Detectable ,-glucuronidase activity in transgenic cranberry is affected by endogenous inhibitors and plant development

Serres, R.; McCown, Brent H.; Zeldin, Eric L.

doi:10.1007/s002990050294

Cited by 23 publications

(30 citation statements)

References 21 publications

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“…Other plants containing high concentrations of phenolics, such as cranberry, also reduce GUS activity (Serres et al, 1997). However, the 15% decrease which occurred in our experiments was not enough to fully explain the very low (sometimes undetectable) levels of GUS activity observed in most of the transgenic sorghum tissues analyzed.…”

Section: Pluronic-f68 Dramatically Improved Transient Gus Expression contrasting

confidence: 43%

Agrobacterium-mediated transformation of sorghum: factors that affect transformation efficiency

et al. 2004

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The results presented in this work support the hypothesis that Agrobacterium-mediated transformation of sorghum is feasible, analogous to what has been demonstrated for other cereals such as rice, maize, barley and wheat. The four factors that we found most influenced transformation were: the sensitivity of immature sorghum embryos to Agrobacterium infection, the growth conditions of the donor plant, type of explant and co-cultivation medium. A major problem during the development of our protocol was a necrotic response which developed in explants after co-cultivation. Immature sorghum embryos proved to be very sensitive to Agrobacterium infection and we found that the level of embryo death after co-cultivation was the limiting step in improving transformation efficiency. The addition of coconut water to the co-cultivation medium, the use of vigorous and actively growing immature embryos and the removal of excess bacteria significantly improved the survival rate of sorghum embryos and was critical for successful transformation. Hygromycin phosphotransferase (hpt) proved to be a good selectable marker for sorghum. We also found that β-glucuronidase (GUS) activity was low in most of the transgenic plant tissues tested, although it was very high in immature inflorescences. Although promising, the overall transformation efficiency of the protocol is still low and further optimization will require particular attention to be given to the number of Agrobacterium in the inoculum and the selection of sorghum genotypes and explants less sensitive to Agrobacterium infection.

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Section: Pluronic-f68 Dramatically Improved Transient Gus Expression contrasting

confidence: 43%

Agrobacterium-mediated transformation of sorghum: factors that affect transformation efficiency

et al. 2004

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“…Those high inhibition capacities of different doses extracts were possibly due to those secondary compounds characterized by HPLC. Some compounds of plant origin were reported as the potent GUS inhibitor [9] and phenolic compounds were reported as one likely inhibitor group [21,10] …”

Section: Effect Of Crude Extract Concentration On Gus Activitymentioning

confidence: 99%

“…Besides, it was reported that activity of GUS enzyme extracted from the plant tissues was also influenced by interfering secondary plant compounds. Those secondary compounds were phenolic compounds, endogenous flavonols and proanthocyanidins and were reported as likely the inhibitor groups [10]. Thus it is possible that those plants may take a role in cancer prevention by their antioxidant and GUS inhibition capacities.…”

Section: Introductionmentioning

confidence: 99%

Pistacia terebinthus extract: As a potential antioxidant, antimicrobial and possible β-glucuronidase inhibitor

Kavak

Altıok

Bayraktar

et al. 2010

Journal of Molecular Catalysis B: Enzymatic

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a b s t r a c tIn this study, inhibition of Escherichia coli glucuronidase (GUS) by a plant extract of Pistacia terebinthus was investigated. P. terebinthus (from Karaburun region in Turkey) leaf extract exhibits various bioactivities, such as antioxidant, antimicrobial and cytotoxic property because of its flavonoid, phenolic and alkaloid contents. HPLC analyses were performed to evaluate the composition of the extract and main flavonoid compounds such as luteolin, luteolin-7-glucoside and apigenin-7-glucoside were detected in extract. GUS activity and kinetic data of the GUS were obtained by measuring rate of production of resulting fluorescent 4-methylumbelliferone (4-MU) and phenolphthalein fluorometrically and spectrophotometrically, respectively. Plant leaves showed higher antioxidant capacity as 85 TEAC (trolox equivalent antioxidant capacity) value and higher antimicrobial activity towards Staphylococcus aureus. 92.4% GUS enzyme inhibition was achieved with 8.3 g ml −1 crude extract concentration, where IC 50 value was found as 2.1 (±0.2) g ml −1 . Kinetic studies by nonlinear regression revealed that inhibition type caused by plant extract was mixed type (noncompetitive and competitive together). K m and V max values were 96.94 ± 9.68 nM and 7.81 ± 0.16 nmol min −1 mg −1 , respectively. Inhibition constant (˛K i ) was found as 7.00 ± 0.93 nM having˛value 2.17 ± 0.93. Our results may provide useful information regarding the inhibition of microbial GUS activity, preventing deglucuronidation and reducing possible cancer risk by bioactive P. terebinthus crude extract.

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“…Detection of β-glucuronidase (GUS) expression can be used to indicate both transient and stable transformation (Serres et al, 1997). In present study, GUS activity was detected in leaf explants after three days of co-cultivation and in putative transformed callus on the subculture 1,2,3,4, and 5.…”

Section: Expression Of β-Glucuronidase (Gus) In Transformed Tissuementioning

confidence: 76%

Agrobacterium-Mediated Stable Transformation of Medicinal Plant Andrographis paniculata Callus Expressing β-glucuronidase (GUS) Gene

Marwani¹,

Tangapo²,

Dwivany³

2015

IJBiotech

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This study was carried out to establish a stable genetic transformation in callus culture of Andrographis paniculata mediated by Agrobacterium tumefaciens. The leaf disks of A. paniculata were infected with A. tumefaciens LBA4404 carrying a binary vector pCAMBIA1304 that contain β-glucuronidase (GUS) and hygromycin phosphotransferase (hpt) genes. The infection was conducted by dipping method for one hour, followed by co-cultivation in the dark for three days. To examine transient GUS expression, the co-cultivated leaf disks were assayed for β-glucuronidase activity and to obtain stable transformed callus, the co-cultivated leaf disks were selected on the callus induction medium which contain 20 mg/l hygromycin for selection. The transformed callus was periodically subcultured every three weeks into the fresh selection medium over the 15 weeks period. To test a stable transformation, the callus was subjected to PCR analysis for GUS gene detection. The results indicated that the co-cultivated leaf disks expressed GUS activity and proliferated to produce callus on the selective medium. Analysis of PCR on the transformed callus indicated the presence 976 bp fragment that confi rmed the presence of β-glucuronidase gene. These fi ndings imply that the β-glucuronidase was stably integrated into A. paniculata callus culture.

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Detectable ,-glucuronidase activity in transgenic cranberry is affected by endogenous inhibitors and plant development

Cited by 23 publications

References 21 publications

Agrobacterium-mediated transformation of sorghum: factors that affect transformation efficiency

Agrobacterium-mediated transformation of sorghum: factors that affect transformation efficiency

Pistacia terebinthus extract: As a potential antioxidant, antimicrobial and possible β-glucuronidase inhibitor

Agrobacterium-Mediated Stable Transformation of Medicinal Plant Andrographis paniculata Callus Expressing β-glucuronidase (GUS) Gene

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