A cell counting procedure applied to embryogenesis in cell suspension cultures of anise (Pimpinella anisum L.)

1978 ◽  
Vol 12 (3-4) ◽  
pp. 209-215 ◽  
Author(s):  
J Huber ◽  
F Constabel ◽  
O.L Gamborg
2021 ◽  
Vol 6 (3) ◽  
pp. 5
Author(s):  
Muhammad Akram ◽  
Faheem Aftab

Jojoba (Simmondsia chinensis L.) oil is also known as liquid wax or fixed oil. It is an important metabolite of jojoba having commercial importance in cosmetics as well as a potential biofuel source. We presented an efficient system for in vitro establishment of cell suspension cultures (CSC) from proliferating friable calluses. For this purpose, cotyledon, internode, and leaf explants were cultured on MS medium + 1, 2, 4, 6, 8 or 10 µM 2, 4-Dichlorophenoxyacetic acid (2, 4-D), α-Naphthalene acetic acid (NAA) alone or in combination with 1 or 2 µM N6-benzylaminopurine (BAP) or Kinetin. Results demonstrated that 100% healthy, friable and variegated calluses were obtained on 8 µM, 10 µM 2, 4-D or 2, 4-D 10 µM + 2 µM BAP and represented as callus lines (CL) CL-1, CL-2 or CL-3, respectively, after 38 days. One-gram callus tissue per CL was then immersed in the respective liquid medium and agitated on an orbital shaker at 60-70 rpm under the growth room conditions (25 ± 2 °C, 16 h light period) for the preparation of CSC. After 15 days, CSC was sieved and large clumps were removed. Growth measurement of CSC was determined by cell counting, packed cell volume (PCV) and cell viability. The highest number of viable cells was obtained at 2.57 OD with CL-3, where PCV was highest (0.35 ml) on CL-1 of 38 days old calluses. 2,3,5-Triphenyltetrazolium chloride was a reliable approach for the determination of cell viability of CSC.


1991 ◽  
Vol 46 (1-2) ◽  
pp. 58-66 ◽  
Author(s):  
Werner Gunia ◽  
Walter Hinderer ◽  
Uta Wittkampf ◽  
Wolfgang Barz

Abstract A yeast glucan elicitor causes the accumulation of the pterocarpan phytoalexins medicarpin and maackiain in chickpea (Cicer arietinum) cell suspension cultures established from seeds. A cell culture line from a chickpea cultivar resistant against its main fungal pathogen Ascochyta rabiei accumulates large amounts (944 nm ol/g fr. wt.) whereas a cell culture line from a susceptible cultivar accumulates only low amounts (38 nm ol/g fr. wt.) of the phytoalexins. This is consistent with differential accumulation of pterocarpan phytoalexins in intact plants [1], The first reactions in the pterocarpan-specific branch of biosynthesis are hydroxylation of the isoflavone intermediate form ononetin in position 2′ or 3′, catalyzed by microsomal cytochrome P-450 monooxygenases. Upon elicitation form ononetin 2′-hydroxylase undergoes a strong transient induction in the cell suspension culture of the resistant cultivar, whereas in the cell culture from the susceptible cultivar it is only slightly induced. In both cell suspension cul­tures the induction of cinnamic acid 4-hydroxylase and of form ononetin 3′-hydroxylase does not show a clear correlation with phytoalexin accumulation. Experiments with different elici­tor concentrations confirm that formononetin 2′-hydroxylase is much more induced in cell cul­tures from the resistant cultivar than from the susceptible one. It is concluded that the massive difference in phytoalexin accumulation between cell suspension cultures from the resistant and susceptible cultivar is determined mainly by the differential induction of form ononetin 2′-hydroxylase activity.


1986 ◽  
Vol 41 (1-2) ◽  
pp. 187-192 ◽  
Author(s):  
A. Baumert ◽  
G. Schneider ◽  
D. Gröger

A cell-free extract of Ruta graveolens cell suspension cultures, strain R-20, was prepared which synthesizes 1,3-dihydroxy-N-methylacridone from N-methylanthranilic acid and malonylcoenzyme A. The reaction is strongly dependent on the presence of ATP. After adding of coenzyme A to the assay mixtures a second compound X2 is accumulated to a considerable extent. The structure of this substance has not yet been clarified. Apparently X2 is a precursor of 1,3-dihydroxy-N-methylacridone, the key intermediate in the pathway leading to more complex acridones.


Author(s):  
Mariana Sánchez-Ramos ◽  
Laura Alvarez ◽  
Antonio Romero-Estrada ◽  
Antonio Bernabé-Antonio ◽  
Silvia Marquina-Bahena ◽  
...  

Many species of the Asteraceae family are used in traditional Mexican medicine for possessing healing properties. Ageratina pichinchensis (Asteraceae) is a plant used for the treatment of gastric ulcers, deep wounds and for its antifungal effects. The aim of this study was to establish a cell suspension culture of A. pichinchensis, quantify the anti-inflammatory constituents 2,3-dihydrobenzofuran and 3-epilupeol, to evaluate the anti-inflammatory potential of its extracts and perform a phytochemical analysis. Cell suspension cultures were established in MS culture medium supplemented with 30 g L-1 sucrose and 1.0 g L-1 α-naphthaleneacetic acid (NAA) plus 0.1 mg L-1 6-furfurylaminopurine (KIN). The ethyl acetate extracts of cell suspension cultures analyzed by GC revealed that the maximum production of compounds The anti-inflammatory activity of these extracts exhibited significant inhibition of NO production. Furthermore, the phytochemical study of EtOAc and MeOH extracts of the biomass on day 20 led to the identification of 17 known compounds. The structures of compounds were assigned by analysis of 1D and 2D NMR data and the remainder by GC-MS. This is the first report of the production of the (-)-Artemesinol, (-)-Artemesinol glucoside, Encecalin and 3,5-diprenyl-acetophenone compounds by a cell suspension cultures of A. pichinchensis.


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