Chapter title |
In Situ Detection of Programmed Cell Death in Senescing Nicotiana tabacum Leaves Using TUNEL Assay
|
---|---|
Chapter number | 21 |
Book title |
Plant Senescence
|
Published in |
Methods in molecular biology, January 2018
|
DOI | 10.1007/978-1-4939-7672-0_21 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7670-6, 978-1-4939-7672-0
|
Authors |
Branka Uzelac, Dušica Janošević, Snežana Budimir |
Abstract |
Leaf senescence constitutes a highly regulated final phase of leaf development, leading to cell death that is recognized as a type of programmed cell death (PCD). Degradation of nuclear DNA into oligonucleosomal fragments (DNA ladder) and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay are methods commonly used to detect PCD-specific DNA cleavage. TUNEL reaction in situ labels free 3'-OH DNA strand breaks (nicks), thus allowing histological localization of nuclear DNA degradation during PCD. Here we describe in situ labeling of PCD-specific nuclear DNA fragmentation on conventional histological sections of senescing tobacco leaves. Incorporation of fluorescein-labeled dUTPs is detected by fluorescence microscopy, which enables in situ visualization of PCD at the single-cell level in the leaf mesophyll tissues undergoing senescence. |
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