scholarly journals Identification of Stevioside Using Tissue Culture-Derived Stevia (Stevia rebaudiana) Leaves

2015 ◽  
Vol 8s2 ◽  
pp. BCI.S30378 ◽  
Author(s):  
Ziaul Karim ◽  
Daisuke Uesugi ◽  
Noriyuki Nakayama ◽  
M. Monzur Hossain ◽  
Kohji Ishihara ◽  
...  
Keyword(s):  
Tissue Culture ◽  
In Vitro Culture ◽  
Large Scale ◽  
Stevia Rebaudiana ◽  
Potential Method ◽  
Modern Technology ◽  
Commercial Production ◽  
Sugar Levels ◽  
Disease Free

Stevioside is a natural sweetener from Stevia leaf, which is 300 times sweeter than sugar. It helps to reduce blood sugar levels dramatically and thus can be of benefit to diabetic people. Tissue culture is a very potential modern technology that can be used in large-scale disease-free stevia production throughout the year. We successfully produced stevia plant through in vitro culture for identification of stevioside in this experiment. The present study describes a potential method for identification of stevioside from tissue culture-derived stevia leaf. Stevioside in the sample was identified using HPLC by measuring the retention time. The percentage of stevioside content in the leaf samples was found to be 9.6%. This identification method can be used for commercial production and industrialization of stevia through in vitro culture across the world.

scholarly journals A REVISED METHOD FOR SUCKER STERILIZATION TO SUPPORT THE IN VITRO PROPAGATION OF SAGO PALM (Metroxylon sagu Rottb.)

2014 ◽  
Vol 1 (1) ◽  
pp. 21
Author(s):  
Teuku Tajuddin ◽  
Karyanti . ◽  
Tati Sukarnih ◽  
Nadirman Haska
Keyword(s):  
Tissue Culture ◽  
In Vitro Culture ◽  
In Vitro Propagation ◽  
Large Scale ◽  
Culture Method ◽  
Large Area ◽  
Sago Palm ◽  
Superior Genotypes ◽  
Metroxylon Sagu

Hutan sagu (Metroxylon sagu Rottb.) dapat ditemukan dalam area yang cukup luas di wilayah Maluku dan Papua. Besarnya keanekaragaman hayati dari pohon sagu dapat dilihat di areal ini. Pohon sagu tumbuh secara alami terutama di daerah dataran atau rawa dengan sumber yang air melimpah. Tanaman sagu dapat diperbanyak dengan metode generatif melalui biji, dan vegetatif melalui tunas anakan. Dalam rangka mendukung perbanyakan pohon induk yang unggul secara in vitro dalam skala besar, perbaikan metode sterilisasi tunas anakan mutlak diperlukan. Tunas anakan muda (15-20 cm) yang diperoleh dari Propinsi Papua digunakan sebagai eksplan. Tujuan percobaan sterilisasi ini dilakukan untuk mendukung perbanyakan pohon sagu secara in vitro. Pada percobaan ini antibiotik digunakan untuk membersihkan jaringan internal eksplan dari jamur dan bakteri. Hasil percobaan ini menunjukkan bahwa campuran alkohol dan antibiotik dapat menekan pertumbuhan kontaminan.Kata kunci: Antibiotik, kontaminan jamur dan bakteri, kultur in vitro, metode sterilisasi, sagu ABSTRACTNatural sago (Metroxylon sagu Rottb.) forest can be found in large area in Maluku and Papua regions. There are wide genetic diversities of sago palm found in these areas. This palm grows along riverbanks and in swampy areas which are not suitable for other crops. Sago palm is propagated generatively by seed and vegetatively by suckers. With the purpose of establishing the in vitro culture method for a large-scale of mass clonally propagation of superior genotypes of sago palm, generating sterilized explants are very important. Young suckers (15-20 cm) obtained from areas of Papua Province were used as explants. The sterilization experiments were carrying out to support the tissue culture of sago palm. Sterilization was conducted using antibiotics in order to get rid of fungi and bacteria from inner part of explants tissues. The results showed that from all sterilization methods tested, the best result was treatment using alcohol and antibiotic as disinfectant agents.Keywords: Antibiotics, fungi and bacteria contaminants, in vitro culture, sterilization method, sago palm

scholarly journals Salinity Effects on Gene Expression, Morphological, and Physio-Biochemical Responses of Stevia rebaudiana Bertoni In Vitro

Plants ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 820
Author(s):  
Clara Azzam ◽  
Sudad Al-Taweel ◽  
Ranya Abdel-Aziz ◽  
Karim Rabea ◽  
Alaa Abou-Sreea ◽  
...  
Keyword(s):  
Salt Tolerance ◽  
Biochemical Parameters ◽  
Large Scale ◽  
Stevia Rebaudiana ◽  
Multiple Shoots ◽  
High Salt ◽  
Stress Factors ◽  
Stevia Rebaudiana Bertoni ◽  
Nacl Concentration

