As a series of studies on the nucleic acids and their related substances 210 samples were collected from 76 places such as farm soil, compost of heap, nuruk and meju to obtain microbial strains which produce 5'-phosphodiesterase. From these samples total of 758 strains were isolated by the use of dilution pour plate method. For all isolated strains primary screening of the productivity of RNA depolymerase was performed and useful strains with regard to 5'-phosphodiesterase productivities were identified. For these useful strains optimum condition, the effect of various compounds on the activity of 5'-phosphodiesterase, and the optimum condition for enzyme reaction were discussed. The quantitative of 5'-mononucleotides produced by the action of 5'-phosphodiesterase was performed using anion-exchange column chromatography and their identified was done by paper chromatography, thinlayer chromatography, ultra violet spectrophotometry, and characteristic color reaction using carbazole and schiff's reagent. (1) Penicillium citreo-viride PO 2-11 and Streptomyces aureus SOA 4-21 from soil were identified as a potent 5'-phosphodiesterase producing strains. (2) Optimum culture conditions for Penicillium citreo-viride PO 2-11 strain isolated were found to be pH 5.0 and $30^{\circ}C$, and the optimum conditions for enzyme action of 5'-phosphodiesterase were pH 4.2 and $60^{\circ}C$. Best carbon source for the production of 5'-phosphodiesterase was found to be sucrose and ammonium nitrate for nitrogen source. Addition of 0.01% corn steep liquor or yeast extract exhibited 20% increase in the amount of 5'-phosphodiesterase production compared to the control. 5'-phosphodiesterase produced by this strain was activated by $Mg^{++},\;Ca^{++},\;Zn^{++},\;Mn^{++}$ and was inhibited by EDTA, citrate, $Cu^{++},\;CO^{++}$. 5'-phosphodiesterase produced 5'-mononucleotide from RNA at a rate of 65.81%, and among the 5'-mononucleotides accumulated 5'-GMP only was found to have flavorous and the strain was also found lack of 5'-AMP deaminase. Productivity of flavorous 5'-GMP was found to be 186.7mg per gram of RNA. (3) Optimum culture canditions for the isolated Streptomyces aureus SOA 4-21 strain were pH 7.0 and $28^{\circ}C$, and the optimum conditions for the action of 5'-phosphodiesterase were pH 7.3 and $50^{\circ}C$. The best carbon source for 5'-phosphodiesterase production was found to be glucose and that of nitrogen was asparagine. Addition of 0.01% yeast extract exhibited increased productivity of 5'-phosphodiesterase by 40% compared to the non-added control. 5'-phosphodiesterase produced by this strain was activated by $Ca^{++},\;Zn^{++},\;Mn^{++}$ and was inhibited by citrate, EDTA, $Cu^{++}$. It was also found that the strain produce 5'-AMP deaminase in addition to 5'-phosphodiesterase. For this reason although decomposition rate was 63.58% the accumulation of 5'-AMP, 5'-CMP, 5'-GMP and 5'-UMP occurred by the breakdown of RNA. In the course of these reaction 5'-AMP deaminase converted 60% of 5'-AMP thus produced into 5'-IMP and flavorous 5'-mono nucleotide production was significantly increased by this strain over the above mentioned one. Production rates were found to be 171.8mg per grain of RNA for 5'-IMP and 148.2mg per gram of RNA for 5'-GMP, respectively.
Carrot cultured cells are able to respond to a temperature increase by inducing a set of new proteins, heat shock proteins (HSP). Such an induction of the HS gene was known to be achieved mainly at the level of transcription. However there has been an increasing number of evidences showing that a translational control was involved in the regulation of the HS gene expression. A comparison of HSP synthesized in vivo to in vivo (represent for mRNA level since the amount of the proteins produced by in vivo translation system will be proportional to an amount of the corresponding mRNA)showed no correlation between the amount of HS mRNA and the amount of the corresponding HSP at $30^{\circ}C$, It appears that a translational control may exert a major role in the expression of HS gene in carrot callus cells at $30^{\circ}C$.
