scholarly journals Callus Production Protocol for Aegle marmelos (L.) Corr.: As a Tool for Extraction of Secondary Metabolites

2020 ◽  
Author(s):  
D.L.C.K. Fonseka ◽  
H.N. Aluthgamage
Keyword(s):  
Secondary Metabolites ◽  
Seed Coat ◽  
Callus Induction ◽  
Dry Weight ◽  
Ms Medium ◽  
Aegle Marmelos ◽  
Fresh Weight ◽  
Randomized Design ◽  
Optimized Protocol ◽  
Callus Production

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).

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
Keyword(s):  
Survival Rate ◽  
Callus Induction ◽  
Indole Acetic Acid ◽  
Casein Hydrolysate ◽  
Dry Weight ◽  
Ms Medium ◽  
Fresh Weight ◽  
Benzyl Adenine ◽  
Mature Embryos ◽  
Survival Percentage

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.

The growth and reserpine content of callus culture of Rauvolfia verticillata on the variation of sucrose concentration in MS medium

2007 ◽  
Vol 5 (1) ◽  
pp. 38-46
Author(s):  
IRMAWATI IRMAWATI ◽  
SOLICHATUN SOLICHATUN ◽  
ENDANG ANGGARWULAN
Keyword(s):  
Callus Culture ◽  
Quantitative Data ◽  
Sucrose Concentration ◽  
Culture Method ◽  
Dry Weight ◽  
Ms Medium ◽  
Fresh Weight ◽  
Significance Level ◽  
Randomized Design ◽  
Two Stages

Irmawati, Solichatun, Anggarwulan E. 2007. The growth and reserpine content of callus culture of Rauvolfia verticillata on the variation of sucrose concentration in MS medium. Biofarmasi 5: 38-46. The aim of this research was to study the effect of the variation of sucrose concentration on the growth and reserpine content on callus culture of Rauvolfia verticillata (Lour.) Baillon. The research was conducted with callus culture method consisted of two stages. The first stage was callus initiation to induce callus from leaf explant of R. verticillata, and the second stage was the reserpine production on treatment medium. This research used a Completely Randomized Design (CRD) by one factor, i.e. the variation of sucrose concentration. The sucrose concentration consisted of five levels, i.e. 0 g/L, 10 g/L, 20 g/L, 30 g/L and 40 g/L, each concentration in five replicates. The collected data consisted of qualitative and quantitative data. The qualitative data, callus morphology, was presented descriptively. The quantitative data, included fresh weight callus, dry weight callus and reserpine content, were analyzed by using ANOVA and followed by DMRT at 5% significance level. The result of research showed that the variation of sucrose concentration influenced fresh weight callus, dry weight callus and reserpine content. The increasing of sucrose concentration tended to raise callus growth, which could be seen from the fresh and dry weight callus. The highest fresh weight callus was found in medium with sucrose concentration of 20 g/L, while the highest dry weight callus was found in medium with sucrose concentration of 40 g/L. The increasing of sucrose concentration until 30 g/L raised reserpine content, but the sucrose concentration over 30 g/L decreased the reserpine content.

scholarly journals Sterilization protocols and the effect of plant growth regulators on callus induction and secondary metabolites production in in vitro cultures Melia azedarach L.

AMB Express ◽  
2022 ◽  
Vol 12 (1) ◽  
Author(s):  
Fatemeh Ahmadpoor ◽  
Nasser Zare ◽  
Rasool Asghari ◽  
Parisa Sheikhzadeh
Keyword(s):  
Secondary Metabolites ◽  
Plant Growth ◽  
Plant Growth Regulators ◽  
Growth Regulators ◽  
Callus Induction ◽  
Callus Cultures ◽  
Melia Azedarach ◽  
Ms Medium ◽  
Fresh Weight ◽  
Metabolites Production

AbstractMelia azedarach L. is a valuable source of antioxidants and secondary metabolites. This study is a first extensive report about the effect of different serialization protocols and plant growth regulators (PGRs) on explant disinfection efficiency, callus induction and secondary metabolites production and accumulation in callus cultures of M. azedarach L. In this regard, the effect of plant growth regulators on callus induction and secondary metabolites production were examined. In addition, different sterilization agents were evaluated for disinfection of chinaberry leaf explants. The results showed that the lowest percentage of explant contamination and browning with the highest percentage of callus induction and callus growth obtained with explants pretreated with benomyl (2 g/L) for 2 h and sterilized with 7% H2O2 for 10 min and NaOCl 2% (without pH adjustment) for 12 min. Although adjusting the pH of NaOCl to pH  = 7 and 10 significantly reduced the microbial contamination and increased the percentage of contamination-free cultures of M. azedarach L., adversely influenced the explant viability and callus induction and growth. The highest percentage of callus induction obtained on the MS medium containing 3 mg/L NAA/2,4-D and 1 or 3 mg/L Kin/BAP, and the highest callus yield (1804.833 mg/explant) belonged to the MS medium supplemented with 5 mg/L 2,4-D and 5 mg/L Kin. The callus cultures grown on the MS medium supplemented with 3 mg/L NAA and 1 mg/L Kin produced the highest amount of Quercetin (2.06 mg/g fresh weight), Rutin (5.56 mg/g fresh weight) and Kaempferol (1.84 mg/g fresh weight).

