Catalase isozymes as biochemical markers of different developmental stages in cypress (Cuptessussempervirens)

1996 ◽  
Vol 26 (9) ◽  
pp. 1629-1633 ◽  
Author(s):  
M.L. Racchi ◽  
A.P. Chiusi ◽  
R. Giannini
Keyword(s):  
Biochemical Markers ◽  
Temporal Pattern ◽  
Developmental Stages ◽  
Polyacrylamide Gel Electrophoresis ◽  
Developmental Time ◽  
Mature Plant ◽  
Development Stages ◽  
Young Embryo ◽  
Native Polyacrylamide Gel Electrophoresis ◽  
Different Tissues

The expression of catalase isozymes has been investigated to define the different developmental stages of cypress (Cupressussempervirens L). To examine the temporal pattern of catalase expression during development, catalases in dry seeds, young embryos, and different tissues of the mature plant were characterized by native polyacrylamide gel electrophoresis. The data indicated the presence of two patterns of expression: one specific to the seed and young embryo and another for the mature plant. Defined isoforms were specific to each developmental time. The results indicated that different catalase isozymes are developmentally expressed. Consequently the changes in their expression pattern can be used to define the development stages of the tree.

scholarly journals Analysis of Several Popular Cultivars of Madagascar Periwinkle (Catharanthus roseus (L.) G. Don.) using Biochemical Markers

2013 ◽  
Vol 5 (4) ◽  
pp. 458-461
Author(s):  
Owk ANIEL KUMAR ◽  
Sape S. TATA ◽  
Kancharla PAVAN KUMAR
Keyword(s):  
Catharanthus Roseus ◽  
Gel Electrophoresis ◽  
Biochemical Markers ◽  
Polyacrylamide Gel Electrophoresis ◽  
Cultivar Differences ◽  
Madagascar Periwinkle ◽  
Large White ◽  
Electrophoretic Patterns ◽  
Native Polyacrylamide Gel Electrophoresis ◽  
Isozyme Polymorphism

Band designs of esterase (EST), peroxidase (PO) and polyphenol oxidase (PPO) isozymes in several selected cultivars of Catharanthus roseus by using native polyacrylamide gel electrophoresis (PAGE) were investigated in this study. It was confirmed that cultivar differences in isozyme polymorphism can be revealed by applied electrophoretic patterns. Three isozyme systems produced a total of 16 bands with polymorphism ranged from 66.6-100%. Considering the patterns of isozyme variations in the five cultivars of Catharanthus roseus, it is evident that the cultivar ‘First kiss coral’ displayed crimson red petal with large white eye’ displayed demarked profiles of EST, PO and PPO isozymes than other cultivars. This is the first report on isozyme polymorphism in members of the Cathanarathus roseus (L.) G. Don.

scholarly journals Validation of Reference Genes via qRT-PCR in Multiple Conditions in Brandt’s Voles, Lasiopodomys brandtii

Animals ◽  
2021 ◽  
Vol 11 (3) ◽  
pp. 897
Author(s):  
Lin Tian ◽  
Yan Chen ◽  
Da-Wei Wang ◽  
Xiao-Hui Liu
Keyword(s):  
Reference Gene ◽  
Reference Genes ◽  
Developmental Stages ◽  
Development Stages ◽  
Qrt Pcr ◽  
Polymerase Chain ◽  
Experimental Treatments ◽  
Different Tissues ◽  
Lasiopodomys Brandtii ◽  
Multiple Conditions

The choice of optimal reference gene is challenging owing to the varied expression of reference genes in different organs, development stages, and experimental treatments. Brandt’s vole (Lasiopodomys brandtii) is an ideal animal to explore the regulatory mechanism of seasonal breeding, and many studies on this vole involve gene expression analysis using quantitative real-time polymerase chain reaction (qRT-PCR). In this study, we used the method of the coefficient of variation and the NormFinder algorithm to evaluate the performance of nine commonly used reference genes Gapdh, Hprt1, β-actin, PPIA, Rpl13a, Tbp, Sdha, Hmbs, and B2M using qRT-PCR in eight different tissues, five developmental stages, and three different photoperiods. We found that all nine genes were not uniformly expressed among different tissues. B2M and Rpl13a were the optimal reference genes for different postnatal development stages in the hypothalamus for males and females, respectively. Under different photoperiods in the hypothalamus, none of the selected genes were suitable as reference genes at 6 weeks postnatal; β-actin and PPIA were the optimal reference genes at 12 weeks postnatal; Hprt1, β-actin, PPIA, Hmbs, and B2M were excellent reference genes at 24 weeks postnatal. The present study provides a useful basis for selecting the appropriate reference gene in Lasiopodomys brandtii.

