scholarly journals In Vitro Callus Induction from Leaf Explants of Vanda sp Stimulated by 2,4-D

2017 ◽  
Vol 9 (3) ◽  
pp. 492 ◽  
Author(s):  
Iman Budisantoso ◽  
Nurul Amalia ◽  
Kamsinah Kamsinah
Keyword(s):  
Success Factors ◽  
Callus Formation ◽  
Leaf Explants ◽  
Callus Growth ◽  
Growth Time ◽  
Critical Success ◽  
Randomized Design ◽  
The Difference ◽  
Treatment Concentration

<p class="IsiAbstrakIndo"><span lang="EN-GB">The addition of growth regulator is one of the critical success factors in in vitro cultures. 2,4-D as a plant regulator in media can stimulate the cell division, enlargement of the explants and promotes the formation and growth of callus. The purpose of this study was to determine the time of callus formation and to determine the best concentration of 2.4-D in inducing the growth of callus from leaf explants of </span><em><span lang="EN-GB">Vanda</span></em><span lang="EN-GB"> sp. This research was conducted by experiment with completely randomized design, which consists of six levels of treatment concentration of 2,4-D i.e. 0 ppm; 1 ppm; 1.5 ppm; 2 ppm; 2.5 ppm; and 3 ppm. The parameters observed were the percentage of callus formation and the form of callus from </span><em><span lang="EN-GB">Vanda</span></em><span lang="EN-GB"> sp leaf explants. The results were statistically analyzed by using MINITAB program version 17. Analysis of variance (ANOVA) was performed and the difference between means score/value was separated by F test at p &lt; 0.05. The results showed that 2,4-D treatment give significant effect (F 5,12 = 3,20; p = 0,046 &lt; 0,05) on the callus growth time and its percentage. Application of 2 ppm 2.4-D was the best concentration for accelerating the callus growth time (14.3 days after planting) and increasing the percentage of callus formation (83.3%). Most of callus type were proliferative callus (36.11%) and senescence callus (11.11%). The results of this research are very important to grow the callus from Vanda leaves orchid explant because it is very diffucult to grow.</span></p>

scholarly journals The effect of DL-tryptophan on callus growth and alkaloid-reserpin production of pule pandak [Rauvolfia serpentina (L.) Bentham ex Kurz] in vitro

2005 ◽  
Vol 3 (2) ◽  
pp. 52-56 ◽  
Author(s):  
HENI ARYATI ◽  
ENDANG ANGGARWULAN ◽  
SOLICHATUN SOLICHATUN
Keyword(s):  
Amino Acid ◽  
Quantitative Data ◽  
Callus Formation ◽  
Leaf Explants ◽  
Treatment Phase ◽  
Optimum Concentration ◽  
Callus Growth ◽  
Second Phase ◽  
Rauvolfia Serpentina

The purposes of this research were to study the effect of amino acid DL-tryptophan at various concentrations on culture callus growth and production of alkaloid-reserpine of Rauvolfia serpentina and to determine optimum concentration of DL-tryptophan to yield maximum alkaloid reserpine of the callus. This research was consisted of three phases. First phase was to determine compatible sterilan for R. serpentina leaf explants. Second phase was to initiate/ to induce callus formation from the explants. Third phase was treatment phase to know the influence of DL-tryptophan addition on growth and alkaloid-reserpine production of the callus. Experimental design that used was Completely Randomized Design (CRD). The treatment was concentration of the amino acid DL- tryptophan with three levels concentration three restating for each level. The levels were 0 mg/L, 10 mg/L and 20 mg/L. Obtained data were analyzed quantitatively and qualitatively. Quantitative data were wet weight, dry weight, growth rate and alkaloid content of the callus at each level of concentration. Qualitative data have been measured were compatible sterilant test and callus morphology (color and texture). Quantitative data then have been analyzed by analysis of variance (ANOVA) and continued with DMRT test at level 95%. The result indicated that addition of DL- tryptophan precursor had an effect on the reduction of the callus growth of R. serpentina in media MS in vitro. Addition of DL-tryptophan precursor to produce of alkaloid-reserpine of the callus of R. serpentina in media MS in vitro and optimum concentration of DL-tryptophan precursor that must be added to yield of alkaloid-reserpine could not be determined yet, it caused by the alkaloidreserpine was not detected yet by the thin layer chromatography (TLC).

FACTORS AFFECTING THE REGENERATION OF PEPPER (CAPSICUM ANNUUM L.)

