scholarly journals Effect of N-methyl-N-nitro-N-nitrosoguanidine (NTG) on callus induction and survival rate on mature embryos of beans (Phaseolus vulgaris)

2009 ◽  
Vol 3 (2) ◽  
pp. 91-98
Author(s):  
Sattar A. Shlahi ◽  
Zahra N. Hashim Al- Hattab

This research was conducted to study the effect of the chemical mutagen N-methyl-N-nitro-N-nitrosoguanidine on the percentage of callus induction and survival from mature beans embryos harvester cultivar. Seeds were treated with (0.2 or 0.4) millimolar of the mutagen NTG in combination with 0.0, 4 or 8% of ethanol, pH 5 ±2 0. for 24 h. Calli were induced on mature embryos by using MS medium with 0.5 mg/l of Benzyl adenine (BA), 1 mg/l Indole acetic acid (IAA) and 100 mg/l from each of Casein hydrolysate, Glycine, Asparagine, Tyrosine, and Myo-Inositol. Results showed that the hypocotyl surpassed the radical and the plume significantly in terms of survival reached 56.3%. Mutagen treatments showed asignificant effect on calli survival. Treatment with 8% Ethanol was lethal for all explants. While treatment with 0.4 mM NTG without Ethanol gaved the highest survival rate. The interaction between the treatments and the explants showed that the lowest survival percentage was which 8.8% that was for shoots treated with 0.2 mM of 4% Ethanol. Calli induced on hypocotyls treated with 0.4 mM NTG without Ethanol gave the highest fresh weight (347.2) mg while the lowest was (60) mg for calli induced on the radical treated with 0.4 mM NTG with 4% Ethanol. Moreover the highest dry weight was 22.5 mg for calli induced from hypocotyls treated with 0.4 millimolar NTG without Ethanol that was higher than the control 17.2 mg.The lowest dry weight obtained from calli induced on the radical treated with 0.4 mM NTG with 4% Ethanol was 3 mg. In conclusion the results showed that 0.4 mM NTG without Ethanol gave the highest survival rate and the highest fresh and dry weight for calli induced on the hypocotyl.

2017 ◽  
Vol 15 (10) ◽  
pp. 753-763
Author(s):  
Thi Linh HO ◽  
Sompong TE-CHATO ◽  
Sureerat YENCHON

This study was conducted to determine the optimum concentrations of 2, 4-D, L-proline and casein hydrolysate (CH) for efficient callus induction and plantlet regeneration from culturing mature embryos of Sangyod, an economically important indica rice variety in Thailand. The highest frequency of callus induction (73.08 ± 2.65 %) was obtained from MS medium supplemented with 2 mg/L 2,4-D, 750 mg/L CH and 200 mg/L L-proline. The combination of 1 mg/L BA, 0.5 mg/L Kn and 0.5 mg/L NAA containing solidified MS medium gave the maximum mean fresh weight of callus (938.9 ± 44 mg), green spot formation (64.17 ± 7.08 %), shoot induction frequency (66.25 ± 6.80 %) and mean number of shoots/explant (6.12 ± 0.36 shoots). The greatest mean number of shoots/explant (14.93 ± 0.97 shoots) and root formation percentage (82.71 ± 3.03 %) was observed in liquified MS medium supplemented with 0.5 mg/L NAA and 1 mg/L BA.


2014 ◽  
Vol 8 (4) ◽  
pp. 79-84
Author(s):  
Sattar A. Shlahi ◽  
Zahra N. Al- Hattab

This research was conducted to study the effect of the chemical mutagen N-methyl-N-nitro-N-nitrosoguanidine (NTG)on the chromosomes of callus induced from mature bean embryos, Harvester cultivar. Seeds were treated with 0.2 or o.4 mM of the mutagen that mixed with different percentages of ethanol for 24 hrs. Calli were induced on MS medium in the presence of 0.5 mg/L of Benzyl adenine (BA), 1 mg/L Indole acetic acid (IAA) and 100 mg/L from each of Casein hydrolysate, Glycine, Asparagine, Tyrosine, and Myo-Inositol. Samples were pretreated with 1, 2- benzene dichloride, Para –dichlorobenzene, or Colchicine. Two different staining methods were used to stain the chromosomes from root tips and calli.The results showed that Para –dichlorobenzene is the best pretreatment for both root tips and callicells. However, the stain acetoorcein was the best for the root tips while Feulgen stain was the best for calli cells. Chromosome count showed that there were 22 chromosomes in all the cells of bean root tips (control). While a wide range of chromosome numbers were obtained from calli cells with or without mutagen treatment. Ninety six percent of the non treatedcalli gave the normal number of chromosomes while only 60% of calli treated with (0.4 mM+4% ethanol) gave the normal number of chromosomes. Calli cells from all the treatments showed chromosome multiplication except in the presence of ethanol.


