scholarly journals Understanding Alstroemeria pallida Flower Colour: Links between Phenotype, Anthocyanins and Gene Expression

Plants ◽  
2020 ◽  
Vol 10 (1) ◽  
pp. 55
Author(s):  
Amanda Donoso ◽  
Constanza Rivas ◽  
Alan Zamorano ◽  
Álvaro Peña ◽  
Michael Handford ◽  
...  
Keyword(s):  
High Performance ◽  
Chalcone Synthase ◽  
Ornamental Plants ◽  
Flower Colour ◽  
Anthocyanin Content ◽  
Anthocyanidin Synthase ◽  
Visual Evaluation ◽  
Royal Horticultural Society ◽  
Polymerase Chain ◽  
Colour Chart

Flower colour is mainly due to the accumulation of flavonoids, carotenoids and betalains in the petals. Of these pigments, flavonoids are responsible for a wide variety of colours ranging from pale yellow (flavones, flavonols and flavanodiols) to blue-violet (anthocyanins). This character plays a crucial ecological role by attracting and guiding pollinators. Moreover, in the ornamental plants market, colour has been consistently identified as the main feature chosen by consumers when buying flowers. Considering the importance of this character, the aim of this study was to evaluate flower colour in the native Chilean geophyte Alstroemeria pallida, by using three different approaches. Firstly, the phenotype was assessed using both a colour chart and a colourimeter, obtaining CIELab parameters. Secondly, the anthocyanin content of the pigmented tepals was evaluated by high-performance liquid chromatography (HPLC), and finally, the expression of two key flavonoid genes, chalcone synthase (CHS) and anthocyanidin synthase (ANS) was analysed using real-time polymerase chain reaction (PCR). Visual evaluation of A. pallida flower colour identified 5 accessions, ranging from white (Royal Horticultural Society (RHS) N999D) to pink (RHS 68C). Moreover, this visual evaluation of the accessions correlated highly with the CIELab parameters obtained by colourimetry. An anthocyanidin corresponding to a putative 6-hydroxycyanidin was identified, which was least abundant in the white accession (RHS N999D). Although CHS was not expressed differentially between the accessions, the expression of ANS was significantly higher in the accession with pink flowers (RHS 68C). These results suggest a correlation between phenotype, anthocyanin content and ANS expression for determining flower colour of A. pallida, which could be of interest for further studies, especially those related to the breeding of this species with ornamental value.

scholarly journals Expression Analysis of the Anthocyanin Genes in Pink Spathes of Anthurium with Different Color Intensities

2015 ◽  
Vol 140 (5) ◽  
pp. 480-489 ◽  
Author(s):  
David Gopaulchan ◽  
Adrian M. Lennon ◽  
Pathmanathan Umaharan
Keyword(s):  
Chalcone Synthase ◽  
Developmental Stages ◽  
Genetic Model ◽  
Color Space ◽  
Negative Relationship ◽  
Anthocyanin Content ◽  
Anthocyanidin Synthase ◽  
Strong Negative Relationship ◽  
Anthurium Andraeanum ◽  
Different Color

Anthurium (Anthurium andraeanum) is a tropical ornamental valued for its colorful spathe (modified bract) that subtends the inflorescence. The present genetic model for spathe color inheritance in anthurium does not account for differences among the red- and pink-spathed cultivars or for differences in the shades of pink among pink cultivars. To identify the mechanisms responsible for the variation in color and intensity, five genetically defined pink-spathed cultivars, with respect to the O, R, and M loci, with varying shade intensities, along with a genetically defined red-spathed cultivar (control), were analyzed at the mRNA, protein, chemical, and phenotypic levels at different spathe development stages. Spathe color values were recorded based on CIE L*a*b* system. Intensity of color (L*, which represents lightness) correlated with the anthocyanin content, with L* showing a strong negative relationship with anthocyanin abundance. Additionally, the red spathe accumulated anthocyanin throughout the spathe developmental stages, whereas the pinks either produced anthocyanin at early stages of development, which decreased as the spathe matured or showed a marked delay in anthocyanin accumulation. The level of anthocyanin closely mirrored flavonoid 3′-hydroxylase (F3′H) expression but did not correspond with the expression of any of the other genes assayed, chalcone synthase (CHS), flavanone 3-hydroxylase (F3H), dihydroflavonol 4-reductase (DFR), and anthocyanidin synthase (ANS). It was found that earlier the expression and higher the rate of expression of F3′H during spathe development, the greater the accumulation of anthocyanin in the spathe. Differences in the a* color space parameter among cultivars also suggests that qualitative differences in color could be mediated through F3′H. Other ancillary mechanisms that down regulate F3H, ANS, and DFR expression levels, evident in some pink cultivars, are discussed.