HortScience ◽  
1990 ◽  
Vol 25 (9) ◽  
pp. 1120G-1120
Author(s):  
J. L. Jacobs ◽  
C. T. Stephens
Keyword(s):  
Growth Hormone ◽  
Silver Nitrate ◽  
Callus Formation ◽  
Leaf Explants ◽  
Regeneration System ◽  
Factors Affecting ◽  
Nitrate Concentrations ◽  
Leaf Differentiation ◽  
Whole Plants

Several growth hormone combinations and silver nitrate concentrations were examined for their effect on regeneration of different pepper genotypes. Primary leaf explants from in vitro seedlings were cultured on a revised Murashige and Skoog medium supplemented with auxin, cytokinin and 1.6% glucose. Combinations of different concentrations of indole-3-acetic acid (IAA), 0-5 mg/l, and 6-benzylaminopurine (BAP), 0-5 mg/l, were tested to determine the most effective medium for shoot primordium formation. Experiments with IAA and BAP did not result in a specific growth hormone combination appropriate for regeneration of all genotypes tested. Of the silver nitrate concentrations tested, 10 mg/l resulted in the best shoot and leaf differentiation and reduced callus formation. Differences in organogenic response of individual genotypes were evaluated on a single regeneration medium. Whole plants were regenerated from 11 of 63 genotypes examined. Based on these experiments, a reproducible regeneration system for pepper was developed with a total of 500 plants regenerated to date.

scholarly journals Induction and growth pattern of callus from Piper permucronatum leaves

2016 ◽  
Vol 18 (1) ◽  
pp. 142-148 ◽  
Author(s):  
M.R.A. SANTOS ◽  
M.C.M. GUIMARÃES ◽  
E.S. PAZ ◽  
G.M.O. MAGALHÃES ◽  
C.A. SOUZA ◽  
...  
Keyword(s):  
Growth Pattern ◽  
Callus Induction ◽  
Callus Formation ◽  
Leaf Explants ◽  
Cell Suspension Cultures ◽  
Callus Growth ◽  
Deceleration Phase ◽  
Stomach Pain ◽  
Perennial Shrub ◽  
A Cell

ABSTRACT Piper permucronatum is a perennial shrub, a medicinal plant native to the Amazon Rainforest. Traditionally, the tea of its leaves is used to combat menstrual and intestinal cramps, stomach pain, digestive problems, diarrhea, hemorrhage, and nausea. Its leaf’s essential oil is effective against Aedes aegypti larvae; its flavones and flavanones have a fungicidal effect against Clamidosporium cladosporioides and C. sphaerospermum; its hexanic extract is effective against Leishmania amazonensis. The objective of this study was to provide a protocol for callus induction from P. permucronatum leaves and an identification of the callus growth pattern, focusing on the deceleration phase, when the callus cells must be subcultured into liquid medium in order to produce a cell suspension cultures. Leaf explants were inoculated in a solid MS medium supplemented with factorial combinations of 2,4-D, BA, NAA and GA3. Callus formation was evaluated weekly until the 49th day. Subsequently, new explants were inoculated at the hormonal combination that resulted in the highest callus cell proliferation and, every seven days during a period of 70 days, samples were dried and weighed to determine the callus growth pattern. NAA and GA3 were not effective for callus induction. Combinations of 2,4-D and BA resulted in callus induction and proliferation. The highest percentage of callus induction was observed with the combination of 4.52 µM 2,4-D and 4.44 µM BA. The calluses thereby produced were friable and whitish. The callus growth pattern followed a sigmoid shape. The deceleration phase started on the 56th day of culture.

scholarly journals Callus Induction of Leaves and Stems in Krisan (Chrysanthemum morifolium Ramat cv Dewi ratih) with Alternative Foliar Fertilizers Media

2021 ◽  
Vol 9 (2) ◽  
pp. 109-115
Author(s):  
Ahmad Saifun Naser ◽  
Muhammad Wisnu
Keyword(s):  
Tissue Culture ◽  
Callus Formation ◽  
Leaf Explants ◽  
Chrysanthemum Morifolium ◽  
Stem Explants ◽  
Appearance Time ◽  
Randomized Design ◽  
The Media ◽  
Completely Randomized Design ◽  
Chrysanthemum Morifolium Ramat

