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Determination of Reduced and Total Glutathione Content in Extremophilic Microalga Galdieria phlegrea

Authors: Giovanna Salbitani
Giovanna SalbitaniAffiliation: Dipartimento di Biologia, Università di Napoli Federico II, Via Foria 223, I-80139 Napoli, Italy
Bio-protocol author page: a4677
, Claudia Bottone
Claudia BottoneAffiliation: Dipartimento di Biologia, Università di Napoli Federico II, Via Foria 223, I-80139 Napoli, Italy
Bio-protocol author page: a4765
 and Simona Carfagna
Simona CarfagnaAffiliation: Dipartimento di Biologia, Università di Napoli Federico II, Via Foria 223, I-80139 Napoli, Italy
For correspondence: simcarfa@unina.it
Bio-protocol author page: a4678
,  date: 7/5/2017, 96 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2372.

[Abstract] Glutathione is an important molecule involved in the primary and secondary metabolism of all organisms. The Glutathione redox status is an indicator of the cellular redox state. Therefore, it is important to have precise methods on hand to determine the glutathione redox status in the cell. In this protocol, we describe an improved spectrophotometric ...

Acetyl Bromide Soluble Lignin (ABSL) Assay for Total Lignin Quantification from Plant Biomass

Authors: William J. Barnes
William J. BarnesAffiliation 1: Department of Biology, The Pennsylvania State University, University Park, PA, USA
Affiliation 2: Center for Lignocellulose Structure and Formation, The Pennsylvania State University, University Park, PA, USA
Bio-protocol author page: a4185
 and Charles T. Anderson
Charles T. AndersonAffiliation 1: Department of Biology, The Pennsylvania State University, University Park, PA, USA
Affiliation 2: Center for Lignocellulose Structure and Formation, The Pennsylvania State University, University Park, PA, USA
For correspondence: cta3@psu.edu
Bio-protocol author page: a4143
,  date: 3/5/2017, 867 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2149.

[Abstract] Lignin is the second most abundant biopolymer on Earth, providing plants with mechanical support in secondary cell walls and defense against abiotic and biotic stresses. However, lignin also acts as a barrier to biomass saccharification for biofuel generation (Carroll and Somerville, 2009; Zhao and Dixon, 2011; Wang et al., 2013). For these reasons, ...

Rice Root Organic Acid Efflux Measurement by Using Ion Chromatography

Authors: Chun-quan Zhu
Chun-quan ZhuAffiliation: State Key Laboratory of Soil and Sustainable Agriculture, Institute of Soil Science, Chinese Academy of Sciences, Nanjing, China
Bio-protocol author page: a4131
, Xiao-fang Zhu
Xiao-fang Zhu Affiliation: State Key Laboratory of Soil and Sustainable Agriculture, Institute of Soil Science, Chinese Academy of Sciences, Nanjing, China
Bio-protocol author page: a4132
 and Ren-fang Shen
Ren-fang ShenAffiliation: State Key Laboratory of Soil and Sustainable Agriculture, Institute of Soil Science, Chinese Academy of Sciences, Nanjing, China
For correspondence: rfshen@issas.ac.cn
Bio-protocol author page: a4133
,  date: 2/20/2017, 853 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2141.

[Abstract] Organic acids secreted from plant roots play important roles in various biological processes including nutrient acquisition, metal detoxification, and pathogen attraction. The secretion of organic acids may be affected by various conditions such as plant growth stage, nutrient deficiency, and abiotic stress. For example, when white lupin (Lupinus albus ...

Chromatographic Separation of the Codonocarpine Type Alkaloids from the Root Bark of Capparis decidua

Authors: Yvonne Forster
Yvonne ForsterAffiliation: Department of Chemistry, University of Zurich, Zurich, Switzerland
Bio-protocol author page: a4136
, Abdul Ghaffar
Abdul GhaffarAffiliation: Department of Chemistry, Baghdad-ul-Jadeed Campus, The Islamia University of Bahawalpur, Bahawalpur, Pakistan
Bio-protocol author page: a4135
 and Stefan Bienz
Stefan BienzAffiliation: Department of Chemistry, University of Zurich, Zurich, Switzerland
For correspondence: stefan.bienz@chem.uzh.ch
Bio-protocol author page: a4137
,  date: 2/20/2017, 711 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2144.

[Abstract] Various parts of the caper tree Capparis decidua have found application in traditional medicine. The isolation and structural elucidation of the codonocarpine type alkaloids contained in the root bark, however, is not trivial and has probably led to misinterpretation in the past. This protocol describes the extraction and chromatographic separation ...