High performance Nanogold™-in situ hybridzation and its use in the detection of hybridized and PCR-amplified microscopical preparations

1996 ◽  
Vol 54 ◽  
pp. 896-897
Author(s):  
G. W. Hacker ◽  
I. Zehbe ◽  
J. Hainfeld ◽  
A.-H. Graf ◽  
C. Hauser-Kronberger ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Messenger Rna ◽  
High Performance ◽  
Detection System ◽  
Direct Methods ◽  
Genetic Alterations ◽  
Chain Reaction ◽  
Protein Encoding ◽  
Polymerase Chain

In situ hybridization (ISH) with biotin-labeled probes is increasingly used in histology, histopathology and molecular biology, to detect genetic nucleic acid sequences of interest, such as viruses, genetic alterations and peptide-/protein-encoding messenger RNA (mRNA). In situ polymerase chain reaction (PCR) (PCR in situ hybridization = PISH) and the new in situ self-sustained sequence replication-based amplification (3SR) method even allow the detection of single copies of DNA or RNA in cytological and histological material. However, there is a number of considerable problems with the in situ PCR methods available today: False positives due to mis-priming of DNA breakdown products contained in several types of cells causing non-specific incorporation of label in direct methods, and re-diffusion artefacts of amplicons into previously negative cells have been observed. To avoid these problems, super-sensitive ISH procedures can be used, and it is well known that the sensitivity and outcome of these methods partially depend on the detection system used.

scholarly journals Effect of Allium senescens Extract on Sorafenib Resistance in Hepatocarcinoma Cells

2021 ◽  
Vol 11 (8) ◽  
pp. 3696
Author(s):  
Sohyeon Park ◽  
Yoonjin Park ◽  
Heejong Shin ◽  
Boyong Kim ◽  
Seunggwan Lee
Keyword(s):  
Drug Resistance ◽  
Hepg2 Cells ◽  
High Performance ◽  
Quantitative Polymerase Chain Reaction ◽  
Mitochondrial Apoptosis ◽  
Coumaric Acid ◽  
Allium Species ◽  
Hepatocarcinoma Cells ◽  
Polymerase Chain ◽  
Resistant Cells

Although Allium species are involved in bioactivity, to the best of our knowledge, there is no research on the effects of Allium senescens on drug resistance in hepatocarcinoma. Ultra-high performance liquid chromatography was used to determine the concentration of several bioactive compounds in A. senescens extract; flow cytometry, reverse transcription–quantitative polymerase chain reaction, and siRNA-mediated knockdown to estimate the levels of different markers in HepG2 cells. The quantity of p-coumaric acid in the extract was 4.7291 ± 0.06 μg/mL, and the protein of relevant evolutionary and lymphoid interest (PRELI) in the resistant cells decreased 2.1 times in the presence of p-coumaric acid. The resistant cells strongly downregulated the efflux transporters (ABCB1, ABCC2, and ABCG2) when exposed to the extract or p-coumaric acid and when PRELI was knocked down, in contrast to the influx proteins (OCT-1). Additionally, the extract induced mitochondrial apoptosis and suppressed autophagy. Consequently, the extract and p-coumaric acid attenuated drug resistance of HepG2 cells through the downregulation of PRELI, a key protein associated with the modulation of drug transporter expression, the activation of autophagy, and mitochondrial apoptosis. Our results indicate that A. senescens extract is beneficial in protecting cancer cells against drug resistance and sustaining the efficacy of sorafenib against liver cancer.

