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Root and Root Endophytes from the Eyes of an Electron Microscopist

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Root Engineering

Part of the book series: Soil Biology ((SOILBIOL,volume 40))

Abstract

Electron microscopes, because of their higher resolutions and magnifications, can be employed in many scientific areas where light microscopes have limited utility. SEM can achieve resolution better than 1 nm. Specimens can be observed in high and low vacuum and in wet conditions.

As a case study, SEM of Piriformospora indica was performed. The micrographs showed young growing hyphae. They were straight and parallel to each other, and the surface was smooth. As the sporulation commenced, hyphae became more nodulated and highly branched. Mature chlamydospores which were pear shaped were clearly observed. The junction of hypha and chlamydospores was broad. Confocal microscopy revealed that the fungus colonizes the root surface, enters the root cortex, and establishes inter- and intracellularly. Mycelium showed feeble autofluorescence. The young spores showed fluorescence, and at maturity the wall was thick and emitted strong fluorescence. Normally, it takes several hours for conventional staining, but it takes only few minutes by employing confocal microscopy in order to observe root colonization by the fungus. Future research on gene expression, microRNA, and the degradome sequencing to understand the plant–microbe interaction is warranted. For this target ultrastructure microscopes may play an important role.

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Acknowledgments

The authors thank Dr. A. Morte for her useful comments to the manuscript.

Author information

Authors and Affiliations

  1. Institute of Biotechnology, Zhejiang University, Hangzhou, 310058, China

    Binggan Lou & Bing Peng

  2. Center of Electron Microscope, Zhejiang University, Hangzhou, 310058, China

    Nianhang Rong & Yunqin Li

  3. Equipment and Technology Service Platform, College of Life Sciences, Zhejiang University, Hangzhou, 310058, China

    Hanmin Chen

  4. Amity Institute of Microbial Technology, Amity University Uttar Pradesh, Gautam Budhnagar, Noida, UP, India

    K. Sowjanya Sree & Ajit Varma

  5. Analysis Center of Agrobiology and Environmental Sciences, Zhejiang University, Hangzhou, 310058, China

    Qikang Gao

Authors
  1. Binggan Lou
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  2. Bing Peng
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  4. Yunqin Li
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  5. Hanmin Chen
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  6. K. Sowjanya Sree
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  7. Qikang Gao
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  8. Ajit Varma
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Corresponding author

Correspondence to Ajit Varma .

Editor information

Editors and Affiliations

  1. Facultad de Biología, Universidad de Murcia, Murcia, Spain

    Asunción Morte

  2. Amity Institute of Microbial Technology, Amity University Uttar Pradesh, Noida, Uttar Pradesh, India

    Ajit Varma

Appendix

Appendix

Hill and Käfer medium (2001)

Composition

(g/L)

Glucose

20.0

Peptone

2.0

Yeast extract

1.0

Casein hydrolysate

1.0

Vitamin stock solution

1.0 mL

Macroelements from stock

50.0 mL

Microelements from stock

2.5 mL

Agar

0.8 % (w/v)

CaCl2

0.1 M, 1.0 mL

FeCl3

0.1 M, 1.0 mL

pH

5.8

Macroelements stock

(g/L)

NaNO3

120.0

KCl

10.4

MgSO4 ∙ 7H2O

10.4

KH2PO4

30.4

Minor elements stock

(g/L)

ZnSO4

22.0

H3BO3

11.0

MnCl2â‹…4H2O

5.0

CoCl2â‹…6 H2O

1.6

CuSO4â‹…5 H2O

1.6

(NH4)6Mo7O24â‹…7H2O

1.1

Na2EDTA

50.0

Vitamins

% (w/v)

Biotin

0.05

Nicotinamide

0.5

Pyridoxal phosphate

0.1

Amino benzoic acid

0.1

Riboflavin

0.25

  1. The pH was adjusted to 5.8 with 1 N HCl. All stocks were stored at 4 °C except the vitamins which were stored at −20 °C

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Lou, B. et al. (2014). Root and Root Endophytes from the Eyes of an Electron Microscopist. In: Morte, A., Varma, A. (eds) Root Engineering. Soil Biology, vol 40. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-54276-3_22

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