scholarly journals The Development of Gametophyte Sterilization Method for Liverworts Acrolejeunea fertilis (Reinw., Blume and Nees) Schiffn. In vitro Culture

2021 ◽  
Vol 28 (1) ◽  
pp. 39
Author(s):  
Mouleidi Dwi Putri ◽  
Windri Handayani ◽  
Astari Dwiranti ◽  
Andi Salamah ◽  
Niarsi Merry Hemelda ◽  
...  
Keyword(s):  
In Vitro Culture ◽  
Exposure Time ◽  
Common Type ◽  
Internal Contamination ◽  
Sterilization Method ◽  
Combination Treatments ◽  
Sterilization Process ◽  
Bacteria And Fungi ◽  
High Level

In vitro culture gametophytes of leafy liverworts often have problems in their sterilization process. These problems due to the high level of contamination and the fragile structure of the gametophyte leafy liverworts. The structures can be easily to damage after exposure to disinfectant. This study aimed to observe the concentration and the exposure time of “Bayclin” commercial bleach to suppress contamination with the viability of Acrolejeunea fertilisgametophytic explants. This research was conducted using control and 6 combination treatments with “Bayclin” concentration (1.00%, 1.25%, and 1.50%) and exposure time 60 and 120 seconds, then accompanied by the addition of Tetracycline 2.5 mg/ml. The qualitative parameters observed were the explant color, the type and location of contamination, and the growth of explants. The quantitative parameters were the percentage of contamination, the percentage of growth, and the number of new branches. The results showed that “Bayclin” 1.25% and 1.50% with 60 seconds exposure time has the lowest percentage of contamination which is 70% until the 7th days after planting. The most common type of internal contamination from the explant is bacteria and fungi. However, the growth of the new branch still occurs in some explants even though it has been contaminated and browned.

scholarly journals The Development of In Vitro Culture Sterilization Method of Gametophyte Explant Lopholejeunea sp.

2021 ◽  
Vol 28 (2) ◽  
pp. 110
Author(s):  
Anna Widyastuti ◽  
Afiatry Putrika ◽  
Astari Dwiranti ◽  
Andi Salamah ◽  
Niarsi Merry Hemelda ◽  
...  
Keyword(s):  
In Vitro Culture ◽  
Exposure Time ◽  
Shoot Formation ◽  
Common Type ◽  
In Vitro Cultures ◽  
Sterilization Process ◽  
Bacteria And Fungi ◽  
High Level ◽  
Viable Culture

In vitro cultures of leafy liverworts are still facing significant challenges due to high-level of explant contamination. The sterilization process can easily damage the structure of liverwort after exposure to the disinfectant. This study was to determine the concentration and time exposure of commercial bleach as a disinfectant to suppress contamination using the gametophyte culture of Lopholejeunea sp. The experiment consisted of control and six treatment combinations of commercial bleach with concentration 0.5, 0.75, and 1% (v/v), and exposure time (60 and 90 seconds). The type and location of contamination, the color of the explants after sterilization, and response after 30 days were observed. The results showed that the 0.75% bleach with 60 and 90 seconds exposure time had a lower contamination until the 7th day of culture. The most common type of contamination is bacteria and fungi that arise from the explant. Despite the contamination, it did not inhibit shoot formation. Further studies still needed to determine the type of fungicides and antibiotics with the most potent concentration and exposure time should be tested to obtain an axenic and viable culture of liverworts Lopholejeunea sp.

scholarly journals EFEKTIVITAS MERKURI KLORIDA (HgCl2) PADA STERILISASI TUNAS SAMPING JATI (Tectona grandis) IN VITRO

2017 ◽  
Vol 4 (2) ◽  
pp. 25
Author(s):  
Yusuf Sigit Ahmad Fauzan ◽  
. Supriyanto ◽  
Teuku Tajuddin
Keyword(s):  
In Vitro Culture ◽  
Tectona Grandis ◽  
In Vitro Cultures ◽  
Mercury Chloride ◽  
Main Obstacle ◽  
Sterilization Process ◽  
High Level ◽  
Growth Of Fungi ◽  
Tectona Grandis L.F

