No abstract

Clavibacter michiganensis subsp. michiganensis (Cmm) is a quarantine organism in Europe and in many other countries. It is one of the most severe bacterial pathogens affecting tomato. Screening tomato plants for their resistance level to Cmm requires a large amount of space under quarantine conditions and is therefore costly. This project developed a new inoculation protocol on in vitro tomato plants to facilitate a more economic and higher throughput disease screening. A new method using the PathoScreen system was tested to localize green fluorescent protein-tagged Cmm in planta and to quantify the pathogen based on the percentage of corrected GFP (cGFP%). The system was sensitive in detecting the GFP-tagged Cmm in the shoots, but in the roots a high autofluorescence masked detection and thus sensitivity of the assay. The in vitro protocol was tested on several wild relatives of tomato, which were previously screened in a greenhouse assay. The correlation between wilt symptoms in vitro and wilt symptoms in the greenhouse was overall moderate (r = 0.6462). The protocol worked well in differentiating the two parents that were used in the mapping studies. This study shows that the in vitro protocol can be efficiently used for resistance breeding in many tomato genotypes.

O crestamento bacteriano comum, causado por Xanthomonas axonopodis pv. phaseoli (Xap), é a principal bacteriose do feijoeiro no Brasil, sendo transmitida principalmente por sementes. O presente trabalho teve como objetivo aperfeiçoar uma técnica, por meio de diferentes métodos de preparação do extrato, para a detecção de Xap, bem como sua detecção simultânea com Curtobacterium flaccumfaciens pv. flaccumfaciens (Cff) nos extratos de sementes de feijão,via PCR. A partir de amostras de sementes de feijão inoculadas artificialmente com Xap e lotes comerciais, foram avaliados: extrato bruto obtido diretamente das sementes, extrato concentrado por filtração em membrana de 0,22µM de diâmetro, extrato concentrado por centrifugação e extrato plaqueado em meio semi-seletivo XCP1 com e sem antibióticos (BIO-PCR). Avaliou-se a presença simultânea de Xap e Cff em 10 lotes comerciais de sementes de feijão através da reação multiplex, utilizandos-se os primers X4c, X4e, CffFOR2 e CffREV4. A partir do extrato bruto, do extrato concentrado por centrifugação e por filtragem em membrana Millipore® não foi possível a detecção de Xap nas sementes de feijão artificialmente contaminadas nem nos 47 lotes comerciais de sementes/ grãos de feijão. A técnica de BIO-PCR permitiu a detecção de Xap a partir de extratos de sementes de feijão artificialmente contaminadas e em 18 dos 47 lotes comerciais. A técnica de detecção simultânea de Xap e Cff no mesmo gel é viável, por amplificar fragmentos de DNA típicos de cada fitobactéria. O uso do meio de cultura XCP1 sem adição de antibióticos permitiu detectar Xap com período de incubação menor em um dia em comparação à detecção utilizando-se o meio de cultura com antibióticos

No abstract

FLOWERING OF 'PONKAN' MANDARIN (Citrus reticulata

Ramulosis (Colletotrichum gossypii var. cephalosporioides) is an important fungal disease of cotton in Brazil, exclusively controlled by fungicide application. Therefore, sustainable management of ramulosis is essential. This work aimed to evaluate the potential of three bacterial strains, Bacillus amyloliquefaciens (UFLA285), Bacillus velezensis (UFLA401), and Paenibacillus lentimorbus (MEN2), for the biocontrol of ramulosis in cotton and their effects on yield and fiber quality. Seed treatment (ST), foliar spray, and soil drenching application methods were used (separately or combined) under greenhouse and field conditions. Chemical treatments recommended against ramulosis and water were used as controls. Under greenhouse conditions all strains reduced the disease incidence. While B. velezensis UFLA401 and P. lentimorbus MEN2 reduced the incidence by 56.6% and 45.7%, respectively, independent of the application method, B. amyloliquefaciens UFLA285 reduced the disease by about 60% when applied as a foliar spray or ST + foliar spray. Two field trials were performed and all bacterial strains reduced ramulosis incidence. In the first year, B. velezensis UFLA401 sprayed on the plants reduced incidence by 22.3% and ST + two foliar sprays resulted in the best performance, decreasing ramulosis by 57%. In both seasons the yield increased by using either bacterial or chemical treatments compared to the water control. The combination B. velezensis UFLA401 and P. lentimorbus MEN2 sprays provided better fiber quality than chemical treatment. Therefore, Bacillus sp. (UFLA285 and UFLA401) and P. lentimorbus MEN2 are potential tools to reduce ramulosis, increase cotton yield and fiber quality.