Phloem

Methods and Protocols
1. Auflage 2019
ISBN: 978-1-4939-9564-6
Verlag: Springer

Buch, Englisch, 473 Seiten, Format (B × H): 178 mm x 254 mm, Gewicht: 909 g

Reihe: Methods in Molecular Biology

This detailed book presents methods that enable the determination of parameters relevant to phloem research in the most efficient ways, putting plant scientists with access to adequate instrumentation in the position to answer any phloem-related question. The collection explores techniques that have been used for decades, such as tracing phloem transport with carbon isotopes, as well as recent developments, such as esculin-based assays of phloem transport and super-resolution microscopy of phloem proteins. As such, the book presents the state-of-the-art in phloem research and, at the same time, a starting point for the development of new methods that will fill the remaining gaps in the phloem researcher’s toolbox in the future. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and tips on troubleshooting and avoiding known pitfalls.

 Authoritative and comprehensive, Phloem: Methods and Protocols is an ideal guide for all researchers exploring this challenging area of plant biology.

Zielgruppe

Professional/practitioner

Autoren/Hrsg.

Liesche, Johannes

Fachgebiete

• Naturwissenschaften Biowissenschaften Botanik

Inhaltsverzeichnis

Investigating phloem structure

1.       Three-dimensional visualization of phloem cells by whole-mount staining

2.       How to minimize artifacts in transmission electron microscopy of the phloem

3.       Scanning electron microscopy of sieve elements

4.       Non-invasive investigation of phloem structure by 3D synchrotron X-ray microtomography

5.       Optimal preparation of phloem tissue sections for light-microscopic analysis

6.       Quantification of foliar phloem infrastructure with light and electron microscopy

Identification, localization and characterization of phloem proteins

7.       Super-resolution microscopy of phloem proteins

8.       Live-cell imaging of fluorescently-tagged phloem proteins with confocal microscopy

9.       Proteomic analysis of proteins in the phloem sap

10.   Proteomic analysis of membrane bound and -associated proteins in the phloem

11.   Electrophysiological characterization of sugar transporters in Xenopus oocytes and membrane vesicles

12.   Investigating the regulation of sucrose transporters by heterologous expression in yeast

13.   Resolving the transport cycle of sugar transporters by voltage clamp fluorometry

14.   Correlative imaging of sieve elements and proteins localized within

15.   Immunohistochemical localization of proteins in the phloem

Sampling of phloem sap

16.   A critical view on studying phloem loading by the EDTA-facilitated exudation technique

17.   Sampling phloem sap with aphids

18.   Phloem sap metabolomics

19.   Quantification of non-structural carbohydrates in the secondary phloem of trees

Non-invasive measurement of phloem transport

20.   Extracting the phloem-generated turgor signal from stem diameter measurements to analyze phloem transport in the field

21.   Using 13C to quantify phloem transport in the field

22.   Detecting rapid changes in carbon transport and partitioning with 11C

23.   Measuring phloem transport speed and volume with magnetic resonance imaging

24.   Visualizing phloem transport of ions and photoassimilates with a real-time radioisotope imaging system

25.   Studying phloem transport with small fluorescent tracers

26.   Non-invasive determination of phloem transport speed with 14C

27.   Studying transport routes and translocation dynamics of recently fixed photoassimilates with combined magnetic resonance imaging (MRI) and positron emission tomography (PET)

Phloem loading and unloading

28.   How to determine the phloem loading type of a plant

29.   Quantification of symplasmic phloem loading and unloading capacity with photoactivation microscopy

30.   Determination of sub-cellular sugar concentrations in relation to phloem loading and unloading

31.   Testing the influence of candidate genes on phloem loading in monocots

32.   Determining the mechanism of phloem unloading in fruit

33.   Overview: Techniques for the genetic manipulation of phloem loading

Phloem physiology

34.   Measuring pressure inside the phloem

35.   Non-invasive measurement of phloem sap viscosity

36.   Evaluating the influence of water stress on phloem transport capacity

37.   Overview: How to assess the role of phloem transport in the plant’s response to environmental factors

Theoretical models and microfluidics of phloem function

38.   Gaining insights on tree physiology from a theoretical model that integrates phloem and xylem function

39.   Using a multi-compartmental metabolic model to predict carbon allocation in Arabidopsis thaliana

40.   Making microfluidic devices that simulate phloem transport

41.   Using a resistance model of phloem transport to evaluate anatomical optimization for pressure-driven flow and predicting transport speed

42.   A mechanistic model to predict distribution of carbon among multiple sinks

43.   Using a mathematical model of phloem transport to optimize strategies for crop improvement

Detecting signals in the phloem

44.   A strategy to avoid false-positives when searching for phloem-mobile RNAs

45.   Detecting Ca signals along the phloem pathway in intact plants

46.   Assessing the redox status of phloem proteins

47.   Identification of herbivore-induced electrical signals that propagate along the phloem

48.   Efficient establishment of homo- and heterografts

49.   Identification of phloem-mobile mRNAs in Arabidopsis thaliana

50.   Analyzing and predicting the phloem mobility of macromolecules with an online database

51.   Camera-based high-resolution measurement of electrical activity in the phloem

Phloem development

52.   Overview: the tools to study phloem development

53.   Identification of molecular processes during phloem formation through transcriptomics

54.   A vascular cell induction culture system to study sieve elements differentiation in vitro

55.   Measuring phloem transport of auxin and cytokinin in relation to phloem development

56.   Investigating the role of phospholipid signaling in phloem cell differentiation with the help of radiolabeled lipids

57.   Bioinformatic classification of RNAseq reads to investigate the development of phloem connections between parasitic and host plants

58.   Studying phloem development in nematode-induced feeding sites

Miscellaneous

59.   The AtSUC2 promoter: Powerful tool to study phloem physiology and development

60.   How to assess the importance of transcription factors for sieve element evolution