Stevia rebaudiana Bertoni is a little bush, which is cultivated on a large scale in many countries for medicinal purposes and used as a natural sweetener in food products. The present work aims to conduct a protocol for stevia propagation in vitro to produce and introduce Stevia rebaudiana plants as a new sweetener crop to Egyptian agriculture. To efficiently maximize its propagation, it is important to study the influence of stress factors on the growth and development of Stevia rebaudiana grown in vitro. Two stevia varieties were investigated (Sugar High A3 and Spanti) against salt stress. Leaves were used as the source of explants for callus initiation, regeneration, multiplication and rooting. Some stress-related traits, i.e., photosynthetic pigments, proline contents, and enzyme activity for peroxidase (POD), polyphenol oxidase (PPO), and malate dehydrogenase (MDH) were studied. Murashig and Skoog (MS) medium was supplemented with four NaCl concentrations: 500, 1000, 2000, and 3000 mgL−1, while a salt-free medium was used as the control. The data revealed that salinity negatively affected all studied characters: the number of surviving calli, regeneration%, shoot length, the number of multiple shoots, number of leaf plantlets−1, number of root plantlets−1, and root length. The data also revealed that Sugar High A3 is more tolerant than Spanti. The total chlorophyll content decreased gradually with increasing NaCl concentration. However, the opposite was true for proline content. Isozyme’s fractionation exhibited high levels of variability among the two varieties. Various biochemical parameters associated with salt tolerance were detected in POD. Namely, POD4, POD6, POD 9 at an Rf of 0.34, 0.57, and 0.91 in the Sugar High A3 variety under high salt concentration conditions, as well as POD 10 at an Rf of 0.98 in both varieties under high salt concentrations. In addition, the overexpression of POD 5 and POD 10 at Rf 0.52 and 0.83 was found in both varieties at high NaCl concentrations. Biochemical parameters associated with salt tolerance were detected in PPO (PPO1, PPO2 and PPO4 at an Rf of 0.38, 0.42 and 0.62 in the Sugar High A3 variety under high salt concentrations) and MDH (MDH 3 at an Rf of 0.40 in both varieties at high salt concentrations). Therefore, these could be considered as important biochemical markers associated with salt tolerance and could be applied in stevia breeding programs (marker-assisted selection). This investigation recommends stevia variety Sugar High A3 to be cultivated under salt conditions.

scholarly journals Influence of Indole acetic acid and Tryptophan on production of Vinblastine and Vincristine of Catharanthus roseous callus cells in the accumulation media of In Vitro tissue culture

2010 ◽  
Vol 7 (3) ◽  
pp. 1113-1119
Author(s):  
Baghdad Science Journal
Keyword(s):  
Tissue Culture ◽  
High Performance Liquid Chromatography ◽  
Acetic Acid ◽  
Liquid Chromatography ◽  
In Vitro Culture ◽  
High Performance ◽  
Indole Acetic Acid ◽  
Culture Technique

This study on the plant of Ain –AL Bason Catharanthus roseous showed the ability of callus cells that is produced by In Vitro culture technique and transformed to the accumulated media (MS 40gm/L sucrose ,2gm/L IAA Indole acetic acid , 0.5gm/L Tryptophan) to produce Vinblastine and Vincristine compounds. Extraction, purification and quantitive determination of Vinblastine and Vincristine compounds using High performance liquid chromatography technique (HPLC)were carried out. The results showed that the highest concentration of Vinblastine and Vincristine compounds were ( 4.653,12.5 (ppm /0.5 dry Wight respectively from transformed callus cells from MS 40 gm /L sucrose , 2 gm / L NAA Naphthaline acetic acid .

scholarly journals Transplanting Space Effect on In-vitro Raised Sugarcane

2018 ◽  
Vol 10 (01) ◽  
pp. 12-16
Author(s):  
Sonali Gangwar ◽  
Saurabh Pathak ◽  
Maya Datt Joshi
Keyword(s):  
Tissue Culture ◽  
Plant Growth ◽  
Growth And Yield ◽  
Cane Yield ◽  
Sugarcane Variety ◽  
Rapid Multiplication ◽  
Space Effect ◽  
Healthy Seed ◽  
Disease Free

Tissue cuture is considered to be a best technique for rapid multiplication and production of disease free, healthy seed cane. With a view to studying the effect of transplanting spacing on growth and yield of micropropagated crop of sugarcane, an experiment was carried out at Tissue Culture Laboratory, Shobhit University, Meerut. Tissue culture raised plantlets of sugarcane variety. In vitro cultured sugarcane were transplanted at various spacing of 90 x 45, 90 x 60, 90 x 90 and 120 x 60 cm. Among the four spacings, the highest plant growth, number of tillers, number of malleable canes, cane height and cane yield were recorded at 90 x 60 cm. Thus, a spacing of 90 x 60 cm was found most suitable for transplantation of tissue culture raised plantlets of sugarcane.

scholarly journals Cannabis sativa: From Therapeutic Uses to Micropropagation and Beyond