Nitrate Assay for Plant Tissues

Authors: Lufei Zhao
Lufei ZhaoAffiliation: State Key Laboratory of Crop Biology, College of Life Sciences, Shandong Agricultural University, Tai’an, China
For correspondence: lufeizh@163.com
Bio-protocol author page: a3770
 and Yong Wang
Yong WangAffiliation: State Key Laboratory of Crop Biology, College of Life Sciences, Shandong Agricultural University, Tai’an, China
Bio-protocol author page: a3771
,  date: 1/20/2017, 1482 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2029.

[Abstract] Nitrogen is an essential macronutrient for plant growth and nitrate content in plants can reflect the nitrogen supply of soil. Here, we provide the salicylic acid method to evaluate the nitrate content in plant tissues. The method is reliable and stable, thus it can be a good choice for measurement of nitrate in plant tissues....

Cation (Ca2+ and Mn2+) Partitioning Assays with Intact Arabidopsis Chloroplasts

Authors: Anna Harms
Anna HarmsAffiliation: Biozentrum der LMU München, Department Biologie I, Munich, Germany
Bio-protocol author page: a3963
, Iris Steinberger
Iris SteinbergerAffiliation: Biozentrum der LMU München, Department Biologie I, Munich, Germany
Bio-protocol author page: a2369
 and Anja Schneider
Anja SchneiderAffiliation: Biozentrum der LMU München, Department Biologie I, Munich, Germany
For correspondence: anja.schneider@lrz.uni-muenchen.de
Bio-protocol author page: a2371
,  date: 1/5/2017, 966 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2094.

[Abstract] Determination of the relative distribution of Ca2+ and Mn2+ is an important tool for analyzing mutants showing altered levels of calcium and/or manganese transporters in the chloroplast envelope or thylakoid membrane. The method described in this protocol allows quantitative analyses of the relative distribution of calcium and manganese ions between ...

Quantitative Determination of Ascorbate from the Green Alga Chlamydomonas reinhardtii by HPLC

Authors: László Kovács
László KovácsAffiliation: Institute of Plant Biology, Biological Research Centre of the Hungarian Academy of Sciences, Szeged, Hungary
Bio-protocol author page: a3895
, André Vidal-Meireles
André Vidal-MeirelesAffiliation: Institute of Plant Biology, Biological Research Centre of the Hungarian Academy of Sciences, Szeged, Hungary
Bio-protocol author page: a3896
, Valéria Nagy
Valéria NagyAffiliation: Institute of Plant Biology, Biological Research Centre of the Hungarian Academy of Sciences, Szeged, Hungary
Bio-protocol author page: a3897
 and Szilvia Z. Tóth
Szilvia Z. TóthAffiliation: Institute of Plant Biology, Biological Research Centre of the Hungarian Academy of Sciences, Szeged, Hungary
For correspondence: toth.szilviazita@brc.mta.hu
Bio-protocol author page: a3898
,  date: 12/20/2016, 919 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2067.

[Abstract] Ascorbate (Asc, also called vitamin C) is of vital importance to the cellular functions of both animals and plants. During evolution, Asc has become one of the most abundant metabolites in seed plants; however, Asc contents in cyanobacteria, green algae and bryophytes are very low. Here we describe a sensitive and reliable HPLC method for the quantitative ...

Detection of Reactive Oxygen Species in Oryza sativa L. (Rice)

Authors: Navdeep Kaur
Navdeep KaurAffiliation: Department of Biotechnology, Guru Nanak Dev University, Amritsar, Punjab, India
Bio-protocol author page: a3877
, Isha Sharma
Isha SharmaAffiliation: Department of Biotechnology, Guru Nanak Dev University, Amritsar, Punjab, India
Bio-protocol author page: a3878
, Kamal Kirat
Kamal KiratAffiliation: Department of Biotechnology, Guru Nanak Dev University, Amritsar, Punjab, India
Bio-protocol author page: a3879
 and Pratap Kumar Pati
Pratap Kumar PatiAffiliation: Department of Biotechnology, Guru Nanak Dev University, Amritsar, Punjab, India
For correspondence: pkpati@yahoo.com
Bio-protocol author page: a3876
,  date: 12/20/2016, 1500 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2061.

[Abstract] Superoxide ions (O2-) and hydrogen peroxide (H2O2) are the reactive oxygen species (ROS) that play a significant role in regulation of many plant processes. The level of O2- ions is determined qualitatively using nitrobluetetrazolium (NBT) assay while the H2O2 is qualitatively estimated using 3,3-diaminobenzidine (DAB) and 2’,7’-dichlorodihydrofluorescein ...

