Differentiation of Phytopathogenic Agrobacterium spp.
Diferencijacija fitopatogenih vrsta roda Agrobacterium
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Author
Kuzmanovic, Nemanja
Ivanovic, Milan
Calic, Andjelka
Gasic, Katarina
Obradovic, Aleksa
Keywords
Crown gallPlasmid
Agrobacterium
„Duplex“ PCR
Pathogenicity
Bakteriozni rak
plazmid
patogenost
Metadata
Show full item recordAbstract
Due to the difficulties in differentiation of phytopathogenic Agrobacterium spp. and
lack of a standardized protocol, we carried out selection and evaluation of suitable methods
based on the bacterial physiological, genetic and pathogenic properties. Strains of
Agrobacterium tumefaciens, A. rhizogenes and A. vitis were differentiated using standard bacteriological
and molecular methods. The biochemical and physiological tests confirmed
authenticity of the strains. Two duplex PCR methods were conducted with four different
primer pairs. In all strains, presence of plasmid virD2 and virC pathogenicity genes was
detected. Chromosomal pehA gene was determined in A. vitis strain. Pathogenicity was
confirmed on carrot slices and young plants of tomato and sunflower. Strains of A. tumefaciens
and A. vitis were pathogenic on all test plants, while strain of A. rhizogenes induced
characteristic symptoms only on carrot slices. The tests used in this study provided reliable
discrimination between the three species and confirmed their identity as tumorigenic (Ti)
Agrobacterium tumefaciens and A. vitis, and rhizogenic (Ri) A. rhizogenes. Usled poteškoća u razlikovanju vrsta roda Agrobacterium i nedostatka standardizovanog
protokola izvršena je procena i odabir pogodnih metoda u cilju njihove diferencijacije
na osnovu fizioloških, genetskih i patogenih odlika. Diferencirani su sojevi Agrobacterium
tumefaciens, A. rhizogenes i A. vitis primenom standardnih bakterioloških i molekularnih metoda.
Primenom diferencijalnih testova sojevi su ispoljili očekivane biohemijsko-fiziološke
karakteristike. Izvedene su dve „duplex“ PCR metode sa 4 različita tipa prajmera. Kod proučavanih
sojeva detektovano je prisustvo virD2 i virC gena patogenosti, koji se nalaze na plazmidnoj
DNK bakterije. Prisustvo hromozomskog gena pehA, odgovornog za sintezu enzima
poligalakturonaze, utvrđeno je kod soja A. vitis. Patogenost je proverena na kriškama mrkve
i mladim biljkama paradajza i suncokreta. Sojevi A. tumefaciens i A. vitis bili su tumorogeni
na svim test biljkama, dok je soj A. rhizogenes ispoljio patogenost jedino na kriškama mrkve.
Na osnovu dobijenih rezultata, proučavani sojevi diferencirani su kao tumorogeni (Ti) Agrobacterium
tumefaciens i A. vitis, i kao rizogeni (Ri) A. rhizogenes.