scholarly journals In situ detection of Esr proteins secretion during maize microspore embryogenesis and their secretion blockage show effects on the culture progression

2010 ◽  
Vol 37 (10) ◽  
pp. 985 ◽  
Author(s):  
Pilar S. Testillano ◽  
María-José Coronado ◽  
Anne-Marie Thierry ◽  
Elisabeth Matthys-Rochon ◽  
María C. Risueño
Keyword(s):  
Liquid Medium ◽  
Embryo Development ◽  
Secretory Pathway ◽  
Microspore Culture ◽  
Microspore Embryogenesis ◽  
In Situ Monitoring ◽  
Subcellular Localisation ◽  
Embryo Cells

In vitro plant cells in culture release proteins and carbohydrates, but the active molecules responsible for sustaining the switch in embryogenic development and progression have not yet been identified. In maize (Zea mays L.), the Esr genes encode for small hydrophilic proteins and are expressed in the restricted region of maize endosperm surrounding the embryo: the embryo surrounding region (ESR). In the present work, the possible influence of secreted molecules in the liquid medium during microspore-derived embryo development and specifically, the presence of Esr proteins, has been analysed in maize microspore cultures. The study has been conducted with in situ monitoring of the structural and cellular organisation of developing embryos and the subcellular localisation of the Esr proteins by immunofluorescence and immunogold labelling. The results obtained using confocal and electron microscopy revealed that Esr proteins were localised in elements of the secretory pathway and cell walls in microspore-derived embryo cells during early embryogenesis. Esr proteins were also detected in the liquid medium of maize microspore cultures and accumulated at 20 days of culture. Tunicamycin treatment to block protein glycosilation and, therefore, secretion inhibited microspore-derived embryo development, which was subsequently recovered by supplementation with medium containing all the secreted factors from a well developed microspore culture. Esr labelling was not present in non-developing microspore embryos of cultures treated with tunicamycin, whereas labelling was present again in the Golgi elements and secretory vesicles of embryo cells when development was restored. The results indicate that Esr proteins are part of the secreted proteins, which show a nursing or signalling role during in vitro embryo development in maize microspore embryogenesis cultures and provide new evidence for an endosperm-like function of microspore-derived embryo structures during the early stages.

scholarly journals The Influence of pH on Microspore Embryogenesis of White Cabbage (Brassica oleracea L.)

2013 ◽  
Vol 5 (4) ◽  
pp. 485-489 ◽  
Author(s):  
Tina Oana CRISTEA
Keyword(s):  
Brassica Oleracea ◽  
Microspore Culture ◽  
Microspore Embryogenesis ◽  
Critical Factor ◽  
Cellular Level ◽  
Strong Base ◽  
White Cabbage ◽  
In Vitro Microspore Culture ◽  
Influence Of Ph

In vitro microspore culture is one of the top techniques utilised now-a-days for the obtaining of double haploid plants in many plant species, including Brassica. The pH of the medium is a critical factor for the success of In vitro microspore culture as it influences the invertase enzyme activity, translated at cellular level through an acceleration or reduction of sucrose cleavage. The results published until now shows rather contradictory findings, as the response of microspores have been proved to be highly depending on genotypes, most of them being focused on Brassica napus. Thus, in the present study, the effect of different NLN liquid medium pH, ranging between 5.0 to 7.0 were tested in order to establish the most suitable pH for the expression of embryogenic competences of microspores cultivated on medium In vitro and ultimately for the obtaining of microspore-derived embryos. Among the 11 values of pH tested, the best results were obtained on variants with pH 5.8 and 6.0, both in what concern the maintaining of microspores viability and the number of microspore-derived embryos. The findings of the present study provide a strong base for the establishment of an efficient protocol for the In vitro culture of microspore at Brassica oleracea L. genotypes with Romanian origin.

Study of androgenesis and spontaneous chromosome doubling in barley ( Hordeum vulgare L.) genotypes using isolated microspore culture

2009 ◽  
Vol 57 (2) ◽  
pp. 155-164 ◽  
Author(s):  
D. Kahrizi ◽  
R. Mohammadi
Keyword(s):  
Chromosome Doubling ◽  
Microspore Culture ◽  
Doubled Haploids ◽  
Negative Relationship ◽  
Microspore Embryogenesis ◽  
Culture Technique ◽  
Hordeum Vulgare L ◽  
Spontaneous Chromosome ◽  
Barley Genotypes

