Biochemical and Raman spectroscopic insights into plant-mold interactions

Ayse SEN; Muhammad  AHMED; Omar  DARKAZANLI; Duygu  GOKSAY-KADAIFCILER; Feyza GUZELCIMEN

doi:10.15835/nbha53114218

Authors

Ayse SEN Istanbul University, Faculty of Science, Department of Biology, 34134, Istanbul (TR)
Muhammad AHMED Istanbul University, Graduate School of Engineering and Science, 34116, Istanbul (TR)
Omar DARKAZANLI Technical University of Munich (TUM Department of Radiation Oncology and Radiotherapy), Ismaninger Str. 22, 81675 München (DE)
Duygu GOKSAY-KADAIFCILER Istanbul University, Faculty of Science, Department of Biology, Basic and Industrial Microbiology Section, 34134 Istanbul (TR)
Feyza GUZELCIMEN Istanbul University, Faculty of Science, Department of Physics, 34134, Istanbul (TR)

DOI:

https://doi.org/10.15835/nbha53114218

Keywords:

metabolome analysis, mold contaminations, oxidative stress in plants, plant tissue culture, Raman spectroscopy, wheat

Abstract

Plants continuously interact with diverse biotic and abiotic factors in their environment, and understanding these intricate ecological relationships is crucial for advancing sustainable agriculture and biodiversity conservation. This study investigates the interactions between wheat plants and mold contaminants in in vitro cultures using Raman Spectroscopy (RS) and biochemical analyses, offering a novel approach to understanding plant responses at a molecular level. Conventional culturing methods identified Aspergillus versicolor, Penicillium sp., and Nigrospora sp. as the primary mold strains present in the cultures. Biochemical assays revealed that mold contamination led to a marked decrease in chlorophyll and carotenoid contents, along with an increase in thiobarbituric acid reactive substances (TBARS), indicating heightened oxidative stress. Antioxidant enzyme activities, including superoxide dismutase (SOD), peroxidase (POX), and catalase (CAT), were significantly elevated, though proline levels remained unchanged. Raman Spectroscopy further uncovered profound metabolomic shifts, particularly in carotenoid-related peaks, signalling impaired photosynthetic function. Additionally, RS detected increased carbohydrate-associated bands, suggesting that carbohydrate-derived osmolytes may play a more pivotal role than proline in maintaining cellular integrity under mold stress. To validate these findings, advanced multivariate techniques such as Principal Component Analysis (PCA) and Linear Discriminant Analysis (LDA) were employed, achieving high accuracy in distinguishing mold-infected samples from controls. These results highlight the potential of RS as a rapid, non-invasive diagnostic tool for plant health monitoring, offering significant advantages over traditional methods. This approach not only advances our understanding of plant-microbe interactions but also offers practical applications for enhancing agricultural productivity and sustainability in the face of global food security challenges.

References

Aebi H (1984). Catalase in vitro. In: Methods in Enzymology. Academic press, pp 121-126. https://doi.org/10.1016/S0076-6879(84)05016-3

Altangerel N, Ariunbold GO, Gorman C, Alkahtani MH, Borrego EJ, Bohlmeyer D, ... Scully MO (2017). In vivo diagnostics of early abiotic plant stress response via Raman spectroscopy. Proceedings of the National Academy of Sciences 114(13):3393-3396. https://doi.org/10.1073/pnas.1701328114

Barnett HL (1955). Illustrated genera of imperfect fungi. https://www.cabidigitallibrary.org/doi/full/10.5555/19561100378

Bates LS, Waldren RPA, Teare ID (1973). Rapid determination of free proline for water-stress studies. Plant and Soil 39:205-207. https://doi.org/10.1007/BF00018060

Beauchamp C, Fridovich I (1971). Superoxide dismutase: improved assays and an assay applicable to acrylamide gels. Analytical Biochemistry 44(1):276-287. https://doi.org/10.1016/0003-2697(71)90370-8

Beck JJ, Alborn HT, Block AK, Christensen SA, Hunter CT, Rering CC, ... Tumlinson JH (2018). Interactions among plants, insects, and microbes: elucidation of inter-organismal chemical communications in agricultural ecology. Journal of Agricultural and Food Chemistry 66(26):6663-6674. https://doi.org/10.1021/acs.jafc.8b01763

