scholarly journals In vitro regeneration protocol development via callus formation from stem explant of tomato

2016 ◽  
Vol 1 (3) ◽  
pp. 589-599
Author(s):  
Meherunnesa Papry ◽  
SM Ahsan ◽  
Sayeed Shahriyar

The experiment was conducted on in vitro regeneration of tomato at the Plant Biotechnology Laboratory, Department of Horticulture, Patuakhali Science and Technology University, Patuakhali. The objective was to develop an efficient regeneration protocol in tomato through callus induction for subsequent plantlet regeneration. Seeds were inoculated on MS medium where germination rate was 78.4%. The stems of in vitrocultured seedlings were used as explants. Different concentrations and combinations of growth regulators were added to MS medium to observe their efficacy on callus induction, shoot initiation and root formation. Stem explants cultured on MS medium fortified with 2 mg/L BAPgave the highest number of shoots (3.0) at 45 DAC. Among the concentrations of PGRs, 0.25 mg/L IAA produced the highest length (4.064 cm) of plantlets, number (5.0) of leaves and fresh weight (0.663 g) of plantlets with the stem explants at 45 DAC. The concentration of 0.5 mg/L IAA produced the highest number (21.00) of roots/plantlet, length (7.676 cm) of roots at 45 DAC, from the same explants. The highest survival rate of in vitro regenerated plantlets in the pot was 70.00 % with the stem explants. The results of the current study showed significant increase in the growth of callus of Solanumlycopersicon Mill. Indicating a good efficiency of the optimized media composition and the experimental model used in comparison to other studies of similar nature.Asian J. Med. Biol. Res. December 2015, 1(3): 589-599

2021 ◽  
Vol 5 (1) ◽  
pp. 61-67
Author(s):  
Sitti Inderiati ◽  
FNU Yanti ◽  
Eka Ria Mentari

In vitro propagation is a method to produce massive healthy new planting materials quickly. An experiment was carried out for morphogenic callus induction and regeneration of a domestic sugarcane variety. The Explants used was an inert folded leaf and incubated on modified MS medium augmented with 1 mg/l, 2.5 mg/l, and 5 mg/l of 2,4-D for callus induction. The leaf calluses were subcultured on MS medium enriched with different growth regulators for shoot initiation and multiplication. The highest percentage of callus formation was achieved in the medium containing 2.5 mg/l of 2,4-D, while the fastest callus initiation was noticed in MS medium supplemented with 5 mg/l 2,4-D, and maximum proliferation and the morphogenic response of callus were obtained in 3rd subculture. Two types of callus observed on the induction medium were dry nodular friable and smooth compact. This highly morphogenic callus was white to white creamy in color and easy to separate.   The highest shoot proliferation rate was found on the medium containing 2 mg/l Kinetin + 1 mg/l IAA and no growth was noticed on the medium containing Kinetin alone. Therefore, the study suggests that the growth hormone of cytokinin in combination with auxin is necessary for in vitro regeneration of sugarcane callus culture.


2019 ◽  
Vol 8 (4) ◽  
pp. 6934-6938

The present work was carried out to investigate the effects of gamma radiation on regeneration of Capsicum annuum L. var Kulai via in vitro. Seeds of C. annuum were irradiated with various doses of gamma ray (0, 20, 40, 60, 80, 100, 200, 300, 400, 500, and 600 Gy) emitted from the Caesium-137 source at the rate of 4.31 Gy per minute. Irradiated seeds grown on MS medium without hormone for hypocotyl and cotyledon preparation as explant for in vitro regeneration. Seed germination rate revealed significant variation between treatments, and seeds started to germinate between 6 to 17 days. Irradiated seeds between 0-60 Gy were observed to germinate in less than 10 days. All explants including hypocotyl and cotyledon were cultured on MS medium with different concentrations of BAP in combination with AgNO3 to observe the response of these explants to different hormone concentrations. From the observation, calluses were induced in 90% of hypocotyl and cotyledon explants in all treatments. The characteristics of calluses were varied with greenish friable, greenish compact, yellowish watery, yellowish friable and yellowish compact. In other treatments, calluses were found in purple, bright yellow and yellowish orange. On the other hand, shoot regeneration was observed in treatment between 40-100 Gy. In conclusion, gamma radiation gave impact on seed germination, seedling growth performance, in vitro callus formation and shoot regeneration of Capsicum annuum var. Kulai


2017 ◽  
Vol 14 (2) ◽  
pp. 607-614
Author(s):  
Hossein Nazarian ◽  
Maryam Beigi Harchegani ◽  
Mahmoud Otroshy ◽  
Ali Motamedi

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.