scholarly journals Inhibitory Activity of 8 Alkaloids on P-gp and Their Distribution in Chinese Uncaria Species

2020 ◽  
Vol 15 (11) ◽  
pp. 1934578X2097350
Author(s):  
Yanan Gai ◽  
Nannan Yang ◽  
Jian Chen
Keyword(s):  
High Performance ◽  
Positive Control ◽  
Inhibitory Effects ◽  
Glycoprotein P ◽  
Chromatography Analysis ◽  
Chemical Content ◽  
Polymerase Chain ◽  
Chemical Distribution ◽  
And Function ◽  
Mcf 7

P-glycoprotein (P-gp) inhibition conduces to improving the ability of chemicals to cross through blood-brain barrier (BBB). The hook-bearing branch of Uncaria is used as a traditional herbal medicine for the treatment of hypertension, headache, stroke, and Alzheimer’s disease in recent years. In this study, the inhibitory effects of 8 alkaloids sourced from Uncaria plants on P-gp were evaluated. Meanwhile, the content of 8 alkaloids in 8 Chinese Uncaria species was quantified simultaneously by high-performance liquid chromatography analysis. Western blotting, real-time quantitative reverse transcription-polymerase chain reaction, and flow cytometry were used to evaluate the P-gp levels in MCF-7/adriamycin-resistant (ADR) cells. It was shown that hirsuteine and hirsutine, selected from 8 candidate alkaloids, could remarkably suppressed P-gp level and function in MCF-7/ADR cells when using Verapamil as positive control. By tracking their chemical distribution in 8 Chinese Uncaria species, we found that hirsuteine and hirsutine were much more abundant in Uncaria rhynchophylla and Uncaria hirsuta than in any other 6 species. And a chemical content profile of these alkaloids was plotted within 8 Chinese species.

scholarly journals Chalcone Synthase: Method Optimization of Extraction and Activity Assay in Strawberry Fruit

2020 ◽  
Author(s):  
Linda Zoungrana ◽  
Alyssa N. Smith ◽  
M. Cecilia do Nascimento Nunes
Keyword(s):  
High Performance ◽  
Method Development ◽  
Chalcone Synthase ◽  
Biological Tissues ◽  
Spectrophotometric Analysis ◽  
Strawberry Fruit ◽  
Activity Assay ◽  
Plant Materials ◽  
Lack Of Information ◽  
Method Optimization

Method development and optimization play a central role in analytical chemistry and more specifically in food biochemistry. When it comes to research, it is common that analytical methods need to be modified to specific experimental biological tissues. While there are several published works on the activity of the enzyme chalcone synthase (CHS) in plant materials, such as sweet basil using ultra- high-performance liquid chromatography, there is a lack of information regarding extraction and activity of CHS in strawberries. Therefore, the main objective of this work was to optimize existing published methods for extraction and activity of CHS in strawberries, using spectrophotometric analysis. It was done through a literature search, a method dissection was performed, followed by theoretical optimization of the protocol, and finally an experimental optimization

scholarly journals Anthocyanin Accumulation during Potato Tuber Development

1997 ◽  
Vol 122 (1) ◽  
pp. 20-23 ◽  
Author(s):  
Chen-Yi Hung ◽  
John R. Murray ◽  
Sarah M. Ohmann ◽  
Cindy B.S. Tong
Keyword(s):  
High Performance ◽  
Anthocyanin Content ◽  
Mrna Levels ◽  
Biosynthetic Enzyme ◽  
Potato Tubers ◽  
Tuber Development ◽  
Surface Color ◽  
Dihydroflavonol Reductase ◽  
Anthocyanin Concentration ◽  
Tuber Periderm