Analysis of the cytosolic proteins of chick embryo gonads by two-dimensional gel electrophoresis

Development ◽  
1986 ◽  
Vol 94 (1) ◽  
pp. 221-230
Author(s):  
J. Samsel ◽  
B. Lorber ◽  
A. Petit ◽  
J. -P. Weniger
Keyword(s):  
Gel Electrophoresis ◽  
Developmental Stages ◽  
Polyacrylamide Gel Electrophoresis ◽  
Specific Protein ◽  
Chick Embryos ◽  
Two Dimensional ◽  
Two Dimensional Gel Electrophoresis ◽  
Specific Proteins ◽  
Different Tissues

Gonads and mesonephros dissected from normal chick embryos, as well as a blood sample, were labelled in vitro with [35S]methionine and [14C]leucine. The patterns of cytosolic protein synthesis of the different tissues were analysed using two-dimensional (2-D) polyacrylamide gel electrophoresis. Three developmental stages, i.e. 8, 12 and 17 days of incubation, were investigated. Five sex-specific proteins were detected in the male and two in the female. In the testis, only one protein is already present at the 8-day stage. In the ovary, one protein exists since the 8-day stage, but it is also synthesized in the mesonephros. The second ovary-specific protein appeared only at the 12-day stage.

scholarly journals Spatio-temporal selection of reference genes in the two congeneric species of Glycyrrhiza

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Yuping Li ◽  
Xiaoju Liang ◽  
Xuguo Zhou ◽  
Yu An ◽  
Ming Li ◽  
...  
Keyword(s):  
Gene Expression ◽  
Developmental Stage ◽  
Reference Genes ◽  
Developmental Stages ◽  
Individual Factors ◽  
Congeneric Species ◽  
Spatio Temporal ◽  
The Stability ◽  
Different Tissues ◽  
Selection Of

AbstractGlycyrrhiza, a genus of perennial medicinal herbs, has been traditionally used to treat human diseases, including respiratory disorders. Functional analysis of genes involved in the synthesis, accumulation, and degradation of bioactive compounds in these medicinal plants requires accurate measurement of their expression profiles. Reverse transcription quantitative real-time PCR (RT-qPCR) is a primary tool, which requires stably expressed reference genes to serve as the internal references to normalize the target gene expression. In this study, the stability of 14 candidate reference genes from the two congeneric species G. uralensis and G. inflata, including ACT, CAC, CYP, DNAJ, DREB, EF1, RAN, TIF1, TUB, UBC2, ABCC2, COPS3, CS, R3HDM2, were evaluated across different tissues and throughout various developmental stages. More importantly, we investigated the impact of interactions between tissue and developmental stage on the performance of candidate reference genes. Four algorithms, including geNorm, NormFinder, BestKeeper, and Delta Ct, were used to analyze the expression stability and RefFinder, a comprehensive software, provided the final recommendation. Based on previous research and our preliminary data, we hypothesized that internal references for spatio-temporal gene expression are different from the reference genes suited for individual factors. In G. uralensis, the top three most stable reference genes across different tissues were R3HDM2, CAC and TUB, while CAC, CYP and ABCC2 were most suited for different developmental stages. CAC is the only candidate recommended for both biotic factors, which is reflected in the stability ranking for the spatio (tissue)-temporal (developmental stage) interactions (CAC, R3HDM2 and DNAJ). Similarly, in G. inflata, COPS3, R3HDM2 and DREB were selected for tissues, while RAN, COPS3 and CS were recommended for developmental stages. For the tissue-developmental stage interactions, COPS3, DREB and ABCC2 were the most suited reference genes. In both species, only one of the top three candidates was shared between the individual factors and their interactions, specifically, CAC in G. uralensis and COPS3 in G. inflata, which supports our overarching hypothesis. In summary, spatio-temporal selection of reference genes not only lays the foundation for functional genomics research in Glycyrrhiza, but also facilitates these traditional medicinal herbs to reach/maximize their pharmaceutical potential.

