Impedance Flow Cytometry: A Novel Technique in Pollen Analysis

Heidmann, Iris; Schade-Kampmann, Grit; Lambalk, Joep; Ottiger, Marcel; Berardino, Marco Di

doi:10.1371/journal.pone.0165531

Cited by 70 publications

(76 citation statements)

References 66 publications

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“…() described the potential of using impedance flow cytometry (IFC) for evaluating pollen quality by distinguishing between viable and heat‐inactivated pollen of several species. In IFC, cells are passed through a micro‐fluidic chip and their dielectric properties (impedance) are measured in a range of several radio frequencies, which can be used to analyze cell size, shape and membrane properties (Heidmann et al ., ). Although this appears to be a good non‐invasive and label‐free method for evaluating quality of pollen, correlation of IFC viability score with actual pollen germination capacity was not presented.…”

Section: Discussionmentioning

confidence: 99%

Direct analysis of pollen fitness by flow cytometry: implications for pollen response to stress

et al. 2019

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Summary Sexual reproduction in flowering plants depends on the fitness of the male gametophyte during fertilization. Because pollen development is highly sensitive to hot and cold temperature extremes, reliable methods to evaluate pollen viability are important for research into improving reproductive heat stress (HS) tolerance. Here, we describe an approach to rapidly evaluate pollen viability using a reactive oxygen species (ROS) probe dichlorodihydrofluorescein diacetate (i.e. H2DCFDA‐staining) coupled with flow cytometry. In using flow cytometry to analyze mature pollen harvested from Arabidopsis and tomato flowers, we discovered that pollen distributed bimodally into ‘low‐ROS’ and ‘high‐ROS’ subpopulations. Pollen germination assays following fluorescence‐activated cell sorting revealed that the high‐ROS pollen germinated with a frequency that was 35‐fold higher than the low‐ROS pollen, supporting a model in which a significant fraction of a flower's pollen remains in a low metabolic or dormant state even after hydration. The ability to use flow cytometry to quantify ROS dynamics within a large pollen population was shown by dose‐dependent alterations in DCF‐fluorescence in response to oxidative stress or antioxidant treatments. HS treatments (35°C) increased ROS levels, which correlated with a ~60% reduction in pollen germination. These results demonstrate the potential of using flow cytometry‐based approaches to investigate metabolic changes during stress responses in pollen.

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Section: Discussionmentioning

confidence: 99%

Direct analysis of pollen fitness by flow cytometry: implications for pollen response to stress

et al. 2019

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“…To quantify the amount and the viability of the pollen on both stressed and not stressed plants, we used the impedance flow cytometry (IFC), manufactured by Amphasys ® AG (Amphasys ® AG, Root, Switzerland), as a fast and non-invasive technique. It measures electrical capacity (and hence viability) of a cell, utilizing a small microfluidic chip where the pollen grains flow through and the electric charge of the cells is measured via different radio frequencies [18].…”

Section: Pollen Analyses Via Amphasys ® Impedance Flow Cytometrymentioning

confidence: 99%

“…The production of a high pollen number with sufficient vitality also at lower temperature is considered to be the underlying physiological process to ensure seed set under cold stress in sorghum [11] and other tropical crops like chickpea [15] and rice [16,17]. While in the past the screening of pollen vitality traits based on staining techniques was labor-intensive and tedious, nowadays, the impedance flow cytometry [18] provides an efficient novel tool for a fast screening of a higher number of genotypes.…”

Section: Introductionmentioning

confidence: 99%

Reproductive Cold Stress Tolerance in Sorghum F1 Hybrids is a Heterotic Trait

Schaffasz¹,

Windpassinger²,

Snowdon³

et al. 2019

Agronomy

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The sensitivity of sorghum to pre-flowering cold stress, resulting in reduced pollen viability and poor seed set, is a major constraint for expanding growing areas into higher altitudes and latitudes. Nevertheless, compared to juvenile cold tolerance, reproductive cold tolerance in sorghum has received much less attention so far, and very little is known about its inheritance in F1-hybrids. We have composed a representative factorial (n = 49 experimental F1-hybrids) for a comprehensive study on heterosis and combining ability for crucial tolerance traits as spikelet fertility (panicle harvest index), seed yield and pollen viability, using field trials in stress- and control environments in Germany and Mexico as well as climate chamber experiments. Our results indicate a heterotic and rather dominant inheritance of reproductive cold tolerance in sorghum, with strong effects of female general combining ability (GCA) on F1-hybrid performance in our material. These findings, together with the comparatively low contribution of specific combining ability (SCA) effects and high heritability estimates, suggest that robust and efficient enhancement of reproductive cold tolerance is feasible via hybrid breeding.

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“…Pollen viability test. The pollen viability of the pollen and 1:0, 0:1, 1:1, 1:4, 1:16, and 1:1 volume/volume (v/v) ratios of pollen and dust and herbicide-contaminated dust were tested by AmpaZ30 (Amphasys AG, Root, Switzerland) (Heidmann et al, 2016). The 1:1 herbicidecontaminated treatment was the pollen and dust mixture in which the 300°F-processed dust was presoaked with commonly used herbicides GlyStar Ò Plus (Albaugh LLC, Ankeny, IA) and Treevix Ò (BASF, Durham, NC) with labelrecommended concentrations and dried at 75°F under a fume hood.…”

Section: Methodsmentioning

confidence: 99%

Dust Interferes with Pollen–Stigma Interaction and Fruit Set in Pistachio Pistacia vera cv. Kerman

Zhang¹,

Beede²,

Bañuelos³

et al. 2019

horts

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Springtime flail mowing of row middles for weed control in California pistachio (Pistacia vera L.) orchards blows dust into the leafless canopy if it occurs during bloom. The effect of dust on pistachio pollination and fruit set is unknown. Rachises were bagged prebloom and hand pollinated with pollen and dust mixtures at 1:0, 1:1, 1:4, 1:16, and 0:1 volume/volume ratios on five successive days. The 2016 and 2017 trials demonstrated that the inflorescences treated with a high pollen:dust ratio (0:1, 1:4, and 1:16) had significantly lower split nut rates (commercially less profitable) compared with low dust ratio tests (1:0 and 1:1). Our results also showed that dust damaged both pollen viability and stigma quality, particularly if contaminated with herbicide residues (GlyStar® Plus and Treevix®). Decreased yield was a function of decreased fruit set; increased embryo abortion, parthenocarpy, or both; and a lower split nut percentage. The GA3 content in flowers of both the pollen and dust treatments was significantly higher than that in nonpollinated flowers, suggesting dust stimulated parthenocarpy, resulting in empty nutshells, “blanks” at harvest.

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Impedance Flow Cytometry: A Novel Technique in Pollen Analysis

Cited by 70 publications

References 66 publications

Direct analysis of pollen fitness by flow cytometry: implications for pollen response to stress

Direct analysis of pollen fitness by flow cytometry: implications for pollen response to stress

Reproductive Cold Stress Tolerance in Sorghum F1 Hybrids is a Heterotic Trait

Dust Interferes with Pollen–Stigma Interaction and Fruit Set in Pistachio Pistacia vera cv. Kerman

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