Vibrational microspectroscopy enables chemical characterization of single pollen grains as well as comparative analysis of plant species based on pollen ultrastructure

Zimmermann, Boris; Bağcıoğlu, Murat; Sandt, Christophe; Köhler, Achim

doi:10.1007/s00425-015-2380-7

Cited by 48 publications

(64 citation statements)

References 42 publications

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“…Moreover, intact pollen grains can create saturation effects when measured in transmission mode (Zimmermann et al . ). The saturation effect happens when the central part of the grain is opaque for a large part of the infrared spectrum of the light, and the small fraction of light that reaches the detector has passed through the periphery areas of pollen grain.…”

Section: Discussionmentioning

confidence: 97%

“…; Zimmermann & Kohler ; Bağcıoğlu, Zimmermann & Kohler ; Zimmermann et al . , ) as well as multi‐grains (Wolkers & Hoekstra , ; Dell'Anna et al . ; Bağcıoğlu, Zimmermann & Kohler ) and photoacoustic FTIR spectroscopy (Parodi, Dickerson & Cloud ).…”

Section: Introductionmentioning

confidence: 99%

“…Infrared spectra of pollen are precise fingerprints of the overall biochemical composition of a pollen grain, and contain specific signals of lipids, proteins, carbohydrates, cell wall biopolymers and water. The spectra can be obtained by using diverse techniques, such as single- (Gottardini et al 2007;Zimmermann 2010;Lahlali et al 2014;Zimmermann & Kohler 2014; Ba gcıo glu, Zimmermann & Kohler 2015;Jiang et al 2015;Zimmermann et al 2015b) and multi-reflection (Mularczyk-Oliwa et al 2012) attenuated total reflection (ATR) FTIR, diffuse reflectance FTIR (DRIFT) (Pappas et al 2003;Mularczyk-Oliwa et al 2012), transmission FTIR measurements between windows (Crowe, Hoekstra & Crowe 1989;Sowa, Connor & Towill 1991) and in pellets (Pappas et al 2003;Zimmermann 2010;Mularczyk-Oliwa et al 2012;Zimmermann & Kohler 2014;Ba gcıo glu, Zimmermann & Kohler 2015), FTIR microspectroscopy measurements of single pollen grains (Pappas et al 2003;Gottardini et al 2007;Pummer et al 2013;Zimmermann & Kohler 2014;Ba gcıo glu, Zimmermann & Kohler 2015;Zimmermann et al 2015aZimmermann et al , 2016 as well as multi-grains (Wolkers & Hoekstra 1995Dell'Anna et al 2009; Ba gcıo glu, Zimmermann & Kohler 2015) and photoacoustic FTIR spectroscopy (Parodi, Dickerson & Cloud 2013). Recent studies have shown that FTIR spectroscopy achieves simple, economical and rapid identification and classification of pollen by spectral fingerprinting, as well as characterization and biochemical interpretation with respect to environmental stress (Lahlali et al 2014;Zimmermann & Kohler 2014;<...>…”

Section: Introductionmentioning

confidence: 99%

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Monitoring of plant–environment interactions by high‐throughput FTIR spectroscopy of pollen

et al. 2016

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Summary Fourier transform infrared (FTIR) spectroscopy enables chemical analysis of pollen samples for plant phenotyping to study plant–environment interactions, such as influence of climate change or pathogens. However, current approach, such as microspectroscopy and attenuated total reflection spectroscopy, does not allow for high‐throughput protocols. This study at hand suggests a new spectroscopic method for high‐throughput characterization of pollen. Samples were measured as thin films of pollen fragments using a Bruker FTIR spectrometer with a high‐throughput eXTension (HTS‐XT) unit employing 384‐well plates. In total, 146 pollen samples, belonging to 31 different pollen species of Fagaceae and Betulaceae and collected during three consecutive years (2012–2014) at locations in Croatia, Germany and Norway, were analysed. Critical steps in the sample preparation and measurement, such as variabilities between technical replicates, between microplates and between spectrometers, were studied. Measurement variations due to sample preparation, microplate holders and instrumentation were low, and thus allowed differentiation of samples with respect to phylogeny and biogeography. The spectral variability for a range of Fagales species (Fagus, Quercus, Betula, Corylus, Alnus and Ostrya) showed high‐species‐specific differences in pollen's chemical composition due to either location or year. Statistically significant inter‐annual and locational differences in the pollen spectra indicate that pollen chemical composition has high phenotypic plasticity and is influenced by local climate conditions. The variations in composition are connected to lipids, proteins, carbohydrates and sporopollenins that play crucial roles in cold and desiccation tolerance, protection against UV radiation and as material and energy reserves. The results of this study demonstrate the value of high‐throughput FTIR approach for the systematic collection of data on ecosystems. The novel FTIR approach offers fast, reliable and economical screening of large number of samples by semi‐automated methodology. The high‐throughput approach could provide crucial understanding on plant–climate interactions with respect to biochemical variation within genera, species and populations.

