scholarly journals Picloram-induced enhanced callus-mediated regeneration, acclimatization, and genetic clonality assessment of gerbera

Author(s):  
Saikat Gantait ◽  
Manisha Mahanta

Abstract Background Gerbera jamesonii Bolus ex Hooker f. (African daisy) is listed among the top five most important ornamental plants in the global floricultural industry. To satisfy its demand, the floriculture industry relies on reproducible and effective propagation protocol while retaining the genetic uniformity of G. jamesonii. The present study, for the first time, reports the potential of picloram for enhanced induction of organogenic calli from leaves of G. jamesonii and its high-frequency indirect regeneration. Results The fastest induction of calli with maximum fresh and dry weight was recorded in the Murashige and Skoog (MS) semisolid medium supplemented with 1 mg/l picloram. In addition, callus induction was observed in 2,4-dichlorophenoxy acetic acid- and α-napthaleneaceticacid-supplemented media but with delayed response and reduced fresh and dry weight. The proliferated calli were transferred to shoot induction media containing MS salt and 0.5–1 mg/l N6-benzylaminopurine, kinetin, or thidiazuron. A mean number of ~6 shoots per callus were developed after 5 days of culture in the MS medium supplemented with 1 mg/l kinetin, with a mean length of 5.2 cm. Successful rooting of shoots was achieved in the MS medium fortified with 1.5 mg/l indole-3-acetic acid, wherein the earliest root initiation (~5 days), as well as the maximum number (~9) and length (~4.8 cm) of roots, were recorded. Complete plantlets were primarily acclimatized in sand before being transferred to a mixed substrate (of soil, sand, tea leaf waste, and cow urine) that secured >90% survival and further growth of the plantlets. Eventually, clonal fidelity of the in vitro regenerants assessed via inter-simple sequence repeats (ISSR) primers exhibited a monomorphic banding patterns that suggested genetic integrity within the plantlets as well as with their mother plant. Conclusions The results of the present study should be of interest for commercial propagation and mutagenesis- as well as genetic transformation-related research.

2016 ◽  
Vol 47 (3) ◽  
Author(s):  
Al- Khazali & Hamad

This  research  was  conducted  in  the  plant  tissue  culture  Lab. College  of Agriculture / University  of  Baghdad  from  February to  October  2015. The aim  of  the  study  was  investigating  the  influences  of  combinations  of  Naphthalene  acetic  acid (NAA) , Thidiazuron (TDZ) Spermidine  (Spd. ) and 2,4-Dichlorophenoxy  acetic  acid (2,4-D) , Benzyl  adenine (BA) on callus  induction  and  adventitous  shoot  regeneration  originated  from  cotyledon  of  Citrus volkameriana  seeds. Seeds  were  disinfested  with 0.1 % of  HgCl2  for 15 minutes. The MS  medium  supplemented  with  (0.0,1.5 , 3.0 ) mg L-1  NAA in combination with (0.0, 0.05, 0.1) mg L-1  TDZ and (0.0, 0.5 ,1.0) mg L-1 Spd. and MS medium supplemented with (0.0, 1.5 , 3.0) mg L-1  2,4-D in combination with (0.0 ,1.0 , 2.0 )  mg L-1  BA and (0.0 ,0.5 , 1.0) mg L-1  Spd. the  interaction between 1.5 mg L-1  NAA and (0.05 , 0.1)  mg L-1  TDZ and the interaction between 3.0 mg L-1  NAA and (0.05 ,0.1) mg L-1  TDZ with all concentrations of Spd.   gave  the  highest  percentage of  callus  induction  100 % . While  the  MS  medium  supplemented  with  3 mg L-1 of  2,4-D in  combination  with  all  concentrations  of  BA  and  spd.  gave  the  highest  percentage 100 % of  callus induction. Results showed that  MS medium supplemented  with 1.5 mg L-1  NAA in combination  with  0.1 mg L-1 TDZ and  1.0 mg L-1 spd.  gave  the  highest  values  of  fresh  and  dry  weight  of  callus  (668.8, 44.59 ) mg  respectively . While  the  MS  medium  supplemented  with  3 mg L-1 2,4-D  in combination with 1.0 mg  L-1  spd.  And 0.0 mg L-1 BA gave  the  highest  values  of  fresh  and  dry  weight  of  callus  (709.2 , 47.28 ) mg  respectively.