Availability of quality seeds in production of krisan (Chrysanthemum morifolium Ramat cv Dewi ratih) cultivation is still rare, therefore research on seed multiplication through tissue culture is needed. The media used in tissue culture is relatively expensive for home industry. This study aims to determine the respond of leaf and stem explants using foliar fertilizers (Growmore, Gandasil D and Mutiara) as an alternative media for callus inductions. This study used a Completely Randomized Design (CRD) consisted of 4 treatments: P0: ½ MS + 0,25 mg/l BAP, P1 (Growmore + 0,25 mg/l BAP), P2 (Gandasil D + 0,25 mg/l BAP), P3 (Mutiara + 0,25 mg/l BAP). The variables observed in this study included callus appearance time, callus color and callus texture. The result of this study indicated that the use of BAP (6-Benzyl Amino Purine) affected the time of callus formation and callus morphology. Callus was formed on leaf explants 13 days after planting while on stem explants 7 days after planting and compact texture. Growmore + 0,25 mg/l BAP treatment yields the best callus on leaf explant, while Gandasil D + 0,25 mg/l BAP treatment yields the best callus on stem explant.

scholarly journals Callus Induction in Baru (Dipteryx alata Vog.) Explants

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
Keyword(s):  
Ascorbic Acid ◽  
Callus Induction ◽  
Callus Formation ◽  
Leaf Explants ◽  
Naphthalene Acetic Acid ◽  
Ms Medium ◽  
Alternative Technique ◽  
Propagation Methods ◽  
Dipteryx Alata

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.

scholarly journals Comparative analyses of stevioside between fresh leaves and in-vitro derived callus tissue from Stevia rebaudiana Bert. using HPLC

2015 ◽  
Vol 49 (4) ◽  
pp. 199-204 ◽  
Author(s):  
S Mahmud ◽  
S Akter ◽  
IA Jahan ◽  
S Khan ◽  
A Khaleque ◽  
...  
Keyword(s):  
Retention Time ◽  
Callus Tissue ◽  
Callus Formation ◽  
Leaf Explants ◽  
Stevia Rebaudiana ◽  
Poor Performance ◽  
The Other ◽  
Ms Medium ◽  
Green Callus

A protocol was developed to produce large amount of callus in short a period of time from leaf explants of Stevia rebaudiana Bert. The highest amount of white callus was obtained on MS medium supplemented with 2.5 mg/l 2, 4-D and 0.5 mg/l BAP after 3 weeks of inoculating leaf segments. On the other hand, 0.5 mg/l BAP and 1.0 mg/l Kn exhibits poor performance towards callus formation while after using 1.0 mg/l Kn alone did not develop any callus. In this experiment, highest amount of green callus was obtained when MS medium supplemented with 2.5 mg/l NAA and 10% coconut water was used. An improved analytical method HPLC was applied to analyze stevioside extracted from the leaf and callus of Stevia rebaudiana. The stevioside in each sample were analyzed by comparing their retention times with those of the standards. The retention time (RT) of stevioside for leaves were found 14.96 and for callus 13.81 mins. The percentage of stevioside content from leaves and callus was 12.19% and 12.62% respectively DOI: http://dx.doi.org/10.3329/bjsir.v49i4.22621 Bangladesh J. Sci. Ind. Res. 49(4), 199-204, 2014

scholarly journals In vitro adventitious shoot regeneration from leaf explants of some apricot cultivars

2019 ◽  
Vol 43 ◽  
Author(s):  
Olga Vladimirovna Mitrofanova ◽  
Irina Vjacheslavovna Mitrofanova ◽  
Tatyana Nikolaevna Kuzmina ◽  
Nina Pavlovna Lesnikova-Sedoshenko ◽  
Sergey Vladimirovich Dolgov
Keyword(s):  
Callus Formation ◽  
System Development ◽  
Low Frequency ◽  
Leaf Explants ◽  
Adventitious Shoot ◽  
Limiting Factor ◽  
Ms Medium ◽  
Regeneration System ◽  
Propagation Methods

