PMID- 22350735
OWN - NLM
STAT- MEDLINE
DCOM- 20140122
LR  - 20240109
IS  - 1615-6102 (Electronic)
IS  - 0033-183X (Print)
IS  - 0033-183X (Linking)
VI  - 250
IP  - 1
DP  - 2013 Feb
TI  - Determination of ACC-induced cell-programmed death in roots of Vicia faba ssp. 
      minor seedlings by acridine orange and ethidium bromide staining.
PG  - 121-8
LID - 10.1007/s00709-012-0383-9 [doi]
AB  - Fluorescence staining with acridine orange (AO) and ethidium bromide (EB) showed 
      that nuclei of cortex root cells of 1-aminocyclopropane-1-carboxylic acid 
      (ACC)-treated Vicia faba ssp. minor seedlings differed in color. Measurement of 
      resultant fluorescence intensity (RFI) showed that it increased when the color of 
      nuclear chromatin was changed from green to red, indicating that EB moved to the 
      nuclei via the cell membrane which lost its integrity and stained nuclei red. 
      AO/EB staining showed that changes in color of the nuclear chromatin were 
      accompanied by DNA condensation, nuclei fragmentation, and chromatin degradation 
      which were also shown after 4,6-diamidino-2-phenylindol staining. These results 
      indicate that ACC induced programmed cell death. The increasing values of RFI 
      together with the corresponding morphological changes of nuclear chromatin were 
      the basis to prepare the standard curve; cells with green unchanged nuclear 
      chromatin were alive while those with dark orange and bright red nuclei were 
      dead. The cells with nuclei with green-yellow, yellow-orange, and bright orange 
      chromatin with or without their condensation and fragmentation chromatin were 
      dying. The prepared curve has became the basis to draw up the digital method for 
      detection and determination of the number of living, dying, and dead cells in an 
      in planta system and revealed that ACC induced death in about 20% of root cortex 
      cells. This process was accompanied by increase in ion leakage, shortening of 
      cells and whole roots, as well as by increase in weight and width of the apical 
      part of roots and appearance of few aerenchymatic spaces while not by 
      internucleosomal DNA degradation.
FAU - Byczkowska, Anna
AU  - Byczkowska A
AD  - Department of Cytophysiology, University of Lodz, Pomorska 141/143, PL-90236 
      Lodz, Poland.
FAU - Kunikowska, Anita
AU  - Kunikowska A
FAU - Kazmierczak, Andrzej
AU  - Kazmierczak A
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20120215
PL  - Austria
TA  - Protoplasma
JT  - Protoplasma
JID - 9806853
RN  - 0 (Amino Acids, Cyclic)
RN  - 0 (Fluorescent Dyes)
RN  - 0 (Indicators and Reagents)
RN  - 3K9EJ633GL (1-aminocyclopropane-1-carboxylic acid)
RN  - EN464416SI (Ethidium)
RN  - F30N4O6XVV (Acridine Orange)
SB  - IM
MH  - Acridine Orange/*chemistry
MH  - Amino Acids, Cyclic/*pharmacology
MH  - Cell Death/drug effects
MH  - Cell Membrane/drug effects
MH  - Cell Nucleus/drug effects/metabolism
MH  - Ethidium/*chemistry
MH  - Fluorescent Dyes/chemistry
MH  - Indicators and Reagents/chemistry
MH  - Microscopy, Fluorescence/methods
MH  - Plant Roots/drug effects/metabolism
MH  - Vicia faba/*drug effects/metabolism
PMC - PMC3557382
EDAT- 2012/02/22 06:00
MHDA- 2014/01/23 06:00
PMCR- 2012/02/15
CRDT- 2012/02/22 06:00
PHST- 2011/10/21 00:00 [received]
PHST- 2012/01/26 00:00 [accepted]
PHST- 2012/02/22 06:00 [entrez]
PHST- 2012/02/22 06:00 [pubmed]
PHST- 2014/01/23 06:00 [medline]
PHST- 2012/02/15 00:00 [pmc-release]
AID - 383 [pii]
AID - 10.1007/s00709-012-0383-9 [doi]
PST - ppublish
SO  - Protoplasma. 2013 Feb;250(1):121-8. doi: 10.1007/s00709-012-0383-9. Epub 2012 Feb 
      15.