2009 ◽  
Vol 36 (No. 4) ◽  
pp. 140-146 ◽  
Author(s):  
J.K. Kanwar ◽  
S. Kumar

The influence of growth regulators, explants and their interactions on in vitro shoot bud formation from callus was studied in <I>Dianthus caryophyllus</I> L. The leaf and internode explants were cultured on Murashige and Skoog (MS) medium containing different concentrations of growth regulators. The highest callus induction was observed with 2 mg/l 2,4-dichlorophenoxy acetic acid (2,4-D) and 1 mg/l benzyl adenine (BA). Out of twenty seven shoot regeneration media tested, only 2 mg/l thidiazuron (TDZ) and zeatin alone or in combination with naphthalene acetic acid (NAA) and/or indole acetic acid (IAA) could differentiate calli. The highest average number of shoots was observed with 2 mg/l TDZ and 1 mg/l IAA. Significant differences were observed in calli producing shoots and number of shoots per callus in the explants of leaf and internode. The shoots were elongated and multiplied on MS medium supplemented with 1 mg/l BA and solidified with 1% agar. The shoots were rooted and hardened with 76% survival success in pots after six weeks of transfer to the pots.


2016 ◽  
Vol 41 (3) ◽  
pp. 521-528
Author(s):  
MR Kabir ◽  
S Ahmed ◽  
MAY Akhond

Seedling-derived cotyledonary nodes and hypocotyl explants of BARI Dherosh- 1 were cultured in vitro on MS medium supplemented with varying concentrations of 2, 4-Dichlorophenoxy acetic acid (2, 4-D), 6- Benzylaminopurine (BAP), Thidiazuron (TDZ), BAP with 1-Nepthaleneacetic acid (NAA), BAP with Indole 3-butyric acid (IAA) and Zeatin with IAA along with a control. Shooting response (100%) with callus was only observed from cotyledonary nodes on thidiazuron (TDZ) where hypocotyls produced only callus or callus with roots on different concentrations of plant growth regulators. Considering the shooting response, the cotyledonary nodes of BARI Dherosh-1 were cultured on various concentrations of TDZ for regeneration. The highest percentage (64.0) with maximum number (6.8) of shoots per explant were observed in 0.044 ?M TDZ in 8.4 days. The regenerated shoots were rooted on ½ strength MS, MS supplemented with 2.46 ?M IBA and 0.53 ?M NAA. The highest percentage (83.3) and minimum days (9.7) required for root induction were recorded in 2.46 ?M IBA. The rooted plantlets were transferred to soil and hardened in the plastic pots under green house conditions. The rooted shoots grew normally under natural conditions following acclimatization.Bangladesh J. Agril. Res. 41(3): 521-528, September 2016


2016 ◽  
Vol 8 (1) ◽  
pp. 412-415 ◽  
Author(s):  
Archana Rani ◽  
M. Kumar ◽  
Sanjeev Kumar

An efficient protocol for callus induction of Withania somnifera through in vitro culture of shoot apex and leaf explant was standardized. Of the various combinations of phytohormones evaluated, MS media supplemented with 6-furfuryl aminopurine (KIN) 0.5 mg/l + 2,4-dichlorophenoxy acetic acid (2, 4-D) 2.0 mg/l was found to be bestfor mean callus induction (86%) in leaf explants after 6 weeks of culture and in case of shoot apex expant the best response and growth of callusing was observed on MS medium supplemented with 2,4-D 1.0 mg/l + BAP 2.0 mg/l (77%).The response of callus growth increases gradually with the reductions in concentration of KIN in culturemedium of both the explants. This protocol might be used in further research for mass propagation of W. somnifera via indirect regeneration methods.