ABSTRACT Apricot is one of the most valuable commercial fruits. In vitro propagation of apricot is very important for rapid multiplication of cultivars with desirable traits and production of cleaning up and virus-free plants. Low frequency of multiplication is the main limiting factor for traditional propagation methods. In this regard, the objective of our investigation was to study the morphogenetic capacity of apricot leaf explants of the promising cultivars ‘Iskorka Tavridy’, ‘Magister’ and ‘Bergeron’ for regeneration system development and solving some breeding questions. The source of explants was in vitro plants regenerated and cultured on QL medium. Leaves were maintained in the dark at 24±1 °C in thermostat for three-four weeks. Morphogenic callus and structures were mainly formed at the central and proximal parts of leaves on MS, QL and WPM media with 1.5 or 2.0 mg L-1 BAP and 1.5 or 2.0 mg L-1 IAA in different combinations, or TDZ (0.6 and 1.3 mg L-1). Callus with adventive buds was transferred to regeneration medium and placed into a growth chamber at 24±1 °C and 16-hour photoperiod with a light intensity of 37.5 μmol m-2 s-1. The best results were obtained when adaxial leaf surface was in contact with the culture medium. Frequency of leaf callus formation on MS medium with 1.5 mg L-1 BAP and 1.5 mg L-1 IAA was higher in the explants of ‘Iskorka Tavridy’ (80.0%) than in - ‘Bergeron’ (50.0%) and ‘Magister’ (36.7%). The best results of callogenesis for ‘Magister’ was obtained on MS medium with 1.3 mg L-1 TDZ (53.3%). Active microshoot regeneration in ‘Iskorka Tavridy’ cultivar was shown on MS medium with BAP and IAA and in ‘Magister’ cultivar - on MS medium with TDZ. Rhizogenesis was obtained on half strength MS medium with 2.0 mg L-1 IBA.

scholarly journals Pengaruh Komposisi 2,4-D dan BAP Terhadap Pembentukan Kalus Eksplan Pucuk Nilam (Pogostemon Cablin Benth.) secara In Vitro dengan Pemotongan Horizontal dan Vertikal

2021 ◽  
Vol 5 (1) ◽  
pp. 9-20
Author(s):  
Khairan Khairan ◽  
Betty Mauliya Bustam ◽  
Yunita ◽  
Riska Meilinda ◽  
Raudhatul Muna
Keyword(s):  
Acetic Acid ◽  
Callus Formation ◽  
Callus Growth ◽  
Cutting Method ◽  
Highest Weight ◽  
Pogostemon Cablin

This study aims to determine the effect of 2,4-Diclorophenoxy Acetic Acid (2,4-D) and Benzyl Amino Purin (BAP) on the formation of callus of patchouli (Pogostemon cablin Benth.) shoot explants by horizontal and vertical cutting methods. The parameters that observed in this study were the percentage growth of callus, time appearance of callus, weight of callus and the morphology of callus. The results showed that horizontal cutting method was able to induce callus growth with the percentages growth of callus were 18,75%, with the time appearance of callus was at 16 days at P1; P10; P12; P13 dan P14. The highest weight of callus obtained was 0.19 grams at P8. The results also showed that the callus yielded had a yellow and cream color, with a compact and crumb textures. Meanwhile, the vertical cutting method was able to induce callus formation with the percentage growth of callus were 12,5%. The fastest time of callus appearance was obtained in P6 and P8, which was 12 day after planting with the highest weight of callus obtained was 0.05 grams at P12.   The results also showed that vertical cutting method had brown and dark-brown of callus with a compact and crumb textures.

scholarly journals In vitro callogenesis induction of Guarianthe skinneri (Bateman) Dressler & W.E. Higgins (Orchidaceae)

2017 ◽  
Vol 66 (2) ◽  
Author(s):  
Ana Gabriela Coutiño-Cortés ◽  
Vincenzo Bertolini ◽  
Leobardo Iracheta Donjuan ◽  
Lorena Ruíz-Montoya ◽  
Javier Francisco Valle-Mora
Keyword(s):  
Statistical Analysis ◽  
Solid Medium ◽  
Callus Formation ◽  
Natural Habitat ◽  
Leaf Explants ◽  
Lower Percentage ◽  
Production Technique ◽  
Flowering Season ◽  
Semi Solid

Guarianthe skinneri (Bateman) Dressler & W.E. Higgins., is a native orchid from Mexico, considered as threatened species by NOM-ECOL-059-SEMARNAT-2010, mainly due to the disappearance of its natural habitat and the illegal collection during its flowering season. The aim of this research was to induce in vitro callogenesis from different type of explants, using phytoregulators, in order to look for a massive production technique to contribute to its conservation. We evaluated the leaf and pseudobulb marrow explants growing in semi-solid medium MS adding BAP, 2, 4-D, Kin, the interaction of BAP/2, 4-D/Kin/Sad and a control without any type of plant growth regulators. Statistical analysis showed the pseudobulb marrow explants are more suitable for in vitro introduction in comparison to leaf explants, since they perform a lower percentage of contamination (18.8% in marrow and 73.2% in leaves). Likewise, the pseudobulb marrow explants increased callus formation (10.8%) in comparison to leaf explants (7.6%). Regarding the phytoregulators employed, BAP have allowed to increased callus formation (17%) compared to other phytoregulators (7-10%). This is the first report, which proposes the use of pseudobulb marrow as explant for callus induction in G. skinneri.

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