scholarly journals Effect of activated charcoal and ascorbic acid on in vitro morphogenesis and o-dihydroxyphenols content in Paphiopedilum insigne

2022 ◽  
Author(s):  
Monika Poniewozik ◽  
Marzena Parzymies ◽  
Paweł Szot
Keyword(s):  
Ascorbic Acid ◽  
Phenolic Compounds ◽  
Activated Charcoal ◽  
Root Formation ◽  
Multiplication Rate ◽  
Ms Medium ◽  
In Vitro Morphogenesis ◽  
Half Strength ◽  
Positive Effect

Phenolic compounds limit micropropagation of many orchids in vitro. The aim of the study was to estimate the effect of activated charcoal (AC);1, 2 or 4 g/L) or ascorbic acid (AA; 10, 20 or 30 mg/L) added to the half strength MS medium on the growth and o-dihydroxyphenols content in Paphiopedilum insigne in vitro. A positive effect of AC on the shoot and root formation has been found. The highest multiplication rate (5.6 shoots/explant) and rooting frequency were obtained on medium containing 2 g/L of AC. However, AC reduced the leaf number as compared to the control. The lowest content of o-dihydroxyphenols was marked in Paphiopedilum insigne leaves when the shoots were grown on medium with 10 mg/L AA, followed by AC at 1 or 2 g/L.

scholarly journals In Vitro Long-Term Cultures of Papaya (Carica Papaya L.).

2021 ◽  
Author(s):  
Jose Javier Regalado González ◽  
Manuel López Granero ◽  
Carlos Lopez Encina
Keyword(s):  
Morphological Characteristics ◽  
Basal Medium ◽  
Multiplication Rate ◽  
Ms Medium ◽  
High Quality ◽  
Poor Growth ◽  
Average Multiplication ◽  
Half Strength

Abstract We present the data on proliferation corresponding to 10 years of continuous incubation in vitro of papaya shoots, and propose a reliable method for long-term micropropagation for papaya, using two types of explants: Microshoots from somatic embryos, and from axillary buds of papaya. Three different media were assayed. The proliferation medium (PPRM) allowed to maintain papaya shoots under continuous proliferation during 20 years, maintaining a consistent behaviour. Most of the shoots developed in PPRM rooted during the incubation, and after acclimated easily, maintaining the morphological characteristics of the parental plants, flowering and setting fruits normally. The PPRM medium consist in MS medium supplemented with NAA (0.1 mg l-1), BA (0.5 mg l-1), GA3 (0.5 mg l-1) and Adenine sulphate (40 mg l-1). The average multiplication rate was higher than 20 shoots per explant along the long-term assay. The elongation medium (PELM), was designed to recover shoots with a poor growth, and allowed the development of high quality shoots ready for rooting, and consist in a MS basal medium supplemented with NAA (0.1 mg l-1), Kin (0.5 mg l-1) and GA3 (1 mg l-1). The rooting medium (PROM) was designed to induce high quality roots from non-rooted shoots and consist in a half strength MS medium plus IBA (1mg l-1). On PROM, agar can be exchanged for expanded vermiculite. Acclimation took place inside an acclimatization tunnel under progressive hydric stress. After 4 weeks, the plant recovery rate was 90% for plants maintained under continuous proliferation during ten years.

scholarly journals ВМІСТ ФОТОСИНТЕТИЧНИХ ПІГМЕНТІВ У РОСЛИНАХ РОДУ CARLINA L. У ПРИРОДІ ТА КУЛЬТУРІ IN VITRO

2020 ◽  
Vol 78 (4) ◽  
pp. 16-23
Author(s):  
N. B. Kravets ◽  
L. R. Hrytsak ◽  
M. Z. Prokopyak ◽  
O. Yu. Mayorova ◽  
N. M. Drobyk
Keyword(s):  
Acid Solution ◽  
Growth Regulators ◽  
Root Formation ◽  
Cultivated Plants ◽  
Presowing Treatment ◽  
Murashige Skoog ◽  
Half Strength ◽  
Semi Solid ◽  
Positive Effect

