Morphological and Molecular Identification of Colletotrichum acutatum from Tomato Fruit
Morfološka i molekularna identifikacija Colletotrichum acutatum sa ploda paradajza
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Author
Zivkovic, Svetlana
Stojanovic, Sasa
Ivanovic, Zarko
Trkulja, Nenad
Dolovac, Nenad
Aleksic, Goran
Balaz, Jelica
Keywords
AnthracnoseTomato
Colletotrichum acutatum
Identification
Antraknoza
paradajz
Colletotrichum acutatum
identifikacija
Metadata
Show full item recordAbstract
Colletotrichum gloeosporioides, Colletotrichum acutatum, Colletotrichum coccodes, and
Colletotrichum dematium are the four main species of Colletotrichum that cause tomato
anthracnose. In Serbia, the occurrence of anthracnose on tomato fruit has been recorded
during the last several years. Typical fruit symptoms include dark, sunken, and circular
lesion with orange conidial masses. Pathogen isolates were obtained from a diseased
tomato fruits, on PDA medium forming a white to gray colonies. The cultures developed
black acervuli around the center of the colony. Conidia were hyaline, aseptate, and fusiform
or rarely cylindrical. Appressoria were smooth, simple, clavate to ovate, and varied
from light to dark brown. Pathogenicity tests with representative isolates were conducted
on symptomless, detached tomato fruits. All tested isolates caused anthracnose lesions
on tomato fruit after 7 days of incubation. Koch’s postulates were fulfilled by reisolation
from inoculated tomato fruits. PCR analysis (using species-specific primer pair, CaInt2/
ITS4) of genomic DNA from tomato isolates resulted in an amplification product of 490
bp, specific for C. acutatum, further confirming the identity of the pathogen. Based on
morphological and molecular characteristics, the isolates from tomato fruit were determined
as C. acutatum. Četiri glavna prouzrokovača antraknoze paradajza iz roda Colletotrichum su: Colletotrichum
gloeosporioides, Colletotrichum acutatum, Colletotrichum coccodes i Colletotrichum
dematium. U Srbiji je tokom poslednjih godina zabeležena pojava antraknoze ploda paradajza.
Tipični simptomi na plodu su tamne, ulegnute, kružne lezije sa masom narandžastih
konidija. Sa obolelih plodova paradajza dobijeni su izolati patogena koji na KDA podlozi
obrazuju bele do sivo obojene kolonije. Acervuli crne boje formiraju se u kulturi oko centra
kolonije. Konidije su hialinske, neseptirane, fusiformne ili ređe cilindrične. Apresorije su
glatke, jednostavne, okruglastog ili oblika izdužene palice, svetlo do tamno braon boje. Test
patogenosti je obavljen sa reprezentativnim izolatima, na odabranim, zdravim plodovima.
Svi ispitivani izolati prouzrokuju antraknozne lezije na plodu paradajza 7 dana nakon inokulacije.
Kohovi postulati su zadovoljeni reizolacijama sa inokulisanih plodova paradajza. PCR
analiza (korišćenjem para prajmera specifičinog za vrstu, CaInt2/ITS4) iz genoma DNA izolata
sa paradajza rezultirala je amplifikacijom produkta od 490 bp, specifičnog za C. acutatum,
što je potvrdilo identitet patogena. Na osnovu morfoloških i molekularnih karakteristika
izolati sa ploda paradajza determinisani su kao C. acutatum.