1969 ◽  
Vol 83 (3-4) ◽  
pp. 169-173
Author(s):  
Keithley L. Amory ◽  
John M. Gill

Young leaves of Liriope muscari provide an ample source of explants for in vitro propagation in tropical countries where flowering is scarce. Leaves were induced to form calli on a solid medium containing Murashige and Skoog (MS) salts and vitamins, 3% sucrose, 0.7% agar, 1 mg/L 2,4-dichlorophenoxy- acetic acid (2, 4-D) and 1 mg/L 6-furfurylaminopurine (kinetin). Only the proximal segments of the leaves produced calli. These calli were induced to produce multiple plantlets on MS medium, 3% sucrose, 0.7% agar, and 10 mg/L N6 (2-isopentenyl) adenine (2 ip). It is possible to use leaf explants for in vitro mass production of Liriope. However, in variegated varieties, only green or white plants were produced, because of a chimera in the original tissue.


2011 ◽  
Vol 29 (3) ◽  
pp. 359-363 ◽  
Author(s):  
Jean C Cardoso ◽  
Elizabeth O Ono

One of the most important applications of plant tissue culture is mass propagation of ornamental plants. This experiment evaluated the effect of different concentrations of NH4NO3 and KNO3 and BAP on the in vitro growth of orchid hybrid Brassocattleya 'Pastoral'. Seedlings of this orchid hybrid were used as explants and cultivated in medium with mineral salts and vitamins from the MS medium (Murashige & Skoog, 1962), with the macronutrients P, Ca and Mg reduced by half, and with an addition of 25 g L-1 of sucrose, 0.1 g L-1 of myo-inositol and 1.5 g L-1 of activated charcoal. Agar-agar was added (6.5 g L-1) and the pH was adjusted to 5.8. As treatments, four concentrations of the NH4NO3 and KNO3 (2x; 1x; ½ and ¼ MS medium) and three concentrations of BAP (0.0; 0.5 and 1.0 mg L-1) were assayed. The multiplication, growth in height, fresh and dry weight and sugar level in dry weight of sprouts were evaluated. There occurred a higher growth in height with 0.25x NH4NO3 and KNO3 salts concentrations of MS medium and higher rate of multiplication with combination of NH4NO3 and KNO3 reduced by half of the MS medium concentration and 1.0 mg L-1 BAP.


2014 ◽  
Vol 8 (1) ◽  
pp. 66-71
Author(s):  
Abedaljasim M. Aljibouri ◽  
Silva A. Yakoub Zokian ◽  
Ali H. Almusawi

Callus cultures were initiated for four Euphorbia species. Nodule explants cultured on Murashige and Skoog (MS) medium, supplemented with different concentrations 0,0.5,1,1.5,2 mg/l of the auxin 2,4-Dichlorophenoxy acetic acid 2,4-D. Half of the cultures were incubated for16 hrs/day photoperiod, while the other half was incubated under complete darkness. The incubation temperature was 25±1ºC. Observations on number of nodule explants initiated callus were recorded at 2,4,6,8 weeks of culture. For callus maintenance, 50mg of callus produced were re-cultured on MS medium supplemented with different concentrations of 2,4-D 0,0.4,0.8,1.2,1.6 mg/l. Callus fresh and dry weights were recorded after 4 weeks. Results showed that nodule explants of Epeplus and Ehirta incubated under light conditions achieved the highest response to callus initiation 75-100% compared with the other species under experimental conditions. E.Helioscopia incubated under light conditions achieved the lowest response for callus initiation 25-75%. Results also showed significant differences between Euphorbia species in fresh and dry callus weights, E. Ehirta produced the highest fresh and dry weight of callus reaching 1.410 and 0.046 mg respectively. The amount of fresh and dry weight of callus produced under dark conditions was significantly higher than that produced under light conditions.