Metabolite Profiling of Mature Arabidopsis thaliana Seeds Using Gas Chromatography-Mass Spectrometry (GC-MS)

Authors: Hagai Cohen
Hagai CohenAffiliation 1: Laboratory of Plant Science, Migal Galilee Technology Center, Kiryat Shmona, Israel
Affiliation 2: Faculty of Biology, Technion - Israel Institute of Technology, Haifa, Israel
For correspondence: Hagaic@Weizmann.ac.il
Bio-protocol author page: a3644
, Ifat Matityahu
Ifat MatityahuAffiliation: Laboratory of Plant Science, Migal Galilee Technology Center, Kiryat Shmona, Israel
Bio-protocol author page: a3645
 and Rachel Amir
Rachel AmirAffiliation 1: Laboratory of Plant Science, Migal Galilee Technology Center, Kiryat Shmona, Israel
Affiliation 2: Tel-Hai College, Upper Galilee, Israel
For correspondence: Rachel@migal.org.il
Bio-protocol author page: a3646
,  date: 11/5/2016, 1259 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1981.

[Abstract] Metabolite profiling using gas chromatography-mass spectrometry (GC-MS) permits the annotation and quantification of a relatively wide variety of metabolites, covering a wide range of biochemical groups of metabolites. Lisec et al. (2006) established a method for GC-MS profiling in plants. Based on this protocol, we provide here a detailed GC-MS-based ...

Hydrogen Peroxide Measurement in Arabidopsis Root Tissue Using Amplex Red

Authors: Tzvetina Brumbarova
Tzvetina BrumbarovaAffiliation: Institute of Botany, Heinrich-Heine University, Düsseldorf, Germany
Bio-protocol author page: a1670
, Cham Thi Tuyet Le
Cham Thi Tuyet LeAffiliation: Department of Plant Genetics and Breeding, Faculty of Agronomy, Vietnam National University of Agriculture, Trauquy, Gialam, Hanoi, Vietnam
Bio-protocol author page: a3686
 and Petra Bauer
Petra BauerAffiliation: Institute of Botany, Heinrich-Heine University, Düsseldorf, Germany
For correspondence: petra.bauer@uni-duesseldorf.de
Bio-protocol author page: a3687
,  date: 11/5/2016, 1614 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1999.

[Abstract] This protocol describes the measurement of hydrogen peroxide (H2O2) content in Arabidopsis root tissue by using the Amplex® Red Hydrogen Peroxide/Peroxidase Assay Kit. When root tissue is disrupted and resuspended in phosphate buffer, H2O2 is released from the cells. The obtained root extracts containing H2O2 can be mixed with a solution containing ...
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Analysis of Malondialdehyde, Chlorophyll Proline, Soluble Sugar, and Glutathione Content in Arabidopsis seedling

Authors: Zhijin Zhang
Zhijin ZhangAffiliation: Cold and Arid Regions Environmental and Engineering Research Institute, Chinese Academy of Sciences, Lanzhou, China
Bio-protocol author page: a684
 and Rongfeng Huang
Rongfeng HuangAffiliation: Cold and Arid Regions Environmental and Engineering Research Institute, Chinese Academy of Sciences, Lanzhou, China
For correspondence: rfhuang@caas.cn
Bio-protocol author page: a362
,  date: 7/20/2013, 11118 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.817.

[Abstract] The protocol has four sub-protocols, which are about the measurement of malondialdehyde, chlorophyll proline, soluble sugar, and glutathione content, respectively, in Arabidopsis seedling by using spectrophotometer. These methods are simple, effective and reproducible, which will help the researchers ...

Histochemical Detection of Superoxide and H2O2 Accumulation in Brassica juncea Seedlings

Authors: Deepak Kumar
Deepak KumarAffiliation: School of Life Sciences, Jawaharlal Nehru University, New Delhi, India
Bio-protocol author page: a1306
, Mohd Aslam Yusuf
Mohd Aslam YusufAffiliation: School of Life Sciences, Jawaharlal Nehru University, New Delhi, India
Bio-protocol author page: a1307
, Preeti Singh
Preeti SinghAffiliation: Department of Biosciences, Jawaharlal Nehru University, New Delhi, India
Bio-protocol author page: a1308
, Meryam Sardar
Meryam SardarAffiliation: Department of Biosciences, Jamia Millia Islamia, New Delhi, India
Bio-protocol author page: a1309
 and Neera Bhalla Sarin
Neera Bhalla SarinAffiliation: School of Life Sciences, Jawaharlal Nehru University, New Delhi, India
For correspondence: neerasarin@rediffmail.com
Bio-protocol author page: a475
,  date: 4/20/2014, 7763 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1108.

