Ramón Santos

Use of culture-derived Fusarium oxysporum f. sp. cubense, race 1 filtrates for rapid and non-destructive in vitro differentiation between resistant and susceptible clones of field-grown banana

Banana and plantain are among the most important food crops in developing countries but production is threatened by increasing virulent forms of Fusarium oxysporum f. sp. cubense. Chemical control is not economically effective and,therefore, breeding programs are necessary. Traditional field studies of new genotype resistance to this disease are time-consuming and destructive. Therefore,we developed a rapid and non-destructive procedure to differentiate field-grown banana resistant from susceptible clones. This procedure implicates application of culture filtrates of Fusarium oxysporum f. sp. cubense race 1 onto banana leaves. The relationship between duration of the fungal in vitro incubation, and the fungal culture fresh mass, the culture filtrate absorbency, and the Gross Michel (susceptible cultivar)leaf lesion area (after application of the culture filtrate) were similar and at 24day-incubation the highest values of the recorded indicators were observed. A comparison between Gross Michel and FHIA-01(resistant) was also performed. The most relevant differences between cultivars were observed at 48 hours after application of the culture filtrate, and in the middle-aged leaves. The position of the culture filtrate application in the leaf limb (distal, middle, proximal) was not determinant. A wider comparison among banana cultivars confirmed previous results informed by other researchers using different systems to study this plant-fungus interaction. Such a confirmation validates the effectiveness of the procedure described here to select rapid and non-destructively banana resistance to this disease at field level.

Phytotoxic effect of culture filtrate from Fusarium subglutinans the causal agent of fusariose of pineapple (Ananas comosus (L.) Merr

Two pineapple varieties differing in resistance to fusariose were examined for the phytotoxic effect of Fusarium subglutinans culture filtrate. The cultivars were Perolera (more resistant to pathovars of Fusarium subglutinans) and Smooth Cayenne (more susceptible). The phytotoxic effect of culture filtrate was assessed in tissue culture pineapple plantlets (by electrolyte leakage and placing the culture filtrate on wound leaf segments) and callus (inhibition of growth). Smooth Cayenne proved to be the most sensitive cultivar in each test, whereas Perolera showed resistance to the culture filtrate and its callus grew in the presence of high concentrations of culture filtrate that were completely toxic to Smooth Cayenne. These results show that plants can display cellular resistance to the culture filtrate.

Bioassay for in vitro differentiation of pineapple cultivar resistance levels to heart rot disease

SummaryAn in vitro bioassay to differentiate pineapple plant resistance levels to Phytophthora nicotianae var. parasitica (heart rot disease) is deseribed here. Conditions to cause death of in vitro-cultured plants were defined using a cultivar previously found to be susceptible to this fungus in our Field-Grown Pineapple Germplasm Bank (ev. Smooth Cayenne Serrana). The effects of zoospore concentration, inoculation technique, and disease progress during the course of time after infection were evaluated. The highest rates of plant death were observed with the use of 108 zoospores ml−1, and the inoculation technique of needle-mediated leaf base wound. One hundred percent plant death was observed at 144h after infection. Different susceptible varieties along with a resistant pineapple relative were additionally compared. In vitro results confirmed previous observations obtained under field conditions. The protocol described here may be used for early selection (in vitro) of new pineapple genotypes showing resistance to this fungus. At present, this protocol is extensively used in the Biotechnology-assisted Cuban Program for Pineapple Breeding.

Improved technique for rapid and non-destructive in vitro differentiation between resistant and susceptible banana clones of Fusarium oxysporum f. sp. cubense

Culture filtrates of Fusarium oxysporum f. sp. cubense were applied to field-grown banana leaves of susceptible and resistant clones. The difference in leaf lesions, measured after 48 h, varied from 13 to 51 mm2 depending on the composition of the growth medium.