scholarly journals Variation in Genome Size, Ploidy, Stomata, and rDNA Signals in Althea

2019 ◽  
Vol 144 (2) ◽  
pp. 130-140 ◽  
Author(s):  
Jason D. Lattier ◽  
Hsuan Chen ◽  
Ryan N. Contreras
Keyword(s):  
Flow Cytometry ◽  
Genome Size ◽  
Guard Cell ◽  
Stomatal Density ◽  
Anatomical Variation ◽  
Average Density ◽  
Stomata Size ◽  
Significant Difference ◽  
Random Segregation ◽  
Precipitous Decline

Althea (Hibiscus syriacus) is a shrub prized for its winterhardiness and colorful summer flowers. Altheas are tetraploids (2n = 4x = 80); however, breeders have developed hexaploids and octoploids. Previous studies report anatomical variation among polyploids, including stomata size. The purpose of this study was 4-fold. First, identify genome size and ploidy variation in cultivars via flow cytometry and chromosome counts. Second, create a ploidy series consisting of 4x, 5x, 6x, and 8x cytotypes. Third, investigate the ploidy series for variation in stomatal guard cell lengths, stomatal density, and copy number of fluorescent ribosomal DNA (rDNA) signals. Fourth, investigate segregation patterns of rDNA signals in a subset of pentaploid seedlings. Flow cytometry revealed most cultivars to be tetraploid with holoploid 2C genome sizes from 4.55 ± 0.02 to 4.78 ± 0.06 pg. Five taxa (‘Aphrodite’, ‘Pink Giant’, ‘Minerva’, Azurri Satin®, and Raspberry Smoothie™) were hexaploids (6.68 ± 0.13 to 7.05 ± 0.18 pg). Peppermint Smoothie™ was a cytochimera with tetraploid cells (4.61 ± 0.06 pg) and octoploid cells (8.98 ± 0.13 pg). To create pentaploids, reciprocal combinations were made between hexaploid ‘Pink Giant’ and tetraploid cultivars. To create octoploids, seedlings were treated with agar solutions containing 0.2% colchicine or 125 μM oryzalin. Guard cell lengths were significantly different among the four cytotypes: 4x (27.36 ± 0.04 μm), 5x (30.35 ± 1.28 μm), 6x (35.59 ± 0.63 μm), and 8x (40.48 ± 1.05 μm). Measurements of stomatal density revealed a precipitous decline in average density from the 4x cytotype (398.22 ± 15.43 stomata/mm2) to 5x cytotype (194.06 ± 38.69 stomata/mm2) but no significant difference among 5x, 6x, and 8x cytotypes. Fluorescent in situ hybridization (FISH) revealed an increase in 5S and 45S rDNA signals that scaled with ploidy: 4x (two 5S + four 45S), 6x (three 5S + six 45S), and 8x (four 5S + eight 45S). However, pentaploid (5x) seedlings exhibited random segregation of rDNA signals between the 4x and 6x cytotypes, including all six possible combinations (two 5S, three 5S) × (four 45S, five 45S, six 45S).

Genome size variation in Hordeum spontaneum populations

Genome ◽  
1999 ◽  
Vol 42 (6) ◽  
pp. 1094-1099 ◽  
Author(s):  
Timo Turpeinen ◽  
Johanna Kulmala ◽  
Eviatar Nevo
Keyword(s):  
Flow Cytometry ◽  
Genome Size ◽  
Intraspecific Variation ◽  
Wild Barley ◽  
Size Variation ◽  
Hordeum Spontaneum ◽  
Genome Size Variation ◽  
Propidium Iodide Staining ◽  
Significant Difference ◽  
Positive Correlations

Populations of wild barley, Hordeum spontaneum (C. Koch), originating from 10 ecologically and geographically different sites in Israel, were assessed for genome size. Measurements were obtained by flow cytometry using propidium iodide staining. Genome sizes ranged from 9.35 to 9.81 pg. Variance analysis indicated a significant difference between populations. Genome sizes were positively correlated with mean January temperature. Our results corroborate previous findings of intraspecific variation in genome size from different plant species. The positive correlations between climate and genome size suggest that the latter is adaptive and determined by natural selection.Key words: Hordeum spontaneum, genome size, flow cytometry, intraspecific variation, natural selection.

scholarly journals Ploidy level determination of Hedera (Araliaceae) with an emphasis on discussable species (Hedera hibernica)

Caryologia ◽  
2021 ◽  
Vol 74 (1) ◽  
pp. 109-116
Author(s):  
Fahimeh Fallah ◽  
Farrokh Ghahremaninejad
Keyword(s):  
Flow Cytometry ◽  
Genome Size ◽  
Nuclear Dna ◽  
Petroselinum Crispum ◽  
New Taxon ◽  
Internal Reference ◽  
Ploidy Levels ◽  
Internal Reference Standard ◽  
Significant Difference

