Enhanced anthraquinone production in presence of silver nitrate in suspension culture of Gynochthodes umbellata (L.) Razafim. & B. Bremer (Rubiaceae), a traditional medicinal plant mentioned in Hortus Malabaricus

2018 ◽  
Vol 5 (2) ◽  
Author(s):  
Gangaprasad A

Silver nitrate (AgNO3) enhanced production of anthraquinone was standardized in cell suspension cultures of Gynochthodes umbellata, a plant mentioned in the Hortus Malabaricus. The present research investigates the effect of silver nitrate, an abiotic elicitor on production of anthraquinone in in vitro cell suspension cultures of G. umbellata. Friable callus culture was established using in vitro derived leaf segment obtained from the nodal explant culture maintained in Murashige and Skoog (MS) medium containing 2 mg/l benzyl amino purine (BAP) and 3% sucrose. The in vitro derived leaf segments (0.5cm2) were cultured on MS medium containing 1 mg/l 2,4-D and 2% glucose for the production of friable callus. After 30 days of culture, uniform yellow friable callus was inoculated into MS liquid medium containing 1 mg/l 2,4-D and 2 % glucose for raising suspension culture. Uniform cell suspension was transferred to same media constituents and treated with different concentrations of AgNO3 on 25th day of culture. Fresh weight, dry weight and accumulation of anthraquinone content was studied and found that AgNO3 caused a marginal increase in biomass and anthraquinone based on the concentration and duration of AgNO3 treatment. A maximum fresh weight (19.48 g/fwt) dry weight (1.92g/dwt) and highest amount of anthraquinone content (48.62 mg/gdwt) were recorded in MS medium supplemented with 1 mg/l 2,4-D, 2%glucose and 3.5µM AgNO3 after 72 hrs of incubation.

Rodriguésia ◽  
2021 ◽  
Vol 72 ◽  
Author(s):  
Ivan Gonçalves Ribeiro ◽  
Tatiana Carvalho de Castro ◽  
Marsen Garcia Pinto Coelho ◽  
Norma Albarello

Abstract Medicinal plants are an important therapeutic option for a large share of the world’s population. To establish an in vitro culture system for the production of secondary metabolites from Hovenia dulcis, we studied the effect of auxins, cytokinins, absence of light, and silver nitrate on the development of friable callus. Callus cultures were established for the first time and used to obtain cell suspension cultures. Supplementation with KIN (Kinetin) produced calli with both compact and friable areas, while the addition of TDZ (Thidiazuron) only produced compact callus. The maintenance of cultures in the dark induced a slight enhancement on friability when the auxin PIC (Picloram) was present in the culture medium. The addition of silver nitrate promoted the formation of friable calli. Dry weight analysis showed no significant differences in biomass growth, and, therefore, 2.0 mg.L-1 was considered the most suitable treatment. The presence of silver nitrate was not required for the establishment of cell suspension cultures. Dry weight analysis of cell suspensions showed higher biomass production in the absence of silver nitrate. PIC promoted 100% of cell suspension culture formation in the absence of silver nitrate, and higher biomass production was observed with the lowest concentration (0.625 mg.L-1). No morphological differences were observed among the different concentrations of PIC. Phytochemical screening showed the presence of saponins, flavonoids, flavonols and catechins in the extracts obtained from H. dulcis calli. These results show that the cell cultures herein established are potential sources for the production of H. dulcis secondary metabolites of medicinal interest.


2017 ◽  
Vol 16 (1) ◽  
pp. 1-11
Author(s):  
Aryani Leksonowati ◽  
Witjaksono Witjaksono ◽  
Diah Ratnadewi

Aquilaria malaccensis Lam. is a plant species producing fragrant woody material that contains some resin. The compounds can be used as medicine and perfume. Sesquiterpenoid, one group of compounds has been found being synthesized and subsequently extracted from callus and cell suspension culture of Aquilaria species. The aim of this research was to find a method of producing friable calli and cell suspension cultures from leaves or internodes of A. malaccensis in vitro by using suitable plant growth regulators; cell suspension that will suitably serve as material to produce sesquiterpenoid afterwards. Calli were established in almost all treatments of auxin-cytokinin on both leaves and internod explants. The treatment of 10 mg/L IBA induced the highest percentage of callus coverage from leaves with a rather compact structure. The combined treatment of 1–2 mg/L 2.4-D and 0.2–0.3 mg/L BA induced friable callus formation in more than 80% of cultures with 27–32% callus coverage percentage.  The use of 2,4-D induced a better formation of cell suspension than Picloram, with maximum volume up to 7 mL. Cell suspension culture with fine and homogenous aggregate could be established in the medium supplemented with 0.5 –1 mg/L 2,4-D.