scholarly journals Effect of Different Strenght of Medium on Germination and Seedling Growth of Tomato and Sucrose Effect on Biomass of Tomato Callus

Jurnal Biota ◽  
2020 ◽  
Vol 6 (1) ◽  
pp. 12-18
Author(s):  
Arkan Setiaji
Keyword(s):  
Seedling Growth ◽  
Plant Cell Culture ◽  
Sucrose Concentration ◽  
Dry Weight ◽  
Ms Medium ◽  
Fresh Weight ◽  
Cytoprotective Activity ◽  
Callus Production ◽  
Sucrose Effect ◽  
Nutraceutical Products

Bioactive compounds of plant cell culture extracts are currently being looked at for its potential for the development of nutraceutical products. Tomato callus is known to have cytoprotective activity. The availability for explants and its callus production are two important things. This paper reports the effect of different strengths of medium on seed germination and seedling growth of tomato cv. Permata and sucrose effects on their biomass callus. Seeds were grown on 4 kind of germination media. Hypocotyl were taken as explants for callus induction in MS medium supplemented by 2.0 mg/l BA and 0.2 mg/l NAA, 3% sucrose, and subcultured after 20 days on medium with 6 variations of sucrose. The highest values ​​for tomato hypocotyl’s height were obtained from the ½ and ¼ MS, while the highest fresh weight was obtained from the MS 0. Both the fresh weight and dry weight of the callus increased along with increasing sucrose concentration. The explant age and biomass need to be a concern regarding what kind of callus that will be produced.  Application of 5% sucrose concentration is recommended for callus production in this study.

scholarly journals Pengaruh Hormon Indole Butyric Acid (IBA) dan 6-Benzyl Amino Purin (BAP) terhadap Induksi Kalus Piper betle L. var Nigra

2019 ◽  
Vol 4 (2) ◽  
pp. 85-90
Author(s):  
Junairiah . ◽  
Artifa Rachmah ◽  
Yosephine Sri Wulan Manuhara ◽  
Ni’matuzahroh . ◽  
Lilis Sulistyorini ◽  
...  
Keyword(s):  
Butyric Acid ◽  
Growth Regulators ◽  
Callus Induction ◽  
Callus Formation ◽  
Dry Weight ◽  
Piper Betle ◽  
Fresh Weight ◽  
Indole Butyric Acid ◽  
Randomized Design ◽  
Benzyl Amino Purine

ABSTRAKPenelitian ini bertujuan untuk mengetahui pengaruh kombinasi konsentrasi zat pengatur tumbuh IBA (Indole Butyric Acid) dan BAP (6-Benzyl Amino Purine) yang paling baik untuk induksi kalus sirih hitam (Piper betle L.). Penelitian ini menggunakan rancangan acak lengkap dengan 25 perlakuan dan setiap perlakuan memiliki 6 ulangan sehingga terdapat 150 unit ekperimen. Pada tahap kultur kalus dilakukan dengan menambahkan zat pengatur tumbuh IBA dan BAP ke dalam medium Murashige and Skoog (MS). Hasil uji tersebut menunjukkan bahwa zat pengatur tumbuh IBA dan BAP dengan kombinasi konsentrasi berbeda berpengaruh terhadap waktu induksi kalus, berat segar dan berat kering kalus sirih hitam. Hasil penelitian menunjukkan waktu tercepat pembentukan kalus pada IBA 2,0 mg/L dan BAP 2,0 mg/L yaitu 10 hari. Berat segar dan berat kering tertinggi pada IBA 2,0 mg/L dan 2,0 mg/L yaitu 0,8507 gram untuk berat segar dan 0,0769 untuk berat kering. Warna kalus adalah putih kehijauan dengan tekstur kompak dan remah.Kata kunci: Induksi kalus, Piper betle L., Indole Butyric Acid, 6-Benzyl Amino Purine.ABSTRACTThe purpose of this research to determine the effect of the combination concentration of growth regulators IBA (Indole Butyric Acid) and BAP (6-Benzyl Amino Purine) was best for callus induction black betel (Piper betle L.). This research used completely randomized design with 25 treatments and 6 replicates of each treatment, hence there were 150 experimental units. At this stage of callus culture was done by adding the growth regulators IBA and BAP into Murashige and Skoog (MS). The test results showed that plant growth regulators IBA and BAP in combination with different concentrations of influence on callus induction time, fresh weight and dry weight callus Piper betle L. The results showed the fastest time of callus formation at IBA 2,0 mg/L and BAP 2,0 mg/L at 10 days. Fresh weight and dry weight of the highest in the IBA 2,0 mg/L and BAP 2,0 mg/L were 0,8507 grams and 0,0769 grams fresh weight to dry weight. The color of callus was white greenish with compact and friable texture.Keywords: Callus induction, Piper betle L., Indole Butyric Acid, 6-Benzyl Amino Purine.