This research aimed to study the androgenesis and spontaneous chromosome doubling of five barley genotypes using an isolated in vitro microspore culture technique, involving a completely randomized design (CRD) with three replications. Statistical analysis of embryogenesis and cytogenetic results showed that genotype had a significant effect on haploid embryogenesis (P<0.01) and on spontaneous chromosome doubling (P<0.05). The genotype Igri was found to have the highest potential to produce haploid embryos (1577 embryos from 100 anthers), followed by the genotypes Boyer/Rojo, Afzal/Turkman/Kavir, Ashar/Hebo and Agrigashar/Matico with 369, 304, 278 and 150 embryos from 100 anthers, respectively. The highest percentage of spontaneous chromosome doubling (76%) was observed for the genotype which had the lowest embryogenesis (Agrigashar/Matico) and the lowest (65%) for the genotype with the highest androgenic capacity (Igri). Microspore embryogenesis also showed comparatively higher genotypic (109.2) and phenotypic (109.5) coefficients of variation, heritability (99.62) and genetic advance (1206.77), indicating the pre-dominance of additive gene action in the control of this character in the material studied. Estimates of genetic parameters (PCV, GCV and heritability) for microspore embryogenesis were higher than for spontaneous doubled haploids. These results indicated that selection for androgenic capacity would be more effective than for spontaneous doubled haploids. The findings showed a negative relationship (r= −0.68) between embryogenesis and spontaneous chromosome doubling in the barley genotypes studied. All the large embryos used had high regenerability and good plantlet formation.

scholarly journals Adequate UV Exposures for Healthy Life: In Situ Monitoring and Model Calculation of the Vitamin-D-Synthetic Capacity of Sunlight

2012 ◽  
Vol 7 (1) ◽  
pp. 98-103
Author(s):  
Irina Terenetskaya ◽  
Tetiana Orlova ◽  
Pavel Kapinos
Keyword(s):  
Vitamin D ◽  
Human Skin ◽  
Direct Calculation ◽  
Uv Spectra ◽  
In Situ Monitoring ◽  
Solar Uv ◽  
Vitro Model ◽  
Synthetic Capacity

Vitamin D which is formed upon UV solar radiation in human skin is essential in many physiological functions. To estimate beneficial vitamin-D-synthetic capacity of sunlight a bio-equivalent UV dosimeter that is based on the same molecular photochemistry from which vitamin D is photosynthesized in human skin has been developed. The examples of an in situ monitoring of the vitamin-D-synthetic capacity of sunlight using an in vitro model of vitamin D synthesis are presented, and various operational principles of the UV biodosimeter are discussed. In addition, reliable algorithm is presented for direct calculation of previtamin D3 accumulation using the photoreaction mathematical model with solar UV spectra as input data. Critical dependence of previtamin D3 accumulation on cloudiness and aerosols is demonstrated.

scholarly journals Effects of Genotype and Culture Conditions on Microspore Embryogenesis and Plant Regeneration in Brassica Rapa ssp. Rapa L.

Plants ◽  
2020 ◽  
Vol 9 (2) ◽  
pp. 278 ◽  
Author(s):  
Daria Shumilina ◽  
Dmitry Kornyukhin ◽  
Elena Domblides ◽  
Alexey Soldatenko ◽  
Anna Artemyeva
Keyword(s):  
Microspore Culture ◽  
Doubled Haploids ◽  
Genetic Disorder ◽  
Cold Treatment ◽  
Microspore Embryogenesis ◽  
Culture Conditions ◽  
Secondary Embryogenesis ◽  
Isolated Microspore Culture ◽  
Pure Lines

Turnip is a biennial crop and, consequently, the creation of pure lines for breeding is a time-consuming process. The production of pure turnip lines using doubled haploids produced in isolated microspore culture has not been sufficiently developed. The aim of the present work was to determine some key factors inducing embryogenesis in the isolated microspore culture of turnip, as well as investigating the manners of embryo development. It was shown that the acidity of the medium is an important factor in embryo production; different optimal pH levels ranging from 6.2 to 6.6 corresponded to individual genotypes. Such factors as the cold treatment of buds and the addition of activated charcoal to the nutrient medium increased the responsiveness of all genotypes studied. The turnip variety ‘Ronde witte roodkop herfst’ demonstrated a genetic disorder in the development of microspores; namely, non-separation of some microspores from tetrads. In the in vitro culture, each of the daughter microspores developed on its own. This indicates the dependence of the possibility of embryogenesis in the turnip microspore culture on the genotype. Results suggest that the initiation of secondary embryogenesis in primary embryos leads to an increase in the proportion of doubled haploid plants.

scholarly journals Bioreactor Platform for Biomimetic Culture and in situ Monitoring of the Mechanical Response of in vitro Engineered Models of Cardiac Tissue

2020 ◽  
Vol 8 ◽  
Author(s):  
Diana Massai ◽  
Giuseppe Pisani ◽  
Giuseppe Isu ◽  
Andres Rodriguez Ruiz ◽  
Giulia Cerino ◽  
...  
Keyword(s):  
Cardiac Tissue ◽  
Mechanical Response ◽  
In Situ Monitoring

scholarly journals In Situ Monitoring of Streptothricin Production by Streptomyces rochei F20 in Soil and Rhizosphere