Plants ◽  
2021 ◽  
Vol 10 (10) ◽  
pp. 2078
Author(s):  
Tristan K. Adams ◽  
Nqobile A. Masondo ◽  
Pholoso Malatsi ◽  
Nokwanda P. Makunga
Keyword(s):  
Tissue Culture ◽  
Genetic Engineering ◽  
Large Scale ◽  
Gene Editing ◽  
Cannabis Sativa ◽  
Scale Production ◽  
Indirect Organogenesis ◽  
Large Scale Production ◽  
Pharmaceutical Industries

The development of a protocol for the large-scale production of Cannabis and its variants with little to no somaclonal variation or disease for pharmaceutical and for other industrial use has been an emerging area of research. A limited number of protocols have been developed around the world, obtained through a detailed literature search using web-based database searches, e.g., Scopus, Web of Science (WoS) and Google Scholar. This article reviews the advances made in relation to Cannabis tissue culture and micropropagation, such as explant choice and decontamination of explants, direct and indirect organogenesis, rooting, acclimatisation and a few aspects of genetic engineering. Since Cannabis micropropagation systems are fairly new fields, combinations of plant growth regulator experiments are needed to gain insight into the development of direct and indirect organogenesis protocols that are able to undergo the acclimation stage and maintain healthy plants desirable to the Cannabis industry. A post-culture analysis of Cannabis phytochemistry after the acclimatisation stage is lacking in a majority of the reviewed studies, and for in vitro propagation protocols to be accepted by the pharmaceutical industries, phytochemical and possibly pharmacological research need to be undertaken in order to ascertain the integrity of the generated plant material. It is rather difficult to obtain industrially acceptable micropropagation regimes as recalcitrance to the regeneration of in vitro cultured plants remains a major concern and this impedes progress in the application of genetic modification technologies and gene editing tools to be used routinely for the improvement of Cannabis genotypes that are used in various industries globally. In the future, with more reliable plant tissue culture-based propagation that generates true-to-type plants that have known genetic and metabolomic integrity, the use of genetic engineering systems including “omics” technologies such as next-generation sequencing and fast-evolving gene editing tools could be implemented to speed up the identification of novel genes and mechanisms involved in the biosynthesis of Cannabis phytochemicals for large-scale production.

scholarly journals IN VITRO SHOOT MULTIPLICATION OF STEVIA REBAUDIANA (BERT) THROUGH PLANT TISSUE CULTURE.

2016 ◽  
Vol 4 (11) ◽  
pp. 2300-2307
Author(s):  
Vibha Bhingradiya ◽  
◽  
Archana Mankad ◽  
Ruby Patel ◽  
Shivangi Mathur ◽  
...  
Keyword(s):  
Tissue Culture ◽  
Plant Tissue Culture ◽  
Plant Tissue ◽  
Stevia Rebaudiana ◽  
Shoot Multiplication

scholarly journals Introduction of Stevia rebaudiana Bertoni in in vitro culture

2020 ◽  
Author(s):  
S. Sh. Asadova
Keyword(s):  
Cell Culture ◽  
In Vitro Culture ◽  
High Speed ◽  
Stevia Rebaudiana ◽  
Stevia Rebaudiana Bertoni ◽  
A Cell ◽  
Different Levels ◽  
Adult Plants

A cell culture obtained from explants of adult plants and aseptic seedlings of the Stevia rebaudiana Bertoni variety with different levels of ploidy, characterized by high speed, proliferation and ability to morphogenesis.

scholarly journals ISOLATION AND CHARACTERIZATION OF MDR BACTERIA FROM IN VITRO CULTURE OF BACOPA MONNIERA AND SUPPLEMENTATION OF NATURAL EXTRACTS TO CONTROL BACTERIAL CONTAMINATION

2016 ◽  
Vol 8 (11) ◽  
pp. 102
Author(s):  
Yash Sharma ◽  
Manish Bhardwaj ◽  
Anshita Nagar ◽  
Neeta Bhagat
Keyword(s):  
Tissue Culture ◽  
In Vitro Culture ◽  
Plant Tissue Culture ◽  
Plant Tissue ◽  
Bacterial Contamination ◽  
Resistant Bacteria ◽  
Bacopa Monniera ◽  
Natural Extracts ◽  
Isolation And Characterization

Objective: The purpose of the present study was to isolate and characterize bacterial contamination from in vitro culture of Bacopa monniera callus culture.Methods: The two selected isolates (1 and 2) were identified by morphological, biochemical and molecular (16S rRNA gene sequencing) methods. Beside this antibiotic resistance was also determined.Results: The isolates were identified as closely related to Enterobacter cloacae (KU350623) (Isolate 1) and Myroides odoratimimus (KU382740) (Isolate 2). The isolate 1 and 2 were found to be resistant to streptomycin (25 mcg), dapsone (10 mcg), erythromycin (15 mcg), chloroamphenicol (25 mcg) and ampicillin (10 mcg). Supplementation of the natural extract was tested to control the contamination due to these multi drug resistant bacteria. Aqueous and alcoholic leaf extracts of Azadirachta indica was added to plant tissue culture media i.e. MS media in order to control contamination.Conclusion: The present studies suggest using natural extracts supplementation as a promising strategy to control the in vitro bacterial contamination in plant tissue culture.

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