scholarly journals The Native Monomer of Bacillus Pumilus Ribonuclease Does Not Exist Extracellularly

2018 ◽  
Vol 2018 ◽  
pp. 1-7 ◽  
Author(s):  
Olga Ilinskaya ◽  
Vera Ulyanova ◽  
Irina Lisevich ◽  
Elena Dudkina ◽  
Nataliya Zakharchenko ◽  
...  
Keyword(s):  
Conformational Changes ◽  
Protein Concentration ◽  
Bacillus Pumilus ◽  
Polyacrylamide Gel Electrophoresis ◽  
Culture Fluid ◽  
Size Exclusion ◽  
High Protein Concentration ◽  
Exclusion Chromatography ◽  
Native Polyacrylamide Gel Electrophoresis ◽  
Hypochromic Effect

Supported by crystallography studies, secreted ribonuclease of Bacillus pumilus (binase) has long been considered to be monomeric in form. Recent evidence obtained using native polyacrylamide gel electrophoresis and size-exclusion chromatography suggests that binase is in fact dimeric. To eliminate ambiguity and contradictions in the data we have measured conformational changes, hypochromic effect, and hydrodynamic radius of binase. The immutability of binase secondary structure upon transition from low to high protein concentration was registered, suggesting the binase dimerization immediately after translocation through the cell membrane and leading to detection of binase dimers only in the culture fluid regardless of ribonuclease concentration. Our results made it necessary to take a fresh look at the binase stability and cytotoxicity towards virus-infected or tumor cells.

scholarly journals Identity and Substrate Specificity of Reductive Dehalogenases Expressed in Dehalococcoides-Containing Enrichment Cultures Maintained on Different Chlorinated Ethenes

2015 ◽  
Vol 81 (14) ◽  
pp. 4626-4633 ◽  
Author(s):  
Xiaoming Liang ◽  
Olivia Molenda ◽  
Shuiquan Tang ◽  
Elizabeth A. Edwards
Keyword(s):  
Specific Activity ◽  
Enrichment Culture ◽  
Polyacrylamide Gel Electrophoresis ◽  
Peptide Sequencing ◽  
Enzyme Assays ◽  
Content Type ◽  
Substrate Preferences ◽  
Native Polyacrylamide Gel Electrophoresis ◽  
History Of ◽  
Different Strains

ABSTRACTMany reductive dehalogenases (RDases) have been identified in organohalide-respiring microorganisms, and yet their substrates, specific activities, and conditions for expression are not well understood. We tested whether RDase expression varied depending on the substrate-exposure history of reductive dechlorinating communities. For this purpose, we used the enrichment culture KB-1 maintained on trichloroethene (TCE), as well as subcultures maintained on the intermediatescis-dichloroethene (cDCE) and vinyl chloride (VC). KB-1 contains a TCE-to-cDCE dechlorinatingGeobacterand severalDehalococcoidesstrains that together harbor many of the known chloroethene reductases. Expressed RDases were identified using blue native polyacrylamide gel electrophoresis, enzyme assays in gel slices, and peptide sequencing. As anticipated but never previously quantified, the RDase fromGeobacterwas only detected transiently at the beginning of TCE dechlorination. TheDehalococcoidesRDase VcrA and smaller amounts of TceA were expressed in the parent KB-1 culture during complete dechlorination of TCE to ethene regardless of time point or amended substrate. TheDehalococcoidesRDase BvcA was only detected in enrichments maintained on cDCE as growth substrates, in roughly equal abundance to VcrA. Only VcrA was detected in subcultures enriched on VC. Enzyme assays revealed that 1,1-DCE, a substrate not used for culture enrichment, afforded the highest specific activity.trans-DCE was substantially dechlorinated only by extracts from cDCE enrichments expressing BvcA. RDase gene distribution indicated enrichment of different strains ofDehalococcoidesas a function of electron acceptor TCE, cDCE, or VC. Each chloroethene reductase has distinct substrate preferences leading to strain selection in mixed communities.