Belkacem-Hanfi N, Semmar N, Perraud-Gaime I, Guesmi A, Cherni M, Cherif I, ... Roussos S (2013). Spatio-temporal analysis of post-harvest moulds genera distribution on stored durum wheat cultivated in Tunisia. Journal of Stored Products Research 55:116-123. https://doi.org/10.1016/j.jspr.2013.08.008

Bever JD, Westover KM, Antonovics J (1997). Incorporating the soil community into plant population dynamics: the utility of the feedback approach. Journal of Ecology 561-573. https://doi.org/10.2307/2960528

Bhojwani SS, Razdan MK (1996). Plant tissue culture: theory and practice. Elsevier.eBook ISBN: 9780080539096.

Bradford MM (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72(1-2):248-254. https://doi.org/10.1016/0003-2697(76)90527-3

Cassells AC (2012). Pathogen and biological contamination management in plant tissue culture: phytopathogens, vitro pathogens, and vitro pests. Plant Cell Culture Protocols 57-80. https://doi.org/10.1007/978-1-61779-818-4_6

Czolpinska M, Rurek M (2018). Plant glycine-rich proteins in stress response: an emerging, still prospective story. Frontiers in Plant Science 9:302. https://doi.org/10.3389/fpls.2018.00302

Egging V, Nguyen J, Kurouski D (2018). Detection and identification of fungal infections in intact wheat and sorghum grain using a hand-held Raman spectrometer. Analytical Chemistry 90(14):8616-8621. https://doi.org/10.1021/acs.analchem.8b01863

Ellis MB (1971). Dematiaceous hyphomycetes. Commonwealth Mycological Institute, pp 608.

Espina A, Cañamares MV, Jurasekova Z, Sanchez-Cortes S (2022). Analysis of iron complexes of tannic acid and other related polyphenols as revealed by spectroscopic techniques: Implications in the identification and characterization of iron gall inks in historical manuscripts. ACS Omega 7(32):27937-27949. https://doi.org/10.1021/acsomega.2c01679

Gierlinger N, Schwanninger M (2007). The potential of Raman microscopy and Raman imaging in plant research. Spectroscopy 21(2):69-89. https://content.iospress.com/articles/spectroscopy/spe302

Heath RL, Packer L (1968). Photoperoxidation in isolated chloroplasts: I. Kinetics and stoichiometry of fatty acid peroxidation. Archives of Biochemistry and Biophysics 125(1):189-198. https://doi.org/10.1016/0003-9861(68)90654-1

Hoffmann, H. (2015). violin. m-Simple violin plot using matlab default kernel density estimation. INRES (University of Bonn), Katzenburgweg 5:53115.

“HORIBA Scientific.” Accessed: Sep. 22, 2024. https://www.horiba.com/int/scientific/?utm_source=uhw&utm_medium=301&utm_campaign=uhw-redirect

Horner WE (2003). Assessment of the indoor environment: evaluation of mold growth indoors. Immunology and Allergy Clinics 23(3):519-531. https://www.immunology.theclinics.com/article/S0889-8561(03)00063-8/abstract#page-body-id

Hussain A, Qarshi IA, Nazir H, Ullah I (2012). Plant tissue culture: current status and opportunities. Recent Advances in Plant In Vitro Culture 6(10):1-28.

Jehlička J, Edwards HG, Osterrothov K, Novotn J, Nedbalov L, Kopecký J, … Oren A (2014a). Potential and limits of Raman spectroscopy for carotenoid detection in microorganisms: implications for astrobiology. Philosophical Transactions of the Royal Society A: Mathematical, Physical and Engineering Sciences 372(2030):20140199. https://doi.org/10.1098/rsta.2014.0199

Jehlička J, Edwards HG, Oren A (2014b). Raman spectroscopy of microbial pigments. Applied and Environmental Microbiology 80(11):3286-3295. https://doi.org/10.1128/AEM.00699-14

Kadaifciler DG, Demirel R (2018). Fungal contaminants in man-made water systems connected to municipal water. Journal of Water and Health 16(2):244-252. https://doi.org/10.2166/wh.2018.272

