ABSTRACT. Curtobacterium flaccumfaciens pv. flaccumfaciens (Cff), which is a gram-positive bacterium that causes wilting in several cultivated plants, is one of the main disease-causing bacteria in beans (Phaseolus vulgaris L.). Different techniques are employed to detect pathogens in bean plants; however, only a few slow and inefficient methodologies can be used to detect Cff. Hence, there is a need for rapid and efficient techniques to detect Cff for disease management. The objective of our study was to develop a technique to effectively detect Cff in the leaves of infected bean plants by optimizing a previously developed method. We modified the extraction method by reducing the amount of water used, replacing seeds with leaf tissues (obtained from the veins and petioles) and using CffFOR2 and CffREV4 primers specific for Cff. Our optimized method exploits a concentration gradient generated in the plant tissue through osmosis to disrupt the plant cell wall and efficiently isolate bacterial cells. DNA extracted from the isolated bacterial cells was amplified by polymerase chain reaction. This method allowed efficient detection of Cff in leaf tissues.

O cártamo (Carthamus tinctorius) é uma espécie cultivada de importância medicinal e industrial e atualmente é considerado uma planta de grande potencial na produção de biodiesel. O objetivo deste trabalho foi verificar qual o agente causal das manchas observadas em plantas de cártamo presentes no campo experimental do IAPAR em Londrina-PR. Foram realizados isolamentos a partir de lesões foliares de plantas sintomáticas e realizado teste de patogenicidade dos isolados em plantas sadias de cártamo. As plantas inoculadas apresentaram sintomas de antracnose nas folhas, nas hastes e no capítulo, assim como morte das plântulas. A observação e análise de estruturas reprodutivas do fungo em meio de cultura e no tecido do hospedeiro, assim como a PCR táxon-específica permitiu a identificação dos isolados do fungo como pertencentes à espécie Colletotrichum gloeosporioides. Este foi o primeiro relato de antracnose por C. gloeosporioides em cártamo no estado do Paraná.

The safflower (Carthamus tinctorius) is a crop of medicinal and industrial importance and is now considered a plant of great potential to biodiesel production. The objective of this study was to verify the causal agent of the spots observed in safflower plants in the IAPAR experimental field in Londrina-PR. Isolations were made from leaf lesions of symptomatic plants and pathogenicity test of the isolates in healthy plants of safflower. The inoculated plants showed symptoms of anthracnose on leaves, stems and the flower bud , and death of seedlings. The observation and analysis of reproductive structures of the fungus in media culture and in host tissue, as well as taxon-specific PCR allowed the identification of the fungus as belonging to Colletotrichum gloeosporioides specie. This was the first report of anthracnose caused by C. gloeosporioides in safflower in Parana state.