im. The aim of the study was to choose conditions for rooting improvement of in vitro cultivated plants of some species of Carlina L. genus. Methods. For receiving and rooting of aseptic sprouts, seeds of Carlina acaulis L., Carlina cirsioides Klok and Carlina onopordifolia Besser ex Szafer, Kulcz. et Pawl were subjected to presowing treatment with gibberellic acid solution (GA3) or indolebyturic acid solution (IBA). Sterilized seeds were planted in sterile Petri dishes on semi-solid Murashige, Skoog nutrient medium with half-strength concentrations of macro- and microsalts without growth regulators. Results. It was found that with the seed soaking of C. acaulis, C. cirsioides and C. onopordifolia in GA3 solution the percentage of root formation amounted to 33.3 %, 33.3 % and 22.2 % respectively. Presowing treatment of carlina seeds in IBA solution with concentration of 1000 mg for 2–4 hours before sterilization gave a positive effect: the percentage of root formation for C. acaulis, C. cirsioides and C. onorordifolia was 2.4–4.5 times higher compared to the treatment with GA3 solution. Conclusions. To form the root system of carlina plants it is effective to soak the seeds in the solution of IBA. Thus we were able to increase the percentage of rooting of C. sirsioides and C. onorordifolia plants to 100 %, C. acaulis plants – up to 80 % and avoid sprouts’ injury and changes in the concentrations of the IBA, which may occur during sterilization at high temperatures by using non-sterile solution of growth regulators.

scholarly journals Young capitulum as important explant in in vitro mass propagation of gerbera (Gerbera jamesonii)

2020 ◽  
Vol 12 (2) ◽  
pp. 264-276
Author(s):  
Budi WINARTO ◽  
Kurnia YUNIARTO ◽  
Rudy SOEHENDI
Keyword(s):  
Multiple Shoots ◽  
Shoot Formation ◽  
Leaf Length ◽  
Multiplication Rate ◽  
Ms Medium ◽  
Plantlet Production ◽  
Half Strength ◽  
Flower Stage ◽  
The One

A new route of in vitro propagation of gerbera selected clones was successfully established using young capitula in tight buds and buds that were started to unfold stage as explant source. The one-fourth pieces of young capitula of tight flower stage and half-strength MS medium containing 0.25 mg/l BAP was the suitable for initiation and produced higher number of shoots per explant up to 3.8 shoots. The results were improved by culturing the one-fourth piece of 01.092 capitulums on MS medium fortified by 0.2 mg/l BAP and 0.02 mg/l NAA producing the highest shoot formation up to 8.5 shoots per explant with 28.7 leaves per explant and 2.1 cm leaf length. High multiple shoots were determined in third to fourth subculture periods and reduced thereafter with high multiplication rate noted on 01.092 clone. Shoots were easily rooted on half-strength MS medium supplemented with 2 g/l activated charcoal. Plantlets were transferred to ex vitro condition with 96.4% survivability of 03.045 clone using Cycas rumphii bulk and cocopeat (1:1, v/v) under spraying 1 g/l Growmore (32N:10P:10K) solution once week periodically. The route has high potential applied in qualified plantlet production for other Gerbera’s due to high shoots produced up to 35 shoots per whole young capitulum used. 

scholarly journals Nutritional requirements for germination and in vitro development of three Orchidaceceae species in the southern Brazilian Amazon

2018 ◽  
Vol 24 (2) ◽  
pp. 87-94 ◽  
Author(s):  
Viviane Luiza Hunhoff ◽  
Lais Alves Lage ◽  
Ednamar Gabriela Palú ◽  
Willian Krause ◽  
Celice Alexandre Silva
Keyword(s):  
Activated Charcoal ◽  
Culture Media ◽  
Leaf Number ◽  
Alternative Form ◽  
Ms Medium ◽  
Root Number ◽  
Shoot Height ◽  
Full Strength ◽  
Half Strength

