High-performance liquid chromatographic determination and standardization of agmatine

Bencsik, Krisztina; Kremmer, T.; Boldizsár, M.; Tamás, J.; Mák, Marianna; Páldi, Emil

doi:10.1016/s0021-9673(97)00193-3

Cited by 15 publications

(9 citation statements)

References 15 publications

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“…Agmatine is also a potent DON inducer and a precursor of putrescine synthesis but using our extraction methods we were unable to detect agmatine in wheat heads (data not shown). This is most likely due to the instability of agmatine, as observed by others as well [37]. Interestingly, another polyamine and potent inducer of DON production, cadaverine [15], was not detected in the mock-inoculated heads at all but accumulated in infected heads, with significant increases observed at three days post-inoculation, eventually reaching a concentration of 200 nmoles/g fresh weight.…”

Section: Resultsmentioning

confidence: 71%

Early activation of wheat polyamine biosynthesis during Fusarium head blight implicates putrescine as an inducer of trichothecene mycotoxin production

et al. 2010

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BackgroundThe fungal pathogen Fusarium graminearum causes Fusarium Head Blight (FHB) disease on wheat which can lead to trichothecene mycotoxin (e.g. deoxynivalenol, DON) contamination of grain, harmful to mammalian health. DON is produced at low levels under standard culture conditions when compared to plant infection but specific polyamines (e.g. putrescine and agmatine) and amino acids (e.g. arginine and ornithine) are potent inducers of DON by F. graminearum in axenic culture. Currently, host factors that promote mycotoxin synthesis during FHB are unknown, but plant derived polyamines could contribute to DON induction in infected heads. However, the temporal and spatial accumulation of polyamines and amino acids in relation to that of DON has not been studied.ResultsFollowing inoculation of susceptible wheat heads by F. graminearum, DON accumulation was detected at two days after inoculation. The accumulation of putrescine was detected as early as one day following inoculation while arginine and cadaverine were also produced at three and four days post-inoculation. Transcripts of ornithine decarboxylase (ODC) and arginine decarboxylase (ADC), two key biosynthetic enzymes for putrescine biosynthesis, were also strongly induced in heads at two days after inoculation. These results indicated that elicitation of the polyamine biosynthetic pathway is an early response to FHB. Transcripts for genes encoding enzymes acting upstream in the polyamine biosynthetic pathway as well as those of ODC and ADC, and putrescine levels were also induced in the rachis, a flower organ supporting DON production and an important route for pathogen colonisation during FHB. A survey of 24 wheat genotypes with varying responses to FHB showed putrescine induction is a general response to inoculation and no correlation was observed between the accumulation of putrescine and infection or DON accumulation.ConclusionsThe activation of the polyamine biosynthetic pathway and putrescine in infected heads prior to detectable DON accumulation is consistent with a model where the pathogen exploits the generic host stress response of polyamine synthesis as a cue for production of trichothecene mycotoxins during FHB disease. However, it is likely that this mechanism is complicated by other factors contributing to resistance and susceptibility in diverse wheat genetic backgrounds.

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Section: Resultsmentioning

confidence: 71%

Early activation of wheat polyamine biosynthesis during Fusarium head blight implicates putrescine as an inducer of trichothecene mycotoxin production

et al. 2010

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“…The polyamine standards 1,4-diaminobutane, spermidine trihydrochloride, and spermine tetrahydrochloride (Sigma) were prepared in water. The dansylation reaction was carried out by the method of Bencsik et al (1). Briefly, 100 l of saturated sodium carbonate and 200 l of dansyl chloride (5 mg/ml in acetone) were added to 100 l of the tissue supernatant or the polyamine standards and incubated at 56°C for 1 h in the dark.…”

Section: Methodsmentioning

confidence: 99%

The mechanism of opiorphin-induced experimental priapism in rats involves activation of the polyamine synthetic pathway

Kanika

Tar

Tong

et al. 2009

American Journal of Physiology-Cell Physiology

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Intracorporal injection of plasmids encoding opiorphins into retired breeder rats can result in animals developing a priapic-like condition. Microarray analysis demonstrated that following intracorporal gene transfer of plasmids expressing opiorphins the most significantly upregulated gene in corporal tissue was the ornithine decarboxylase gene (ODC). Quantitative RT-PCR confirmed the upregulation of ODC, as well as other genes involved in polyamine synthesis, such as arginase-I and -II, polyamine oxidase, spermidine synthase, spermidine acetyltransferase (SAT), and S-adenosylmethionine decarboxylase. Western blot analysis demonstrated upregulation of arginase-I and -II, ODC, and SAT at the protein level. Levels of the polyamine putrescine were upregulated in animals treated with opiorphin-expressing plasmids compared with controls. A direct role for the upregulation of polyamine synthesis in the development of the priapic-like condition was supported by the observation that the ODC inhibitor 1,3-diaminopropane, when added to the drinking water of animals treated with plasmids expressing opiorphins, prevented experimental priapism. We also demonstrate that in sickle cell mice, another model of priapism, there is increased expression of the mouse opiorphin homologue in corporal tissue compared with the background strain at a life stage prior to evidence of priapism. At a life stage when there is onset of priapism, there is increased expression of the enzymes involved in polyamine synthesis (ODC and arginase-I and -II). Our results suggest that the upregulation of enzymes involved in the polyamine synthetic pathway may play a role in the development of experimental priapism and represent a target for the prevention of priapism.

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“…The quantitative and qualitative analysis of polyamines was carried out using a modified high-performance liquid chromatography (HPLC) method (Bencsik et al, 1998), which allowed the accurate detection of agmatine (Agm). One-gram plant samples were frozen in liquid nitrogen, squashed and homogenised in 5 mL buffer (5 mM phosphate buffer, pH 7.0+85% NaCl), after which the polyamines were extracted with 3 Â 5 mL perchloric acid.…”

Section: Hplc Assaymentioning

confidence: 99%

S-methylmethionine reduces cell membrane damage in higher plants exposed to low-temperature stress

Rácz

Páldi

Szalai

et al. 2008

Journal of Plant Physiology

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High-performance liquid chromatographic determination and standardization of agmatine

Cited by 15 publications

References 15 publications

Early activation of wheat polyamine biosynthesis during Fusarium head blight implicates putrescine as an inducer of trichothecene mycotoxin production

Early activation of wheat polyamine biosynthesis during Fusarium head blight implicates putrescine as an inducer of trichothecene mycotoxin production

The mechanism of opiorphin-induced experimental priapism in rats involves activation of the polyamine synthetic pathway

S-methylmethionine reduces cell membrane damage in higher plants exposed to low-temperature stress

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