Effectiveness of Mercury Chloride (HgCl2) in Sterilization of Teak (Tectona grandis L.f.) In VitroThe main obstacle in obtaining sterile materials in in vitro cultures derived from meristems is high level of surface contamination caused by fungi and bacteria, which often results in explant death. The objective of this study was to obtain an appropriate mercury chloride (HgCl2) concentration for the sterilization of Tectona grandis nodes in in vitro culture. One cm long-sized nodes with 0.2 mm diameter were immersed in HgCl2at concentrations of 0, 100, 200 and 300 mg/L for 3 minutes. The results showed that the higher concentration of HgCl2was able to suppress the growth of fungi and bacteria and increased the percentage of aseptic explants. The best HgCl2concentration was 300 mg/L since it suppressed the growth of fungi and bacteria up to 100% and 75%, respectively, and produced the highest aseptic explants of 85% at 9 days after treatment. The small sized explants supported the sterilization process and reduced browning levels.Keywords: Browning, HgCl2, in vitro, sterilization, T. grandisABSTRAKKendala utama dalam mendapatkan material steril pada kultur in vitro yang berasal dari meristem adalah tingginya tingkat kontaminasi permukaan yang disebabkan oleh jamur dan bakteri, dan sering menyebabkan kematian eksplan. Tujuan penelitian ini adalah untuk memperoleh konsentrasi merkuri klorida (HgCl2) yang tepat untuk sterilisasi eksplan tunas samping tanaman jati (Tectona grandis) pada kultur in vitro. Tunas samping berukuran 1 cm dan diameter 0,2 mm direndam dalam HgCl2 pada konsentrasi 0, 100, 200 dan 300 mg/L selama 3 menit. Hasil penelitian menunjukkan bahwa penambahan konsentrasi HgCl2 yang semakin tinggi mampu menekan pertumbuhan jamur dan bakteri pada eksplan serta meningkatkan persentase eksplan aseptik. HgCl2 dengan konsentrasi 300 mg/L merupakan konsentrasi terbaik karena dapat menekan pertumbuhan jamur hingga 100% dan bakteri mencapai 75%, serta menghasilkan tingkat eksplan aseptik dan hidup tertinggi yaitu sebesar 85% pada 9 hari setelah perlakuan. Ukuran eksplan yang kecil mendukung proses sterilisasi dan mengurangi tingkat browning. Kata kunci: HgCl2,in vitro, pencoklatan jaringan, sterilisasi, T. grandis, Received: 02 November 2017                 Accepted: 14 December 2017                Published: 29 December 2017

scholarly journals Introduction of hyssopus officinalis l . into in vitro culture to optimize the conditions for obtaining callus tissues and microclonal propagation as a promising metod of innovative agrobiotechnologies

2021 ◽  
Vol 30 ◽  
pp. 05006
Author(s):  
Elena Maslova ◽  
Natalya Gulya ◽  
Tatyana Perelugina ◽  
Valeria Semykina ◽  
Elena Kalashnikova
Keyword(s):  
In Vitro Culture ◽  
Exposure Time ◽  
Crop Production ◽  
Callus Tissue ◽  
Nutrient Media ◽  
Sterilization Process ◽  
Callus Tissues ◽  
Active Substances ◽  
Hyssopus Officinalis

The sterilization process of plant explants of H. officinalis was optimized when introduced into an in vitro culture, the most effective sterilization modes, optimal sterilizing agents, their exposure time and concentration were selected. Callus tissues and mini-plants of H. officinalis were obtained in vitro and the most optimal nutrient media were determined both for microclonal propagation and for the induction of callus tissue H. officinalis, which can be further used for mass cultivation of cell and culture and obtaining safe bio-additives with active substances for livestock and crop production as a part of the development of modern agrobiotechnologies.

scholarly journals Photodynamic Therapy Combined with Antibiotics or Antifungals against Microorganisms That Cause Skin and Soft Tissue Infections: A Planktonic and Biofilm Approach to Overcome Resistances