[Abstract] Plant cells continually produce reactive oxygen species (ROS) as a by-product of aerobic metabolism. Increased production of ROS occurs under unfavorable conditions imposed by various abiotic and biotic factors. Accumulation of ROS is damaging to various cellular components and macromolecules including ...

Quantification of Anthocyanin Content

Authors: Masaru Nakata
Masaru NakataAffiliation: Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology, Tsukuba, Ibaraki, Japan
Bio-protocol author page: a1283
 and Masaru Ohme-Takagi
Masaru Ohme-TakagiAffiliation 1: Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology, Tsukuba, Ibaraki, Japan
Affiliation 2: Institute for Environmental Science and Technology, Saitama University, Saitama, Japan
For correspondence: m-takagi@aist.go.jp
Bio-protocol author page: a1284
,  date: 4/5/2014, 7635 views, 1 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1098.

[Abstract] Anthocyanins are a class of flavonoids and important plant pigments. They attract insects to pollinate flowers, protect plants from UV irradiation, and act as antimicrobial agents against herbivores and pathogens. Biosynthesis of anthocyanin is stimulated by diverse developmental signals and environmental ...

Measurement of Endogenous H2O2 and NO and Cell Viability by Confocal Laser Scanning Microscopy

Authors: Mi-Mi Wu
Mi-Mi WuAffiliation: School of Life Sciences, Shaanxi Normal University, Xi’an, China
Bio-protocol author page: a869
, Xian-Ge Ma
Xian-Ge MaAffiliation: School of Life Sciences, Shaanxi Normal University, Xi’an, China
Bio-protocol author page: a870
 and Jun-Min He
Jun-Min HeAffiliation: School of Life Sciences, Shaanxi Normal University, Xi’an, China
For correspondence: hejm@snnu.edu.cn
Bio-protocol author page: a871
,  date: 10/5/2013, 5054 views, 1 Q&A
DOI: https://doi.org/10.21769/BioProtoc.920.

[Abstract] Recently, there is compelling evidence that hydrogen peroxide (H2O2) and nitric oxide (NO) function as signaling molecules in plants, mediating a range of responses including stomatal movement. Thus, the choice of sensitive methods for detection of endogenous H2O2 and NO in guard cells are very important ...

Chlorophyll Fluorescence Measurements in Arabidopsis Plants Using a Pulse-amplitude-modulated (PAM) Fluorometer

Authors: Reiko Motohashi
Reiko MotohashiAffiliation: Agriculture Department, Shizuoka University, Shizuoka, Japan
For correspondence: armotoh@ipc.shizuoka.ac.jp
Bio-protocol author page: a2179
 and Fumiyoshi Myouga
Fumiyoshi MyougaAffiliation: RIKEN Center for Sustainable Resource Science, Yokohama, Japan
Bio-protocol author page: a2181
,  date: 5/5/2015, 4627 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1464.

[Abstract] In this protocol, to analyze PSII activity in photosynthesis, we measure the Fv/Fm (Fv=Fm ± Fo) value (Fo and Fm are the minimum and maximum values of chlorophyll fluorescence of dark-adapted leaves, respectively). Fv/Fm is a reliable marker of photo- inhibition (Krause et al., 1988). Chlorophyll fluorescence ...

Detection and Measurement of ROS in Tobacco Leaves

Authors: Shunhua Ding
Shunhua DingAffiliation: The Key Lab of Photobiology, Institute of Botany, Chinese Academy of Sciences, Beijing, China
Bio-protocol author page: a433
 and Congming Lu
Congming LuAffiliation: The Key Lab of Photobiology, Institute of Botany, Chinese Academy of Sciences, Beijing, China
For correspondence: lucm@ibcas.ac.cn
Bio-protocol author page: a304
,  date: 3/5/2013, 4594 views, 1 Q&A
DOI: https://doi.org/10.21769/BioProtoc.407.

[Abstract] Leaf metabolism produces hydrogen peroxide (H2O2) at high rates, high level H2O2 accumulation can cause oxidative stress. This protocol describes a method for determining H2O2 concentration in tobacco leaves. In this method all extractions were performed with HClO4, neutralized, and pretreated with ...

Quantification of Total and Soluble Inorganic Phosphate

Authors: Vicki Knowles
Vicki KnowlesAffiliation: Department of Biology, Queen's University, Kingston, Canada
Bio-protocol author page: a811
 and William Plaxton
William PlaxtonAffiliation: Department of Biology, Queen's University, Kingston, Canada
For correspondence: plaxton@queensu.ca
Bio-protocol author page: a812
,  date: 9/5/2013, 4572 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.890.