The fluctuations of biochemical and molecular activities III the harmful dinoflagellate, Cochlodinium polykrikoides, depending on water temperatures, were studied. In genomic DNA concentration, a similar value of 0.6 was shown at $12^{\circ}C$ and $15^{\circ}C$, but significantly increasing DNA from $18^{\circ}C$ (p<0.05), with a maximum of 1.8 at $24^{\circ}C$. After$24^{\circ}C$, the DNA significantly decreased to 0.6. Likely, the concentrations of RNA and total protein were at their highest values of 1.7 and 0.07 g $mL^1$ at $24^{\circ}C$, respectively. In contrast to ONA, RNA and total protein began to increase at $15^{\circ}C$. Oxygen availability between lower and higher temperatures was significantly different and increased from $18^{\circ}C$ according to light intensity, regardless of wavelengths (p<0.05). At $24^{\circ}C$, the highest value of the maximum electron transport rate (ETRmax), ranging from 537.9 (Ch 1) to 602.5 mol electrons $g^{-1}$ Ch1 a $s^{-1}$ (Ch 4), was also shown. Nitrate reductase (NR) and ATPase activities were at their highest values of 0.11 mol $NO_2^-g^{-1}$ Ch1 a $h^{-1}$ and 0.78 pmol 100 $mg^{-1}$$at^2$$4^{\circ}C$, respectively. When the cells cultured at $15^{\circ}C$, NR and ATPase activities significantly increased compared to $12^{\circ}C$ (p<0.05). In an analysis of CHN, the concentration of C and N also significantly increased (p<0.05). However, at $27^{\circ}C$, most of the molecular and biochemical movements were much lower, compared to $24^{\circ}C$. These results suggest that C. polykrikoides is very sensitive biochemical and molecular activities depending on water temperatures. Possibly, it is desirable to estimate at $18^{\circ}C$ the initiation of the massive blooming development of C. polykrikoides. In nature, it will be very difficult to maintain the massive blooms after $24^{\circ}C$ because of the possibility of significantly decreasing the molecular movement and activity of C. polykrikoides.
Chung, Bong Nam;Koh, Sang Wook;Choi, Kyung San;Joa, Jae Ho;Kim, Chun Hwan;Selvakumar, Gopal
The Plant Pathology Journal
/
v.33
no.5
/
pp.522-527
/
2017
We determined the effects of atmospheric temperature ($10-30{\pm}2^{\circ}C$ in $5^{\circ}C$ increments) and carbon dioxide ($CO_2$) levels ($400{\pm}50ppm$, $540{\pm}50ppm$, and $940{\pm}50ppm$) on the infection of Solanum tuberosum cv. Chubaek by Potato leafroll virus (PLRV). Below $CO_2$ levels of $400{\pm}50ppm$, the PLRV infection rate and RNA content in plant tissues increased as the temperature increased to $20{\pm}2^{\circ}C$, but declined at higher temperatures. At high $CO_2$ levels ($940{\pm}50ppm$), more plants were infected by PLRV at $30{\pm}2^{\circ}C$ than at 20 or $25{\pm}2^{\circ}C$, whereas PLRV RNA content was unchanged in the $20-30{\pm}2^{\circ}C$ temperature range. The effects of atmospheric $CO_2$ concentration on the acquisition of PLRV by Myzus persicae and accumulation of PLRV RNA in plant tissues were investigated using a growth chamber at $20{\pm}2^{\circ}C$. The M. persicae PLRV RNA content slightly increased at elevated $CO_2$ levels ($940{\pm}50ppm$), but this increase was not statistically significant. Transmission rates of PLRV by Physalis floridana increased as $CO_2$ concentration increased. More PLRV RNA accumulated in potato plants maintained at 540 or $940{\pm}50ppm$$CO_2$, than in plants maintained at $400{\pm}50ppm$. This is the first evidence of greater PLRV RNA accumulation and larger numbers of S. tuberosum plants infected by PLRV under conditions of combined high $CO_2$ levels ($940{\pm}50ppm$) and high temperature ($30{\pm}2^{\circ}C$).