2012 ◽  
Vol 15 (2) ◽  
pp. 69-77
Author(s):  
Tram Thi My Pham ◽  
Tien Thi Thuy Le

Cell suspension cultures were initated from calli derived from in vitro strawberry leaves on MS medium (Murashige and Skoog, 1962) supplemented with 30 g/l sucrose, 1.0 mg/l 2,4-D and 0.3 mg/l kinetin. There were many factors affected on cell suspension cultures growth (it was found that …). Cell suspension cultures grew better on MS medium with 30 g/l sucrose. 1 g (fresh weight) of cells in 20 ml of medium was the best initial inoculum cell density for cell suspension cultures to grow. A shaking speed of 100 rpm on rotary shaker was suitable for the cells. The growth of cell suspension in dark was better than that under light condition. Anthocyanin in the cells was determined by pH differential method.


2013 ◽  
Vol 8 (4) ◽  
pp. 1934578X1300800 ◽  
Author(s):  
Tharita Kitisripanya ◽  
Jukrapun Komaikul ◽  
Nirachara Tawinkan ◽  
Chuennapha Atsawinkowit ◽  
Waraporn Putalun

The highest dicentrine content (19.5 ± 0.3 mg/g dry weight) from callus culture of Stephania venosa was achieved from stem segments cultured on MS medium supplemented with TDZ 0.5 mg/L and NAA 1.0 mg/L. Cell suspension cultures were established from callus cultured on MS liquid medium with the same plant growth regulators. Dicentrine production from S. venosa cell suspension cultures was obtained in the range of 15–26 mg/g dry weight. Elicitation in cell suspension cultures by chitosan (50 mg/L) and salicylic acid (2 mg/L) for 6 days significantly increased dicentrine content. Our findings indicate that callus and cell suspension cultures of S. venosa can produce high levels of dicentrine as an alternative source of plant materials.


2018 ◽  
Vol 42 (5) ◽  
pp. 464-473 ◽  
Author(s):  
Poornananda Madhava Naik ◽  
Jameel Mohammed Al-Khayri

ABSTRACT Date palm accumulates a wide range of secondary metabolites high in nutritional and therapeutic value. In the present study, date palm (Phoenix dactylifera L., cv. Shaishi) shoot-tip-induced callus was used to establish cell suspension cultures in Murashige and Skoog (MS) liquid medium containing 1.5 mg L-1 2-isopentenyladenine (2iP) and 10 mg L-1 naphthaleneacetic acid (NAA). To study the growth kinetics, cultures were maintained for 12 weeks during which weekly measurements were carried out to determine the biomass accumulation based on packed cell volume (%), fresh weight and dry weight (g). In addition, weekly determination of polyphenols (catechin, caffeic acid, kaempferol, and apigenin) was carried out using high performance liquid chromatography (HPLC). The 11-week-old culture was found highest in the production of biomass (62.9 g L-1 fresh weight and 7.6 g L-1 dry weight) and polyphenols (catechin-155.9 µg L-1, caffeic acid-162.7 µg L-1, kaempferol-89.7 µg L-1, and apigenin-242.7 µg L-1) from the cell suspension cultures. This is the first report on the production of polyphenols from the cell suspension culture of date palm. This study facilitates further development of large-scale production of polyphenols and the utilization of bioreactors.


1970 ◽  
Vol 48 (6) ◽  
pp. 1119-1130 ◽  
Author(s):  
Deng-Fong Liau ◽  
W. G. Boll

A solid medium was developed for callus cultures originating from explants of root, hypocotyl, and cotyledon of seedlings of bush bean, and a liquid medium was developed for the growth of cell suspension cultures derived from the callus cultures. Some unsatisfactory media are recorded. Concentrations of mineral salts for cell suspension cultures are lower than for callus cultures. Both coconut milk and other organic substances are required for maximum growth. With cell suspensions the effect of deproteinized coconut milk is the same as that of raw coconut milk but, with callus cultures, deproteinized coconut milk gives greater yield. There are no obvious differences in yield of callus derived from root, hypocotyl, or cotyledon. Few differences in yield were obtained between cell suspension cultures from root, hypocotyl, and cotyledon but those from root gave the highest yield in dry weight. However, in the same medium, cells from the three origins are very similar in form and appearance. Some effects of different media on cell form and clumping are described. The yield in suspension culture is very high. Increase in cell number, fresh weight, and dry weight is about 100-fold in 12 days involving about six to seven divisions per cell.


