Effects of sampling methods on starch granule size measurement of potato tubers under a light microscope

Light-microscopic measurement of starch granule size is the preferred approach in most laboratories because it is simple, rapid and visual and because it can study both size and shape. However, potato juice consists of starch granules with very different sizes and precipitation/movement speeds, which causes differences when sampling the juice and taking the microscopic images. The previously described method is to scrape and transfer some juice from potatoes using a razor blade directly to a slide with some water for microscopic observation. In this study we used a tape-hole chamber on the microscopic slide to reduce the cover-slip-induced shifting of small and medium granules. We improved the starch measurement reproducibility by testing various juice sampling methods. The reproducibility between repeated experiments using 10 cultivars was increased from a correlation efficient r = 0.815 in the razor-blade-scraping method to r = 0.923 in a squeezing-juice method. The largest starch granule detected was 151 µm in length. Sampling methods (using a razor-blade or a garlic press) strongly influenced the granule length values measured from the same potato tuber. The results indicated that 1) The squeezing-juice approach is more reproducible, and 2) The average length of starch granules is one of the most reproducible scores but varies according to juice-sampling methods.

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Starch Granule Size and Morphology of Arabidopsis thaliana Starch-Related Mutants Analyzed during Diurnal Rhythm and Development

Molecules ◽

10.3390/molecules26195859 ◽

2021 ◽

Vol 26 (19) ◽

pp. 5859

Author(s):

Qingting Liu ◽

Yuan Zhou ◽

Joerg Fettke

Keyword(s):

Arabidopsis Thaliana ◽

Starch Granule ◽

Granule Size ◽

High Temporal Resolution ◽

Starch Granules ◽

Transitory Starch ◽

Young Leaves ◽

Size And Morphology ◽

Safranin O

Transitory starch plays a central role in the life cycle of plants. Many aspects of this important metabolism remain unknown; however, starch granules provide insight into this persistent metabolic process. Therefore, monitoring alterations in starch granules with high temporal resolution provides one significant avenue to improve understanding. Here, a previously established method that combines LCSM and safranin-O staining for in vivo imaging of transitory starch granules in leaves of Arabidopsis thaliana was employed to demonstrate, for the first time, the alterations in starch granule size and morphology that occur both throughout the day and during leaf aging. Several starch-related mutants were included, which revealed differences among the generated granules. In ptst2 and sex1-8, the starch granules in old leaves were much larger than those in young leaves; however, the typical flattened discoid morphology was maintained. In ss4 and dpe2/phs1/ss4, the morphology of starch granules in young leaves was altered, with a more rounded shape observed. With leaf development, the starch granules became spherical exclusively in dpe2/phs1/ss4. Thus, the presented data provide new insights to contribute to the understanding of starch granule morphogenesis.

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A carbohydrate-binding protein, B-GRANULE CONTENT 1, influences starch granule size distribution in a dose-dependent manner in polyploid wheat

Journal of Experimental Botany ◽

10.1093/jxb/erz405 ◽

2019 ◽

Vol 71 (1) ◽

pp. 105-115 ◽

Cited By ~ 5

Author(s):

Tansy Chia ◽

Marcella Chirico ◽

Rob King ◽

Ricardo Ramirez-Gonzalez ◽

Benedetta Saccomanno ◽

...

Keyword(s):

Size Distribution ◽

Starch Granule ◽

Starch Synthesis ◽

Granule Size ◽

Starch Granules ◽

Grain Development ◽

Polyploid Wheat ◽

Gene Dose ◽

Dose Dependent ◽

B Granules

Abstract In Triticeae endosperm (e.g. wheat and barley), starch granules have a bimodal size distribution (with A- and B-type granules) whereas in other grasses the endosperm contains starch granules with a unimodal size distribution. Here, we identify the gene, BGC1 (B-GRANULE CONTENT 1), responsible for B-type starch granule content in Aegilops and wheat. Orthologues of this gene are known to influence starch synthesis in diploids such as rice, Arabidopsis, and barley. However, using polyploid Triticeae species, we uncovered a more complex biological role for BGC1 in starch granule initiation: BGC1 represses the initiation of A-granules in early grain development but promotes the initiation of B-granules in mid grain development. We provide evidence that the influence of BGC1 on starch synthesis is dose dependent and show that three very different starch phenotypes are conditioned by the gene dose of BGC1 in polyploid wheat: normal bimodal starch granule morphology; A-granules with few or no B-granules; or polymorphous starch with few normal A- or B-granules. We conclude from this work that BGC1 participates in controlling B-type starch granule initiation in Triticeae endosperm and that its precise effect on granule size and number varies with gene dose and stage of development.

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Starch granule size distribution in wheat grain in relation to phosphorus fertilization

The Journal of Agricultural Science ◽

10.1017/s0021859611000475 ◽

2011 ◽

Vol 150 (1) ◽

pp. 45-52 ◽

Cited By ~ 14

Author(s):

Y. NI ◽

Z. WANG ◽

Y. YIN ◽

W. LI ◽

S. YAN ◽

...