This study was conducted to determine the change in freezing tolerance of Chinese cabbage (Brassica campestris L. ssp. penkinensis). Four-week old seedlings grown in a greenhouse at $25^{\circ}C$$\pm$0.5 were transferred to 5, 10 or 15$^{\circ}C$ condition for acclimation for one day, and then transferred to a lethal temperature of -3$^{\circ}C$ condition. Optimal acclimation temperatures were 5$^{\circ}C$ and 1$0^{\circ}C$. Freezing tolerance of leaf tissues was investigated during a week-long cold stress. Development of freezing tolerance was shown by survival ratio, and amounts of total RNA, soluble protein and soluble sugar. LT$_{50}$ value increased from -3 to -4$^{\circ}C$ after 5 days acclimation at 5 or 1$0^{\circ}C$, and this was considered to be the first indication of enhanced freezing tolerance. Plants, cold acclimated for five days, reached to a survival ratio of 60%. This increase in freezing tolerance was found to be associated with the increased levels of total RNA, soluble sugar and soluble protein. These metabolic changes imply the association of adjustment of growth and cell metabolism with low temperatures at the beginning of cold acclimation in chinese cabbage.e.
Cho Young-Sun;Lee Sang-Yoon;Bang In-Chul;Kim Dong-Soo;Nam Yoon-Kwon
Journal of Aquaculture
/
v.19
no.3
/
pp.157-165
/
2006
Expression of major antioxidant enzyme (AOE) including Cu/Zn superoxide dismutase (Cu/Zn-SOD), catalase (CAT), glutathione-S-transferase (GST) and 3 glutathione peroxidase isotypes (GPXs) at mRNA levels during heat stress was examined in mud loach (Misgurnus mizolepis) liver. Based on the semi-quantitative RT-PCR, real-time RT-PCR and/or northern dot blot hybridization, the antioxidant enzyme genes were generally up-regulated during elevation of water temperature from $23^{\circ}C$ up to $32^{\circ}C$. GPXs and SOD displayed the most significant elevation of mRNA levels (up to 3 and 2 folds, respectively) while CAT showed the steady-state expression irrespective of thermal conditions. GST represented the relatively moderate response (1.3-fold increase) in its transcription to thermal stress. The transcriptional activation of AOE genes was not significant at the treatment temperature lower than $29^{\circ}C$. Increased mRNA levels of GPX (extracellular form) and SOD genes in the fish exposed to $32^{\circ}C$ was readily detectable 1 day after exposure to heat stress.
Water temperature influences on various key biological events in fish, but the internal pathway of the temperature effects are not well understood. Heat shock proteins (HSPs), known to respond in the level of cells to many environmental factors including temperature, could improve our understanding on the pathway. Some biological processes such as gonadal development and sex differentiation in the Nile tilapia Oreochromis niloticus is particularly sensitive to water temperature. In this study, we have investigated the expressions of HSP70 and HSP90 genes in young tilapia at an ordinary temperature ($28^{\circ}C$) and elevated water temperature ($36^{\circ}C$). The distribution of the expressions of HSP70 and HSP90 mRNA in this species were found to be almost ubiquitous, being detected in all tissues studied here (brain, gonad, liver and muscle), suggesting the house keeping functions of these genes. Heat shock by elevating temperature from $28^{\circ}C$ to $36^{\circ}C$ significantly increased the expression of HSP70 mRNA in the gonad, liver and muscle for several hours (P<0.05) (brain tissue was not examined for this). The increased level of HSP70 gene expression recovered to the level at control temperature ($28^{\circ}C$) when fish were kept continuously at high temperature ($36^{\circ}C$) for 24 hours. Contrary to this, expression of HSP90 mRNA did not show significant increase in the gonad and muscle by the same heat shock (P>0.05), except in the liver where the expression of HSP90 mRNA increased continuously for 24 hours at $36^{\circ}C$. The results obtained in this study suggest that response to temperature change in different tissue or organ may utilize different heat shock proteins, and that HSP70 may have some importance in temperature-sensitive gonadal event in the Nile tilapia.
