Differentiation of Phytopathogenic agrobacterium spp.

Abstract

REZIMEUsled 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.

In-Depth Characterization of Crown Gall Disease of Tobacco in Serbia

“…Furthermore, using a set of specific primers [19], the tobacco isolates were identified at the species/biovar level (A. tumefaciens, bv1), indicating that these tobacco isolates were members of this species. Two duplex PCR methods conducted with four different primer pairs confirmed the presence of the plasmid virD2 and virC pathogenicity gene, indicating the presence of Ti or Ri plasmids in the tested isolates [16,22,23]. Kuzmanović et al [36] stated that VCF3/VCR3 primers (targeting virC1-virC2 gene) are applicable in a wide variety of tumorigenic agrobacteria and are therefore considered a reliable tool for detecting crown gall pathogens.…”

“…Two duplex PCR analyses were carried out using the A/C (virD2) and CYT/CYT (ipt) primers [20] for the first reaction, and PGF/PGR (pehA) [21] and VCF3/VCR3 (virC) [22] primers for the second reaction (Table 1), adopting the amplification conditions outlined by Kuzmanović et al [23]. The PCR products were separated by electrophoresis using 2% agarose gel, stained with Midori Green Advance.…”