2016 ◽  
Vol 73 (1) ◽  
Author(s):  
. SUMARYONO ◽  
Imron RIYADI

SummaryIn vitro technology of plants can be used topropagate plants and to produce secondarymetabolites with a short and continuousproduction cycle. Callus cultures of cinchona(Cinchona ledgeriana Moens) on solid media andcell cultures in liquid media have beenestablished. Callus could be easily initiated fromvarious explants of cinchona clone CB5, GA22and QRC312. The best callus initiation andproliferation were obtained on a Woody Plant(WP) solid medium supplemented with 15 µMpicloram,0.5 µM BAP and 1 µM phloroglucinol.In this medium the fresh weight of callusincreased by 12 to 14-fold within 5 to 6 weeks.Callus that constantly grew fast was selected as amaterial source for cell suspension cultures. InWP liquid medium with the same composition,the cells remained to grow fast where cell volumeafter sedimentation (CVS) increased by almost4-fold in two weeks. However, repeated sub-cultures decreased cell growth rate. The cellsuspension culture was then scaled-up in a 5-Lbioreactor. The culture medium was the same asin Erlenmeyer flasks. Cells in a bioreactor grewvery slowly, the cell biomass fresh weight andpacked cell volume (PCV) increased by 34% and50% respectively after 21 days of culture,although most of the cells remained viable.RingkasanTeknologi in vitro tanaman dapat digunakanuntuk memperbanyak tanaman dan memproduksisenyawa sekunder dengan siklus sangat singkatdan berkelanjutan. Biak kalus tanaman kina(Cinchona ledgeriana Moens) pada mediumpadat dan biak sel di medium cair telahdikembangkan. Kalus dengan mudah dapatdiinduksi dari berbagai jenis eksplan tanamankina klon CB5, GA22 dan QRC312. Inisiasi danproliferasi kalus terbaik diperoleh pada mediaWoody Plant (WP) padat dengan pikloram 15µM, BAP 0,5 µM dan floroglusinol 1 µM. Padamedium ini bobot basah kalus meningkat 12-14kali lipat dalam waktu 5-6 minggu. Kalus yangtetap tumbuh cepat dipilih sebagai sumber bahanuntuk biak suspensi sel. Dalam medium cair WPdengan komposisi yang sama, sel tetap tumbuhdengan pesat, volume sel setelah pengendapan(CVS) meningkat hampir empat kali lipat dalamwaktu dua minggu. Namun subkultur berulangmenurunkan laju pertumbuhan sel. Skala biaksuspensi sel kemudian diperbesar dalam bio-reaktor kapasitas 5 L. Medium kultur yangdigunakan sama dengan medium pada labuErlenmeyer. Pertumbuhan sel dalam bioreaktorsangat lambat, bobot basah sel dan packed cellvolume (PCV) hanya bertambah berturut-turutsebesar 34% dan 50% setelah 21 hari dalamkultur, walaupun sebagian besar sel tetap viabel.