Tissue culture is an alternative form of producing healthy, vigorous and regular plants on a large scale. The purpose of this study was to evaluate the most efficient culture medium for in vitro plantlet germination and development of three Orchidaceae species. Seeds disinfested of three species were dispersed in distilled water and dripped into basic Murashige and Skoog (MS) medium. The experimental design was completely randomized in a factorial 3 x 4 (three species x four culture media), with 5 replications. Four treatments were established: (1) full-strength MS medium, with the full nutrient concentration (MSØ), (2) full-strength MS medium plus 0.3% activated charcoal (MSØ ACh), (3) half- strength MS medium (½ MS) and (4) half- strength MS medium with 0.3 % activated charcoal (½ MS ACh). Germination was evaluated after 15, 20, 25, 30, and 60 days. The shoot height, leaf number and length, root number and length of plantlets of the three studied species were assessed. In A. variegata, 73% germinated after 60 days in ½ MS ACh medium. In the same period, 100% of E. viparum and S. gloriosa seeds germinated in MSØ ACh medium. The plant height, leaf number and length, root number and length were significantly higher for the species A. variegata and E. viviparum in MSØ ACh medium. The culture media ½ MS and MSØ with addition of activated charcoal favored in vitro germination for the three orchid species of this study.

INFLUENCE DES CONCENTRATIONS HORMONALES, DU MILIEU DE CULTURE ET D’UN ANTI-OXYDANT SUR LE TEMPS DE DOUBLAGE DES TIGES DE FRAMBOISIERS ’MADAWASKA’ CULTIVES IN VITRO

1987 ◽  
Vol 67 (3) ◽  
pp. 863-869 ◽  
Author(s):  
Y. DESJARDINS ◽  
A. GOSSELIN
Keyword(s):  
Ascorbic Acid ◽  
Chlorophyll Content ◽  
Butyric Acid ◽  
Doubling Time ◽  
Rubus Idaeus ◽  
Multiplication Rate ◽  
Ms Medium ◽  
Indole Butyric Acid ◽  
Doubling Times

Our results show that the shoot doubling time is a useful and accurate parameter to characterize the effect of media on the multiplication rate of tissue-cultured raspberry (Rubus idaeus L. ’Madawaska’). We have determined that 1 mg L−1 of BA (benzyladenine) and 0–0.05 mg L−1 IB A (indole butyric acid) resulted in the highest rates of proliferation, with shoot doubling times of 19 and 18 d for MS (Murashige and Skoog) and Anderson medium, respectively. A reduction of 10 d in the shoot doubling time was obtained with the addition of ascorbic acid (50 mg L−1) to both MS and Anderson media. However, this treatment did not reduce yellowing of cultures since the chlorophyll content was not significantly different in presence or absence of ascorbic acid. Generally, the MS medium was superior to Anderson medium.Key words: Raspberry, doubling time, multiplication in vitro, ascorbic acid, Rubus idaeus L.

scholarly journals Micropropagation and Polyploid Induction of Acer platanoides ‘Crimson Sentry

2013 ◽  
Vol 31 (4) ◽  
pp. 246-252 ◽  
Author(s):  
Jason D. Lattier ◽  
Darren H. Touchell ◽  
Thomas G. Ranney ◽  
Jeremy C. Smith
Keyword(s):  
In Vitro Rooting ◽  
Naphthaleneacetic Acid ◽  
Multiplication Rate ◽  
Ms Medium ◽  
Norway Maple ◽  
Future Improvement ◽  
Half Strength ◽  
Improvement Programs ◽  
Indole 3 Acetic Acid

Protocols were developed for micropropagation and induction of autopolyploids in a fastigiate cultivar of Norway maple (A. platanoides L. ‘Crimson Sentry’). Murashige and Skoog (MS) medium, woody plant medium (WPM), and Quoirin and Lepoivre medium were supplemented with 2 μM 6-benzylaminopurine (BA), meta-Topolin, 6-(γ,γ-dimethylallylamino) purine, kinetin, or thidiazuron to evaluate microshoot proliferation. Murashige and Skoog medium with 2 μM BA yielded the most microshoots (3.2) and longest microshoots (30.6 mm) per subsample after 5 weeks. The influence of BA concentration on proliferation was evaluated at 0, 2, 4, 8, or 16 μM. Optimal multiplication rate was achieved at 2 or 4 μM BA producing approximately 2.8 microcuttings per subsample after 5 weeks. To induce in vitro rooting, half-strength WPM was supplemented with 0, 5, 10, 20, 40, or 80 μM indole-3-butyric acid (IBA). Optimal in vitro rooting (70%), number of roots (2.5), and root length (15 mm) per subsample were achieved with 10 μM IBA after 8 weeks. To induce polyploidy, microcuttings were pretreated for 7 days on MS medium with 4 μM BA alone or combined with 1 μM IBA, indole-3-acetic acid (IAA), or 1-naphthaleneacetic acid prior to treatment in liquid MS medium containing 15 μM oryzalin for 3 days. Homogenous tetraploids were only obtained from shoots pretreated with IAA. This research provides optimized protocols for micropropagation and autopolyploid induction of A. platanoides ‘Crimson Sentry’ and demonstrates the development of tetraploid lines for use in future improvement programs.