Keyword(s):

Size Distribution ◽

Starch Granule ◽

Amylose Content ◽

Granule Size ◽

High Yield ◽

Phosphorus Fertilization ◽

Equivalent Diameter ◽

Starch Granules ◽

P Fertilization ◽

Volume Proportion

SUMMARYStarch granule size distribution of wheat is an important characteristic that can affect its chemical composition and functionality. Phosphorus (P) fertilization has been studied extensively; however, little is known about its impact on starch granule size distribution in wheat. In the present study, two high-yield winter wheat cultivars were grown under different P fertilization conditions to evaluate its effect on starch granule size distribution and starch components in wheat grains at maturity. P fertilization resulted in a significant increase in the proportions (both by volume and by surface area) of B-type (<9·9 μm equivalent diameter (e.d.)) starch granules, with a reduction in those of A-type (>9·9 μm e.d.) starch granules. The P fertilization also increased starch content, amylose content and amylopectin content at maturity. However, P fertilization conditions significantly reduced the ratio of amylose to amylopectin, which showed a significant positive relationship with the volume proportion of granules 22·8–42·8 μm e.d. but was negatively related to the volume proportion of granules 2·8–9·9 μm e.d.

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Manipulation of starch granule size distribution in potato tubers by modulation of plastid division

Plant Biotechnology Journal ◽

10.1111/j.1467-7652.2005.00163.x ◽

2006 ◽

Vol 4 (1) ◽

pp. 123-134 ◽

Cited By ~ 23

Author(s):

Sylvia Pater ◽

Martien Caspers ◽

Marijke Kottenhagen ◽

Henk Meima ◽

Renaldo ter Stege ◽

...

Keyword(s):

Size Distribution ◽

Starch Granule ◽

Granule Size ◽

Plastid Division ◽

Potato Tubers

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Starch Granule Size and Morphology as a Proxy for Water Regime Influence on Zea mays

Ethnobiology Letters ◽

10.14237/ebl.12.1.2021.1725 ◽

2021 ◽

Vol 12 (1) ◽

pp. 35-43

Author(s):

Stefania Wilks ◽

Lisbeth A. Louderback ◽

Shannon Boomgarden

Keyword(s):

Zea Mays ◽

Starch Granule ◽

Granule Size ◽

Plant Domestication ◽

Starch Granules ◽

Morphological Attributes ◽

Essential Parameter ◽

Size And Morphology ◽

Human Subsistence

A wealth of information on the patterns of human subsistence and plant domestication has been generated from studies on maize (Zea mays) starch granules. However, very little work has been conducted on how the size and morphology of those granules might change as a function of water stress during the growing season. In the arid Southwest, the role of irrigation in growing maize is an essential parameter in many foraging models. Our study seeks to determine if there are significant changes in the size and other morphological attributes of starch granules from maize planted at Range Creek Canyon under two different irrigation regimes ranging from little water (once every three weeks) to ample water (once a day). Our results provide data on the effects of irrigation on Z. mays starch granules and, therefore, have implications for identifying archaeological maize and possibly determining past water regimes at Range Creek Canyon.

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Starch granule size distribution from twelve wheat cultivars in east China’s Huaibei region

Canadian Journal of Plant Science ◽

10.1139/cjps-2015-0048 ◽

2016 ◽

Vol 96 (2) ◽

pp. 176-182 ◽

Cited By ~ 4

Author(s):

Wenyang Li ◽

Suhui Yan ◽

Xiaqing Shi ◽

Congyu Zhang ◽

Qingqin Shao ◽

...

Keyword(s):

Size Distribution ◽

Starch Granule ◽

Amylose Content ◽

Granule Size ◽

Volume Distribution ◽

Starch Granules ◽

Wheat Cultivars ◽

Volume Percentage ◽

Area Distribution ◽

Two Populations

The granule size distribution of starch strongly influences its physicochemical properties and the functionality of wheat. Twelve wheat cultivars grown in east China’s Huanghuai region were used for investigating the granule size distribution, amylose content, and their interrelationship. The results showed that the volume distribution of starch granules show the typical bimodal with peak values in the ranges of 4.44–5.36 µm and 21.7–23.82 µm, respectively. Surface area distribution of granules was also bimodal with peak values in the ranges of 2.53–3.06 µm and 19.8–21.7 µm, respectively. The limits between the two populations both occurred at 10 µm. A typical population of number distribution of granules with peak values in the range of 0.52–0.67 µm. Proportions of granules <2.6 µm, 2.6–10 µm, and 10–40 µm were in the range of 10.06–13.63%, 28.54–41.6%, and 45.4–61.3% of total volume, respectively. Proportions of granules <10 µm were in the range of 99.9% of the total number. The amylose content was significant and negatively correlated to volume percentage of granule <10 µm, and significant and positively correlated to the volume percentage of granule 22–40 µm.

