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71.
72.
Wessel Van Leeuwen Marc J. M. Hagendoorn Tom Ruttink Remco Van Poecke Linus H. W. Van Der Plas Alexander R. Van Der Krol 《Plant Molecular Biology Reporter》2000,18(2):143-144
The properties of the firefly luciferase (LUC) make it a very good nondestructive reporter to quantify and image transgene
promoter activity in plants. The short half-life of the LUC mRNA and protein, and the very limited regeneration of the LUC
protein after reacting with luciferin, enables monitoring of changes in gene activity with a high time resolution. However,
the ease at which luciferase activity is measuredin planta, using a light sensitive camera system (2D-luminometer), contrasts sharply with the complications that arise from interpreting
the results. A variegated pattern of luciferase activity, that is often observed inin planta measurements, might either be caused by differences in influx, availability of the substrates (luciferin, oxygen, ATP) or
by local differences in reporter gene activity. Here we tested the possible contribution of differences in the availability
of each substrate to the variegatedin planta luciferase activity, and we show whenin planta luciferase activity is measured under substrate equilibrium conditions and can be related to the promoter activity of the
reporter gene. Furthermore, we demonstrate the effects of protein stability, apparent half-life of luciferase activity, regeneration
of luciferase and pH on thein vivo andin vitro luciferase measurements. The combined results give the prerequisites for the correct utilisation of the luciferase reporter
system, especially forin vivo gene expression studies in plant research. 相似文献
73.
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75.
Structural and functional integrity of Sulla carnosa photosynthetic apparatus under iron deficiency conditions 下载免费PDF全文
A. Elkhouni M. Rabhi A. G. Ivanov M. Krol W. Zorrig A. Smaoui C. Abdelly N. P. A. Huner 《Plant biology (Stuttgart, Germany)》2018,20(3):415-425
- The abundance of calcareous soils makes bicarbonate‐induced iron (Fe) deficiency a major problem for plant growth and crop yield. Therefore, Fe‐efficient plants may constitute a solution for use on calcareous soils.
- We investigated the ability of the forage legume Sulla carnosa (Desf.) to maintain integrity of its photosynthetic apparatus under Fe deficiency conditions. Three treatments were applied: control, direct Fe deficiency and bicarbonate‐induced Fe deficiency.
- At harvest, all organs of deficient plants showed severe growth inhibition, the effect being less pronounced under indirect Fe deficiency. Pigment analysis of fully expanded leaves revealed a reduction in concentrations of chlorophyll a, chlorophyll b and carotenoids under Fe deficiency. Electron transport rate, maximum and effective quantum yield of photosystem II (PSII), photochemical quenching (qP), non‐photochemical quenching (qN) as well as P700 activity also decreased significantly in plants exposed to direct Fe deficiency, while qN was not affected. The effects of indirect Fe deficiency on the same parameters were less pronounced in bicarbonate‐treated plants. The relative abundances of thylakoid proteins related to PSI (PsaA, Lhca1, Lhca2) and PSII (PsbA, Lhcb1) were also more affected under direct than indirect Fe deficiency.
- We conclude that S. carnosa can maintain the integrity of its photosynthetic apparatus under bicarbonate‐induced Fe deficiency, preventing harmful effects to both photosystems under direct Fe deficiency. This suggests a high capacity of this species not only to take up Fe in the presence of bicarbonate (HCO3?) but also to preferentially translocate absorbed Fe towards leaves and prevent its inactivation.
76.
CYCLOHEXIMIDE AS A MEDIA AMENDMENT FOR ENUMERATING BACTERIAL POPULATIONS IN ANIMAL FEEDS 总被引:1,自引:0,他引:1
S.D. HA S.D. PILLAI K.G. MACIOROWSKI S.C. RICKE 《Journal of Rapid Methods and Automation in Microbiology》1995,4(2):95-105
The present study was designed to evaluate cycloheximide as a potential media amendment for differential bacterial and fungal enumeration of animal feeds. The objectives of this study were to determine the effect of cycloheximide on bacterial growth rates and to evaluate its efficacy for the reduction of indigenous spreading fungi when enumerating bacterial populations in three types of feeds and after short or long-term storage. Escherichia coli, Bacillus cereus, Staphylococcus aureus, Listeria monocytogenes and Pseudomonas fluorescens were grown in tryptic soy broth containing cycloheximide to determine its effect on bacterial specific growth rates. Growth rates of B. cereus and S. aureus were significantly decreased by the addition of 600 and 1000 mg/L cycloheximide respectively, but other pure cultures were not significantly influenced by cycloheximide addition. Intrinsic bacterial populations from feed were not significantly affected by cycloheximide additions at concentrations from 10 to 300 mg/L, but the indigenous spreading molds from feeds were significantly decreased by these cycloheximide concentrations and were decreased below detection levels by 300 mg/L of cycloheximide. The addition of 300 mg/L of cycloheximide effectively eliminates fungal growth for accurate enumeration of bacterial populations in feeds. 相似文献
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78.