1970 ◽  
Vol 35 (1) ◽  
pp. 125-134 ◽  
Author(s):  
MA Rahman ◽  
MA Alam ◽  
MR Hossain ◽  
A Hossain ◽  
R Afroz

Regeneration ability of five Nicotiana varieties viz., Virginia, Jati, Motihari, CC Bengal and Sumatra were investigated via callus induction using leaf discs. Explants were cultured on MS medium supplemented with different concentrations and combinations of plant growth regulators. Callus formation frequency was 67.20%. Among the varieties used, Motihari induced the highest percentage (97.50%) of callus followed by Jati (92.50%) in 2.0 rng/L Kinetin and 2.0 mg/L IAA. Shoots were induced from calli cultured on the same medium. Maximum shoot formation from leaf discs was 82.50% on medium supplemented with 2.0 mg/L Kinetin and 2.0 mg/L IAA. It was also revealed from this study that Motihari was the best variety for callus formation and subsequent plantlet regeneration which is a pre-requisite for vector mediated transformation for varietal improvement of Nicotiana species. The rooting response of regenerated shoots was observed by using ½ MS medium with IBA (0.0, 0.5, and 1.0 mg/L). The highest root formation was found in Motihari (90%) with ½ MS medium supplemented with 0.5 mg/L IBA. After that regenerated plantlets with plenty of roots were transferred successfully to pots and subsequently to the field. Keywords: Tobacco; Nicotiana; in vitro regeneration; callus induction; plantlet regeneration; leaf disc; phytohormone. DOI: 10.3329/bjar.v35i1.5873Bangladesh J. Agril. Res. 35(1) : 125-134, March 2010


2012 ◽  
Vol 40 (2) ◽  
pp. 140 ◽  
Author(s):  
Hafiz Mamoon REHMAN ◽  
Iqrar Ahmad RANA ◽  
Siddra IJAZ ◽  
Ghulam MUSTAFA ◽  
Faiz Ahmad JOYIA ◽  
...  

Dalbergia sissoo Roxb. ex DC. (Sissoo) is a native forest tree species in Pakistan. Many ecological and economical uses are associated with this premier timber species, but dieback disease is of major concern. The objective of this study was to develop a protocol for in vitro regeneration of Sissoo that could serve as target material for genetic transformation, in order to improve this species. Callus formation and plantlet regeneration was achieved by culturing cotyledons, immature seeds, and mature embryos on a modified Murashige and Skoog (1962) (MS) medium supplemented with plant growth regulators. Callus induction medium containing 2.71 ?M 2, 4-dichlorophenoxyacetic acid (2,4-D) and 0.93 ?M kinetin produced better callus on all explants tested compared to other treatments, such as 8.88 ?M 6-benzylaminopurine (BA) and 2.69 ?M ?-naphthalene acetic acid (NAA), or 2.71 ?M 2, 4-D and 2.69 ?M NAA. Shoot regeneration was best on MS medium containing 1.4 ?M NAA and 8.88 ?M BA compared to other treatments, such as 1.4 ?M NAA and 9.9 ?M kinetin, or 2.86 ?M indole-3-acetic acid and 8.88 ?M BA. Murashige and Skoog medium containing 1.4 NAA ?M and 8.88 ?M BA was better in general for regeneration regardless of callus induction medium and the type of explant used. Rooting was best on half-strength MS medium with 7.35 ?M indole-3-butyric acid. Regenerated plantlets were acclimatized for plantation in the field. Preliminary genetic transformation potential of D. sissoo was evaluated by particle bombardment of callus explants with a pUbiGus vector. The bombarded tissue showed transient Gus activity 1week after bombardment. Transformation of this woody tree is possible provided excellent regeneration protocols. The best combination for regeneration explained in this study is one of such protocols.