scholarly journals Protokol Perbanyakan Masal Dendrobium ‘Balithi CF22-58’ secara In Vitro Melalui Embriogenesis Somatik Tidak Langsung (In Vitro Propagation Protocol of Dendrobium ‘Balithi CF22-58’ via Indirect Somatic Embryogenesis)

2020 ◽  
Vol 29 (2) ◽  
pp. 137
Author(s):  
Fitri Rachmawati ◽  
Dewi Permanik ◽  
Ronald Bunga Mayang ◽  
Budi Winarto
Keyword(s):  
Somatic Embryogenesis ◽  
Activated Charcoal ◽  
Embryogenic Callus ◽  
Culture System ◽  
In Vitro Propagation ◽  
Clonal Propagation ◽  
Ms Medium ◽  
Indirect Somatic Embryogenesis ◽  
Half Strength

<p>Protokol perbanyakan klonal yang efektif dan efisien sangat diperlukan untuk produksi benih berkualitas pada komersialisasi produk unggulan hasil pemuliaan. Penelitian bertujuan untuk mendapatkan protokol perbanyakan klonal Dendrobium ‘Balithi CF22-58’ melalui embriogenesis tidak langsung. Percobaan dilakukan di Laboratorium Kultur Jaringan Balai Penelitian Tanaman Hias dari bulan Januari hingga Desember 2017. Penelitian ini menekankan pada penggunaan  jenis eksplan, media, dan sistem kultur. Jenis eksplan yang diuji adalah tunas pucuk, tunas lateral, dan pangkal plantlet dengan tiga media inisiasi [½ Murashige and Skoog (MS) dikombinasikan dengan 1,5 mg/l thidiazuron (TDZ) dan 0,5 mg/l 6-benzylaminopurine (BAP) (MI-1), 2,5 mg/l metathopolin (mT) dan 0,05 mg/l BAP (MI-2), dan 5 mg/l mT dan 0,05 mg/l BAP (MI-3)]; empat media proliferasi, yaitu ½ MS dengan kombinasi: MP-1 (0,75 mg/l TDZ + 0,25 mg/l BAP), MP-2 (1,5 mg/l TDZ + 0,5 mg/l BAP), MP-3 (2,5 mg/l mT + 0,05 mg/l BAP), dan MP-4 (5,0 mg/l+ 0,05 mg/l BAP); dua sistem kultur (padat dan cair); dan tiga media regenerasi MPP-1 (½ MS dengan vitamin penuh (1/2 MS-FV) + 2% charcoal); MPP-2 (½ MS-FV); dan MPP-3 (2 g/l Rosasol 18:18:18 TE). Percobaan disusun menggunakan rancangan acak kelompok faktorial dengan lima ulangan. Hasil penelitian menunjukkan bahwa inisiasi kalus embriogenik (KE) tertinggi, yaitu 38,3% dengan waktu inisiasi 16,8 hari dihasilkan dari eksplan pangkal plantlet pada medium MI-1. Medium MP-2 dan sistem kultur cair mampu mempertahankan proliferasi KE sampai 83,1% dengan rasio penggandaan 3,23 kali. Perkecambahan embrio terbaik sampai 86,9% embrio berkecambah dengan 18,2 kecambah per rumpun dalam waktu 21,3 hari, ditunjukkan pada medium MPP-1, sedangkan pembesaran plantlet terbaik mencapai tinggi plantlet sampai 5 cm, jumlah daun hingga 4,9 helai, dan jumlah akar  2,8, dengan  2,6 cm panjang akar dan 0,27 g bobot basah plantlet, diperoleh pada medium MPP-3. Perbanyakan anggrek dengan protokol ini diperkirakan dapat menghasilkan sekitar 3.000–4.000 plantlet/eksplan/tahun. Protokol hasil penelitian ini sangat potensial diaplikasikan pada perbanyakan klonal Dendrobium melalui kultur jaringan. </p><p><strong>Keywords</strong></p><p><em>Dendrobium</em>; Embriogenesis