Genome size is a helpful tool for circumscribing taxa at diverse taxonomic degrees (mostly species) and resolving intricate low-level taxonomies. The correct genome size in Hedera (Araliaceae) has long been discussed, and the ploidy levels of some taxa are still unclear. Twelve accessions of Hedera were measured via flow cytometry. Flow cytometry is a relatively rapid, inexpensive, and credible tool. Fresh leaves of Hedera samples and internal reference standard parsley (Petroselinum crispum) were stained with propidium iodide (PI). Flow cytometry measurements showed that for the accessions of 2CV (3.09 - 6.40 pg), the lowest amount of nuclear DNA was 3.09 pg for Hedera crebrescens (So), while the highest amount was 6.40 pg for H. hibernica “Hamilton,” representing a statistically significant difference. According to this study, the new taxon (H. crebrescens) is a diploid, though this taxon was previously considered H. hibernica (tetraploid).

scholarly journals POS0369 ELEVATED EXPRESSION OF TIM-3 ON NEUTROPHILS CORRELATES WITH DISEASE ACTIVITY AND SEVERITY OF ANKYLOSING SPONDYLITIS

2021 ◽  
Vol 80 (Suppl 1) ◽  
pp. 414.2-415
Author(s):  
X. Huang ◽  
T. W. Li ◽  
J. Chen ◽  
Z. Huang ◽  
S. Chen ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Ankylosing Spondylitis ◽  
Disease Activity ◽  
Immune Cells ◽  
Clinical Manifestations ◽  
Innate Immune ◽  
Innate Immune Cells ◽  
Bacterial Killing ◽  
Markers Of Inflammation ◽  
Significant Difference

Background:Ankylosing spondylitis (AS) is a type of common, chronic inflammatory disease that compromises the axial skeleton and sacroiliac joints, causing inflammatory low back pain and progressive spinal stiffness, over time some patients develop spinal immobility and ankylosis which can lead to a decrease in quality of life. The last few decades, evidence has clearly indicated that neutrophil also plays key roles in the progression of AS. However, the immunomodulatory roles and mechanisms of neutrophils in AS are poorly understood. T-cell immunoglobulin and mucin domain-containing protein 3 (Tim-3) has been reported as an important regulatory molecule, expressed and regulated on different innate immune cells, plays a pivotal role in several autoimmunity diseases. Recent study indicates that Tim3 is also expressed on neutrophils. However, the frequency and roles of Tim3-expressing neutrophils in AS was not clear.Objectives:In this study, we investigated the expression of Tim3 on neutrophils in AS patients and explored the correlation between the level of Tim3-expressing neutrophils and the disease activity and severity of AS.Methods:Patients with AS were recruited from Guangdong Second Provincial General Hospital (n=62). Age/sex-matched volunteers as Healthy controls (HC) (n=39). The medical history, clinical manifestations, physical examination, laboratory measurements were recorded. The expression of costimulatory molecules including programmed death 1 (PD-1), Tim-3 on neutrophils were determined by flow cytometry. The mRNA expression of PD-1 and Tim-3 was determined by real-time PCR. The levels of Tim3-expressing neutrophils in AS patients were further analyzed for their correlation with the markers of inflammation such as ESR,CRP,WBC and neutrophil count(NE), as well as disease activity and severity of AS. The expression of Tim3 on neutrophils was monitored during the course of treatment (4 weeks).Results:The expression of Tim3 on neutrophils in patients with AS was increased compared to the HC (Figure 1A). However, significant difference was observed in the frequency of PD-1-expressing neutrophils between AS patients and HC (Figure 1B). The expression analysis of Tim-3 mRNA, but not PD-1, confirmed the results obtained from flow cytometry (Figure 1C). The level of Tim3-expressing neutrophils in patients with AS showed an positive correlation with ESR, CRP and ASAS-endorsed disease activity score (ASDAS) (Figure 1D). Moreover, the frequency of Tim3-expressing neutrophils in active patients(ASDAS≥1.3) was increased as compare with the inactive patients (ASDAS<1.3) (Figure 1E). As shown in Figure 1F, the frequency of Tim3-expressing neutrophils decreased after the treatment.Conclusion:Increased Tim-3 expression on neutrophils may be a novel indicator to assess disease activity and severity in AS, which may serves as a negative feedback mechanism preventing potential tissue damage caused by excessive inflammatory responses in AS patients.References:[1]Han, G., Chen, G., Shen, B. & Li, Y., Tim-3: an activation marker and activation limiter of innate immune cells. FRONT IMMUNOL 4 449 (2013).[2]Vega-Carrascal, I. et al., Galectin-9 signaling through TIM-3 is involved in neutrophil-mediated Gram-negative bacterial killing: an effect abrogated within the cystic fibrosis lung. J IMMUNOL 192 2418 (2014).Figure 1.(A,B)The expression of Tim3 and PD-1 on neutrophils in AS and HC were determined by flow cytometry.(C) The expression of Tim3 and PD-1 on neutrophils in AS and HC were determined by RT-PCR.(D)The correction between Tim3-expressing neutrophils and ESR,CRP,ASDAS.(E) The expression of Tim3 on neutrophils in active and inactive patients.(F) Influence of treatment on the frequency of Tim3-expressing neutrophils.Disclosure of Interests:None declared