scholarly journals INFLUENCE OF AUXIN ON IN VITRO SECONDARY METABOLITES PRODUCTION FROM BALANITES AEGYPTIACA CALLUS CULTURES

2021 ◽  
Vol 21 (2) ◽  
Author(s):  
Asmaa A.A. Abdel- Kareem ◽  
H.A. El- Shamy ◽  
A.K. Dawh ◽  
S.G. Gwiefel
Keyword(s):  
Secondary Metabolites ◽  
Callus Cultures ◽  
Total Phenolic ◽  
Total Flavonoids ◽  
Ms Medium ◽  
Total Phenolic Compounds ◽  
Fresh Weight ◽  
Balanites Aegyptiaca ◽  
Metabolites Production

The present work was conducted in order to investigate the effect of auxin type (2,4-D and NAA) and concentration (0.00, 0.25, 0.50, 1.00 and 2.00 mg/l) on Balanites aegyptiaca callus cultures growth and production of secondary metabolites. Obtained results demonstrated that supplementation MS medium with 2,4-D at 2.0 mg/l could enhanced and recorded the ultimate values of callus fresh weight, antioxidant activity (%), total flavonoids, total phenolic compounds and total saponins contents and yields of Balanites aegyptiaca L. callus.

scholarly journals INDUCING SOME SECONDARY METABOLITES FROM CALLUS CULTURES DERIVED FROM Plantago psyllium AND Plantago major EXPOSED TO COBALT STRESS

2020 ◽  
Vol 51 (3) ◽  
pp. 938-943
Author(s):  
S. I. Neamah
Keyword(s):  
Secondary Metabolites ◽  
Dry Weight ◽  
Callus Cultures ◽  
Cobalt Metal ◽  
Plantago Major ◽  
Fresh Weight ◽  
Skoog Medium ◽  
Flavonoid Compounds ◽  
Metal Treatment ◽  
Plantago Psyllium

This experiment was conducted to study the influence of cobalt concentrations on the production of seven flavonoid compounds in callus derived from Plantago psyllium L. and Plantago major L. Results showed that the best combination of 2,4-D and  kinetin concentrations add to Muroshige and Skoog medium to obtain the highest fresh weight of 541.0 mg was 3.0 and 1.0 mg.L-1 respectively. psyllium stimulated callus produced the highest fresh weight of 365.7 mg. The addition of 75 ppm of cobalt resulted in a significantly lower fresh weight of P. psyllium callus (139.8 mg). The interaction between Plantago species and cobalt concentrations was significant. The callus inducted from P. major had significant increases of the scutallarein, apigenin, nepetin and luteolin compounds with 26.40, 22.64, 14.93 and 26.20 µg.100mg-1 dry weight, respectively. The production of the hispidulin compound was increased in P. psyllium at 29.40 µg.100mg-1 dry weight. Also, the addition of cobalt metal stimulated the production of flavonoids at 50 ppm cobalt producing the highest amounts of hispidulin and luteolin at 40.30 and 41.60 µg.100mg-1 dry weight, respectively. Meanwhile, 75 ppm cobalt treatment produced the highest amount of scutallarein, apigenin, nepetin and aucubin at 25.61, 23.25, 15.90 and 13.70 µg.100mg-1 dry weight, respectively. The callus inducted from P. major treated with 50 ppm of cobalt showed the highest production of scutallarein, apigenin and luteolin at 30.33, 32.26 and 51.90 µg.100mg-1 dry weight respectively. Baicalein reached 16.46 µg.100mg-1 dry weight, at 75 ppm of cobalt metal treatment in callus inducted from P. psyllium.

scholarly journals PEMANFAATAN PUPUK ORGANIK CAIR PAITAN (Tithonia diversifolia) PADA BERBAGAI DOSIS DAN RAGAM APLIKASINYA TERHADAP PERTUMBUHAN KAILAN (Brassica oleracea Var. Acephala)

