Ciro Hideki Sumida

Métodos de preservação in vitro de urediniósporos de Puccinia kuehnii

With the aim of evaluating preservation methods of Puccinia kuehnii urediniospores, two bioassays were conducted: the first one (B1) with different methods of dehydration and the second one (B2) with different methods rehydration. B1 and B2 differed in the inclusion of silica gel granule in B1 for preservation in microcentrifuge tubes. Leaves with symptoms of orange rust were harvested from the sugarcane cultivar SP89 1115. P. kuehnii urediniospores were extracted from the leaves with the aid of a vacuum bomb. Later, they were placed in microcentrifuge tubes. Treatments for B1 were: l: dehydration in silica gel, lyophilization and without dehydration, ll: room temperature (20oC), refrigerator (5oC), freezer (-20oC) and deep-freezer (- 80oC). Treatments for B2 were: l: dehydration in silica gel and without dehydration, ll: room temperature (20oC), refrigerator (5oC), freezer (-20oC) and deep-freezer (-80oC), lll: with and without rehydration in the evaluations. The initial germination was carried out for both experiments, other assessments were made at 15 and 30 days of storage and then at every 30 days until 180 days. Urediniospore suspensions were prepared in water and a 0.1-mL aliquot was transferred to Petri dishes containing agar-water (15 g L-1). They remained at 20oC in the dark. To assess viability, the count of 200 urediniospores/plate was performed. The data were subjected to nonparametric Kruskal-Wallis analysis of variance and complemented by Dunns test. Results showed that viability decreased with time, and the best treatments reached 27.6% and 6.6% at 30 days, and 12.0% and 1.9% at 60 days for B1 and B2, respectively. Dehydration method in silica gel followed by storage at -80oC was the only treatment that presented viable urediniospores (1.2%) at 180 days for B1. For B2, the best method was preservation with dehydration, followed by storage at 5oC, without rehydration. This method showed the best germination percentage at 120 days (0.4%). Influence of the factor thermal shock was not observed in the recovery of viable urediniospores in B2. Inclusion of silica gel granules in the microcentrifuge tube allowed the recovery of viability of urediniospores at 180 days.

Trichoderma asperelloides antagonism to nine Sclerotinia sclerotiorum strains and biological control of white mold disease in soybean plants

ABSTRACT Sclerotinia sclerotiorum is an important plant pathogen with worldwide distribution that causes severe economic losses of various agricultural crops such as soybean. This fungus is normally controlled with synthetic chemical fungicides that pose risks to the environment, and can be harmful to human health, and they can also induce resistance in pests. The aim of this study was to investigate the potential of Trichoderma asperelloides as a biocontrol agent towards white mold disease on soybeans crops. The antagonism of two strains of T. asperelloides (T25 and T42) isolated from soil samples was determined in-vitro by dual-culture confrontation testing on nine S. sclerotiorum strains obtained from sclerotia collected on diseased soybean plants. The mycelial growth and inhibition of carpogenic and ascospore germination by T. asperelloides extracts, as well as the efficacy of these on white mold control in soybeans were evaluated. Both strains of T. asperelloides exhibited high potential of antagonism. Methanolic and ethyl acetate extracts of the two T. asperelloides strains showed excellent growth inhibition (60–100%) on all of the pathogens tested. The ethyl acetate extracts of both T. asperelloides strains exhibited the highest efficacy against carpogenic germination, decreasing by 20–30% the number of ascospores per apothecium. Strains of T. asperelloides tested were more efficient in controlling white mold than two commercial products made from Trichoderma harzianum. The new strains of T. asperelloides have potential for successful biological control of white mold disease of soybean crops in the field.

Inibição micelial in vitro de Sclerotinia sclerotiorum por fungicidas

Departamento de Agronomia - Universidade Estadual de Londrina (UEL), Rod. Celso Garcia Cid. (PR 445), Km 380, CP 6001, CEP 86051-990. Londrina, PR. Autor para correspondencia: Ciro Hideki Sumida (cirosumida@hotmail.com)Data de chegada: 23/08/2013. Aceito para publicacao em: 18/02/2014. 1916