Plant and Food Carotenoids, 1st ed. 2020 Methods and Protocols Methods in Molecular Biology Series, Vol. 2083

This volume provides a comprehensive compilation of techniques and protocols used in plant and food carotenoid research. Chapters guide readers through seven major areas on core enzyme activities, analysis of carotenoid profiles, new imaging tools, synthesis and degradation dynamics, biotechnology, nutrition, and health. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Part I: Introduction

1. Pathways for Carotenoid Biosynthesis, Degradation, and Storage

Tianhu Sun, Yaakov Tadmor, and Li Li

Part II: Activities

2. A Simple in vitro Assay to Measure the Activity of Geranylgeranyl Diphosphate Synthase and Other Short-chain Prenyltransferases

M. Victoria Barja and Manuel Rodriguez-Concepcion

Maurizio Camagna and Ralf Welsch

4. Improved Expression and Purification of the Carotenoid Biosynthetic Enzyme Z-ISO

Eleanore T. Wurtzel and Jesús Beltrán

5. Determination of In Vitro and In Vivo Activities of Plant Carotenoid Cleavage Oxygenases

Lourdes Gómez-Gómez, Gianfranco Diretto, Oussama Ahrazem, Salim Al-Babili

6. Expression and Characterization of Mammalian Carotenoid Cleavage Dioxygenases

Linda Thomas, Srinivasagan Ramkumar, and Johannes von Lintig

7. Transportomics for the Characterization of Plant Apocarotenoid Transmembrane Transporters

Olivia Costantina Demurtas, Rita de Brito Francisco, Enrico Martinoia, and Giovanni Giuliano

Part III: Analytics

Paula Mapelli-Brahm, Francisco J. Rodríguez-Pulido, Carla M. Stinco; Francisco J. Heredia, and Antonio J. Meléndez-Martínez

9. A Routine Method for the Extraction and HPLC-DAD Profiling of Major Plant and Food Carotenoids

Dámaso Hornero-Méndez

10. Acquisition of Mass Spectrometry Data of Carotenoids: A Focus on Big Data Management

Yagiz Alagoz, Namraj Dhami, Chris Mitchell, and Christopher I Cazzonelli

12. Determination of Plant Volatile Apocarotenoids

José L. Rambla and Antonio Granell

13. A Method for Extraction and LC-MS Based Identification of Carotenoid-Derived Dialdehydes in Plants

Jianing Mi, Kun-Peng Jia, Aparna Balakrishna, and Salim Al-Babili

14. Fractionation of Tomato Fruit Chromoplasts

Karel De Pourcq and Albert Boronat

15. Quantification of Strigolactones

Carlos Rial, Rosa M. Varela, José M. G. Molinillo, Alexandra G. Durán and Francisco A. Macías

16. Carotenoid and Apocarotenoid Analysis by SFE-SFC-QqQ/MS

Daniele Giuffrida, Mariosimone Zoccali, and Luigi Mondello

Part IV: Imaging

Maria Shumskaya, Rena F. Quinlan, and Eleanore T. Wurtzel

18. Visualization of Carotenoid-storage Structures in Fruits by Transmission Electron Microscopy

19. Light Microscopy and Raman Imaging of Carotenoids in Plant Cells in situ and in Released Carotene Crystals

Tomasz Oleszkiewicz, Marta Z. Pacia, Ewa Grzebelus, and Rafal Baranski

Part V: Dynamics

20. Carotenoid Isotopologue Profiling in ¹³C-labeled Leaf Extracts by LC-MS and LC-FTICR-MS

Björn Thiele and Shizue Matsubara

21. Quantification of Carotenoid Pathway Flux in Green and Non-green Systems

Julian Koschmieder and Ralf Welsch

22. Histochemical and Molecular Quantification of Arbuscular Mycorrhiza Symbiosis

Juan M. García, María J. Pozo, and Juan A. López-Ráez

Part VI: Biotechnology

23. Use of Potyvirus Vectors to Produce Carotenoids in Plants

José-Antonio Daròs

24. Agrobacterium tumefaciens Mediated Stable Transformation of Daucus carota

Christian Gonzalez-Calquin and Claudia Stange

25. CRISPR-Cas-Mediated Gene Knockout in Tomato

26. Modulation of Activity of a Carotenoid Pathway Through the use of the TET-on Regulatory System: Application in the Fungus Fusariumfujikuroi

Part VII: Nutrition and Health

27. Intervention Studies in Humans

Begoña Olmedilla-Alonso

28. In vitro Digestion for Control and Monitoring of Food Effects In Relation To Micellarization Index of Carotenoids

Antonio Pérez-Gálvezand Javier Fontecha

29. Evaluation of Carotenoids Protection against Oxidative Stress in the Animal Model Caenorhabditis elegans

Ramón, Lorenzo Zacarías, and María Jesús Rodrigo

Joan Ribot, Bojan Stojnic, Andreu Palou, and M. Luisa Bonet

Includes cutting- edge methods and protocols

Provides step-by-step detail essential for reproducible results

Contains key notes and implementation advice from the experts