somatik; Perbanyakan masal; Proliferasi;  Sistem kultur  </p><p><strong>Abstract</strong></p><p>The effective and efficient clonal propagation protocol is significantly needed for producing qualified seedling for commercialization of superior breeding products. The objective of the study was to establish clonal propagation protocol for Dendrobium ‘Balithi CF22-58’ via indirect somatic embryogenesis. The study was conducted at the Tissue Culture Laboratory in Indonesian Ornamental Crops Research Institute from January to December 2017. The study emphasized to utilize explant source, culture media, and culture system. Explant types were shoot tip, lateral shoot, and basal part of plantlets; three initiation media [half strength Murashige and Skoog (MS) medium containing 1.5 mg/l thidiazuron (TDZ) and 0.5 mg/l 6-benzylaminopurine (BAP) (MI-1), 2.5 mg/l metathopolin (mT) and 0.05 mg/l BAP (MI-2), and 5 mg/l mT and 0.05 mg/l BAP (MI-3)]; four proliferation media (half strength MS medium supplemented with: MP-1 (0.75 mg/l TDZ and 0.25 mg/l BAP), MP-2 (1.5 mg/l TDZ and 0.5 mg/l BAP), MP-3 (2.5 mg/l mT and 0,05 mg/l BAP), and MP-4 (5.0 mg/l and 0.05 mg/l BAP); two culture system were solid and liquid; and three  regeneration media viz, MPP-1 (half strength MS medium with full vitamin and 2% activated charcoal); MPP-2 (MR-1 activated charcoal free), and MPP-3 (2 g/l Rosasol 18:18:18 TE). These experiments were arranged using a factorial randomized complete block design with five replications. Results of the study revealed that the highest initiation rate of embryogenic callus (EC) was up to 38.3% in 16.8 days after culture. The EC was regenerated from a basal part on MI-1 medium,  MP-2 medium and liquid culture system were able to maintain proliferation of embryogenic callus up to 83.1% with 3.23 multiplication rate. The best embryo germination up to 86.9% with 18.2 germinated embryos per clump within 21.3 days was determined on MPP-1 medium. While the best plantlet performances with 5 cm height of plantlets, 4.9 number of leaves, 2.8 number of roots, 2.6 cm root length, and 0.27 g plantlet fresh weight was obtained MPP-3 medium. With this propagation protocol, 3,000 - 4,000 plantlets/explant/year can be produced. Results of the study have high potential to be applied for in vitro propagation of Dendrobiums.</p>

scholarly journals Phenolic Exudation Control and Establishment of In vitro Strawberry (Fragaria × Ananassa) cv. Chandler

2019 ◽  
pp. 1-5
Author(s):  
Hidayatullah Mir ◽  
Ruby Rani ◽  
Feza Ahmad ◽  
Awadh Kishor Sah ◽  
Shashi Prakash ◽  
...  
Keyword(s):  
Ascorbic Acid ◽  
Activated Charcoal ◽  
Large Scale ◽  
Shoot Proliferation ◽  
Conventional Technique ◽  
Nodal Segments ◽  
Nodal Segment ◽  
Ms Medium ◽  
Planting Material