Kadaifçiler DG (2017). Airborne fungi in the atmosphere in Beyazıt Square, Istanbul, Turkey. Celal Bayar University Journal of Science 13(2):343-351. https://dergipark.org.tr/en/pub/cbayarfbe/issue/29779/319820

Kecoglu I, Sirkeci M, Unlu MB, Sen A, Parlatan U, Guzelcimen F (2022). Quantification of salt stress in wheat leaves by Raman spectroscopy and machine learning. Scientific Reports 12(1):7197. https://www.nature.com/articles/s41598-022-10767-y

Khan IA, Alam SS, Jabbar A (2004). Purification of phytotoxin from culture filtrates of Fusarium oxysporum f. sp. ciceris and its biological effects on chickpea. Pakistan Journal of Botany 36(4):871-880. https://www.pakbs.org/pjbot/PDFs/36(4)/PJB36(4)871.pdf

Klich MA (2002). Identification of common Aspergillus species. Centraalbureau voor Schimmelcultures, Utrecht, The Netherlands.

Kuhar N, Sil S, Umapathy S (2021). Potential of Raman spectroscopic techniques to study proteins. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 258:119712. https://doi.org/10.1016/j.saa.2021.119712

Kuska MT, Heim RH, Geedicke I, Gold KM, Brugger A, Paulus S (2022). Digital plant pathology: A foundation and guide to modern agriculture. Journal of Plant Diseases and Protection 129(3):457-468. https://doi.org/10.1007/s41348-022-00600-z

Leifert C, Woodward S (1998). Laboratory contamination management: the requirement for microbiological quality assurance. Plant Cell, Tissue and Organ Culture 52:83-88. https://link.springer.com/chapter/10.1007/978-94-015-8951-2_30

Leifert C, Cassells AC (2001). Microbial hazards in plant tissue and cell cultures. In Vitro Cellular & Developmental Biology-Plant 37:133-138. https://link.springer.com/article/10.1007/s11627-001-0025-y

Lichtenthaler HK, Wellburn AR (1983). Determinations of total carotenoids and chlorophylls a and b of leaf extracts in different solvents. Biochemical Society Transactions 11(5):591-592. https://doi.org/10.1042/bst0110591

Machida M, Gomi K (2010). Aspergillus: molecular biology and genomics. Caister Academic Press. https://doi.org/10.21775/9781910190159

Morey R, Farber C, McCutchen B, Burow MD, Simpson C, Kurouski D, Cason J (2021). Raman spectroscopy‐based diagnostics of water deficit and salinity stresses in two accessions of peanut. Plant Direct 5(8):e342. https://doi.org/10.1002/pld3.342

Msogoya TJ, Kanyagha H, Mutigitu J, Kulebelwa M, Mamiro D (2012). Identification and management of microbial contaminants of banana in vitro cultures. Journal of Applied Biosciences 55:3987-3994 https://m.elewa.org/JABS/2012/55/5.pdf

Murashige T, Skoog F (1962). A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiologia Plantarum 15(3). https://doi.org/10.1111/j.1399-3054.1962

Najafi M, Esfahani MN, Vatandoost J, Hassanzadeh-Khankahdani H, Moeini MJ (2024). Antioxidant enzymes activity associated with resistance to Phytophthora melonis-pumpkin blight. Physiological and Molecular Plant Pathology 129:102192. https://doi.org/10.1016/j.pmpp.2023.102192

Nikbakht AM, Tavakkoli HT, Malekfar R, Gobadian B (2011). Nondestructive determination of tomato fruit quality parameters using Raman spectroscopy. Journal of Agricultural Science and Technology 13:517-526.

Payne WZ, Kurouski D (2021). Raman spectroscopy enables phenotyping and assessment of nutrition values of plants: a review. Plant Methods 17(1):78. https://link.springer.com/article/10.1186/s13007-021-00781-y

Piot O, Autran JC, Manfait M (2002). Assessment of cereal quality by micro-Raman analysis of the grain molecular composition. Applied Spectroscopy 56(9):1132-1138. https://opg.optica.org/as/abstract.cfm?uri=as-56-9-1132

Pitt JI (2000). A laboratory guide to common Penicillium species, 3th ed., Food Science Australia, North Ryde NSW, Australia.