scholarly journals L-DOPA Trends in Different Tissues at Early Stages of Vicia faba Growth: Effect of Tyrosine Treatment

2018 ◽  
Vol 8 (12) ◽  
pp. 2431 ◽  
Author(s):  
Claudia Oviedo-Silva ◽  
Mhartyn Elso-Freudenberg ◽  
Mario Aranda-Bustos
Keyword(s):  
Amino Acid ◽  
Vicia Faba ◽  
High Performance ◽  
Developmental Stages ◽  
Growth Effect ◽  
Guaiacol Peroxidase ◽  
Nonprotein Amino Acid ◽  
Early Seedling ◽  
Gpx Activity ◽  
Different Tissues

The nonprotein amino acid Levo-3,4-dihydroxyphenylalanine (L-DOPA) has insecticidal, allelochemical, and antiparkinsonian effects. The aim of this research was to assess L-DOPA content in different tissues of Vicia faba (cv. Super Agua Dulce), and to verify if treatment with the phenolic amino acid L-4-hydroxyphenylalanine (tyrosine) had an effect on such content. Under light germination, control and tyrosine-treated early seedling stages of V. faba were studied and L-DOPA was quantified spectrophotometrically (Arnow’s method) and by high-performance thin-layer chromatography (HPTLC), as well. Additionally, tyrosinase (TYROX) and guaiacol peroxidase (GPX) activities (considered markers of a phenolic compounds metabolism) were quantified as germination proceeded. Different organs (roots, sprouts, and seeds) and different developmental stages were considered. Steady high L-DOPA concentrations were found in untreated sprouts and roots compared to seeds, as time progressed. While TYROX activity was not detected in these experiments, GPX had diverse trends. In control tissues, GPX increased in seed tissue as germination progressed, whereas in roots and sprouts, a decreasing GPX activity was observed. Tyrosine exposure decreased L-DOPA content, and decreased or did not change GPX activity (depending on the organ). Both Arnow’s and HPTLC methods were consistent in terms of tendencies, except for the scarce contents found in seeds, in which HPTLC was more sensitive. The richest source of L-DOPA was found in shoots (untreated), reaching as high as 125 mg g−1 DW (12% in DW) (the highest content reported in fava bean seedlings until now), whereas the smallest L-DOPA content was found in seeds. The importance of light germination conditions is discussed in terms of L-DOPA yield and from a physiological perspective. It is concluded that V. faba (cv. Super Agua Dulce) shoots are a good source of L-DOPA and that tyrosine addition (0.55 mM) decreases L-DOPA content in actively growing tissues (shoots and roots).

scholarly journals Comparative study on five kinds of cytokines in the tissues of Chinese giant salamander (Andrias davidianus) of different developmental stages

2021 ◽  
Vol 869 (1) ◽  
pp. 012067
Author(s):  
J Qian ◽  
X Y Zhai ◽  
L Guo ◽  
W G Chen ◽  
J J Fu ◽  
...  
Keyword(s):  
Comparative Study ◽  
Blood Serum ◽  
Developmental Stages ◽  
Distribution Area ◽  
Andrias Davidianus ◽  
Tnf Α ◽  
Sandwich Method ◽  
Different Ages ◽  
Chinese Giant Salamander ◽  
Different Tissues

Abstract By using of the double antibody sandwich method of ELISA, the activities of five cytokines including IL-2, IL-4, IFN-α, IFN-β and TNF-α from the blood serum, liver, intestine and spleen at two developmental stages of Chinese giant salamander (Andrias davidianus) were determined to analyze the distribution of the cytokines. The result indicated that five cytokines were found in these four tissues, while their activities were different in different tissues and different ages. The highest activity of IL-2 and IL-4 was all present in blood serum of two different ages. The activity of IFN-α was the highest in blood serum of 1-year-old and in spleen of 2-year-old, respectively. The activity of IFN-β was also highest in blood serum of two different ages. The activity of TNF-α was highest in liver of two different ages. Thus, this study provides convincing reference for blood serum and liver as the most important distribution area of Chinese giant salamander.

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