2021 ◽  
Vol 14 (7) ◽  
pp. 603
Author(s):  
Vanesa Pérez-Laguna ◽  
Isabel García-Luque ◽  
Sofía Ballesta ◽  
Antonio Rezusta ◽  
Yolanda Gilaberte
Keyword(s):  
Photodynamic Therapy ◽  
Soft Tissues ◽  
Combined Treatment ◽  
Resistance Pattern ◽  
Antimicrobial Photodynamic Therapy ◽  
Antimicrobial Drugs ◽  
Bacteria And Fungi ◽  
High Level ◽  
Selection Of

The present review covers combination approaches of antimicrobial photodynamic therapy (aPDT) plus antibiotics or antifungals to attack bacteria and fungi in vitro (both planktonic and biofilm forms) focused on those microorganisms that cause infections in skin and soft tissues. The combination can prevent failure in the fight against these microorganisms: antimicrobial drugs can increase the susceptibility of microorganisms to aPDT and prevent the possibility of regrowth of those that were not inactivated during the irradiation; meanwhile, aPDT is effective regardless of the resistance pattern of the strain and their use does not contribute to the selection of antimicrobial resistance. Additive or synergistic antimicrobial effects in vitro are evaluated and the best combinations are presented. The use of combined treatment of aPDT with antimicrobials could help overcome the difficulty of fighting high level of resistance microorganisms and, as it is a multi-target approach, it could make the selection of resistant microorganisms more difficult.

scholarly journals Optimasi sterilisasi permukaan eksplan stek mikro tanaman karet Optimization of surface sterilization on rubber microcutting explan

2016 ◽  
Vol 81 (1) ◽  
Author(s):  
Irfan MARTIANSYAH ◽  
Deden Dewantara ERIS ◽  
. NURHAIMI-HARIS ◽  
Darmono TANIWIRYONO
Keyword(s):  
Primary Culture ◽  
Second Step ◽  
Contamination Level ◽  
Surface Sterilization ◽  
Major Constraint ◽  
Culture Stage ◽  
Sterilization Process ◽  
High Level

AbstractAn increasing number of explants is necessary toobtain plantlets in large quantities, for mass propagationof rubber plants. However, high level of contamination atthe primary culture stage is still a major constraint in invitro microcutting of rubber. The aim of this study was tooptimize surface sterilization procedures to reduce micro-bial contamination at the primary culture. Sterilizationexperiment was conducted in two step., The first step wasto determine the effect of washing the explants withrunning water prior to sterilization and then using Deso-germe, ethanol or H 2 O 2 , while the second step was toidentify the suitable sterilization process on reducing thelevel of contamination. The results showed that the surfacesterilization with only one type of sterilization agent couldnot reduce contamination level caused either by bacteriaor fungi, while sterilization with three types of sterilizingagents increased the number of dead explants. The besttreatment for surface sterilization was the directsterilization of explants using 70% ethanol for one minuteand 17.6% H 2 O 2 for 20 minutes without washing with tapwater (A-CD treatment). The percentage of viable andaseptic explantsof this treatment was 76.7%, which wassignificantly higher than those of other treatments. Thistreatment reduced contamination level to 21.7%.AbstrakPeningkatan jumlah eksplan sangat diperlukan untukmemperoleh planlet dalam jumlah besar pada perbanyakanmassal tanaman karet secara in vitro. Namun, tingginyatingkat kontaminasi pada tahap kultur primer masih me-rupakan kendala utama dalam kultur stek mikro tanamankaret. Tujuan penelitian adalah mengoptimasi prosedursterilisasi permukaan eksplan untuk mengurangi jumlaheksplan yang terkontaminasi mikroba pada tahap kulturprimer. Percobaan sterilisasi dilaksanakan dalam duatahap, tahap pertama untuk mengetahui pengaruh pen-cucian eksplan dengan air mengalir pada awal sterilisasiserta penggunaan Desogerme, etanol dan H 2 O 2 , sedang-kan tahap kedua untuk mendapatkan proses sterilisasi yangpaling sesuai dalam menurunkan tingkat kontaminasi.Hasil penelitian menunjukkan bahwa perlakuan sterilisasipermukaan yang menggunakan satu jenis bahan sterilantidak dapat mengurangi kontaminasi, baik oleh bakterimaupun cendawan. Perlakuan sterilisasi eksplan dengantiga jenis bahan sterilan meningkatkan kematian eksplan.Perlakuan sterilisasi permukaan terbaik adalah sterilisasilangsung eksplan menggunakan etanol 70% selama satu  menit dan H 2 O 2 17,6% selama 20 menit, tanpa pencuciandengan air mengalir (perlakuan A-CD). Persentase eksplansteril yang hidup sebesar 76,7%, berbeda nyata dibanding-kan dengan perlakuan lainnya. Perlakuan tersebut dapatmengurangi kontaminasi menjadi sebesar 21,7%.