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Starch granule size distribution in wheat grain in relation to shading after anthesis

The Journal of Agricultural Science ◽

10.1017/s0021859609990554 ◽

2009 ◽

Vol 148 (2) ◽

pp. 183-189 ◽

Cited By ~ 16

Author(s):

W. LI ◽

S. YAN ◽

Y. YIN ◽

Z. WANG

Keyword(s):

Light Intensity ◽

Grain Yield ◽

Size Distribution ◽

Starch Granule ◽

Grain Filling ◽

Granule Size ◽

Wheat Starch ◽

High Yield ◽

Starch Accumulation ◽

Starch Granules

SUMMARYGranule size distribution of wheat starch is an important characteristic that may affect the functionality of wheat products. Light intensity is one of the main factors affecting grain yield and quality. Two high-yield winter wheat cultivars were grown under shade to evaluate the effect of low light intensity after anthesis on starch granule size distribution and starch components in wheat grains at maturity. Shading caused a marked drop in both grain yield and starch yield and led to a significant reduction in the proportion (both by volume and by surface area) of B-type starch granules (⩽9·9 μm), with an increase in those of A-type starch granules (>9·9 μm). This would suggest that the production of B-type starch granules was more sensitive to shading than that of A-type starch granules. It was also found that the proportion by volume of A-type starch granules was significantly increased and that of B-type starch granules was significantly decreased by shading at different grain filling stages, especially at middle and late grain-filling stages. However, shading had little effect on the proportional number of B-type starch granules. The present results suggested that, under dim light conditions, the limited substrate for starch accumulation was mainly partitioned towards hypertrophy (larger granules) not hyperplasia (more) of starch granules.

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Intra-Sample Heterogeneity of Potato Starch Reveals Fluctuation of Starch-Binding Proteins According to Granule Morphology

Plants ◽

10.3390/plants8090324 ◽

2019 ◽

Vol 8 (9) ◽

pp. 324

Author(s):

Stanislas Helle ◽

Fabrice Bray ◽

Jean-Luc Putaux ◽

Jérémy Verbeke ◽

Stéphanie Flament ◽

...

Keyword(s):

Starch Granule ◽

Potato Starch ◽

Granule Size ◽

Starch Granules ◽

Label Free ◽

Granule Morphology ◽

Sample Heterogeneity ◽

Amylopectin Structure ◽

Unimodal Pattern

Starch granule morphology is highly variable depending on the botanical origin. Moreover, all investigated plant species display intra-tissular variability of granule size. In potato tubers, the size distribution of starch granules follows a unimodal pattern with diameters ranging from 5 to 100 µm. Several evidences indicate that granule morphology in plants is related to the complex starch metabolic pathway. However, the intra-sample variability of starch-binding metabolic proteins remains unknown. Here, we report on the molecular characterization of size-fractionated potato starch granules with average diameters of 14.2 ± 3.7 µm, 24.5 ± 6.5 µm, 47.7 ± 12.8 µm, and 61.8 ± 17.4 µm. In addition to changes in the phosphate contents as well as small differences in the amylopectin structure, we found that the starch-binding protein stoichiometry varies significantly according to granule size. Label-free quantitative proteomics of each granule fraction revealed that individual proteins can be grouped according to four distinct abundance patterns. This study corroborates that the starch proteome may influence starch granule growth and architecture and opens up new perspectives in understanding the dynamics of starch biosynthesis.

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Brewing trials with spring and winter barley varieties

Czech Journal of Food Sciences ◽

10.17221/145/2010-cjfs ◽

2012 ◽

Vol 30 (No. 1) ◽

pp. 27-34

Author(s):

A. Mikyška ◽

V. Psota ◽

M. Hrabák

Keyword(s):

Size Distribution ◽

Starch Granule ◽

Granule Size ◽

Starch Granules ◽

Spent Grains ◽

Significant Difference ◽

Brewing Process ◽

Decoction Process ◽

Beer Production

The effects of a set of barley varieties on the brewing process and quality of beer production intermediaries were studied in trial brews (40 l) prepared using the two-mash decoction process. The varieties included in the trial were selected based on the starch granule size distribution determined previously. A significant effect of the varieties on the saccharification time of both mashes was determined. The highest saccharification rate in brews was achieved with the variety Jersey; the saccharification time of the 1st and 2nd mash with the variety Tiffany was markedly longer. The varieties with a greater fraction of large starch granules (Tiffany and Luxor) exhibited a higher haze with sweet wort as well as hopped wort compared to the varieties with a low fraction of large starch granules (Jersey and Tolar). The effect on the lautering time was not demonstrated. Pronounced varietal differences were determined in the extract balance of the brewing process. The varieties Tiffany andLuxor exhibited significantly lower extract yields. The malts from these varieties had lower laboratory extracts and higher extract losses in spent grains. The effect of the variety on the sacharide composition in hopped wort was confirmed. The proportion of fermentable saccharides in hopped wort extract rose from the variety Tiffany (66.9%) to the varietyJersey (83.6%). A significant difference in the final attenuation was also determined (76% in beers prepared from the varieties Tiffany andLuxor compared to 81.5% from theJersey variety).

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