scholarly journals Induction, identification and genetics analysis of tetraploid Actinidia chinensis

2019 ◽  
Vol 6 (11) ◽  
pp. 191052 ◽  
Author(s):  
Shengxing Li ◽  
Xiaozhen Liu ◽  
Huiming Liu ◽  
Xianang Zhang ◽  
Qinxia Ye ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Transcriptome Analysis ◽  
Stress Resistance ◽  
Quantitative Polymerase Chain Reaction ◽  
Colchicine Treatment ◽  
Actinidia Chinensis ◽  
Flow Cytometry Analysis ◽  
Commercially Important ◽  
Polymerase Chain ◽  
Gene Expression Levels

Actinidia chinensis is a commercially important fruit, and tetraploid breeding of A. chinensis is of great significance for economic benefit. In order to obtain elite tetraploid cultivars, tetraploid plants were induced by colchicine treatment with leaves of diploid A. chinensis ‘SWFU03’. The results showed that the best treatment was dipping leaves 30 h in 60 mg l −1 colchicine solutions, with induction rate reaching 26%. Four methods, including external morphology comparison, stomatal guard cell observation, chromosome number observation and flow cytometry analysis were used to identify the tetraploid of A. chinensis . Using the induction system and flow cytometry analysis methods, 187 tetraploid plants were identified. Three randomly selected tetraploid plants and their starting diploid plants were further subjected to transcriptome analysis, real-time quantitative polymerase chain reaction (RT-qPCR) and methylation-sensitive amplification polymorphism (MSAP) analysis. The transcriptome analysis results showed that there were a total of 2230 differentially expressed genes (DEG) between the diploid and tetraploid plants, of which 660 were downregulated and 1570 upregulated. The DEGs were mainly the genes involved in growth and development, stress resistance and antibacterial ability in plants. RT-qPCR results showed that the gene expression levels of the growth and stress resistance of tetraploid plants were higher than those of diploid ones at the transcriptome level. MSAP analysis of DNA methylation results showed that tetraploid plants had lower methylation ratio than diploid ones. The present results were valuable to further explore the epigenetics of diploid and tetraploid kiwifruit plants.

scholarly journals Monitoring the efficacy of specific treatment in chronic Chagas disease by polymerase chain reaction and flow cytometry analysis

Parasite ◽  
2005 ◽  
Vol 12 (4) ◽  
pp. 353-357 ◽  
Author(s):  
G. Sánchez ◽  
X. Coronado ◽  
I. Zulantay ◽  
W. Apt ◽  
M. Gajardo ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Polymerase Chain Reaction ◽  
Chagas Disease ◽  
Specific Treatment ◽  
Flow Cytometry Analysis ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Chronic Chagas Disease

scholarly journals NOTCH Receptors and DLK Proteins Enhance Brown Adipogenesis in Mesenchymal C3H10T1/2 Cells

Cells ◽  
2020 ◽  
Vol 9 (9) ◽  
pp. 2032
Author(s):  
María-Milagros Rodríguez-Cano ◽  
María-Julia González-Gómez ◽  
Beatriz Sánchez-Solana ◽  
Eva-María Monsalve ◽  
María-José M. Díaz-Guerra ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Quantitative Polymerase Chain Reaction ◽  
Mature Adipocyte ◽  
Chain Reaction ◽  
Notch Receptors ◽  
Multipotent Cells ◽  
Numerous Cell ◽  
Cellular Context ◽  
Polymerase Chain ◽  
Gene Expression Levels

The NOTCH family of receptors and ligands is involved in numerous cell differentiation processes, including adipogenesis. We recently showed that overexpression of each of the four NOTCH receptors in 3T3-L1 preadipocytes enhances adipogenesis and modulates the acquisition of the mature adipocyte phenotype. We also revealed that DLK proteins modulate the adipogenesis of 3T3-L1 preadipocytes and mesenchymal C3H10T1/2 cells in an opposite way, despite their function as non-canonical inhibitory ligands of NOTCH receptors. In this work, we used multipotent C3H10T1/2 cells as an adipogenic model. We used standard adipogenic procedures and analyzed different parameters by using quantitative-polymerase chain reaction (qPCR), quantitative reverse transcription-polymerase chain reaction (qRT-PCR), luciferase, Western blot, and metabolic assays. We revealed that C3H10T1/2 multipotent cells show higher levels of NOTCH receptors expression and activity and lower Dlk gene expression levels than 3T3-L1 preadipocytes. We found that the overexpression of NOTCH receptors enhanced C3H10T1/2 adipogenesis levels, and the overexpression of NOTCH receptors and DLK (DELTA-like homolog) proteins modulated the conversion of cells towards a brown-like adipocyte phenotype. These and our prior results with 3T3-L1 preadipocytes strengthen the idea that, depending on the cellular context, a precise and highly regulated level of global NOTCH signaling is necessary to allow adipogenesis and determine the mature adipocyte phenotype.

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