Saletnik A, Saletnik B, Puchalski C (2022). Raman method in identification of species and varieties, assessment of plant maturity and crop quality—A review. Molecules 27(14):4454. https://doi.org/10.3390/molecules27144454

Samson RA, Houbraken J, Thrane U, Frisvad JC, Andersen B (2010). Food and indoor fungi: CBS-KNAW fungal biodiversity centre. CBS-KNAW Fungal Biodiversity Centre, Utrecht, the Netherlands. https://doi.org/10.3920/9789086867226_006

Schenk PM, Carvalhais LC, Kazan K (2012). Unraveling plant–microbe interactions: can multi-species transcriptomics help?. Trends in Biotechnology 30(3):177-184.

Schulz H, Baranska M (2007). Identification and quantification of valuable plant substances by IR and Raman spectroscopy. Vibrational Spectroscopy 43(1):13-25. https://doi.org/10.1016/j.vibspec.2006.06.001

Sen A, Kecoglu I, Ahmed M, Parlatan U, Unlu MB (2023). Differentiation of advanced generation mutant wheat lines: Conventional techniques versus Raman spectroscopy. Frontiers in Plant Science 14:1116876. https://doi.org/10.3389/fpls.2023.1116876

Singla P, Bhardwaj RD, Kaur S, Kaur J, Grewal SK (2020a). Metabolic adjustments during compatible interaction between barley genotypes and stripe rust pathogen. Plant Physiology and Biochemistry 147:295-302. https://doi.org/10.1016/j.plaphy.2019.12.030

Singla P, Bhardwaj RD, Kaur S, Kaur J (2020b). Stripe rust induced defence mechanisms in the leaves of contrasting barley genotypes (Hordeum vulgare L.) at the seedling stage. Protoplasma 257:169-181. https://link.springer.com/article/10.1007/s00709-019-01428-5

Singla P, Bhardwaj RD, Sunidhi SS (2024). Plant–fungus interaction: a stimulus–response theory. Journal of Plant Growth Regulation 43(2):369-381. https://link.springer.com/article/10.1007/s00344-023-11100-1

Skoczowski A, Troć M (2013). Isothermal calorimetry and Raman spectroscopy to study response of plants to abiotic and biotic stresses. Molecular Stress Physiology of Plants 263-288. https://doi.org/10.1007/978-81-322-0807-5_11

Smith SE, Read DJ (2010). Mycorrhizal symbiosis. Academic press.

Synytsya A, Čopı́ková J, Matějka P, Machovič VJCP (2003). Fourier transform Raman and infrared spectroscopy of pectins. Carbohydrate Polymers 54(1):97-106. https://doi.org/10.1016/S0144-8617(03)00158-9

Tatagiba SD, DaMatta FM, Rodrigues FÁ (2015). Leaf gas exchange and chlorophyll a fluorescence imaging of rice leaves infected with Monographella albescens. Phytopathology 105(2):180-188. https://doi.org/10.1094/PHYTO-04-14-0097-R

Wang M, Liu F, Crous PW, Cai L (2017). Phylogenetic reassessment of Nigrospora: ubiquitous endophytes, plant and human pathogens. Persoonia-Molecular Phylogeny and Evolution of Fungi 39(1):118-142. https://doi.org/10.3767/persoonia.2017.39.06

Whitham TG, Gehring CA, Evans LM, LeRoy CJ, Bangert RK, Schweitzer JA, ... Bailey JK (2010). A community and ecosystem genetics approach to conservation biology and management. Molecular approaches in natural resource conservation and management. Cambridge Univ. Press, Cambridge, UK, pp 50-73.

Zar JH (1984). Biostatistical analysis. Prentice-Hall Inc, Englewood Cliffs, Jersey.

Zhang H, Zhao Y, Zhu JK (2020). Thriving under stress: how plants balance growth and the stress response. Developmental Cell 55(5):529-543. https://doi.org/10.1016/j.devcel.2020.10.012

Zhu G, Zhu X, Fan Q, Wan X (2011). Raman spectra of amino acids and their aqueous solutions. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 78(3):1187-1195. https://doi.org/10.1016/j.saa.2010.12.079