scholarly journals INFLUENCE OF THE STERILIZATION METHOD AND THE TIME OF INTRODUCTION INTO IN VITRO CULTURE ON THE VIABILITY OF EXPLANTS OF THE CLONAL APPLE STOCK 54-118

2020 ◽  
Vol 30 (4) ◽  
pp. 411-416
Author(s):  
A.V. Nikitina ◽  
A.M. Lentochkin ◽  
T.G. Lekontseva ◽  
A.V. Fedorov
Keyword(s):  
Hydrogen Peroxide ◽  
In Vitro Culture ◽  
Ethyl Alcohol ◽  
Shoot Growth ◽  
Sterile Culture ◽  
Sterilization Method ◽  
Clonal Micropropagation

The stage of introducing explants into a sterile culture is difficult in the technology of clonal micropropagation of plants. The article shows the possible ways of sterilization and the introduction terms of explants of the clonal apple stocks 54-118 into in vitro culture in order to reduce planting infection and increase the yield of sterile viable explants. The best time for introducing clonal apple stocks 54-118 into sterile in vitro culture is the period of active shoot growth. Sterilization of explants with ethyl alcohol (70,0 %, 1 min) in combination with hydrogen peroxide (33,0 %) for 7 minutes and ethyl alcohol (70,0 %, 1 min) in combination with diacide (0,1 %) within 6 minutes contributed to the production of 63,0 % and 60,0 % of viable sterile explants.

scholarly journals Optimasi sterilisasi permukaan eksplan stek mikro tanaman karet Optimization of surface sterilization on rubber microcutting explan

2016 ◽  
Vol 81 (1) ◽  
Author(s):  
Irfan MARTIANSYAH ◽  
Deden Dewantara ERIS ◽  
. NURHAIMI-HARIS ◽  
Darmono TANIWIRYONO
Keyword(s):  
Primary Culture ◽  
Second Step ◽  
Contamination Level ◽  
Surface Sterilization ◽  
Major Constraint ◽  
Culture Stage ◽  
Sterilization Process ◽  
High Level