2011 ◽  
Vol 5 (2) ◽  
pp. 25-31
Author(s):  
A. A. M. Al-Salihy ◽  
Z. A. H. Al- Hussaini ◽  
Muhmmed. A. K.

hoots of two sugarcane genotypes (Co.J.64, Co.J.86) were rooted in vitro on MS medium supplemented with different concentration of sucrose (0, 30, 50, 60, 70) gm/l or NAA (0, 2.5, 5.0, 7.5, 10.0) mg/l in separate experiments. All cultures were incubated at 25±2C◦with 16 h/d light (1000 lux) for 60 days. Data of roots per plant, length, dry and fresh weights were taken after 8 weeks. Results showed that 70g/l of sucrose gave higher mean in length and fresh weight of roots reached 1.70 cm /plant, 19.80 mg/root, respectively, while 60g/l sucrose gave higher mean in number and dry weight reached 9.95 root, 4.60mg, respectively. NAA concentrations were significantly effected in root length reached 1.23 cm/plant at 5 mg/l NAA while a significant effect in root number, fresh and dry weight. The genotype Co.J.64 was the best in number of roots in sucrose experiment, while genotype Co.J.86 surpassed in number of root, fresh and dry weight in NAA experiment.


2016 ◽  
Vol 11 (3) ◽  
pp. 341
Author(s):  
Djadja Siti Hazar Hoesen

Vegetative propagation from bulb excised of Eleutherine sp. (Iridaceae) were cultured in Murashige and Skoog (MS) medium supplemented with plant growth regulator (PGR) Benzyl adenine (BA) 1 mg/l at initiation stage, BA (2 and 5) mg/l for induced shoot buds formation at multiplication stage. In this study also BA 2 mg/l and BA 2 mg/l combined with naphthalein acetic acid (NAA) 0.5 mg/l were treated for rooting planlets formation. Calli formation were induced with auxin PGR picloram 1 mg/l combined with 2,4 dichlorophenoxy acetic acid (2,4-D) 0.5 and 1 mg/l in concentrations. The media contained 2 mg/l cytokinin (BA) without auxin (NAA), resulted the highest shoot buds formation. Rooting planlets were produced in MS medium combined with BA and NAA.MS medium contains Picloram 1 mg/l and 2,4-D 1 mg/l was optimal for frequency oncalli initiation (100%) and the largest diameter of calli also represented in cultured MS medium with picloram 1 mg/l and 2,4-D 1mg/l. In acclimatization stage, 100% of planlets survived and successfully transplanted to soil medium in the field.Key words: in vitro, Eleutherine sp.


2009 ◽  
Vol 24 ◽  
pp. 82-88 ◽  
Author(s):  
Saraswoti Aryal ◽  
Sanu Devi Joshi

Rauvolfia serpentina (L.) ex. Kurz is an important medicinal plant. Callus induction and regeneration was studied from stem explant of in-vitro grown plant of Rauvolfia serpentina(L.) Benth. ex Kurz (Apocynaceae) on Murashige Skoog (1962) medium supplemented with 1mg/l 2,4-Dichlorophenocy acetic acid (2,4-D) and 1mg/l Kinetin (Kn). Vigorous growth of callus occurs after 4 weeks of culture. Callus was sub-cultured on Murashige and Skoog (MS) medium supplemented with different concentration of 2, 4-D (0.5-3.0 mg/l) and 10% coconut milk. Regeneration of plantlets occurred on MS medium containing 3 mg/1 of 2, 4-D and 10% coconut milk. These plantlets were rooted on MS medium supplemented with 1 mg/l IAA .The regenerated plantlets were able to grow on soil after short period ofacclimatization. Key words: Explant; In-vitro culture; MS medium;  2, 4 Dichlorophenoxy acetic acid; Kinetin; Callus; Tissue culture; Coconut milk. Journal of Natural History Museum Vol. 24, 2009 Page: 82-88


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