The color of red potato tubers is due to an accumulation of anthocyanins in periderm and peripheral cortex tissues. The objective of this study was to characterize changes in anthocyanin content and tuber surface color during tuber development. Using the red tuber-producing potato (Solanum tuberosum L.) cultivar Norland, we observed that chroma (intensity of redness) and anthocyanin content per unit of surface area of greenhouse-grown tubers decreased as tuber weight increased. There was no increase in hue (tint) during the same developmental periods. Using high-performance liquid chromatography (HPLC), we determined that pelargonidin and peonidin are the major anthocyanidins (aglycones of anthocyanins) in the tuber periderm. Northern blot analyses indicated that steady-state mRNA levels of dihydroflavonol reductase (DFR), an anthocyanin biosynthetic enzyme, continued throughout tuber development. These results suggest that anthocyanins are synthesized throughout tuber development, and that cell division and/or enlargement contribute to a decline in chroma and anthocyanin concentration.

scholarly journals Effect of colour and size grading of China aster (Callistephus chinensis Nees) seeds on their germination

2012 ◽  
Vol 62 (2) ◽  
pp. 187-191 ◽  
Author(s):  
Agnieszka Rosińska ◽  
Grzegorz Rosiński ◽  
Roman Hołubowicz
Keyword(s):  
Seed Testing ◽  
Royal Horticultural Society ◽  
China Aster ◽  
Colour Chart ◽  
Size Grading ◽  
Better Than

Seeds of 3 commercial China aster (<i>Callistephus chinensis</i> Nees) lots were divided by hand into 3 grades with different colours: dark brown, brown and light brown, and 2 grades with a different size: length below 3.9 mm (small) and above 3.9 mm (large). The colour grading was done based on the Royal Horticultural Society Colour Chart and size grading was done by hand for each seed. Then, seeds were routinely germinated based on the International Seed Testing Association (ISTA) rules. The size of seeds had no effect on their germination. The dark brown seeds germinated better than the light brown ones. Removing light brown seeds from the China aster seed lot improved their germination.

scholarly journals Genetic polymorphism of thiopurine S-methyltransferase in Argentina

2003 ◽  
Vol 40 (4) ◽  
pp. 388-393 ◽  
Author(s):  
L. E. Laróvere ◽  
R. Dodelson de Kremer ◽  
L. H. J. Lambooy ◽  
R. A. De Abreu
Keyword(s):  
Polymerase Chain Reaction ◽  
Genetic Polymorphism ◽  
Latin American ◽  
High Performance ◽  
Normal Activity ◽  
Tpmt Activity ◽  
Allele Frequency Distribution ◽  
Chain Reaction ◽  
Chromatography Method ◽  
Polymerase Chain

Background: Thiopurine methyltransferase (TPMT) catalyses the S-methylation of 6-thiopurine drugs, which are commonly used in the treatment of autoimmune diseases, leukaemia and organ transplantation. TPMT activity is polymorphic as a result of gene mutations. Ethnic variations in phenotype and genotype have been identified in previous population studies, but no information was available within Latin-American populations. Aim: To establish the genetic polymorphism of TPMT in an Argentine population. Methods: TPMT enzymatic activity of 147 healthy Argentine subjects was measured using a high-performance liquid chromatography method. The genotyping assay for nine defective alleles (TPMT*2 - *8) was based on restriction fragment length polymorphism polymerase chain reaction and allele-specific polymerase chain reaction methods. Results: All subjects had detectable TPMT activity. Twelve individuals with low to intermediate activity were heterozygous for one of the mutant alleles: nine were TPMT*1/*3A, two TPMT*1/*2 and one TPMT*1/*4. All examined subjects with normal activity had wild-type genotype (TPMT*1/*1). Conclusion: Variant TPMT alleles were present in 8·2% of the examined subjects, which is in accordance with other studies. The frequency of TPMT*3A, TPMT*2 and TPMT*4 was 3·1%, 0·7% and 0·3%, respectively. TPMT*3A was the most prevalent allele, which is in accordance with results from Caucasian populations. This study provides the first analysis of TPMT activity and allele frequency distribution in Argentina, South America.

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