2004 ◽  
Vol 70 (9) ◽  
pp. 5222-5228 ◽  
Author(s):  
Usanee Anukool ◽  
William H. Gaze ◽  
Elizabeth M. H. Wellington
Keyword(s):  
Stationary Phases ◽  
In Situ Monitoring ◽  
Dependent Manner ◽  
Rt Pcr ◽  
Transcript Accumulation ◽  
Reverse Transcription Pcr ◽  
Lower Population ◽  
Lower Population Density

ABSTRACT The onset of streptothricin (ST) biosynthesis in Streptomyces rochei F20 was studied by using reverse transcription-PCR (RT-PCR) to detect transcripts of ST genes during growth in liquid medium, soil, and the rhizosphere. In situ results correlated with those obtained in vitro, illustrating the growth phase-dependent manner of ST production by F20. Maximal transcription of ST resistance (sttR) and biosynthesis (sttA) genes occurred during the transition between the exponential and stationary phases of growth, when the specific growth rate (μ) started to decline. A higher level of gene expression of sttR versus sttA was observed in all experiments. In liquid culture, maximal transcript accumulation of the sttA gene was only ca. 40% that of the sttR gene. sttA and sttR mRNAs were detected in soil containing approximately 106 CFU of growing cells g of soil−1. sttR mRNA was detected in sterile and nonsterile rhizosphere colonized with growing mycelium of F20 at 1.2 × 106 and 4.0 × 105 CFU g of soil−1, respectively. However, neither sttR nor sttA transcripts were detected by RT-PCR in the rhizoplane, which supported a lower population density of F20 than the rhizosphere.

scholarly journals Application of image flow cytometry and photoacoustics for the characterization of red blood cell storage lesions

2021 ◽  
Author(s):  
Ruben N. Pinto
Keyword(s):  
Flow Cytometry ◽  
Structural Changes ◽  
Blood Gas Analysis ◽  
Gas Analysis ◽  
In Situ Monitoring ◽  
Image Flow ◽  
Morphology Characterization

Significant functional/structural changes of red blood cells (RBCs) have been documented during its in vitro storage. Collectively termed as RBC storage lesions, changes include an increase in RBC oxygen saturation (SO2) and an increase in irreversibly damaged RBCs (spheroechinocytes). In this work, novel optical techniques are presented for determining the spheroechinocyte population as a function of storage time via automated image flow cytometry (IFC) morphology characterization, and the acquisition of RBC SO2 via an in situ photoacoustic (PA) method. Blood gas analysis (BGA) was used as the gold standard SO2 measure. Over the lifespan of seven blood bags, the IFC spheroechinocyte population – PA SO2 correlation was found to be strong (0.600.95) shows high potential for in situ monitoring of RBC storage lesions.

scholarly journals Chromosome doubling effects of selected antimitotic agents in Brassica napus microspore culture

2008 ◽  
Vol 44 (No. 1) ◽  
pp. 30-36 ◽  
Author(s):  
M. Klíma ◽  
M. Vyvadilová ◽  
V. Kučera
Keyword(s):  
Embryo Development ◽  
Doubled Haploid ◽  
Chromosome Doubling ◽  
Microspore Culture ◽  
Antimitotic Drugs ◽  
Winter Rapeseed ◽  
The Mean ◽  
Whole Plants

Effects of microspore culture treatment with antimitotic agents colchicine, trifluralin and oryzalin on the frequency of embryo formation, embryo development, plant regeneration and diploidization rate in three F<sub>1</sub> hybrids of winter rapeseed cultivars were compared. The ploidy level analysis of 1709 flowering microspore-derived plants showed that in vitro applications of all antimitotic drugs increased the rate of doubled haploid (DH) plants significantly. The mean rate of DH plants from the trifluralin treatment was 85.7%, from colchicine 74.1% and 66.5% in the case of oryzalin, while only 42.3% in the untreated control variant whereas in vivo additional application of colchicine at the plantlet stage did not significantly increase the mean rate of DH plants (55.6%). Although there were no significant differences in diploidization efficiency between the in vitro applications of particular antimitotic agents, trifluralin showed to be the most suitable because of its positive effect on embryo development and conversion into whole plants. In addition, the diploidization rate was sufficient and stable in all genotypes tested. The results indicate that the trifluralin treatment of microspore cultures could provide efficient chromosome doubling for the production of doubled haploid lines from winter oilseed rape breeding materials.

scholarly journals Towards in situ monitoring of in vitro 3D bone models

Bone Reports ◽  
2020 ◽  
Vol 13 ◽  
pp. 100373
Author(s):  
Donata Iandolo ◽  
Mikhael Hadida ◽  
Guénaëlle Bouët Chalon ◽  
Rόisín M. Owens ◽  
Laurence Vico ◽  
...  
Keyword(s):  
In Situ Monitoring
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