scholarly journals Ozone Response of Leaf Physiological and Stomatal Characteristics in Brassica juncea L. at Supraoptimal Temperatures

Land ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 357
Author(s):  
Jong Kyu Lee ◽  
Myeong Ja Kwak ◽  
Sang Hee Park ◽  
Han Dong Kim ◽  
Yea Ji Lim ◽  
...  
Keyword(s):  
Brassica Juncea ◽  
Agricultural Production ◽  
Stomatal Density ◽  
Anthropogenic Activities ◽  
Interactive Effects ◽  
Target Concentration ◽  
Stomatal Characteristics ◽  
Elevated O3 ◽  
Significant Difference ◽  
Supraoptimal Temperatures

Plants are affected by the features of their surrounding environment, such as climate change and air pollution caused by anthropogenic activities. In particular, agricultural production is highly sensitive to environmental characteristics. Since no environmental factor is independent, the interactive effects of these factors on plants are essential for agricultural production. In this context, the interactive effects of ozone (O3) and supraoptimal temperatures remain unclear. Here, we investigated the physiological and stomatal characteristics of leaf mustard (Brassica juncea L.) in the presence of charcoal-filtered (target concentration, 10 ppb) and elevated (target concentration, 120 ppb) O3 concentrations and/or optimal (22/20 °C day/night) and supraoptimal temperatures (27/25 °C). Regarding physiological characteristics, the maximum rate of electron transport and triose phosphate use significantly decreased in the presence of elevated O3 at a supraoptimal temperature (OT conditions) compared with those in the presence of elevated O3 at an optimal temperature (O conditions). Total chlorophyll content was also significantly affected by supraoptimal temperature and elevated O3. The chlorophyll a/b ratio significantly reduced under OT conditions compared to C condition at 7 days after the beginning of exposure (DAE). Regarding stomatal characteristics, there was no significant difference in stomatal pore area between O and OT conditions, but stomatal density under OT conditions was significantly increased compared with that under O conditions. At 14 DAE, the levels of superoxide (O2-), which is a reactive oxygen species, were significantly increased under OT conditions compared with those under O conditions. Furthermore, leaf weight was significantly reduced under OT conditions compared with that under O conditions. Collectively, these results indicate that temperature is a key driver of the O3 response of B. juncea via changes in leaf physiological and stomatal characteristics.

First estimates of genome size in ribbon worms (phylum Nemertea) using flow cytometry and Feulgen image analysis densitometry

2014 ◽  
Vol 92 (10) ◽  
pp. 847-851 ◽  
Author(s):  
Kelly L. Mulligan ◽  
Terra C. Hiebert ◽  
Nicholas W. Jeffery ◽  
T. Ryan Gregory
Keyword(s):  
Flow Cytometry ◽  
Image Analysis ◽  
Body Size ◽  
Genome Size ◽  
Positive Relationship ◽  
Nuclear Dna ◽  
Nuclear Dna Content ◽  
Size Estimation ◽  
Size Diversity ◽  
Size Estimates

Ribbon worms (phylum Nemertea) are among several animal groups that have been overlooked in past studies of genome-size diversity. Here, we report genome-size estimates for eight species of nemerteans, including representatives of the major lineages in the phylum. Genome sizes in these species ranged more than fivefold, and there was some indication of a positive relationship with body size. Somatic endopolyploidy also appears to be common in these animals. Importantly, this study demonstrates that both of the most common methods of genome-size estimation (flow cytometry and Feulgen image analysis densitometry) can be used to assess genome size in ribbon worms, thereby facilitating additional efforts to investigate patterns of variability in nuclear DNA content in this phylum.