2020 ◽  
Vol 11 (1) ◽  
pp. 25
Author(s):  
AHMAD TAOFIK ◽  
SOFIYA HASANI ◽  
AYU CAHYANINGTYAS ◽  
BUDY FRASETYA
Keyword(s):  
Leaf Area ◽  
Plant Height ◽  
Analysis Of Variance ◽  
Growth Parameters ◽  
Organic Fertilizer ◽  
Dry Weight ◽  
Fresh Weight ◽  
Application Method ◽  
Randomized Design ◽  
Two Factors

The paitan plant contains allelopathic compounds for other plants so that the utilize of paitan plant as a source of liquid organic fertilizer (LOF) has not been widely applied. This research aimed to study the interaction between LOF dosage and appropriate application method of LOF to increase the growth of kailan plants. The research conducted at experimental garden of UIN Bandung from June-August 2016. The experimental design was a completely randomized design with two factors and replicated four times. The first factor was application methods (A), i.e. a1=direct application to the soil, and a2= spraying. The second factor was LOF dosage (T), i.e. t0: 0 ml plant-1, t1: 20 ml plant-1, t2: 40 ml plant-1, t3: 60 ml plant-1, t4: 80 ml plant-1. The growth parameters observed i.e. plant height (14, 21, 28, 35 and 42 Day after Transplanting (DAT)), leaf area 42 DAT, plant dry weight 42 DAT, and plant fresh weight 42 DAT. The data then analyzed with analysis of variance at a significant level of 5%. If the analysis of variance was significant, the Duncan multiple range test was used at α=5%. The research results showed there was no interaction between method of application with a various dosage of paitan LOF to all of the growth parameters. The application paitan LOF showed similar growth in terms of plant height, leaf area, plant dry, and plant fresh weight. The application method of paitan LOF direct to the soil or spraying to leaves can not increase kailan plant growth.

scholarly journals Plant Regeneration Through Nodal Explant Derived Callus in Wood Apple (Aegle marmelos L.)

1970 ◽  
Vol 44 (4) ◽  
pp. 415-420 ◽  
Author(s):  
Ranjoy Das ◽  
M Faruk Hasan ◽  
Harunar Rashid ◽  
Motiur Rahman
Keyword(s):  
Plant Regeneration ◽  
Callus Induction ◽  
Nodal Segments ◽  
Nodal Segment ◽  
Ms Medium ◽  
Nodal Explant ◽  
Aegle Marmelos ◽  
Half Strength ◽  
Subsequent Regeneration

This study reports on an improved protocol for callus induction and subsequent regeneration from nodal segment of wood apple (Aegle marmelos L.) Creamish friable competent callus was achieved from nodal segments on MS medium augmented with 4.0 mg1-1 2,4-D within two weeks of inoculation. The callus produced large number of shoots when cultured on MS medium fortified with 2.0 mgl-1 BAP+0.1 mgl-1 NAA within ten days of culture. In vitro raised shoots were rooted on half strength MS medium enriched with 1.0 mgl-1 IBA within fifteen days of culture. The rooted plantlets were successfully established with 80% survival. Key words: Plant regeneration; Callus induction; Nodal explant; Aegle marmelos. DOI: 10.3329/bjsir.v44i4.4590 Bangladesh J. Sci. Ind. Res. 44(4), 415-420, 2009

scholarly journals Pengaruh Campuran Pasir dan Serat Aren pada Kailan (Brassica oleraceae) dengan Hidroponik Substrat

2014 ◽  
Vol 16 (1) ◽  
pp. 19
Author(s):  
Samanhudi Samanhudi ◽  
Dwi Hardjoko ◽  
Riandy Adhitya
Keyword(s):  
Wood Fiber ◽  
Dry Weight ◽  
Fresh Weight ◽  
Randomized Design ◽  
The Media ◽  
Number Of Leaves ◽  
Completely Randomized Design ◽  
Brassica Oleraceae ◽  
Media Mix ◽  
Leaf Greenness

<p>Wood fiber waste of arenga produced from starch palm industry. It is not handled properly so that could be potentially cause problems for the surrounding community. Wood fiber waste of arenga is potentially used as a medium for hydroponics. The waste can be utilized to get a new composition of hydroponic media. This study aims to determine the effect of a mixture of sand with arenga fiber waste. This research was carried out by using a mixture of sand and arenga wood fiber waste as a medium. This is an experimental study using a completely randomized design. This study aims to determine the effect of a mixture of sand and arenga fiber waste for kailan growth. The results showed that the mixture of sand and arenga wood fiber waste has no effect on kailan growth. The combination of mixtures of sand and arenga fiber waste showed no interaction. Observations on plants such as plant height, number of leaves, leaf greenness levels, the fresh weight of crop stover and stover dry weight of the plant, showed a row of the highest results in the media mix washed sand, washed sand and rice husk</p>

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