Author(s):  
D.L.C.K. Fonseka ◽  
H.N. Aluthgamage

Background: Aegle marmelos (L.) Corr. is an important medicinal and a fruit tree belongs to the family Rutaceae possessing numerous valuable secondary metabolites. The growing commercial importance for secondary metabolites has led to a great demand in the pharmaceutical industry in recent years. Therefore, an efficient callus production protocol was developed as a tool for extracting valuable secondary metabolites from Aegle marmelos.Methods: For seeds, callus induction was observed under three conditions as with seed coat, after removing seed coat and split into two halves after removing seed coat. For callus multiplication, 1cm2 pieces of initiated calli were used. These explants were established in MS medium supplemented with combinations of 2, 4 D either with BAP or Kinetin. All experiments were arranged according to the completely randomized design (CRD) with 20 replicates at the Tissue Culture Laboratory, Department of Crop Science, Faculty of Agriculture, University of Ruhuna, Sri Lanka, for a period of 1 year. Percentage of fungal and bacterial contaminations and percentage of bleached explants were observed to select the best explant/s. Percentage of responded explants were observed to select the best condition for callus induction and quality of callus. Growth of callus was observed visually by giving a score. Best hormonal combination for callus multiplication was observed as fresh weight and dry weight of callus produced under each treatment.Result: High quality callus with higher growth was observed in all combinations of BAP and 2, 4 D tested: ranging from 0.5 mgL-1 to 1.5 mgL-1 BAP and 1.0 mgL-1 to 2.0 mgL-1 2,4 D in Murashige and Skoog (MS) medium. Initiated calli were further multiplied in MS medium supplemented with 2,4 D combined with either BAP or Kinetin. Highest amount of callus biomass was recorded in the MS medium with 0.5 mgL-1 2, 4 D and 1.0 mgL-1 Kinetin (132.58 gL-1 fresh weight). The optimized protocol could be used to produce higher amount of callus in order to extract secondary metabolites from Aegle marmelos (L).


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.


2012 ◽  
Vol 60 (1) ◽  
pp. 47-55 ◽  
Author(s):  
A. Ali ◽  
T. Yossef ◽  
A. El-Banna

The present study was carried out for developing an efficient in vitro callus induction and plant regeneration system in four different tomato genotypes (Solanum lycopersicum Mill., previous name: Lycopersicon esculentum), Advantage II, Edkawy, Castle Rock and Super Strain B, using hypocotyl and cotyledon explants. The effects of two cytokinins, BA (benzyl adenine) and Kin (kinetin), on callus induction and plant regeneration frequency were investigated when added to MS medium in combination at varying concentrations. All concentrations of the two cytokinins were suitable for callus induction and plant regeneration. The frequency of callus induction and plant regeneration from both cotyledon and hypocotyl explants reached 100% for all tested genotypes. Cotyledons produced a higher average number of shoots per explants than hypocotyls for all the genotypes in the five concentrations of combined cytokinins. The average number of shoots per explant in Super Strain B was found to be the highest (42 and 60 for the hypocotyl and cotyledon explants, respectively). Supplementing MS medium with 1.0 mg L−1 kinetin and 1.0 mg L−1 benzyl adenine was found to be optimum for producing the highest number of shoots per explant from hypocotyls and cotyledons in the tomato genotypes investigated. The proposed medium showed a significant superiority over the reference media.


2019 ◽  
Vol 7 (4.14) ◽  
pp. 107
Author(s):  
D S M Soder ◽  
D N A A Khalid ◽  
A Saleh ◽  
F Pardi ◽  
N J Sidik

Barringtonia racemosa is mangroves type of plant which had been extensively utilized in conventional practices for relieving ailments of pain and inflammation. Many studies have been done on ethnobotanical profiles, pharmacological activities and chemical compounds in Barringtonia racemosa. However, there is a limited study on callogenesis of this plant particularly from different maturity stage of fruits. The present study is to identify the callogenesis of Barringtonia racemosa from endosperm explants of immature and mature fruits in MS medium supplemented with different concentrations of hormones 2,4-Dichlorophenoxyacetic acid (2,4-D) (0, 0.5, 1.0, 1.5 and 2.0 mg/L) and Kinetin (KIN) (0, 0.5, 1.0, 1.5 and 2.0 mg/L). The optimum hormone combination was found in callus grown on endosperm of immature fruits in MS medium supplemented with 1.5 mg/L 2,4-D and 1.0 mg/L KIN. It was also found that the callus in this treatment grew profusely with highest fresh weight (0.513 ± 0.022 g), 100% callus induction and friable callus texture. The callus fresh weight on endosperm explants was higher in immature fruits compared to mature fruits for all the hormone combinations. Therefore, callogenesis were found more efficient from endosperm explant of immature fruits in Barringtonia racemosa species.   