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Section: Discussionmentioning

confidence: 97%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Monitoring of plant–environment interactions by high‐throughput FTIR spectroscopy of pollen

et al. 2016

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“…For that reason, FTIR spectroscopy is widely used for the rapid differentiation and identification of microorganisms [16–19]. The main advantages of FTIR spectroscopy are that (a) several compounds can be measured simultaneously, (b) the method is rapid since little or no sample preparation is required for spectral acquisition, (c) it is chemical-free, (d) it can be used for HTS and for real-time bioprocess monitoring, (e) and even spatial information can be obtained by the use of FTIR microspectroscopy systems [20, 21]. Since infrared spectra are highly complex, with many overlapping signals, multivariate data analysis is required to gain useful information [17, 22].…”

Section: Introductionmentioning

confidence: 99%

Microtiter plate cultivation of oleaginous fungi and monitoring of lipogenesis by high-throughput FTIR spectroscopy

et al. 2017

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BackgroundOleaginous fungi can accumulate lipids by utilizing a wide range of waste substrates. They are an important source for the industrial production of omega-6 polyunsaturated fatty acids (gamma-linolenic and arachidonic acid) and have been suggested as an alternative route for biodiesel production. Initial research steps for various applications include the screening of fungi in order to find efficient fungal producers with desired fatty acid composition. Traditional cultivation methods (shake flask) and lipid analysis (extraction-gas chromatography) are not applicable for large-scale screening due to their low throughput and time-consuming analysis. Here we present a microcultivation system combined with high-throughput Fourier transform infrared (FTIR) spectroscopy for efficient screening of oleaginous fungi.ResultsThe microcultivation system enables highly reproducible fungal fermentations throughout 12 days of cultivation. Reproducibility was validated by FTIR and HPLC data. Analysis of FTIR spectral ester carbonyl peaks of fungal biomass offered a reliable high-throughput at-line method to monitor lipid accumulation. Partial least square regression between gas chromatography fatty acid data and corresponding FTIR spectral data was used to set up calibration models for the prediction of saturated fatty acids, monounsaturated fatty acids, polyunsaturated fatty acids, unsaturation index, total lipid content and main individual fatty acids. High coefficients of determination (R2 = 0.86–0.96) and satisfactory residual predictive deviation of cross-validation (RPDCV = 2.6–5.1) values demonstrated the goodness of these models.ConclusionsWe have demonstrated in this study, that the presented microcultivation system combined with rapid, high-throughput FTIR spectroscopy is a suitable screening platform for oleaginous fungi. Sample preparation for FTIR measurements can be automated to further increase throughput of the system.Electronic supplementary materialThe online version of this article (doi:10.1186/s12934-017-0716-7) contains supplementary material, which is available to authorized users.

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“…Alternative analyses, particularly regarding chemical composition of pollen such as measurement of proteins (Roulston, Cane, & Buchmann, 2000), carbohydrates (Speranza et al, 1997), and lipids (Piffanelli, Ross, & Murphy, 1998), are rarely conducted because they require complex sample preparation and laborious analysis. Recently, Fourier transform infrared spectroscopy (FTIR) has emerged as a significant breakthrough in pollen analysis as a precise fingerprint of the overall biochemical composition of pollen grain is provided (Bağcıoğlu, Zimmermann, & Kohler, 2015;Gottardini, Rossi, Cristofolini, & Benedetti, 2007;Jiang et al, 2015;Lahlali et al, 2014;Pappas, Tarantilis, Harizanis, & Polissiou, 2003;Zimmermann, 2010;Zimmermann, Bagcioglu, Sandt, & Kohler, 2015a;Zimmermann & Kohler, 2014;Zimmerman, Tafintseva, Bagcioglu, Hoegh Berdahl, & Kohler, 2016;Zimmermann, Tkalcec, Mesic, & Kohler, 2015b). Infrared spectra of pollen contain specific signals (i.e., vibrational frequencies of molecular bonds) that can be directly related to molecular functional groups and indirectly to biomolecules, such as lipids, proteins, carbohydrates, cell wall biopolymers, and other biochemical constituents.…”

Section: Introductionmentioning

confidence: 99%

A high‐throughput FTIR spectroscopy approach to assess adaptive variation in the chemical composition of pollen

Zimmermann

Bağcıoğlu

Tafinstseva

et al. 2017

Ecology and Evolution

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The two factors defining male reproductive success in plants are pollen quantity and quality, but our knowledge about the importance of pollen quality is limited due to methodological constraints. Pollen quality in terms of chemical composition may be either genetically fixed for high performance independent of environmental conditions, or it may be plastic to maximize reproductive output under different environmental conditions. In this study, we validated a new approach for studying the role of chemical composition of pollen in adaptation to local climate. The approach is based on high‐throughput Fourier infrared (FTIR) characterization and biochemical interpretation of pollen chemical composition in response to environmental conditions. The study covered three grass species, Poa alpina, Anthoxanthum odoratum, and Festuca ovina. For each species, plants were grown from seeds of three populations with wide geographic and climate variation. Each individual plant was divided into four genetically identical clones which were grown in different controlled environments (high and low levels of temperature and nutrients). In total, 389 samples were measured using a high‐throughput FTIR spectrometer. The biochemical fingerprints of pollen were species and population specific, and plastic in response to different environmental conditions. The response was most pronounced for temperature, influencing the levels of proteins, lipids, and carbohydrates in pollen of all species. Furthermore, there is considerable variation in plasticity of the chemical composition of pollen among species and populations. The use of high‐throughput FTIR spectroscopy provides fast, cheap, and simple assessment of the chemical composition of pollen. In combination with controlled‐condition growth experiments and multivariate analyses, FTIR spectroscopy opens up for studies of the adaptive role of pollen that until now has been difficult with available methodology. The approach can easily be extended to other species and environmental conditions and has the potential to significantly increase our understanding of plant male function.

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Vibrational microspectroscopy enables chemical characterization of single pollen grains as well as comparative analysis of plant species based on pollen ultrastructure

Cited by 48 publications

References 42 publications

Monitoring of plant–environment interactions by high‐throughput FTIR spectroscopy of pollen

Monitoring of plant–environment interactions by high‐throughput FTIR spectroscopy of pollen

Microtiter plate cultivation of oleaginous fungi and monitoring of lipogenesis by high-throughput FTIR spectroscopy

A high‐throughput FTIR spectroscopy approach to assess adaptive variation in the chemical composition of pollen

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