◽  
Philip M. Gilmartin
Keyword(s):  
Flower Development ◽  
In Vitro Regeneration ◽  
Leaf Explants ◽  
Callus Cultures ◽  
Indirect Organogenesis ◽  
Horticultural Crops ◽  
Donor Material ◽  
Primula Vulgaris ◽  
Inbred Parent

Efficient micropropagation of Primula species is important both for fundamental scientific studies and commercial applications. Primula vulgaris (Huds), along with other Primulaceae species, exhibits floral heteromorphy with two distinct forms of hermaphroditic flower. Studies to identify genes that control heteromorphic flower development require propagation of floral mutants, and efficient regeneration is a key requirement for plant transformation. Several species, including P. vulgaris cultivars and P. ×polyantha hybrids, are important horticultural crops in Europe, United States, and Japan and semidouble/double Primula varieties offer a high-end product. Vegetative propagation of sterile double forms, and as a means to increase numbers of inbred parent plants for F1 seed production is, however, slow. Micropropagation offers the most efficient way of increasing these varieties quickly and efficiently. To date, most Primula micropropagation protocols require explant material derived from in vitro grown seedlings or use floral parts as donor material with seasonal limitations. Therefore, an effective and efficient protocol was developed for in vitro regeneration of P. vulgaris via indirect organogenesis from adult leaf–derived explants. Exposure of leaf explants of P. vulgaris to media containing synthetic cytokinin, thidiazuron (TDZ), and auxin [1-naphthylacetic acid (NAA) or 2,4-dichlorophenoxyacetic acid (2,4-D)] resulted in undifferentiated cell proliferation and followed by differentiated growth as shoot organogenesis. Silver nitrate improved in vitro callus growth and increased shoot regeneration further, with up to 72% of explants producing shoots. Regenerated plants developed normally and produced normal fertile flowers within 7 months. The system was also successfully applied for the micropropagation of sterile double-flowered P. vulgaris ‘Sue Jervis’. The protocol reported here enables propagation of P. vulgaris without seasonal limitation or destruction of valuable parent donor material. The protocol, with further development, has the potential to underpin development of a transformation system for Primula, which would be of value in studies on flower development and disease resistance in laboratory grown 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

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