2016 ◽  
Vol 73 (1) ◽  
Author(s):  
. SUMARYONO ◽  
Imron RIYADI

SummaryIn vitro technology of plants can be used topropagate plants and to produce secondarymetabolites with a short and continuousproduction cycle. Callus cultures of cinchona(Cinchona ledgeriana Moens) on solid media andcell cultures in liquid media have beenestablished. Callus could be easily initiated fromvarious explants of cinchona clone CB5, GA22and QRC312. The best callus initiation andproliferation were obtained on a Woody Plant(WP) solid medium supplemented with 15 µMpicloram,0.5 µM BAP and 1 µM phloroglucinol.In this medium the fresh weight of callusincreased by 12 to 14-fold within 5 to 6 weeks.Callus that constantly grew fast was selected as amaterial source for cell suspension cultures. InWP liquid medium with the same composition,the cells remained to grow fast where cell volumeafter sedimentation (CVS) increased by almost4-fold in two weeks. However, repeated sub-cultures decreased cell growth rate. The cellsuspension culture was then scaled-up in a 5-Lbioreactor. The culture medium was the same asin Erlenmeyer flasks. Cells in a bioreactor grewvery slowly, the cell biomass fresh weight andpacked cell volume (PCV) increased by 34% and50% respectively after 21 days of culture,although most of the cells remained viable.RingkasanTeknologi in vitro tanaman dapat digunakanuntuk memperbanyak tanaman dan memproduksisenyawa sekunder dengan siklus sangat singkatdan berkelanjutan. Biak kalus tanaman kina(Cinchona ledgeriana Moens) pada mediumpadat dan biak sel di medium cair telahdikembangkan. Kalus dengan mudah dapatdiinduksi dari berbagai jenis eksplan tanamankina klon CB5, GA22 dan QRC312. Inisiasi danproliferasi kalus terbaik diperoleh pada mediaWoody Plant (WP) padat dengan pikloram 15µM, BAP 0,5 µM dan floroglusinol 1 µM. Padamedium ini bobot basah kalus meningkat 12-14kali lipat dalam waktu 5-6 minggu. Kalus yangtetap tumbuh cepat dipilih sebagai sumber bahanuntuk biak suspensi sel. Dalam medium cair WPdengan komposisi yang sama, sel tetap tumbuhdengan pesat, volume sel setelah pengendapan(CVS) meningkat hampir empat kali lipat dalamwaktu dua minggu. Namun subkultur berulangmenurunkan laju pertumbuhan sel. Skala biaksuspensi sel kemudian diperbesar dalam bio-reaktor kapasitas 5 L. Medium kultur yangdigunakan sama dengan medium pada labuErlenmeyer. Pertumbuhan sel dalam bioreaktorsangat lambat, bobot basah sel dan packed cellvolume (PCV) hanya bertambah berturut-turutsebesar 34% dan 50% setelah 21 hari dalamkultur, walaupun sebagian besar sel tetap viabel.


2016 ◽  
Vol 5 (2) ◽  
pp. 84 ◽  
Author(s):  
Sri Hutami

<p>Cell suspension culture could be defined as a<br />process that allows rapidly dividing homogenous suspension<br />of cells to grow in liquid nutrient media. There are two main<br />types of suspension cultures: (1) Batch cultures in which<br />cells are nurtured in a fixed volume of medium until growth<br />ceases and (2) Continuous cultures in which cell growth is<br />maintained by continuous replenishment of sterile nutrient<br />media. Plant cell suspension cultures are mostly used for the<br />biochemical investigation of cell physiology, growth, metabolism,<br />protoplast fusion, transformation and for large scale<br />production of seed by bioreactor and production of secondary<br />metabolites. Contamination is one of the largest problems<br />when dealing with cell cultures. Differences between<br />the products of cell suspension culture and whole plant are<br />frequently observed. These phenomena’s may be resulted<br />from lack of differentiation and organization and cell cultureinduced<br />variation. Utilization of cell suspension culture in<br />Indonesia is still limited, some of them for mass production<br />of plantation seed with bioreactor system and for production<br />of secondary metabolites. The success of this study give the<br />opportunity for mass production of seeds from other plants<br />and also production of secondary metabolites.</p>


2018 ◽  
pp. 01-07 ◽  
Author(s):  
Samrin Shaikh ◽  
Varsha Shriram ◽  
Tushar Khare and Vinay Kumar

Helicteres isora L. (Malvaceae) is a medicinal plant highly used in traditional therapeutic practices. It has shown wide-spectrum therapeutic activities including anti-plasmodial, hypoglycemic, hypolipidemic, hepatoprotective, antinociceptive, antioxidant and anti-HIV. Present investigation was undertaken with an objective of establishment of cell suspension cultures of this plant which can be further used for in vitro production of desired secondary metabolites and their further scale-up. Seed dormancy was broken using sulfuric acid and seedlings were raised in vitro. MS medium supplemented with 2,4-D (0.5 mg/L) produced maximum callus from the nodal explants. The callus produced was used as an explant for the establishment of suspension cultures. MS medium without any supplement was proved best for the establishment of cell suspension cultures of H. isora. To the best of our knowledge, this is the first report on H. isora cell suspension culture establishment.    


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