SPORULATION OF BACILLUS POPILLIAE ON SOLID MEDIA

1965 ◽  
Vol 11 (5) ◽  
pp. 779-783 ◽  
Author(s):  
R. A. Rhodes ◽  
M. S. Roth ◽  
G. R. Hrubant

Spores of the insect pathogen Bacillus popilliae Dutky have been formed in vitro from vegetative cultures. The procedure results reproducibly in 0.1 to 0.3% spore formation in cells of colonies grown on a solid medium under strictly denned conditions. Sporulation requires a selected strain of the organism, NRRL B-2309S, a relatively large and specific concentration of certain yeast extracts, a specific type of agar, the complete absence of glucose, the presence of acetate, and a pH within the range 7.2 to 7.5. Spore formation occurs slowly during 2- to 4-week incubation periods in surface colonies present in limited numbers on agar plates. Some of the spores formed in this manner survive heating for 15 minutes at 80 °C, and vegetative cultures derived from such spores are pathogenic via injection for larvae of the Japanese beetle, Popillia japonica Newman.

HortScience ◽  
1991 ◽  
Vol 26 (6) ◽  
pp. 728E-728
Author(s):  
Roberto A. Rivas ◽  
P.T. Evans ◽  
D.R. LaBonte

Methodology is presented for organogenesis of `Beauregard sweetpotato, a cultivar released in 1987 that is rapidly increasing in commercial use in the U.S. Regeneration of `Beauregard' sweetpotato plantlets was obtained when complete leaves and 10 mm internode explants were cultured in liquid and solid media respectively over a period of 8 weeks. Leaves in liquid Murashige and Skoog medium containing 2 mg/l of IAA placed on a shaker under dark conditions produced white callus at the cut end of the petiole and roots underneath the callus in 4 weeks. Leaves were subsequently transferred to MS medium containing 500 mg/l of Chlorocholine chloride (CCC) and 0, 1, 5 and 10 mg/l of BAP for 4 more weeks. Shoots were regenerated from callus using 1 mg/l of BAP. The effect of NAA auxin and various concentrations of the cytokinin Thidiazuron on internodes was examined under 16 hr. light and 8 hr dark photoperiod using MS solid medium. Explants on 0.05 mg/l NAA alone produced roots and shoots. The most plantlets however, were regenerated using 0.05 mg/l of NAA and 0.01 mg/l Thidiazuron. Regeneration of plants from leaves and internodes may be a useful system for a clean and rapid propagation of `Beauregard' sweetpotato.


Author(s):  
Sulistyani Pancaningtyas

Research  aimed  to  study  the  effect  of  solid  culture,  liquid  culture,  and sequence  subculture  on  quantity  and  hyperhydricity  of  somatic  embryo  wascarried  out  at  Laboratory  of  Biotechnology,  Indonesian  Coffee  and  Cocoa Research  Institute.  Materials  used  in  this  study  were  embryogenic  callitransferred  on  somatic  embryos  expression  both  in  solid  and  liquid  media with  the  same  media  composition,  namely  MS  medium  with  the  addition  ofAdenine  (0.025  mg/L).  Gelling  agent  used  in  solid  media  was  gelrite  (3  g/L). Clones used in this study was Sca 6. This research consisted of two trials, namely1)  effect  of  medium  type  (solid  and  liquid),  and  2)  sequence  subculture  (four subcultures).  This  results  showed  that  the  production  of  somatic  embryosin  liquid  medium  was  higher  than  in  the  solid  medium.  Regeneration  of somatic  embryos  on  solid  medium  culture  showed  the  highest  percentage  of abnormality  embryos  due  to  hyperhydricity  at  the  cotyledonary  phase  60%. Meanwhile,  the  regeneration  of  somatic  embryos  in  liquid  culture  showed the  highest  percentage  of  abnormality  embryos  due  to   hyperhydricity  at the  globular  and  cotyledonary  phase  37%.  Frequent  subculture  increased abnormal embryos  and  decreased  the  number of  somatic  embryos.Key words: Cacao, hyperhydricity,  somatic embryos,  solid  culture,  liquid  culture,  subculture,  in  vitro.


2020 ◽  
Vol 65 (1) ◽  
pp. e01948-20
Author(s):  
Dalin Rifat ◽  
Si-Yang Li ◽  
Thomas Ioerger ◽  
Keshav Shah ◽  
Jean-Philippe Lanoix ◽  
...  