HortScience ◽  
1990 ◽  
Vol 25 (12) ◽  
pp. 1648-1650 ◽  
Author(s):  
Steven R. Turner ◽  
Suman Singha

Shoots of `Almey' crabapple [Malus baccata (L.) Borkh. × M. pumila var. niedzwetzkyana (Dieck) Schneid.], `Seckel' pear (Pyrus communis L.), and `Mrs. Bradshaw' geum (Geum quellyon Sweet.) were cultured on Murashige and Skoog (MS) medium supplemented with 8.8 μm BA and containing 0.1% to 0.4% Gelrite. Comparative shoot proliferation and vitrification were determined on Phytagar-solidified medium. Shoot proliferation, culture fresh weight, and vitrification declined in crabapple and geum with increasing Gelrite concentration. Pear proliferation and fresh weight increased with increasing Gelrite levels, but all shoots were vitrified. There were differences in the vitrification response between pear and the other two genera. The percent dry weight of vitrified cultures on Gelrite-containing media was generally higher than that of nonvitrified cultures on medium containing Phytagar. Vitrification precludes using low Gelrite concentrations for propagating these plants. Chemical name used: N-(phenylmethyl) -1H-purin-6-amine (BA).


2014 ◽  
Vol 8 (2) ◽  
pp. 27-34
Author(s):  
Emad H. Jassim ◽  
Sami K. M. Ameen

An experiment on the effect of sucrose and mannitol on leave induced callus of Catharanthus roseus was conducted from February 2011 to May 2012. Callus induction was achieved by culturing leaves of the plant on MS medium supplemented with 0.5 mg /L 2,4-D and 1mg / L Kin, The best medium to maintain callus was on MS medium modified with 0.5 mg /L 2,4-D and 1.5 mg / L Kin. when different levels were added to MS medium for each Mannitol 0, 6000 ,8000 ,10000 mg /L and Sucrose 40, 60 ,80, 100 g /L in split experimental and control treatment was MS medium supplemented with 30 g/L sucrose. The results showed that medium supplemented with 100 gL of sucrose gave the highest quantity of Ajmalicine 32.27 µg/100 mg fresh weight of callus,as well as medium supplemented with 8000 mgL of Mannitol gave the highest value of Ajmalicine 120.19 µg/100 mg fresh weight of callus. The concentrations of Ajmalicine, derived from the leaves of the plants grown in pot, were lowest than the concentrations produced by the callus grown in vitro it was 0.047 µg/100 fresh weight of the leaves.


Author(s):  
Dwi Kusuma Wahyuni ◽  
Putri Andriani ◽  
Arif Nur Muhammad Ansori ◽  
Edy Setiti Wida Utami

<p class="IsiAbstrakIndo"><em>Justicia gendarussa </em>Burm.f., a medicinal plant, is Acanthaceae that has many functions. Furthermore, the compounds in gendarussa must be produced in high quantity and quality by applying callus culture method. Accordingly, it is important to study the effects of plant growth regulators (2,4-D, IBA, and BAP) on callus induction of gendarussa leaves. This research design utilized a factorial design with two factors (2,4-D and IBA: 0.5, 1, 1.5 mg/L and BAP: 0.5, 1, 1.5, 2 mg/L). The experiment consisted of 24 treatments, each of which was repeated 3 times. Observation was carried out in 6 weeks. Data on the time of callus formation, percentage of explants formed callus, and callus morphology were analyzed descriptively,while data on fresh and dry weight were analyzed by Two-Way ANOVA (<span>α</span> = 0.5). Interestingly, the results showed that various concentration of plant growth regulators (2,4-D, IBA, and BAP) affected callus induction from leaf explants of gendarussa. We concluded that the most optimal treatment combination of concentration of plant growth regulators in inducing callus from leaf explants of gendarussa is 1.5 mg/L 2,4-D and 2 mg/L BAP with a relatively long period of callus formation at the earliest, i.e. on day 5, 2.247 g of fresh weight, 0.108 gof dry weight, white callus translucent, and friable. <span lang="EN-GB">Moreover, t</span>he optimum treatment will be used to produce secondary metabolite and seed s<span lang="EN-GB">y</span>nthetic by cell suspension culture.</p>


1970 ◽  
Vol 36 (3) ◽  
pp. 397-406 ◽  
Author(s):  
BP Ray ◽  
L Hassan ◽  
KM Nasiruddin

The effect of different explants and concentrations of BAP and NAA on induction of callus and plant regeneration of brinjal cv. Jhumki were investigated. The treatment combinations were BAP (0. 2.0. 3.0, and 4.0 mg/l) and NAA (0. 0.1, 0.5, and 1.0 mg/l). The rate of callus formation varied in different treatments. The highest amount of callus (48.66%) was produced on MS medium containing 2.0 mg/l BAP and 0.5 mg/l NAA from stem, and 8.2 days required for callus induction. The highest fresh weight of callus was 1.12g from stem and 0.48g from root. The number of shoot regenerated through callus from stem containing 2.0 mg/l BAP and 0.5 mg/l NAA was 3.4 (23.287%) and days required for 38.8 days. All regenerated plantlets survived in normal environment. Keywords: NAA; BAP; regeneration; brinjal. DOI: http://dx.doi.org/10.3329/bjar.v36i3.9268 BJAR 2011; 36(3): 397-406


Sign in / Sign up

Export Citation Format

Share Document