[Abstract] A simple, rapid, and sensitive colorimetric microassay for inorganic phosphate (Pi) relies upon the absorption at 660 nm of a molybdenum blue complex that forms upon reduction of an ammonium molybdate-Pi complex in acid. The method for determination of total Pi uses plant tissues that have been ashed ...

Extraction of Coumarins from Leaves, Petioles, Stems and Roots of Ruta graveolens and Nicotiana benthamiana

Authors: Alain Hehn
Alain HehnAffiliation: UMR1121 Laboratoire Agronomie et Environnement, Université de Lorraine-INRA, Vandoeuvre les Nancy, France
For correspondence: alain.hehn@univ-lorraine.fr
Bio-protocol author page: a153
, Guilhem Vialart
Guilhem VialartAffiliation: UMR1121 Laboratoire Agronomie et Environnement, Université de Lorraine-INRA, Vandoeuvre les Nancy, France
Bio-protocol author page: a154
, Alexandre Olry
Alexandre OlryAffiliation: UMR1121 Laboratoire Agronomie et Environnement, Université de Lorraine-INRA, Vandoeuvre les Nancy, France
Bio-protocol author page: a155
 and Frederic Bourgaud
Frederic BourgaudAffiliation: UMR1121 Laboratoire Agronomie et Environnement, Université de Lorraine-INRA, Vandoeuvre les Nancy, France
Bio-protocol author page: a156
,  date: 11/20/2012, 4545 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.288.

[Abstract] This method describes the extraction of coumarins and furanocoumarins from leaves of Ruta graveolens (a natural furanocoumarin) producer, and Nicotiana benthamiana....

Citrus Fruit Ascorbic Acid Extraction and Quantification by HPLC

Authors: Enriqueta Alós
Enriqueta AlósAffiliation: Postharvest Laboratory, Food Science Department, Instituto de Agroquímica y Tecnología de Alimentos (IATA), Consejo Superior de Investigaciones Científicas (CSIC), Valencia, Spain
Bio-protocol author page: a2042
, Joanna Lado
Joanna LadoAffiliation: Postharvest Laboratory, Food Science Department, Instituto de Agroquímica y Tecnología de Alimentos (IATA), Consejo Superior de Investigaciones Científicas (CSIC), Valencia, Spain
Bio-protocol author page: a2043
, María Jesús Rodrigo
María Jesús RodrigoAffiliation: Postharvest Laboratory, Food Science Department, Instituto de Agroquímica y Tecnología de Alimentos (IATA), Consejo Superior de Investigaciones Científicas (CSIC), Valencia, Spain
Bio-protocol author page: a2044
 and Lorenzo Zacarías
Lorenzo ZacaríasAffiliation: Postharvest Laboratory, Food Science Department, Instituto de Agroquímica y Tecnología de Alimentos (IATA), Consejo Superior de Investigaciones Científicas (CSIC), Valencia, Spain
For correspondence: lzacarias@iata.csic.es
Bio-protocol author page: a2045
,  date: 3/5/2015, 4024 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1416.

[Abstract] Citrus are among the most relevant sources of vitamin C (ascorbic acid + dehydroascorbic acid). Recent studies have revealed that it increases in the peel as fruit ripens and remains constant or even decreases in the pulp tissue. Moreover, important differences on ascorbic acid content exist among citrus ...

Determination of Nectar Nicotine Concentration in N. attenuata

Authors: Eva Rothe
Eva RotheAffiliation: Department of Molecular Ecology, Max Planck Institute for Chemical Ecology, Jena, Germany
Bio-protocol author page: a578
, Matthias Schöttner
Matthias SchöttnerAffiliation: Department of Molecular Ecology, Max Planck Institute for Chemical Ecology, Jena, Germany
Bio-protocol author page: a579
, Danny Kessler
Danny KesslerAffiliation: Department of Molecular Ecology, Max Planck Institute for Chemical Ecology, Jena, Germany
Bio-protocol author page: a580
 and Ian T. Baldwin
Ian T. BaldwinAffiliation: Department of Molecular Ecology, Max Planck Institute for Chemical Ecology, Jena, Germany
For correspondence: baldwin@ice.mpg.de
Bio-protocol author page: a581
,  date: 4/20/2013, 3992 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.451.

[Abstract] In this protocol, the determination of the nicotine concentration in nectar of Nicotiana attenuata is described. This method is applicable for the investigation of small amounts of nectar (above 1 μl). It is a high-throughput protocol optimized and streamlined for one skilled person to process approximately ...
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