ABSTRACTThe nitroimidazole prodrugs delamanid and pretomanid comprise one of only two new antimicrobial classes approved to treat tuberculosis (TB) in 50 years. Prior in vitro studies suggest a relatively low barrier to nitroimidazole resistance in Mycobacterium tuberculosis, but clinical evidence is limited to date. We selected pretomanid-resistant M. tuberculosis mutants in two mouse models of TB using a range of pretomanid doses. The frequency of spontaneous resistance was approximately 10−5 CFU. Whole-genome sequencing of 161 resistant isolates from 47 mice revealed 99 unique mutations, of which 91% occurred in 1 of 5 genes previously associated with nitroimidazole activation and resistance, namely, fbiC (56%), fbiA (15%), ddn (12%), fgd (4%), and fbiB (4%). Nearly all mutations were unique to a single mouse and not previously identified. The remaining 9% of resistant mutants harbored mutations in Rv2983 (fbiD), a gene not previously associated with nitroimidazole resistance but recently shown to be a guanylyltransferase necessary for cofactor F420 synthesis. Most mutants exhibited high-level resistance to pretomanid and delamanid, although Rv2983 and fbiB mutants exhibited high-level pretomanid resistance but relatively small changes in delamanid susceptibility. Complementing an Rv2983 mutant with wild-type Rv2983 restored susceptibility to pretomanid and delamanid. By quantifying intracellular F420 and its precursor Fo in overexpressing and loss-of-function mutants, we provide further evidence that Rv2983 is necessary for F420 biosynthesis. Finally, Rv2983 mutants and other F420H2-deficient mutants displayed hypersusceptibility to some antibiotics and to concentrations of malachite green found in solid media used to isolate and propagate mycobacteria from clinical samples.


Author(s):  
Kiptiyah Kiptiyah ◽  
Widodo Widodo ◽  
Gatot Ciptadi ◽  
Aulanni’am Aulanni’Am ◽  
Mohammad A. Widodo ◽  
...  

AbstractBackgroundWe investigated whether 10-gingerol is able to induce oxidative stress in cumulus cells.MethodsFor the in-vitro research, we used a cumulus cell culture in M199, containing 10-gingerol in various concentrations (0, 12, 16, and 20 µM), and detected oxidative stress through superoxide dismutase (SOD) activity and malondialdehyde (MDA) concentrations, with incubation periods of 24, 48, 72, and 96 h. The obtained results were confirmed by in-silico studies.ResultsThe in-vitro data revealed that SOD activity and MDA concentration increased with increasing incubation periods: SOD activity at 0 µM (1.39 ± 0.24i), 12 µM (16.42 ± 0.35ab), 16 µM (17.28 ± 0.55ab), 20 µM (17.81 ± 0.12a), with a contribution of 71.1%. MDA concentration at 0 µM (17.82 ± 1.39 l), 12 µM (72.99 ± 0.31c), 16 µM (79.77 ± 4.19b), 20 µM (85.07 ± 2.57a), with a contribution of 73.1%. Based on this, the in-silico data uncovered that 10˗gingerol induces oxidative stress in cumulus cells by inhibiting HTR1A functions and inactivating GSK3B and AKT˗1.Conclusions10-gingerol induces oxidative stress in cumulus cells through enhancing SOD activity and MDA concentration by inhibiting HTR1A functions and inactivating GSK3B and AKT˗1.


Molecules ◽  
2021 ◽  
Vol 26 (10) ◽  
pp. 2835
Author(s):  
Anna Stochmal ◽  
Bartosz Skalski ◽  
Rostyslav Pietukhov ◽  
Beata Sadowska ◽  
Joanna Rywaniak ◽  
...  

Although the major components of various organs of sea buckthorn have been identified (particularly phenolic compounds), biological properties of many of these phytochemicals still remain poorly characterized. In this study, we focused on the chemical composition and biological activity of preparations that were obtained from sea buckthorn twigs and leaves. The objective was to investigate cytotoxicity of these preparations against human fibroblast line HFF-1, using MTT reduction assay, their anti- or pro-oxidant activities against the effects of a biological oxidant -H2O2/Fe—on human plasma lipids and proteins in vitro (using TBARS and carbonyl groups as the markers of oxidative stress). Antimicrobial activity of the tested preparations against Gram-positive (Staphylococcus aureus, S. epidermidis, Enterococcus faecalis) and Gram-negative bacteria (Escherichia coli, Pseudomonas aeruginosa), as well as against fungi (Candida albicans, C. glabrata) by the EUCAST-approved broth microdilution method, followed by growth on solid media, were also assessed. Our analysis showed significant differences in chemical composition and biological properties of the tested preparations (A–F). All tested preparations from sea buckthorn twigs (D–F) and one preparation from sea buckthorn leaves (preparation C) may be a new source of phenolic antioxidants for pharmacological and cosmetic applications.


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