2017 ◽  
Vol 4 (2) ◽  
pp. 52-56
Author(s):  
Mallika Devi T

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.


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

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.


2011 ◽  
Vol 3 (3) ◽  
pp. 93-96
Author(s):  
Ayobola M.A. SAKPERE ◽  
Ejeoghene R. AYISIRE ◽  
Olufemi I. ABIOYE

This study investigated the potential of Launea taraxacifolia for in vitro regeneration. Stem and leaf explants were inoculated on full strength Murashige and Skoog (MS) medium supplemented with varying concentrations of 2, 4-dichlorophenoxyacetic acid (2,4-D). Leaf explants responded to all concentrations of 2,4-D used while stem explants responded to only two of the 2, 4-D concentrations suggesting that leaf explants might be a better source of explants. Leaf explants generated shoots on medium supplemented with 0.5 mg/l kinetin and 0.1 mg/l 2, 4-D. This study is the first report on in vitro regeneration of Launea taraxacifolia.


1970 ◽  
Vol 8 (2) ◽  
pp. 227-232 ◽  
Author(s):  
Sonia B Shahid ◽  
SZ Khan ◽  
L Hassan

The experiment was conducted to optimize a suitable protocol for in vitro regeneration in cole crops. Callus initiation was excellent in the variety Early Tropical. Highest percentage of callus proliferation was observed in Early Tropical (75.0%) followed by Tangail Special Pauslali (55.0%) and the lowest in Tara (40.0% ). Maximum callus proliferation (68.5%) was observed in MS + 3.0 mgL-1 BAP + 0.1 mgL-1 2,4-D + 2.0 mgL-1 AgNO3. Callus proliferation was lowest (40.0%) in MS + 2.5 mgL-1 BAP + 0.1 mgL-1 2,4-D + 2.0 mgL-1 AgNO3. MS medium supplemented with 3.0 mgL-1 BAP + 1.0 mgL-1 2,4-D + 2.0 mgL-1 AgNO3 was the best for shoot initiation & plantlet regeneration. The highest number of shoots per vial was 7.20 and the lowest number of shoots per vial was 4.40. Among the concentration MS + 3.0 mgL-1 BAP + 0.1 mgL-1 2,4-D + 2.0 mgL-1 AgNO3 showed the highest performance of shoots per vial. The variety Tangail Special Pauslali was the best for root initiation. Keywords: Brassica; Cole crops; Callus; In vitro; Regeneration DOI: 10.3329/jbau.v8i2.7930 J. Bangladesh Agril. Univ. 8(2): 227-232, 2010


2017 ◽  
Vol 20 (2) ◽  
pp. 81-87
Author(s):  
HN Barman ◽  
ME Hoque ◽  
RK Roy ◽  
PL Biswas ◽  
MAI Khan ◽  
...  

The study was conducted at Biotechnology Division of Bangladesh Rice Research Institute (BRRI) to investigate the effects of plant growing medium and plant growth regulator (PGR) for the callus induction and high frequency plantlets regeneration of indica rice. Ten indica rice varieties viz. BR5, BR11, BRRI dhan28, BRRI dhan29, BRRI dhan33, BRRI dhan41, BRRI dhan47, BRRI dhan48, BRRI dhan49 and BRRI dhan50 were cultured on MS, N6 and LS media. The MS medium was found better for callus induction as compared to N6 and LS media. Among the tested varieties BRRI dhan48 induced the highest percent and best quality callus. Interaction effects of BRRI dhan48 to MS medium yielded 71.55% callus induction. The regeneration efficiency of BRRI dhan48 was tested on MS medium supplemented with different combination of NAA plus BAP and NAA plus kinetin. MS medium supplemented with 2.0 mg L-1 NAA and 2.0 mg L-1 Kn was found the best in respect of percent regenerated (76.67%) plantlet as well as for the growth of plantlets in vitro.Bangladesh Rice j. 2016, 20(2): 81-87


Sign in / Sign up

Export Citation Format

Share Document