Downregulation of ovule-specific MADS box genes from petunia results in maternally controlled defects in seed development. 总被引:17,自引:7,他引:10 下载免费PDF全文
L Colombo J Franken A R Van der Krol P E Wittich H J Dons G C Angenent 《The Plant cell》1997,9(5):703-715
A maternally determined seed defect has been obtained by downregulation of the petunia MADS box genes Floral Binding Protein 7 (FBP7) and FBP11. These genes have been previously shown to play central roles in the determination of ovule identity. Aberrant development of the seed coat and consequent degeneration of the endosperm have been observed in transgenic plants in which these two genes are downregulated by cosuppression. Analysis of the expression pattern of FBP7 and FBP11 and genetic analysis confirmed the maternal inheritance of the phenotype. The FBP7 promoter was cloned and fused to reporter genes. One of these reporter genes was the BARNASE gene for targeted cell ablation. Our results indicate that FBP7 promoter activity is restricted to the seed coat of developing seeds and that it is completely silent in the gametophytically derived tissues. The mutants used in this study provided a unique opportunity to investigate one of the poorly understood aspects of seed development: the interaction of embryo, endosperm, and maternal tissues. 相似文献
79.
S.D. HA K.G. MACIOROWSKI Y.M. KWON S.C. RICKE 《Journal of Rapid Methods and Automation in Microbiology》1997,5(4):309-319
The present study was designed to compare ethyl alcohol with buffered propionic acid feed treatment on the survival of indigenous poultry feed bacteria and fungi. The aerobic bacterial poultry feed populations were not substantially reduced by either ethyl alcohol or buffered propionic acid treatments. Likewise, indigenous poultry feed fungal populations also were not markedly reduced by buffered propionic acid treatment of the feed but fungal poultry feed populations exposed to ethyl alcohol treatments were significantly lower (P<0.05) than fungal populations recovered from either control and buffered propionic acid treated feeds. Ethyl alcohol treatment may have potential for reducing fungal contamination in poultry feed. 相似文献
80.
A novel RNA structural motif in the selenocysteine insertion element of eukaryotic selenoprotein mRNAs. 总被引:11,自引:2,他引:9 下载免费PDF全文
In eukaryotes, co-translational insertion of selenocysteine into selenoproteins necessitates the participation of the selenocysteine insertion sequence (SECIS), an element lying in the 3'-untranslated region of selenoprotein mRNAs. We report a detailed experimental study of the secondary structures of the SECIS elements of three selenoprotein mRNAs, the rat and human type I iodothyronine deiodinase (5'DI) and rat glutathione peroxidase (GPx). Based on RNase and chemical probing, a new secondary structure model is established. It is characterized by a stem-loop structure, comprising two helices (I and II) separated by an internal loop, with an apical loop surmounting helix II. Sequence comparisons of 20 SECIS elements, arising from 2 5'DI, 13 GPx, 2 selenoprotein P, and 1 selenoprotein W mRNAs, confirm the secondary structure model. The most striking finding of the experimental study concerns a set of conserved sequences in helix II that interact to form a novel RNA structural motif consisting of a quartet composed of non-Watson-Crick base pairs 5'UGAY3': 5'UGAU3'. The potential for forming the quartet is preserved in 15 SECIS elements, but three consecutive non-Watson-Crick base pairs can nevertheless form in the other five SECIS, the central G.A tandem being invariant in all cases. A 3D model, derived by computer modeling with the use of the solution data, suggests that the base pairing interactions in the G.A tandem are of the type found in GNRA loops. The 3D model displays the quartet lying in an accessible position at the foot of helix II, which is bent at the internal loop, suggesting that the non-Watson-Crick base pair arrangement provides an unusual pattern of chemical groups for putative ligand interaction. 相似文献