The rate of strawberry propagation through conventional technique is quite low and it is difficult to maintain planting material during the summer months under Bihar condition. Further, importing mother plants adds to the production cost. In vitro micro propagation has emerged as a potential alternative for supplying planting material for strawberry. Two type of explants viz., runner tip and nodal segment were used for the study. Phenol exudation was the major problem during establishment which caused death of majority explants. In our experiment, almost no phenolic exudation (+) and maximum percent regeneration for runner tip (55.2 ± 0.52%) and nodal segment (58.1 ± 0.54%) was observed when MS medium was supplemented with ascorbic acid 200 mg per liter. Phenolic exudation was recorded highest (++++) under control when no antioxidants were supplemented. Minimum number of days for runner tips (8.4 ± 0.23) and nodal segments (10.3 ± 0.33) taken for shoot proliferation was observed when MS medium was supplemented with activated charcoal 300 mg and 200 mg per liter, respectively. Though all other antioxidants used in our study including citric acid, PVP and activated charcoal significantly reduced oxidative browning, ascorbic acid was found to be most effective antioxidant in controlling lethal browning during in vitro establishment of strawberry. This protocol has a potential for allowing a large scale multiplication of this important crop.

scholarly journals IN VITRO MORPHOGENETIC REACTION OF MELISSA OFFICINALIS L.

2020 ◽  
Vol 1 (1) ◽  
Author(s):  
Ana Maria Radomir ◽  
Ramona Stan
Keyword(s):  
In Vitro Regeneration ◽  
Clonal Plants ◽  
Multiplication Rate ◽  
Ms Medium ◽  
Melissa Officinalis ◽  
Lemon Balm ◽  
Normal Morphology ◽  
Half Strength ◽  
Micropropagated Plants

Lemon balm (Melissa officinalisL.) is a medicinal plant with a long history in traditional medicine. Classical propagation of this species is inefficient for establishing a good quality clonal plants. The aim of this work was to elaborate an in vitro propagation protocol for M. officinalis using apexes and uninodal fragments as explants. The highest multiplication rate (4.7 shoots/explant) was obtained on a MS medium supplemented with 3 mg/L BAP. A half strength MS medium supplemented with 1 mg/L NAA was the most effective for in vitro rooting of lemon balmmicroshoots. Micropropagated plants transferred ex vitro showed normal morphology and 95% survival rate during acclimatization. The results obtained throughout the in vitro regeneration phases confirm that in vitro tissue culture is an efficient method for multiplication of M. officinalis.

scholarly journals Clonal Propagation of Phalaenopsis amabilis (L.) BL. cv. 'Golden Horizon' Through In vitro Culture of Leaf Segments

1970 ◽  
Vol 46 (2) ◽  
pp. 163-168 ◽  
Author(s):  
P Sinha ◽  
MAA Jahan
Keyword(s):  
In Vitro Culture ◽  
Natural Environment ◽  
Activated Charcoal ◽  
Clonal Propagation ◽  
Ms Medium ◽  
Huge Amount ◽  
Leafy Shoots ◽  
Half Strength ◽  
Leaf Segments

A protocol was established for mass clonal propagation of Phalaenopsis amabilis cv. 'Golden horizon' through in vitro culture of young leaf segments from mature plant. Explants were cultured on half strength Murashige and Skoog (1/2 MS) medium supplemented with N6-benzyladenine (2.0 mg l-1), a-naphthaleneaceetic acid (0.5 mg l-1), 2% (w/v) sucrose, 10% (v/v) coconut water, 2 g l-1 peptone and 1 g l-1 activated charcoal. Each section of explant produced 15 protocorm-like bodies (PLBs) after 12 weeks of culture. When phytohormone was omitted from the medium and 150 mg l-1 L-glutamone was added PLBs were found to be enlarged with leafy shoots and new PLBs were induced from the base of the old ones. Leafy shoots rooted on half strength MS medium supplemented with 2 g l-1 peptone, 2% (w/v) sucrose, 10% (v/v) CW and 1 g l-1 activated charcoal, where 100% explants were developed into plantlets with roots within 8 weeks. The addition of 2.5 g l-1 banana pulp powder enhanced the number and length of roots. Within the first 32 weeks after initiation of culture about 1500 plantlets as well as a huge amount of PLBs were achieved from a single explant section. The plantlets were acclimatized in natural environment. Key words: Phalaenopsis orchid; Leaf segments; Protocorm-like bodies; Micropropagation DOI: http://dx.doi.org/10.3329/bjsir.v46i2.8182 Bangladesh J. Sci. Ind. Res. 46(2), 163-168, 2011

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