AbstractAn increasing number of explants is necessary toobtain plantlets in large quantities, for mass propagationof rubber plants. However, high level of contamination atthe primary culture stage is still a major constraint in invitro microcutting of rubber. The aim of this study was tooptimize surface sterilization procedures to reduce micro-bial contamination at the primary culture. Sterilizationexperiment was conducted in two step., The first step wasto determine the effect of washing the explants withrunning water prior to sterilization and then using Deso-germe, ethanol or H 2 O 2 , while the second step was toidentify the suitable sterilization process on reducing thelevel of contamination. The results showed that the surfacesterilization with only one type of sterilization agent couldnot reduce contamination level caused either by bacteriaor fungi, while sterilization with three types of sterilizingagents increased the number of dead explants. The besttreatment for surface sterilization was the directsterilization of explants using 70% ethanol for one minuteand 17.6% H 2 O 2 for 20 minutes without washing with tapwater (A-CD treatment). The percentage of viable andaseptic explantsof this treatment was 76.7%, which wassignificantly higher than those of other treatments. Thistreatment reduced contamination level to 21.7%.AbstrakPeningkatan jumlah eksplan sangat diperlukan untukmemperoleh planlet dalam jumlah besar pada perbanyakanmassal tanaman karet secara in vitro. Namun, tingginyatingkat kontaminasi pada tahap kultur primer masih me-rupakan kendala utama dalam kultur stek mikro tanamankaret. Tujuan penelitian adalah mengoptimasi prosedursterilisasi permukaan eksplan untuk mengurangi jumlaheksplan yang terkontaminasi mikroba pada tahap kulturprimer. Percobaan sterilisasi dilaksanakan dalam duatahap, tahap pertama untuk mengetahui pengaruh pen-cucian eksplan dengan air mengalir pada awal sterilisasiserta penggunaan Desogerme, etanol dan H 2 O 2 , sedang-kan tahap kedua untuk mendapatkan proses sterilisasi yangpaling sesuai dalam menurunkan tingkat kontaminasi.Hasil penelitian menunjukkan bahwa perlakuan sterilisasipermukaan yang menggunakan satu jenis bahan sterilantidak dapat mengurangi kontaminasi, baik oleh bakterimaupun cendawan. Perlakuan sterilisasi eksplan dengantiga jenis bahan sterilan meningkatkan kematian eksplan.Perlakuan sterilisasi permukaan terbaik adalah sterilisasilangsung eksplan menggunakan etanol 70% selama satu  menit dan H 2 O 2 17,6% selama 20 menit, tanpa pencuciandengan air mengalir (perlakuan A-CD). Persentase eksplansteril yang hidup sebesar 76,7%, berbeda nyata dibanding-kan dengan perlakuan lainnya. Perlakuan tersebut dapatmengurangi kontaminasi menjadi sebesar 21,7%.

Rhizome Buds Disinfection for Preparation of Red Ginger (Zingiber officinale Roxb. var. rubrum Rosc.) In Vitro Culture

2020 ◽  
Vol 3 (1) ◽  
pp. 19-26
Author(s):  
Popy Hartatie Hardjo ◽  
Alfian Hendra Krisnawan
Keyword(s):  
In Vitro Culture ◽  
Axenic Culture ◽  
Zingiber Officinale ◽  
Culture Initiation ◽  
Surface Sterilization ◽  
Sterilization Method ◽  
Ginger Rhizome ◽  
Red Ginger ◽  
Rhizome Buds

The success of culture initiation depends on explant surface sterilization techniques. Suitable concentration, combinations, and duration of exposure of sterilizing agents are important to raise in vitro culture successfully. The aim of this work is to obtain the suitable sterilization method for explant buds of red ginger rhizome to get the axenic culture. Four sterilizing agents, fungicide, bactericide, Cefotaxime antibiotic, and NaOCl were tested for sterilization by various concentration and duration of exposure. The results showed that sterilizing agents 200 mg/L Cefotaxime and 100 mg/L Benomyl combined with NaOCl decreased the contamination of explants, and achieved 20% axenic culture.

The Use of Nanostructured Gibberellic Acid to Optimize the Cultivation of Echinacea purpurea (L.) Moench In Vitro

2020 ◽  
Vol 17 (9) ◽  
pp. 4718-4724
Author(s):  
Elena V. Maslova ◽  
Alexander A. Krolevets ◽  
Polina A. Gaidai ◽  
Elena A. Kalashnikova ◽  
Mikhail Y. Cherednichenko ◽  
...  
Keyword(s):  
Seed Germination ◽  
Gibberellic Acid ◽  
Exposure Time ◽  
Effective Action ◽  
Active Substance ◽  
Callus Tissue ◽  
Echinacea Purpurea ◽  
Action Time ◽  
Sterilization Process

The sterilization process of plant explants of E. purpurea was optimized when introduced into an In Vitro culture, the most effective sterilizers, their exposure time and concentration were selected. An approach to increase the yield of plant explants seedlings from Echinacea purpurea (L.) Moench seeds is proposed. During the study an effective action time of 1 hour and a concentration of 1% nanocapsulated gibberellic acid were selected for more successful seed germination and cultivation of E. purpurea in vitro. This mode is at the same time the most economical both in terms of exposure time and in the minimum consumption of the active substance. The most optimal media compositions were determined, both for microclonal propagation and for the formation of E. purpurea callus tissue in in vitro culture.

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