The mitotic cycle duration and component plase periods of rice (Oryza sativa L. 'Sumjinbyeo and Seokwangbyeo') root merisem cells at 15, 20, and 30$^{\circ}C$ were determined the use of tritiated thymidine, In this work, the time interval between the maxima of sequential mitotic appearances of marked cells was used to estimate the mitotic cycle duration (MCD) of rice, The MCD of rice of the cultivar 'Sumjinbyeo' and 'Seokwangbyeo' at 20. and 30$^{\circ}C$ was 12. and 20hr, respectively. But the MCD of 'Sumjinbyeo' and 'Seokwangbyeo' was 18 and 20hr, at 15$^{\circ}C$. respectively. The MCD decreased with increasing temperature, The duration of component phase of rice cultivar 'Seokwangbyeo' were essentially the same ratio at 20$^{\circ}C$ and 30$^{\circ}C$. but in 'Sumjinbyeo' cultivar the ratio of $G_1$ period was almost doubled while those of $G_2$ and M were decreased by almost two times at 20$^{\circ}C$ and 30$^{\circ}C$. Deoxyribonucleic acid (DNA). Ribonucleic acid (RNA), and protein synthesis were reduced with increasing temperature from 15$^{\circ}C$ to 30$^{\circ}C$ while the MCD was decreased, This result suggest that DNA, RNA and protein synthesis may not affect the MCD from 15$^{\circ}C$ to 30$^{\circ}C$ in rice.
Hepatocellular carcinoma (HCC) is a prevalent malignant tumor with high fatality. It has yet to be reported whether circ-SNX27 can affect the progression of HCC. This study attempted to analyze circ-SNX27's precise role and underlying mechanisms in HCC. HCC cell lines and tumor specimens from HCC patients were analyzed using quantitative real-time PCR and Western blotting to quantify the expressions of circ-SNX27, miR-375, and ribophorin I (RPN1). Cell invasion and cell counting kit 8 experiments were conducted for the evaluation of HCC cell invasion and proliferation. Caspase-3 Activity Assay Kit was utilized to gauge the caspase-3 activity. Luciferase reporter and RNA immunoprecipitation assays were executed to ascertain the relationships among miR-375, circ-SNX27, and RPN1. To determine how circ-SNX27 knockdown affects the growth of HCC xenografts in vivo, tumor-bearing mouse models were constructed. Elevated expressions of circ-SNX27 and RPN1 as well as a reduced miR-375 expression were observed among HCC cells and HCC patient tumor specimens. Knocking-down circ-SNX27 in HCC cells abated their proliferative and invasive abilities but raised their caspase-3 activity. Moreover, the poor levels of circ-SNX27 inhibited HCC tumor growth among the mice. Circ-SNX27 enhanced RPN1 by competitively binding with miR-375. Silencing miR-375 in HCC cells promoted their malignant phenotypes. Nonetheless, the promotive effect of miR375 silencing was reversible via the knockdown of circ-SNX27 or RPN1. This research demonstrated that circ-SNX27 accelerated the progression of HCC by modulating the miR-375/RPN1 axis. This is indicative of circ-SNX27's potential as a target for the treatment of HCC.
Viroid-like RNA molecules were detected from the low molecular weight RNAs isolated from the Korean peonies which showed typical viroid symptoms of epinasty and dwarfing. Low molecular weight RNAs including viroid RNA molecules were purified by the Qiagen anion exchange minicolumns. Viroid-like RNA molecules showed a single viroid specific band in the native polyacrylamide gel. They were separated into two bands in the denaturing gel conditions. The band of circular form of viroid-like RNAs was crossed over the horizontal band of the linear form of viroid-like RNA molecules in 0~8 M urea gradient gel under the denaturing conditions of 37$^{\circ}C$. The two circular forms of viroid-like RNA molecules were detected in the reverse polyacrylamide gel electrophoresis. The viroid-like RNA molecules purified from the peonies were supposed to be unidentified viroid RNA molecules.
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