Comparative genome size estimation of different life stages of grey mullet, Mugil cephalus Linnaeus, 1758 by flow cytometry

2021 ◽  
Author(s):  
Jani Angel J. Raymond ◽  
Mudagandur Shashi Shekhar ◽  
Vinaya Kumar Katneni ◽  
Ashok Kumar Jangham ◽  
Sudheesh Kommu Prabhudas ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Genome Size ◽  
Mugil Cephalus ◽  
Life Stages ◽  
Size Estimation ◽  
Grey Mullet ◽  
Comparative Genome

scholarly journals Coconut genome size determined by flow cytometry: Tall versus Dwarf types

2016 ◽  
Vol 15 (1) ◽  
Author(s):  
M. Freitas Neto ◽  
T.N.S. Pereira ◽  
I.G.C. Geronimo ◽  
A.O.N. Azevedo ◽  
S.R.R. Ramos ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Genome Size

scholarly journals Difference between emergent aquatic and terrestrial monocotyledonous herbs in relation to the coordination of leaf stomata with vein traits

AoB Plants ◽  
2020 ◽  
Vol 12 (5) ◽  
Author(s):  
Wanli Zhao ◽  
Peili Fu ◽  
Guolan Liu ◽  
Ping Zhao
Keyword(s):  
Water Supply ◽  
Aquatic Plants ◽  
Stomatal Density ◽  
Supply And Demand ◽  
Terrestrial Plants ◽  
Major Vein ◽  
Significant Difference ◽  
Leaf Veins ◽  
Vein Diameter ◽  
Stomatal Length

Abstract Emergent aquatic plants mostly occur in shallow waters and root in bottom substrates, but their leaves emerge from the water surface and are thus exposed to air, similar to the leaves of terrestrial plants. Previous studies have found coordination between leaf water supply and demand in terrestrial plants; however, whether such a coordination exists in emergent aquatic plants remains unknown. In this study, we analysed leaf veins and stomatal characteristics of 14 emergent aquatic and 13 terrestrial monocotyledonous herb species (EMH and TMH), with 5 EMH and 8 TMH belonging to Poaceae. We found that EMH had significantly higher mean leaf area, leaf thickness, stomatal density, stomatal number per vein length and major vein diameter, but lower mean major vein length per area (VLA) and total VLA than TMH. There was no significant difference in stomatal length, minor VLA and minor vein diameter between the two groups. Stomatal density and total VLA were positively correlated among the EMH, TMH, as well as the 8 Poaceae TMH species, but this correlation became non-significant when data from both the groups were pooled. Our results showed that the differences in water supply between emergent aquatic and terrestrial plants modify the coordination of their leaf veins and stomatal traits.

scholarly journals A Flow Cytometry-Based Assay for the Measurement of Total Complement Activity in the Serum and Clinical Practice

2022 ◽  
Vol 2022 ◽  
pp. 1-12
Author(s):  
Xuewei Ding ◽  
Shijun Li ◽  
Hui Liu
Keyword(s):  
Flow Cytometry ◽  
Nephrotic Syndrome ◽  
Linear Relationship ◽  
High Volume ◽  
Healthy Individuals ◽  
Serum Samples ◽  
Complement Activity ◽  
Significant Difference ◽  
Cell Groups ◽  
Total Complement

Objective. To develop a novel sensitive and accurate assay suitable for high-volume testing of the total complement activity in the serum for clinical laboratories. Methods. The total complement activity (TCA) to be measured was quantified by detecting the number of fragments produced by erythrocyte lysis and the erythrocyte fragmentation index (EFI), indicating TCA. EFI = M × M 2 / M 1 + M 2 , where M is the number of erythrocyte fragments (removed from the background), M 1 is the number of unagglutinated red cells, M 2 is the number of agglutinated red cell groups, and M 2 / M 1 + M 2 is the agglutination coefficient indicating the degree of erythrocyte agglutination. Mild changes in hemolysin and erythrocyte concentrations were made to optimize the testing conditions. The same serum samples were tested for 10 consecutive days to determine the stability of the experimental results. Serum EFI was detected in both nephrotic syndrome patients and healthy subjects. Results. There was a linear relationship between hemolysin and erythrocyte agglutination ( r = 0.999 , P < 0.001 ). A good linear relationship existed between EFI and TCA ( r = 0.991 , P < 0.001 ). The results were not affected by slight fluctuations in the concentrations of hemolysin or erythrocytes. The interbatch CV = 8.6 % of the test results showed good stability. There was a significant difference in the EFI between nephrotic syndrome patients and healthy individuals, P < 0.001 , and EFI was reduced in nephrotic syndrome patients compared to healthy individuals. Conclusion. The flow cytometry-based assay for TCA was sensitive and accurate and had potential value for clinical application.

Sign in / Sign up

Export Citation Format

Share Document