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A R T I C L E I N F O
A B S T R A C T
Article history:
Received 20 March 2009
Received in revised form 18 June 2009
Accepted 23 June 2009
Available online 2 July 2009
Higher plants are recognized as excellent genetic models to detect environmental mutagens and are
frequently used in monitoring studies. Among the plant species, Alium cepa has been used to evaluate
DNA damages, such as chromosome aberrations and disturbances in the mitotic cycle. Employing the A.
cepa as a test system to detect mutagens dates back to the 40s. It has been used to this day to assess a
great number of chemical agents, which contributes to its increasing application in environmental
monitoring. The A. cepa is characterized as a low cost test. It is easily handled and has advantages over
other short-term tests that require previous preparations of tested samples, as well as the addition of
exogenous metabolic system. Higher plants, even showing low concentrations of oxidase enzymes and a
limitation in the substrate specication in relation to other organism groups, present consistent results
that may serve as a warning to other biological systems, since the target is DNA, common to all
organisms. The A. cepa test also enables the evaluation of different endpoints. Among the endpoints,
chromosome aberrations have been the most used one to detect genotoxicity along the years. The
mitotic index and some nuclear abnormalities are used to evaluate citotoxicity and analyze
micronucleus to verify mutagenicity of different chemicals. Moreover, the A. cepa test system provides
important information to evaluate action mechanisms of an agent about its effects on the genetic
material (clastogenic and/or aneugenic effects). In the face of all the advantages that the A. cepa test
system offers, it has been widely used to assess the impacts caused by xenobiotics, characterizing an
important tool for environmental monitoring studies, where satisfactory results have been reported.
2009 Elsevier B.V. All rights reserved.
Keywords:
Allium cepa
Environmental pollutants
Genotoxicity
Mutagenicity
Contents
1.
2.
3.
4.
5.
6.
7.
Genetic toxicology and environmental monitoring based on the use of higher plants to detect xenobiotics .
Modications of the Allium cepa test system to assess environmental contamination . . . . . . . . . . . . . . . . . .
Analysis of different endpoints by the Allium cepa test . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
3.1.
Mitotic index. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
3.2.
Chromosome aberrations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
3.3.
Nuclear abnormalities. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
3.4.
Micronucleus. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Technical procedure advantages of the Allium cepa test . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
The employment of Allium cepa test to detect different classes of environmental pollutants . . . . . . . . . . . . .
5.1.
Metals . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
5.2.
Pesticides. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
5.3.
Aromatic hydrocarbons . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
5.4.
Textile industry dyes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
5.5.
Products used to disinfect drinking water . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
5.6.
Complex mixtures. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
5.7.
Other agents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Assessment of promutagens by the Allium cepa test system . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Studies of sensitivity and correlation between the Allium test and other test systems . . . . . . . . . . . . . . . . . .
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* Corresponding author at: Av. 24 A, 1515, Bela Vista, 13506-900, Rio Claro, SP, Brazil. Tel.: +55 19 3526 4143; fax: +55 19 3536 0009.
E-mail address: mamm@rc.unesp.br (M.A. Marin-Morales).
1383-5742/$ see front matter 2009 Elsevier B.V. All rights reserved.
doi:10.1016/j.mrrev.2009.06.002
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8.
Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Acknowledgements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
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Fig. 1. A. cepa karyotype, 2n = 16. Arrow indicating the nucleolus organizer region (NOR) located in the terminal portion of the short arm of pair 6.
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Fig. 2. CA observed in A. cepa meristematic cells exposed to chemical agents. (A) Normal metaphase; (A1) Metaphase with chromosome break; (A2) C-metaphases; (A3)
Metaphase with chromosome loss; (A4) Binucleated cell in metaphase; (A5) Metaphase with chromosome adherence; (B) Normal anaphase; (B1) Anaphase with chromosome
bridge; (B2) Anaphase with chromosome breaks; (B3) Anaphase with bridge and chromosome loss; (B4) Anaphase with chromosome loss; (B5) Multipolar anaphase; (C)
Normal telophase; (C1) Telophase with chromosome bridge; (C2) Telophase with bridge and chromosome loss; (C3) Telophase with chromosome break; (C4) Multipolar
telophase; (C5) Multipolar telophase with chromosome bridge.
