Endogenous enzyme activities and polyamine levels in diverse rice cultivars depend on the genetic background and are not affected by the presence of the hygromycin phosphotransferase selectable marker

2002 ◽  
Vol 105 (4) ◽  
pp. 594-603 ◽  
Author(s):  
O. Lepri ◽  
L. Bassie ◽  
P. Thu-Hang ◽  
P. Christou ◽  
T. Capell
2007 ◽  
Vol 6 (11) ◽  
pp. 2066-2071 ◽  
Author(s):  
A. George Smulian ◽  
Reta S. Gibbons ◽  
Jeffery A. Demland ◽  
Deborah T. Spaulding ◽  
George S. Deepe

ABSTRACT The Escherichia coli hygromycin phosphotransferase (hph) gene, which confers hygromycin resistance, is commonly used as a dominant selectable marker in genetically modified bacteria, fungi, plants, insects, and mammalian cells. Expression of the hph gene has rarely been reported to induce effects other than those expected. Hygromycin B is the most common dominant selectable marker used in the molecular manipulation of Histoplasma capsulatum in the generation of knockout strains of H. capsulatum or as a marker in mutant strains. hph-expressing organisms appear to have no defect in long-term in vitro growth and survival and have been successfully used to exploit host-parasite interaction in short-term cell culture systems and animal experiments. We introduced the hph gene as a selectable marker together with the gene encoding green fluorescent protein into wild-type strains of H. capsulatum. Infection of mice with hph-expressing H. capsulatum yeast cells at sublethal doses resulted in lethality. The lethality was not attributable to the site of integration of the hph construct into the genomes or to the method of integration and was not H. capsulatum strain related. Death of mice was not caused by altered cytokine profiles or an overwhelming fungal burden. The lethality was dependent on the kinase activity of hygromycin phosphotransferase. These results should raise awareness of the potential detrimental effects of the hph gene.


Antioxidants ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 1089
Author(s):  
Farwa Basit ◽  
Min Chen ◽  
Temoor Ahmed ◽  
Muhammad Shahid ◽  
Muhammad Noman ◽  
...  

This research was performed to explore the vital role of seed priming with a 0.01 µM concentration of brassinosteroids (EBL) to alleviate the adverse effects of Cr (100 µM) in two different rice cultivars. Seed priming with EBL significantly enhanced the germination attributes (germination percentage, germination energy, germination index, and vigor index, etc.), photosynthetic rate as well as plant growth (shoot and root length including the fresh and dry weight) under Cr toxicity as compared to the plants primed with water. Cr toxicity induced antioxidant enzyme activities (SOD, POD, CAT, and APX) and ROS level (MDA and H2O2 contents) in both rice cultivars; however, a larger increment was observed in YLY-689 (tolerant) than CY-927 (sensitive) cultivar. EBL application stimulatingly increased antioxidant enzyme activities to scavenge ROS production under Cr stress. The gene expression of SOD and POD in EBL-primed rice plants followed a similar increasing trend as observed in the case of enzymatic activities of SOD and POD compared to water-primed rice plants. Simultaneously, Cr uptake was observed to be significantly higher in the water-primed control compared to plants primed with EBL. Moreover, Cr uptake was significant in YLY-689 compared to CY-927. In ultra-structure studies, it was observed that EBL priming relieved the rice plants from sub-cellular damage. Conclusively, our research indicated that seed priming with EBL could be adopted as a promising strategy to enhance rice growth by copping the venomous effect of Cr.


2000 ◽  
Vol 27 (3) ◽  
pp. 201 ◽  
Author(s):  
Narayana M. Upadhyaya ◽  
Brian Surin ◽  
Kerrie Ramm ◽  
Judy Gaudron ◽  
Petra H. D. Schünmann ◽  
...  

We report the first successfulAgrobacterium-mediated transformation of Australian elite rice cultivars, Jarrah and Amaroo, using binary vectors with our improved promoters and selectable markers. Calli derived from mature embryos were used as target tissues. The binary vectors contained hph(encoding hygromycin resistance) or bar (encoding herbicide resistance) as the selectable marker gene and uidA (gus) or sgfpS65T as the reporter gene driven by different promoters. Use of Agrobacterium strain AGL1 carrying derivatives of an improved binary vector pWBVec8, wherein the CaMV35S driven hph gene is interrupted by the castor bean catalase 1 intron, produced a 4- fold higher number of independent transgenic lines compared to that produced with the use of strain EHA101 car-rying the binary vector pIG121-Hm wherein the CaMV35S driven hph is intronless. The Ubiquitin promoter produced 30-fold higher ß-glucuronidase (GUS) activity (derivatives of binary vector pWBVec8) in transgenic plants than the CaMV35S promoter (pIG121-Hm). The two modified SCSV promoters produced GUS activity com-parable to that produced by the Ubiquitin promoter. Progeny analysis (R1) for hygromycin resistance and GUS activ-ity with selected lines showed both Mendelian and non-Mendelian segregation. Lines showing very high levels of GUS activity in T0 showed a reduced level of GUS activity in their T1 progeny, while lines with moderate levels of GUS activity showed increased levels in T1 progeny. Stable heritable green fluorescent protein (GFP) expression was also observed in few transgenic plants produced with the binary vector pTO134 which had the CaMV35S promoter-driven selectable marker gene bar and a modified CaMV35S promoter-driven reporter gene sgfpS65T.


Author(s):  
E. R. Macagno ◽  
C. Levinthal

The optic ganglion of Daphnia Magna, a small crustacean that reproduces parthenogenetically contains about three hundred neurons: 110 neurons in the Lamina or anterior region and about 190 neurons in the Medulla or posterior region. The ganglion lies in the midplane of the organism and shows a high degree of left-right symmetry in its structures. The Lamina neurons form the first projection of the visual output from 176 retinula cells in the compound eye. In order to answer questions about structural invariance under constant genetic background, we have begun to reconstruct in detail the morphology and synaptic connectivity of various neurons in this ganglion from electron micrographs of serial sections (1). The ganglion is sectioned in a dorso-ventra1 direction so as to minimize the cross-sectional area photographed in each section. This area is about 60 μm x 120 μm, and hence most of the ganglion fit in a single 70 mm micrograph at the lowest magnification (685x) available on our Zeiss EM9-S.


Author(s):  
Takuma Saito ◽  
Toshihiro Takizawa

Cells and tissues live on a number of dynamic metabolic pathways, which are made up of sequential enzymatic cascades.Recent biochemical and physiological studies of vision research showed the importance of cGMP metabolism in the rod outer segment of visual cell, indicat ing that the photon activated rhodopsin exerts activation effect on the GTP binding protein, transducin, and this act ivated transducin further activates phosphodiesterase (PDEase) to result in a rapid drop in cGMP concentration in the cytoplasm of rod outer segment. This rapid drop of cGMP concentration exerts to close the ion channel on the plasma membrane and to stop of inward current brings hyperpolarization and evokes an action potential.These sequential change of enzyme activities, known as cGMP cascade, proceeds quite rapidly within msec order. Such a rapid change of enzyme activities, such as PDEase in rod outer segment, was not a matter of conventional histochemical invest igations.


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