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Fig. 3. NA observed in A. cepa meristematic cells exposed to chemical agents. (A) Interphasic cell with nuclear bud and MN; (B) Interphasic nucleus with nuclear bud; (C)
Prophase with synchronic nuclear bud; (D) Nucleus with nuclear point; (E) Lobulated nucleus with MN; (FH) Lobulated nucleus; (I) Interphasic nucleus interconnected by
nuclear bridge and MN.
Fig. 4. A. cepa meristematic cells carrying MN. (A) Normal interphase; (A1) Micronucleated cell in interphase; (B) Normal prophase; (B1) MN in prophase; (C) Normal
metaphase; (C1) MN in metaphase; (D) Normal anaphase; (D1) Anaphase with MN; (E) Normal telophase; (E1) Telophase with MN and chromosome loss.
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Fig. 5. A. cepa meristematic cells carrying MN. (AC) MN of small size in interphase; (D) MN of small size in prophase; (EF) MN of large size in interphase; (GH) MN of large
size in prophase.
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Table 1
A summary of the A. cepa studies reported above.
Tested agents
Endpoint
Cell type
CA
CA, MN
CA, MN
CA, MN, NA
Meristematic
Meristematic
F1 cells
Meristematic
Meristematic
Pesticidetriuralin herbicide
MN, CA
Meristematic cells
MI, CA, MN
Meristematic cells
CA
Meristematic cells
Pesticideatrazine herbicide
MI, CA
Meristematic cells
CA, NA
Meristematic cells
Metal (arsenic)
MI, MN
MI, CA
Meristematic and
F1 cells
Meristematic cells
MI, CA, MN
Meristematic cells
CA
CA
Meristematic cells
Meristematic cells
CA
CA
MI, CA
CA
MN
MI, CA
Meristematic
Meristematic
Meristematic
Meristematic
F1 cells
Meristematic
MI, CA
CA
Meristematic cells
Meristematic cells
MI, CA, MN
Meristematic cells
PAHbenzo[a]pyrene
Oil eld efuent
Efuents of a silk dyeing industry
CA
CA
MI, mitotic
abnormalities
CA
CA
CA
MI, CA
CA
CA, MN
CA,
MI,
CA,
MI,
MN
CA, MN
MN
CA, MN, NA
Results
Reference
cells
and
[8]
[14]
cells
cells
[15]
[16]
cells
cells
cells
cells
[17]
[19]
[20]
[21]
[24]
[26]
Genotoxicity
[27]
Cytotoxicity, genotoxicity
and mutagenicity
Genotoxicity
Genotoxicity
[28]
[31]
[32]
Genotoxicity
Genotoxicity
Cytotoxicity and genotoxicity
Genotoxicity
Mutagenicity
Cytotoxicity and genotoxicity
(aneugenic effect)
Cytotoxicity and genotoxicity
Genotoxicity
[33]
[34]
[35]
[36]
[37]
[38]
[41]
Meristematic cells
Meristematic cells
Meristematic cells
Cytotoxicity, genotoxicity
and mutagenicity
Genotoxicity
Genotoxicity
Cytotoxicity and genotoxicity
[42]
[43]
[44]
Meristematic
Meristematic
Meristematic
Meristematic
Meristematic
Meristematic
cells
cells
cells
cells
cells
cells
Genotoxicity
Genotoxicity
Negative results
Low genotoxicity
Genotoxicity
Genotoxicity and mutagenicity
[45]
[48]
[49]
[50]
[52]
[53]
Meristematic
Meristematic
Meristematic
Meristematic
cells
cells
cells
cells
[54]
[55]
[56]
[57]
cells
Meristematic cells
MI, CA, MN
Meristematic cells
CA
Meristematic cells
Genotoxicity
Genotoxicity and mutagenicity
Genotoxicity
Cytotoxicity and genotoxicity
for the raw sludge
Genotoxicity and mutagenicity
(clastogenic effects for the
raw landfarm soil)
Cytotoxicity, genotoxicity
and mutagenicity
Cytotoxicity and genotoxicity
CA
MI
MI, CA
Meristematic cells
Meristematic cells
Meristematic cells
Genotoxicity
Cytotoxicity
Cytotoxicity and genotoxicity
[39]
[40]
[58]
[59]
[60]
[61]
[62]
[63]
MI: Mitotic index; CA: Chromosome aberrations; MN: Micronuclei, NA: Nuclear abnormalities.
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